Muscle wasting in myotonic dystrophies: a model of premature aging.
Myotonic dystrophy type 1 (DM1 or Steinert's disease) and type 2 (DM2) are multisystem disorders of genetic origin. Progressive muscular weakness, atrophy and myotonia are the most prominent neuromuscular features of these diseases, while other clinical manifestations such as cardiomyopathy, insulin resistance and cataracts are also common. From a clinical perspective, most DM symptoms are interpreted as a result of an accelerated aging (cataracts, muscular weakness and atrophy, cognitive decline, metabolic dysfunction, etc.), including an increased risk of developing tumors. From this point of view, DM1 could be described as a progeroid syndrome since a notable age-dependent dysfunction of all systems occurs. The underlying molecular disorder in DM1 consists of the existence of a pathological (CTG) triplet expansion in the 3' untranslated region (UTR) of the Dystrophia Myotonica Protein Kinase (DMPK) gene, whereas (CCTG)n repeats in the first intron of the Cellular Nucleic acid Binding Protein/Zinc Finger Protein 9 (CNBP/ZNF9) gene cause DM2. The expansions are transcribed into (CUG)n and (CCUG)n-containing RNA, respectively, which form secondary structures and sequester RNA-binding proteins, such as the splicing factor muscleblind-like protein (MBNL), forming nuclear aggregates known as foci. Other splicing factors, such as CUGBP, are also disrupted, leading to a spliceopathy of a large number of downstream genes linked to the clinical features of these diseases. Skeletal muscle regeneration relies on muscle progenitor cells, known as satellite cells, which are activated after muscle damage, and which proliferate and differentiate to muscle cells, thus regenerating the damaged tissue. Satellite cell dysfunction seems to be a common feature of both age-dependent muscle degeneration (sarcopenia) and muscle wasting in DM and other muscle degenerative diseases. This review aims to describe the cellular, molecular and macrostructural processes involved in the muscular degeneration seen in DM patients, highlighting the similarities found with muscle aging.


Beating heart versus conventional mitral valve surgery.
The present study aimed to compare the results of beating heart technique and conventional mitral valve surgery (MVS). Three hundred and nineteen patients who underwent MVS between April 2005 and December 2006 were enrolled in the study. While 125 patients underwent beating heart MVS (group 1), the conventional approach was used for 194 patients (group 2). Of those patients who underwent beating heart MVS, 75 underwent MVS without cross-clamping the aorta. Coronary sinus retroperfusion was used during surgery in the remaining 50 patients. The right anterolateral thoracotomy was performed in nine out of the 29 patients requiring re-operation, while resternotomy was performed in 20. No significant differences were shown between the groups in the preoperative period in terms of the Parsonnet mortality score, Ontario mortality score, and length of intensive care stay. However, there were significant differences with respect to EuroSCORE risk score, EuroSCORE mortality, and Parsonnet risk score, and length of hospital stay according to Ontario risk scoring. It was established that the patients in group 1 had a shorter length of hospital stay [group 1: six days (range, 4-37 days); group 2: 10 days (range, 4-62 days)]. Group 1 was observed to have shorter time periods when the groups were compared regarding operative time [group 1: 130 min (range, 100-270 min); group 2: 240 min (range, 100-360 min)], cross-clamping (XCL) time [group 1: 27.5 min (range, 3-99 min); group 2: 60.5 min (range, 30-163 min)], and cardiopulmonary bypass time [group 1: 57 min (range, 22-150 min); group 2: 90 min (range, 39-388 min)]. There were also significant differences in favor of group 1 in terms of postoperative need for inotropic support [group 1: 26 patients (16%); group 2: 68 patients (35%)]. Although there were no statistically significant differences in the groups in terms of mortality rates according to the Parsonnet scoring system, with the exception of the moderate risk group, it was noted that the mortality rates in the beating heart group were lower. This study concluded that beating heart MVS can be performed successfully, particularly for patients at higher risk which will lead to increased morbidity and mortality in postoperative period.


Dimensionality of Early Adversity and Associated Behavioral and Emotional Symptoms: Data from a Sample of Japanese Institutionalized Children and Adolescents.
Recent approaches have begun to identify common variance across co-occurring childhood adversities (CAs) and their associations with symptoms of psychopathology. However, few studies have investigated these questions in high-risk samples, and in different cultural contexts. This study examined common variance amongst 18 types of CAs and associated symptomatology in 457 children and adolescents living in 24 residential homes in Japan. Principal component analysis identified four significant components that explained 35.1% of the variance: parental abuse, parental psychosocial risks, parental absence, and parental neglect. Path analysis revealed general as well as differential associations with negative outcomes: parental abuse, parental neglect, and parental psychosocial risks significantly associated with conduct problems, whereas parental abuse uniquely associated with peer problems, and parental neglect with hyperactivity/inattention. As well as confirming prior knowledge, these findings also extended understanding of these associations to a new cultural context. Future studies should take into account the multidimensional nature when assessing CAs.


Phase retrieval in digital speckle pattern interferometry by use of a smoothed space-frequency distribution.
We evaluate the use of a smoothed space-frequency distribution (SSFD) to retrieve optical phase maps in digital speckle pattern interferometry (DSPI). The performance of this method is tested by use of computer-simulated DSPI fringes. Phase gradients are found along a pixel path from a single DSPI image, and the phase map is finally determined by integration. This technique does not need the application of a phase unwrapping algorithm or the introduction of carrier fringes in the interferometer. It is shown that a Wigner-Ville distribution with a smoothing Gaussian kernel gives more-accurate results than methods based on the continuous wavelet transform. We also discuss the influence of filtering on smoothing of the DSPI fringes and some additional limitations that emerge when this technique is applied. The performance of the SSFD method for processing experimental data is then illustrated.


Effects of Streptococcus salivarius K12 with nystatin on oral candidiasis-RCT.
To evaluate the efficacy and safety of Streptococcus salivarius K12 as an adjuvant in treating oral candidiasis. A total of 56 patients were participated in the randomized, double-blinded, placebo-controlled clinical trial. The S. salivarius K12 or placebo lozenges plus nystatin tablets were given for up to 4 weeks at 1-week interval and then followed up for 1 week thereafter. We collected and analyzed the mycological and clinical data, treatment course, and safety data. At the end of the treatment, significant differences were found in the mycological cure rates between K12 group and control group (90.48% and 55.56%, respectively, p = 0.008). Survival analysis demonstrated no statistical difference in overall cure rates comprehensively considering mycological cure, clinical improvement, and recurrence (p = 0.078), while statistical difference was found in mycological cure (p = 0.013) between the two groups. The median treatment courses of K12 group and control group were 3 weeks and 4 weeks, respectively. No severe events were reported during the study. Streptococcus salivarius K12 exhibited potential efficacy and safety as an adjuvant in treating oral candidiasis by enhancing mycological cure and shortening the treatment course of conventional antifungal therapy in this randomized, double-blinded, placebo-controlled clinical trial. Further large-scale clinical studies are desired to accumulate more evidence for its clinical applications.


Limitations of surface-color and apparent-color constancy.
Color-constancy mechanisms have been studied and discussed in a number of investigations. However, there has been little attempt to reveal how color constancy deteriorates as the conditions for it become less than optimal. We carried out a series of asymmetric color-matching experiments, using two criteria: surface-color match and apparent-color match. With brief adaptation the degree of color constancy increased as chromatic cues were added in the surround. In the condition of black surround, the test stimuli appeared self-luminous, and chromaticities of the chosen matching stimuli were the same as the physical chromaticities of the test stimulus, indicating a total deficiency of color constancy. With 15 min of preadaptation to the illuminant, the surface-color matches showed almost perfect color constancy under illuminant change. In both adaptation conditions, the chromatic-shift of matches from what would be expected for perfect color constancy increased gradually between 1,700- and 30,000-K illuminant, as chromaticity of the illuminant departed from 6,500-K illuminant. Under 1,000-K illuminant the surface-color appearance became totally achromatic, and color constancy was completely lost. Our results show that, even with brief adaptation to the illuminant, the contribution of the surrounding stimulus is large enough to achieve a fair degree of color constancy, but complete adaptation to the illuminant helps to achieve almost perfect color constancy.


A clinical study of competency to consent to treatment in pediatrics.
A 19-item competency questionnaire for pediatric patients (CQ-Peds) was used to evaluate competency to consent to treatment in pediatric outpatients and inpatients at two university hospitals. Sixty-nine consecutive English-speaking pediatric outpatients were studied at Hospital A, and 23 consecutive English-speaking pediatric inpatients were studied at Hospital B. Demographic data were statistically analyzed using the chi-square test, and there were no significant differences between the competent and incompetent groups (using CQ-Peds scores and cutoffs). CQ-Peds scores correlated highly with age (r = .947, p < .003; Outpatient Hospital A). Using the Child Behavior Checklist (CBCL) and the Pediatric Symptom Checklist (PSC) as a screen for psychopathology, the presence of psychiatric disturbance, per se, did not correlate with low CQ-Peds scores, nor was there a statistical difference between children from Spanish-speaking households and those from English-speaking households (Inpatient Hospital B). Overall, the children scored well on the CQ-Peds and demonstrated a good appreciation for their illnesses and treatment. The CQ-Peds score correlated highly with the that on the Wechsler Intelligence Scale for Children Revised Edition (WISC-R) vocabulary, comprehension, and similarities subtests and also with the Wide-Range Achievement Test-III (WRAT-III) reading assessment score (Inpatient Hospital B).


Genome-wide analysis of ARS (autonomously replicating sequence) binding factor 1 (Abf1p)-mediated transcriptional regulation in Saccharomyces cerevisiae.
Autonomously replicating sequence-binding factor-1 (Abf1p) is an essential sequence-specific transcription factor in Saccharomyces cerevisiae that participates in multiple nuclear events including DNA replication, transcription activation, and gene silencing. Numerous gene-specific analyses have implicated Abf1p in the transcriptional control of genes involved in a diverse range of cellular functions, leading to the notion that Abf1p acts as a global transcriptional regulator. Here we report findings from a genome-wide comparison of the gene expression profiles in the wild-type and abf1-1 temperature-sensitive mutant. The study identifies a total of 86 Abf1p-regulated genes (1.4% of the genome) of which 50 are activated and 36 are repressed by Abf1p. Interestingly, Abf1p binds to its own promoter in vivo and strongly represses its own transcription, suggesting a potential negative regulatory loop in Abf1p-mediated gene regulation. A comparison of our microarray data with the available databases reveals a significant overlap of genes regulated by Abf1p and those by several general transcription factors such as Mot1p and TAFs (TATA-binding protein-associated factors). Different mutant alleles of abf1 affect Abf1p-mediated transcription in a gene-dependent manner. Furthermore, Abf1p in vivo is associated with the promoter region of most Abf1p-activated but not with that of most Abf1p-repressed genes. Taken together, these results strongly suggest distinct underlying mechanisms by which Abf1p regulates gene expression.


Translational control of interleukin 2 messenger RNA as a molecular mechanism of T cell anergy.
T cell stimulation by triggering through the T cell receptor (TCR) in the absence of costimulatory signals or by calcium ionophore induces unresponsiveness in T cells to further stimulation, a phenomenon known as anergy. In freshly isolated T cells, calcium ionophore induces expression of interleukin (IL)-2 messenger (mRNA), but this mRNA is not translated and not loaded with ribosomes. In addition, while plate-bound anti-CD3 stimulation of resting T cells leads to IL-2 mRNA expression and IL-2 secretion, in cells pretreated with calcium ionophore before anti-CD3 stimulation, the IL-2 mRNA remains polysome unloaded and no IL-2 is produced. These observations show that IL-2 expression is controlled at the translational level, by differential ribosome loading. Furthermore, our data suggest that translational control of IL-2 mRNA may be a molecular mechanism by which anergy is attained.


Detection of carcinoembryonic antigen using single-domain or full-size antibodies stained with quantum dot conjugates.
Compact single-domain antibodies (sdAbs) are nearly 13 times smaller than full-size monoclonal antibodies (mAbs) and have a number of advantages for biotechnological applications, such as small size, high specificity, solubility, stability, and great refolding capacity. Carcinoembryonic antigen (CEA) is a tumor-associated glycoprotein expressed in a variety of cancers. Detection of CEA on the tumor cell surface may be carried out using anti-CEA antibodies and conventional fluorescent dyes. Semiconductor quantum dots (QDs) are brighter and more photostable than organic dyes; they provide the possibility for labeling of different recognition molecules with QDs of different colors but excitable with the same wavelength of excitation. In this study, the abilities for specific detection of CEA expressed by tumor cells with anti-CEA sdAbs biotinylated in vitro and in vivo, as well as with anti-CEA mAbs biotinylated in vitro, were compared using flow cytometry and the conjugates of streptavidin with QDs (SA-QDs). The results demonstrated that either in vitro or in vivo biotinylated anti-CEA sdAbs are more sensitive for cell staining compared to biotinylated anti-CEA mAbs. The data also show that simultaneous use of biotinylated sdAbs with highly fluorescent SA-QDs can considerably improve the sensitivity of detection of CEA on tumor cell surfaces.


Vectorcardiographic lead systems for the characterization of atrial fibrillation.
The aim of the study was to design a vectorcardiographic lead system dedicated to the analysis of atrial fibrillation (AF). Body surface potentials during AF were simulated by using a biophysical model of the human atria and thorax. The XYZ components of the equivalent dipole were derived from the Gabor-Nelson equations. These served as the gold standard while searching for an optimal orthogonal lead system for the estimation of the heart vector while using a limited number of electrode positions. Six electrode configurations and their dedicated transfer matrices were tested by using 10 different episodes of simulated AF and 25 different thorax geometries. Root-mean-square-based relative estimation error of the vectorcardiogram using the Frank electrodes was 0.39. An adaptation of 4 of the 9 electrode locations of the standard electrocardiogram, with 1 electrode moved to the back, reduced the error to 0.24. The Frank lead system is suboptimal for estimating the equivalent dipole components (VCG) during AF. Alternative electrode configurations should include at least 1 electrode on the back.


A detailed insight in the high risks of hospitalizations in long-term childhood cancer survivors-A Dutch LATER linkage study.
Insight in hospitalizations in long-term childhood cancer survivors (CCS) is useful to understand the impact of long-term morbidity. We aimed to investigate hospitalization rates and underlying types of diagnoses in CCS compared to matched controls, and to investigate the determinants. We linked 5,650 five-year CCS from the Dutch nationwide Dutch LATER cohort and 109,605 age- and sex-matched controls to the Dutch Hospital Discharge register, which contained detailed information on inpatient hospitalizations from 1995-2016. Relative hospitalization rates (RHRs) were calculated using a Poisson regression model. Adjusting for multiple hospitalizations per person via a Poisson model for generalized estimated equations, we investigated determinants for hospitalizations for all types of underlying diagnoses among CCS. CCS were twice as likely to be hospitalized as reference persons (hospitalization rate 178 and 78 per 1,000 person-years respectively; RHR 2.0, 95% confidence interval (CI) 1.9-2.2). Although CCS had more hospitalizations for 17 types of underlying diagnoses, they were especially more likely to be hospitalized for endocrine conditions (RHR: 6.0, 95% CI 4.6-7.7), subsequent neoplasms (RHR: 5.6, 95% CI 4.6-6.7) and symptoms without underlying diagnoses (RHR: 5.2, 95% CI 4.6-5.8). For those types of underlying diagnoses, female sex and radiotherapy were determinants. This study provides new insights in the high risk of hospitalizations for many types of underlying diagnoses in CCS and treatment related determinants. CCS are especially at high risk for hospitalizations for endocrine conditions, subsequent neoplasms and symptoms without an underlying diagnosis. This new knowledge is important for survivorship care and to identify possible preventable hospitalizations among CCS.


Correlation between ST segment shift and cardiac diastolic function in patients with acute myocardial infarction.
Diastolic dysfunction is the early sign of myocardial ischemia that usually occurs earlier than ECG changes. To determine the existence of a correlation between ST segment shift and diastolic dysfunction among patients with AMI. Fourty six patients with significant ST segment elevation or depression and having symptoms of acute myocardial infarction for <12 h were enrolled in this study. Patients were examined for serial ECG, cardiac enzymes, and echocardiography. There was no significant correlation between ST segment elevation or depression and majority of the diastolic indices. Few diastolic parameters; such as, E/A ratio of the mitral valve and deceleration time of the tricuspid valve showed a direct correlation with the ECG changes. Whereas, the Tei Index of the LV function showed a borderline correlation to the ST segment elevation at discharge. Overall, there was no correlation between either ST segment elevation or depression and the LV or RV diastolic function in patients with acute coronary syndrome (ACS). Improvement of the ST segment total score was associated with improvement of the diastolic function grades at discharge. Moreover, this association has shown an evidence of dose response relationship, the more improvement in total score at discharge the more improvement in diastolic function grade level.


Large-Scale Simulations of Plastic Neural Networks on Neuromorphic Hardware.
SpiNNaker is a digital, neuromorphic architecture designed for simulating large-scale spiking neural networks at speeds close to biological real-time. Rather than using bespoke analog or digital hardware, the basic computational unit of a SpiNNaker system is a general-purpose ARM processor, allowing it to be programmed to simulate a wide variety of neuron and synapse models. This flexibility is particularly valuable in the study of biological plasticity phenomena. A recently proposed learning rule based on the Bayesian Confidence Propagation Neural Network (BCPNN) paradigm offers a generic framework for modeling the interaction of different plasticity mechanisms using spiking neurons. However, it can be computationally expensive to simulate large networks with BCPNN learning since it requires multiple state variables for each synapse, each of which needs to be updated every simulation time-step. We discuss the trade-offs in efficiency and accuracy involved in developing an event-based BCPNN implementation for SpiNNaker based on an analytical solution to the BCPNN equations, and detail the steps taken to fit this within the limited computational and memory resources of the SpiNNaker architecture. We demonstrate this learning rule by learning temporal sequences of neural activity within a recurrent attractor network which we simulate at scales of up to 2.0 × 104 neurons and 5.1 × 107 plastic synapses: the largest plastic neural network ever to be simulated on neuromorphic hardware. We also run a comparable simulation on a Cray XC-30 supercomputer system and find that, if it is to match the run-time of our SpiNNaker simulation, the super computer system uses approximately 45× more power. This suggests that cheaper, more power efficient neuromorphic systems are becoming useful discovery tools in the study of plasticity in large-scale brain models.


Creating individual level air pollution exposures in an anonymised data safe haven: a platform for evaluating impact on educational attainment.
There is a lack of evidence on the adverse effects of air pollution on cognition for people with air quality-related health conditions. We propose that educational attainment, as a proxy for cognition, may increase with improved air quality. This study will explore whether asthma and seasonal allergic rhinitis, when exacerbated by acute exposure to air pollution, is associated with educational attainment. To describe the preparation of individual and household-level linked environmental and health data for analysis within an anonymised safe haven. Also to introduce our statistical analysis plan for our study: COgnition, Respiratory Tract illness and Effects of eXposure (CORTEX). We imported daily air pollution and aeroallergen data, and individual level education data into the SAIL databank, an anonymised safe haven for person-based records. We linked individual-level education, socioeconomic and health data to air quality data for home and school locations, creating tailored exposures for individuals across a city. We developed daily exposure data for all pupils in repeated cross sectional exam cohorts (2009-2015). We have used the SAIL databank, an innovative, data safe haven to create individual-level exposures to air pollution and pollen for multiple daily home and school locations. The analysis platform will allow us to evaluate retrospectively the impact of air quality on attainment for multiple cross-sectional cohorts of pupils. Our methods will allow us to distinguish between the pollution impacts on educational attainment for pupils with and without respiratory health conditions. The results from this study will further our understanding of the effects of air quality and respiratory-related health conditions on cognition. This city-wide study includes longitudinal routinely-recorded educational attainment data for all pupils taking exams over seven years;High spatial resolution air pollution data were linked within a privacy protected databank to obtain individual exposure at multiple daily locations;This study will use health data linked at the individual level to explore associations between air pollution, related morbidity, and educational attainment.


Making the case for uniformity in professional state licensure requirements.
Telehealth, the use of communication and information technologies to deliver health services, was initially envisioned as a way for persons in rural or remote settings to receive otherwise unavailable healthcare services. Now, in addition to overcoming personnel shortages for underserved populations, telehealth shows promise in meeting the needs of a constantly mobile U.S. society and workforce. Fortunately, telerehabilitation can meet the needs of a mobile society and workforce by enabling continuity of care for individuals who are out-of-town, on vacation, in temporary residence as a university student, or on business travel. Unfortunately, outdated legislative and regulatory policies and inhospitable infrastructures currently stand in the way of a seamless continuum of care. In 2010, the American Telemedicine Association's Telerehabilitation Special Interest Group (TR SIG) convened a License Portability Sub-Committee to explore ways to diminish barriers for state licensure portability with a particular focus on physical therapy, occupational therapy, speech therapy, and audiology. In 2011, the Subcommittee published a factsheet (1) that detailed the challenges and potential solutions that surround the difficult issue of licensure portability. Concurrently, the American Telemedicine Association is advocating for national reform of professional licensure. (2) At the heart of all licensure requirements is the ability to determine who should be granted the authority to practice in a particular profession. This is done by focusing on educational, examination and behavioral requirements that are deemed the minimum necessary to protect the public from harm. States, however, with whom authority for licensure of health professionals rests, have independently defined those minimum requirements. This approach has led to a myriad of requirements that vary from state to state. Licensure portability will best succeed when variability between licensure requirements is minimized and an efficient licensure process exists. In this paper, these two critical factors for licensure portability are referred to as "licensure requirements" and "the credentialing process." Currently the variability between both of these factors is different between professions as well as between jurisdictions. To find the best solution to licensure portability, it is critical to determine which of these two elements create significant barriers for licensure mobility. This document outlines a method for the professions to begin collecting data to pinpoint the areas where agreement and variations exist in licensure requirements and processes between states. Such information will inform efforts towards uniformity.


How metal ions affect amyloid formation: Cu2+- and Zn2+-sensitive peptides.
The common feature of proteins involved in many neurodegenerative diseases is their ability to adopt at least two different stable conformations. The conformational transition that shifts the equilibrium from the functional, mostly partially alpha-helical structure, to the beta-sheet rich amyloid can be triggered by numerous factors, such as mutations in the primary structure or changes in the environment. We present a set of model peptides that, without changes in their primary structure, react in a predictable fashion in the presence of transition metal ions by adopting different conformations and aggregate morphologies. These de novo designed peptides strictly follow the characteristic heptad repeat of the alpha-helical coiled-coil structural motif. Furthermore, domains that favor beta-sheet formation have been incorporated to make the system prone to amyloid formation. As a third feature, histidine residues create sensitivity towards the presence of transition metal ions. CD spectroscopy, ThT fluorescence experiments, and transmission electron microscopy were used to characterize peptide conformation and aggregate morphology in the presence of Cu2+ and Zn2+. Furthermore, the binding geometry within peptide-Cu2+ complexes was characterized by electron paramagnetic resonance spectroscopy.


The ability of Anopheles funestus and A. arabiensis to penetrate LLINs and its effect on their mortality.
Background: Variation in mosquito body size and the ability to penetrate long-lasting insecticide-treated nets (LLINs) remains unknown. This study evaluated the ability of Anopheles funestus and A. arabiensis to penetrate commercially available treated and untreated bednets and how this behaviour affects mosquito mortality. Methods: Three types of LLINs; DawaPlus 2.0, PermaNet 2.0, Olyset 2.0, and untreated (Safi Net) were tested inside a semi-field system. One hundred 3-5-day-old and non-starved female A. funestus and A. arabiensis were released in a chamber with a sleeping adult volunteer under a treated or untreated bednet. Mosquitoes that penetrated inside the nets were collected every two hours using a mouth aspirator. Live mosquitoes were put in paper cups, fed on glucose ad libitum and their mortality rate was monitored for 48 h. Results: The ability of A. funestus to penetrate treated and untreated bednets was significantly higher than for A. arabiensis for all three LLIN net types ( P<0.001). For both species the penetration rate was higher for untreated bednets than treated ones except for the Olyset net. Regardless of the assessed mosquito species, all the mosquitoes that penetrated the net, successfully blood-fed on the sleeping volunteer. Compared to A. arabiensis, significant mortality was recorded for A. funestus that were caught inside Olyset nets within 48 hrs of monitoring ( P<0.001). Conclusions: These findings demonstrate the ability of A. funestus and A. arabiensis mosquitoes to penetrate the human-occupied treated and untreated bednets. Despite this ability, mosquitoes that penetrated the bednet succumbed to death within two days.


SU-E-J-157: Simulation and Design of a Real-Time 6D Head Motion Compensation Platform Based on a Stewart Platform Approach.
Real-time sub-millimeter head motion compensation during frameless SRS delivery has the potential to achieve the accuracy of frame-based SRS while being significantly less invasive. Previously, we demonstrated real-time 6D head motion monitoring using an optical camera, however, at the time we were limited to only 3D (x-y-z) of head motion correction due to mechanical restrictions of the head platform. In this work we investigate the feasibility of using a compact 6D robotic Stewart platform (hexapod) placed under the patient's head to perform both translational and rotational motion compensation in real-time. Benefits of a hexapod approach over a conventional serial kinematics stage include less flex, compactness, high force to weight ratio, and fast response times. A hexapod is a parallel robotics device consisting of two platforms connected by six linear actuators oriented at particular angles. To provide accurate motion in 6D, the desired position of the top platform (head) was ascertained using inverse kinematics. MATLAB was used to simulate the six actuator positions for performing motion along x-y-z-phi -theta-psi. Prior recorded 6D human volunteer head motion data was used as an input for simulation of motion compensation. Six Firgelli L12-P linearservo actuators, together with a PCI-7344 motion controller and Labview software, were used for initial construction of a hexapod prototype. The necessary actuator lengths over time were computed for this data, simulating the required 6D movement of the hexapod for motion correction. Simulations on previously collected volunteer data indicate a hexapod system is capable of responding to subject head motion with corrections of precise movements, and solutions to the linear system can be computed at near real-time speeds. Based on simulated results, it was successfully demonstrated that a hexapod device can compensate for small patient head motions along all six degrees of freedom.


80-fs Nd:silicate glass laser pumped by a single-mode 200-mW diode.
A Nd(3+)-doped Schott LG680 silicate glass laser was pumped with a single-mode 200-mW diode. Efficient cw operation was demonstrated with 37.5 mW output power and 36% slope efficiency. Passive mode-locking with a semiconductor saturable absorber mirror yielded 80-fs pulses with a two-prism setup. Alternatively, pulses of approximately 200-fs, tunable over the range 1058-1076 nm, were obtained with either slit-tuning or a single-prism dispersive resonator. Output powers from 6 to 14 mW have been measured.


Risk of suicide and repeat self-harm after hospital attendance for non-fatal self-harm in Sri Lanka: a cohort study.
Evidence from high income countries (HICs) suggests that individuals who present to hospital after self-harm are an important target for suicide prevention, but evidence from low and middle-income countries (LMICs) is lacking. We aimed to investigate the risk of repeat self-harm and suicide, and factors associated with these outcomes, in a large cohort of patients presenting to hospital with self-harm in rural Sri Lanka. In this cohort study, hospital presentations for self-harm at 13 hospitals in a rural area of North Central Province (population 224 000), Sri Lanka, were followed up with a self-harm surveillance system, established as part of a community randomised trial, and based on data from all hospitals, coroners, and police stations in the study area. We estimated the risk of repeat non-fatal and fatal self-harm and risk factors for repetition with Kaplan-Meier methods and Cox proportional hazard models. Sociodemographic (age, sex, and socioeconomic position) and clinical (past self-harm and method of self-harm) characteristics investigated were drawn from a household survey in the study area and data recorded at the time of index hospital presentation. We included all individuals who had complete data for all variables in the study in our primary analysis. Between July 29, 2011, and May 12, 2016, we detected 3073 episodes of self-harm (fatal and non-fatal) in our surveillance system, of which 2532 (82·3%) were linked back to an individual in the baseline survey. After exclusion of 145 ineligible episodes, we analysed 2259 index episodes of self-harm. By use of survival models, the estimated risk of repeat self-harm (12 months: 3· 1%, 95% CI 2·4-3·9; 24 months: 5·2%, 4·3-6·4) and suicide (12 months: 0·6%, 0·4-1·1; 24 months: 0·8%, 0·5-1·3) in our study was considerably lower than that in HICs. A higher risk of repeat self-harm was observed in men than in women (fatal and non-fatal; hazard ratio 2·0, 95% CI 1·3-3·2; p=0·0021), in individuals aged 56 years and older compared with those aged 10-25 years (fatal; 16·1, 4·3-59·9; p=0·0027), and those who used methods other than poisoning in their index presentation (fatal and non-fatal; 3·9, 2·0-7·6; p=0·00027). We found no evidence of increased risk of repeat self-harm or suicide in those with a history of self-harm before the index episode. Although people who self-harm are an important high-risk group, focusing suicide prevention efforts on those who self-harm might be somewhat less important in LMICs compared with HICs given the low risk of repeat self-harm and subsequent suicide death. Strategies that focus on other risk factors for suicide might be more effective in reducing suicide deaths in LMICs in south Asia. A better understanding of the low incidence of repeat self-harm is also needed, as this could contribute to prevention strategies in nations with a higher incidence of repetition and subsequent suicide death. Wellcome Trust.


Evolution of GluN2A/B cytoplasmic domains diversified vertebrate synaptic plasticity and behavior.
Two genome duplications early in the vertebrate lineage expanded gene families, including GluN2 subunits of the NMDA receptor. Diversification between the four mammalian GluN2 proteins occurred primarily at their intracellular C-terminal domains (CTDs). To identify shared ancestral functions and diversified subunit-specific functions, we exchanged the exons encoding the GluN2A (also known as Grin2a) and GluN2B (also known as Grin2b) CTDs in two knock-in mice and analyzed the mice's biochemistry, synaptic physiology, and multiple learned and innate behaviors. The eight behaviors were genetically separated into four groups, including one group comprising three types of learning linked to conserved GluN2A/B regions. In contrast, the remaining five behaviors exhibited subunit-specific regulation. GluN2A/B CTD diversification conferred differential binding to cytoplasmic MAGUK proteins and differential forms of long-term potentiation. These data indicate that vertebrate behavior and synaptic signaling acquired increased complexity from the duplication and diversification of ancestral GluN2 genes.


mTORC2 regulates cardiac response to stress by inhibiting MST1.
The mTOR and Hippo pathways have recently emerged as the major signaling transduction cascades regulating organ size and cellular homeostasis. However, direct crosstalk between two pathways is yet to be determined. Here, we demonstrate that mTORC2 is a direct negative regulator of the MST1 kinase, a key component of the Hippo pathway. mTORC2 phosphorylates MST1 at serine 438 in the SARAH domain, thereby reducing its homodimerization and activity. We found that Rictor/mTORC2 preserves cardiac structure and function by restraining the activity of MST1 kinase. Cardiac-specific mTORC2 disruption through Rictor deletion leads to a marked activation of MST1 that, in turn, promotes cardiac dysfunction and dilation, impairing cardiac growth and adaptation in response to pressure overload. In conclusion, our study demonstrates the existence of a direct crosstalk between mTORC2 and MST1 that is critical for cardiac cell survival and growth.


Side effects of chaperone gene co-expression in recombinant protein production.
Insufficient availability of molecular chaperones is observed as a major bottleneck for proper protein folding in recombinant protein production. Therefore, co-production of selected sets of cell chaperones along with foreign polypeptides is a common approach to increase the yield of properly folded, recombinant proteins in bacterial cell factories. However, unbalanced amounts of folding modulators handling folding-reluctant protein species might instead trigger undesired proteolytic activities, detrimental regarding recombinant protein stability, quality and yield. This minireview summarizes the most recent observations of chaperone-linked negative side effects, mostly focusing on DnaK and GroEL sets, when using these proteins as folding assistant agents. These events are discussed in the context of the complexity of the cell quality network and the consequent intricacy of the physiological responses triggered by protein misfolding.


The semaphorins.
Semaphorins are secreted, transmembrane, and GPI-linked proteins, defined by cysteine-rich semaphorin protein domains, that have important roles in a variety of tissues. Humans have 20 semaphorins, Drosophila has five, and two are known from DNA viruses; semaphorins are also found in nematodes and crustaceans but not in non-animals. They are grouped into eight classes on the basis of phylogenetic tree analyses and the presence of additional protein motifs. The expression of semaphorins has been described most fully in the nervous system, but they are also present in most, or perhaps all, other tissues. Functionally, semaphorins were initially characterized for their importance in the development of the nervous system and in axonal guidance. More recently, they have been found to be important for the formation and functioning of the cardiovascular, endocrine, gastrointestinal, hepatic, immune, musculoskeletal, renal, reproductive, and respiratory systems. A common theme in the mechanisms of semaphorin function is that they alter the cytoskeleton and the organization of actin filaments and the microtubule network. These effects occur primarily through binding of semaphorins to their receptors, although transmembrane semaphorins also serve as receptors themselves. The best characterized receptors for mediating semaphorin signaling are members of the neuropilin and plexin families of transmembrane proteins. Plexins, in particular, are thought to control many of the functional effects of semaphorins; the molecular mechanisms of semaphorin signaling are still poorly understood, however. Given the importance of semaphorins in a wide range of functions, including neural connectivity, angiogenesis, immunoregulation, and cancer, much remains to be learned about these proteins and their roles in pathology and human disease.


Some properties of human platelet monoamine oxidase in iron-deficiency anaemia.
1. Monoamine oxidase activity in platelets prepared from the blood of patients with iron-deficiency anaemia was significantly lowered when compared with that in platelets from normal subjects. 2. The Km values of the platelet enzyme for the substrates dopamine, 5-hydroxytryptamine, phenylethylamine and kynuramine were similar for the platelet enzyme from iron-deficient and normal groups. 3. Heat-in-activation studies showed that the platelet monoamine oxidase from iron-deficient subjects was more labile to this treatment, when compared with the platelet enzyme from normal subjects. 4. The sensitivity of platelet monoamine oxidase to the inhibitors, clorgyline and deprenil, was increased in iron-deficiency anaemia. 5. Binding studies with the 14C-binding irreversible monoamine oxidase inhibitor, deprenil, showed that the amount of enzyme capable of binding this inhibitor was lowered by 48% in platelets from iron-deficient patients when compared with platelets from normal subjects. 6. The results show that there is a lowered amount of active enzyme in platelets from iron-deficient subjects. It is suggested that iron is necessary either for the synthesis of monoamine oxidase apoenzyme or is a cofactor for an enzyme which attaches flavin-adenine dinucleotide covalently to the monoamine oxidase apoenzyme.


Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen.
Detection of anti-Coccidioides complement-fixing (CF) antibody is a valuable diagnostic and prognostic aid in coccidioidomycosis. The CF antibody response is directed against a heat-labile antigen that has chitinase activity, hereafter referred to as the CF/chitinase protein. To identify and clone this immunoreactive enzyme, we constructed a Coccidioides immitis cDNA lambda ZAP expression library from spherule RNA and detected fusion peptides expressing CF epitopes by immunoscreening. A cDNA clone consisting of 1,623 bp was identified, sequenced, and found to contain a single open reading frame that encodes a protein of 47 kDa with 427 amino acids. Deduced amino acid sequence analyses showed that the cloned CF/chitinase cDNA contains a 35-amino-acid region, beginning at Ser-18 and ending at and ending at Arg-52 which has 92% homology with the reported N-terminal amino acid sequence of authentic CF/chitinase protein. The first 17 amino acids in the deduced sequence of the cloned cDNA are not present on the mature CF/chitinase protein, suggesting that it may be a signal peptide. Expression of the CF/chitinase cDNA insert by using the pGEX-4T-3 vector yields a fusion peptide that bears CF-specific epitopes and shows chitinase activity. The CF/chitinase clone will enable large-scale production of the recombinant CF antigen for use in immunoassays and facilitate studies on the role of chitinase in the morphogenesis of C. immitis.


Significant enhancement of the selectivity of propylene epoxidation for propylene oxide: a molecular oxygen mechanism.
As an attractive and environmentally friendly process for propylene oxide (PO) production, direct epoxidation of propylene (DEP) with molecular oxygen catalyzed by metal-based catalysts such as Ag and Cu has drawn much attention, but remains one of the biggest challenges in chemistry. In this work, the crucial competitive reactions of propylene α-H stripping (AHS) versus the oxametallacycle formation (OMMP formation) using adsorbed atomic oxygen (O*) or adsorbed molecular oxygen (O2*) as an oxidant are extensively compared on IB group metal surfaces (Cu, Ag and Au) with varied electronic and structural effects in order to explore the possibility to enhance the PO selectivity by virtue of first-principles calculations. The determining factor for the PO selectivity is quantitatively revealed: it is found that with atomic O*, the AHS pathway was preferred, indicating the reason for low PO selectivity with current catalysts. By contrast, the undissociated molecular O2* species is found to prefer to electrophilically attack the C[double bond, length as m-dash]C double bond of propylene and form a special oxametallacycle intermediate (OOMMP) rather than nucleophilically abstracting the α-H. This OOMMP can readily cleave the O-O bond and transform into OMMP. These results demonstrate that the presence of undissociated O2* can efficiently promote the PO selectivity. Furthermore, the merit of such a molecular O2* mechanism can be rationalized by our quantitative barrier decomposition analyses, which reveal that the lower hydrogen affinity (ΔEH) of the O2* species dominantly contributes to the limited AHS reaction, and boosts the OMMP selectivity. Therefore, ΔEH can be applied as a selectivity descriptor. An efficient strategy to promote PO formation is presented. The insight obtained could pave the way for further development of catalysts for propylene epoxidation.


Depressive symptoms, race, and circulating C-reactive protein: the Coronary Artery Risk Development in Young Adults (CARDIA) study.
To examine the prospective association of depressive symptoms with circulating C-reactive protein (CRP) and to determine the direction of that association. Using data from 2,544 healthy participants in the Coronary Artery Risk Development in Young Adults study (ages, 33-45 years; 55% female; 42% black), we examined the prospective association of depressive symptoms, as measured by the Centers for Epidemiologic Studies Depression Scale, with circulating CRP 5 years later. Depressive symptoms in the Coronary Artery Risk Development in Young Adults study Year 15 predicted CRP at Year 20, independent of demographic characteristics, biological and medical risk factors, health behaviors, and Year 15 CRP. This association, however, was conditional on race such that the increase in CRP with increasing depressive symptoms was present in blacks but not whites. In neither blacks nor whites did Year 15 CRP predict Year 20 depressive symptoms. Among black participants, when examined in separate analyses, higher scores on the depressed affect and somatic symptoms subscales of the Centers for Epidemiologic Studies Depression Scale and lower scores on the positive affect subscale were associated with greater Year 20 CRP. The interpersonal problems subscale was unrelated to CRP. When all four subscale scores were entered simultaneously in the same model, black participants' scores on the positive affect and somatic symptoms subscales emerged as independent predictors of Year 20 CRP, whereas the depressed affect and interpersonal problems subscales did not. Depressive symptoms may be linked more closely to inflammation in blacks than in whites.


Living Arrangements and the Well-being of Single Mothers in Japan.
The goal of this paper is to evaluate the extent to which the well-being of single mothers in Japan is related to coresidence with other adults. Using data from a representative survey of households headed by single mothers, we examine two measures of subjective well-being: perceived economic circumstances and self-rated health. One-fourth of the single mothers surveyed were coresiding with another adult(s) and it is clear that these women fare significantly better than their non-coresiding counterparts on both measures of well-being. Net of several theoretically relevant sociodemographic, family, and employment characteristics, single mothers living with others were significantly less likely to report somewhat difficult/difficult economic circumstances or fair/poor health. Efforts to account for potential endogeneity between well-being and living arrangements suggested that self-rated health, but not subjective economic well-being, is related to selection into coresidence. Single mothers in fair/poor health appear more likely to coreside with others and, accounting for this selection, intergenerational coresidence appears to be very beneficial for self-rated health. We discuss the implications of these findings for processes of stratification in Japan in light of the limited public income support available to single mothers.


Effect of a US National Institutes of Health programme of clinical trials on public health and costs.
Few attempts have been made to estimate the public return on investment in medical research. The total costs and benefits to society of a clinical trial, the final step in testing an intervention, can be estimated by evaluating the effect of trial results on medical care and health. All phase III randomised trials funded by the US National Institute of Neurological Disorders and Stroke before Jan 1, 2000, were included. Pertinent publications on use, cost to society, and health effects for each studied intervention were identified by systematic review, supplemented with data from other public and proprietary sources. Regardless of whether a trial was positive or negative, information on use of tested therapies was integrated with published per-use data on costs and health effect (converted to 2004 US dollars) to generate 10-year projections for the US population. 28 trials with a total cost of 335 million dollars were included. Six trials (21%) resulted in measurable improvements in health, and four (14%) resulted in cost savings to society. At 10 years, the programme of trials resulted in an estimated additional 470,000 quality-adjusted life years at a total cost of 3.6 billion dollars (including costs of all trials and additional health-care and other expenditures). Valuing a quality-adjusted life year at per-head gross domestic product, the projected net benefit to society at 10-years was 15.2 billion dollars. 95% CIs did not include a net loss at 10 years. For this institute, the public return on investment in clinical trials has been substantial. Although results led to increases in health-care expenditures, health gains were large and valuable.


[Reconstruction of isolated axillary nerve lesions: clinical and electrophysiological long-term results].
The purpose of this retrospective study was the clinical and electrophysiological assessment of long-term results following surgical treatment of isolated axillary nerve lesions. 10 patients, who underwent axillary nerve reconstruction using a nerve graft (80%) or a neurolysis alone (20%), with a follow-up period of 6.7 years (1.6-10.8 years) on average, were included in the study. In addition to the clinical examination, we examined reinnervation by electrophysiological methods and analysed their correlation to clinical results. 30% of the patients had muscle strength of M5 and full active range of motion of 180°, 40% reached M4 and averaged 170° (80-180°), 30% of the patients M3 and 65° (40-90°). In patients with M5, muscle compound action potential averaged 40% of the contralateral side (21-62%), in patients with M4 36% (29-58%), and in patients with M3 7% (3-11%). Good results were associated with younger patient age, a short delay prior to the operation and neurolysis alone (indicated by intraoperative electroneurography). The good results confirm our treatment algorithm for isolated axillary nerve lesions by neurolysis alone or reconstruction with autologous nerve transplantation. Measurement of compound muscle action potential is a valuable addition to difficult clinical assessment, as its amplitude enables quantification of axillary nerve recovery.


Overlapping and distinct roles of Aspergillus fumigatus UDP-glucose 4-epimerases in galactose metabolism and the synthesis of galactose-containing cell wall polysaccharides.
The cell wall of Aspergillus fumigatus contains two galactose-containing polysaccharides, galactomannan and galactosaminogalactan, whose biosynthetic pathways are not well understood. The A. fumigatus genome contains three genes encoding putative UDP-glucose 4-epimerases, uge3, uge4, and uge5. We undertook this study to elucidate the function of these epimerases. We found that uge4 is minimally expressed and is not required for the synthesis of galactose-containing exopolysaccharides or galactose metabolism. Uge5 is the dominant UDP-glucose 4-epimerase in A. fumigatus and is essential for normal growth in galactose-based medium. Uge5 is required for synthesis of the galactofuranose (Galf) component of galactomannan and contributes galactose to the synthesis of galactosaminogalactan. Uge3 can mediate production of both UDP-galactose and UDP-N-acetylgalactosamine (GalNAc) and is required for the production of galactosaminogalactan but not galactomannan. In the absence of Uge5, Uge3 activity is sufficient for growth on galactose and the synthesis of galactosaminogalactan containing lower levels of galactose but not the synthesis of Galf. A double deletion of uge5 and uge3 blocked growth on galactose and synthesis of both Galf and galactosaminogalactan. This study is the first survey of glucose epimerases in A. fumigatus and contributes to our understanding of the role of these enzymes in metabolism and cell wall synthesis.


Preservation of donor hearts using hypothermic oxygenated perfusion.
Hypothermic machine perfusion of donor hearts enables continuous aerobic metabolism and washout of toxic metabolic byproducts. We evaluated the effect of machine perfusion on cardiac myocyte integrity in hearts preserved for 4 h in a novel device that provides pulsatile oxygenated hypothermic perfusion (Paragonix Sherpa Perfusion™ Cardiac Transport System). Pig hearts were harvested and stored in Celsior® solution for 4 h using either conventional cold storage on ice (4-h CS, n=6) or the Sherpa device (4-h pulsatile perfusion (PP), n=6). After cold preservation, hearts were evaluated using a non-working heart Langendorff system. Controls (n=3) were reperfused immediately after organ harvest. Biopsies were taken from the apex of the left ventricle before storage, after storage, and after reperfusion to measure ATP content and endothelin-1 in the tissue. Ultrastructural analysis using electron microscopy was performed. Four-hour CS, 4-h PP, and control group did not show any significant differences in systolic or diastolic function (+dP/dt, -dP/dt, EDP). Four-hour PP hearts showed significantly more weight gain than 4-h CS after preservation, which shows that machine perfusion led to myocardial edema. Four-hour CS led to higher endothelin-1 levels after preservation, suggesting more endothelial dysfunction compared to 4-h PP. Electron microscopy revealed endothelial cell rupture and damaged muscle fibers in the 4-h CS group after reperfusion, but the cell structures were preserved in the 4-h PP group. Hypothermic pulsatile perfusion of donor hearts leads to a better-preserved cell structure compared to the conventional cold storage method. This may lead to less risk of primary graft failure after orthotopic heart transplantation.


Challenges of quality improvement in the healthcare of South Africa post-apartheid: A critical review.
There is overwhelming evidence that the quality of health care in South Africa has been compromised by various challenges that impact negatively on healthcare quality. Improvement in quality care means fewer errors, reduced delays in care delivery, improvement in efficiency, increased market share and lower cost. Decline in quality health care has caused the public to lose trust in the healthcare system in South Africa. The purpose of this study was to identify challenges that are being incurred in practice that compromise quality in the healthcare sector, including strategies employed by government to improve the quality of health delivery. Literature search included the following computer-assisted databases and bibliographies: Medline (Medical Literature Online), EBSCOhost, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Google, Google Scholar and ScienceDirect. Furthermore, websites were used to source policy documents of organisations such as the National Department of Health in South Africa and the World Health Organization. Seventy-four articles were selected from 1366 retrieved. These articles quantify problems facing quality care delivery and strategies used to improve the healthcare system in South Africa. The findings revealed that there were many quality improvement programmes that had been initiated, adapted, modified and then tested but did not produce the required level of quality service delivery as desired. As a result, the Government of South Africa has a challenge to ensure that implementation of National Core Standards will deliver the desired health outcomes, because achieving a lasting quality improvement system in health care seems to be an arduous challenge.


Microstructured fiber splicing.
We present experimental results on Microstructured Optical Fiber (MOF) splicing with a simple method relying on conventional electric-arc splicers. The results are presented in terms of fusion losses and tensile strength. An electric-arc splicing system is used to demonstrate its effectiveness in splicing MOFs together as well as splicing MOF with a single mode fiber.


Meningioma mouse models.
Meningiomas, although mostly benign, may sometimes present aggressive features and raise issues concerning alternative treatment options besides surgery. In order to gain new insights in meningioma biology and develop alternative treatments, several meningioma mouse models have been engineered during the past two decades. As rodents very rarely develop spontaneous meningiomas, animal models have been first developed by implanting human meningioma cells derived from a primary tumor and meningioma cell lines subcutaneously into athymic mice. Induction of de novo meningiomas in rodents with mutagens, such as nitrosourea, has also been reported. Advances in our understanding of molecular genetics of meningioma have pinpointed the central role of NF2 tumor suppressor gene in the pathogenesis of those tumors. These discoveries have led to the creation of a genetically engineered model utilizing conditional mutagenesis to specifically inactivate the mouse Nf2 gene in arachnoidal cells, resulting in the formation of intracranial meningothelial hyperplasia and meningiomas and thus reproducing the main mechanism of human meningeal tumorigenesis. This powerful new technology significantly improves on prior models and may open avenues of investigation never before possible in meningioma research. We present here a review of current meningioma mouse models used in translational therapeutics with associated imaging and pre-clinical studies.


Nucleophosmin mediates mammalian target of rapamycin-dependent actin cytoskeleton dynamics and proliferation in neurofibromin-deficient astrocytes.
Neurofibromatosis type 1 (NF1) is a common autosomal dominant tumor predisposition syndrome in which affected individuals develop astrocytic brain tumors (gliomas). To determine how the NF1 gene product (neurofibromin) regulates astrocyte growth and motility relevant to glioma formation, we have used Nf1-deficient primary murine astrocytes. Nf1(-/-) astrocytes exhibit increased protein translation and cell proliferation, which are mediated by Ras-dependent hyperactivation of the mammalian target of rapamycin (mTOR) protein, a serine/threonine protein kinase that regulates ribosomal biogenesis, protein translation, actin cytoskeleton dynamics, and cell proliferation. In this study, we show that Nf1-deficient astrocytes have fewer actin stress fibers and exhibit increased cell motility compared with wild-type astrocytes, which are rescued by pharmacologic and genetic mTOR inhibition. We further show that mTOR-dependent regulation of actin stress fiber formation, motility, and proliferation requires rapamycin-sensitive activation of the Rac1 GTPase but not elongation factor 4E-binding protein 1/S6 kinase. Nf1(-/-) astrocytes also exhibit increased protein translation and ribosomal biogenesis through increased expression of the nucleophosmin (NPM) nuclear-cytoplasmic shuttling protein. We found that NPM expression in Nf1(-/-) astrocytes was blocked by rapamycin in vitro and in vivo and that expression of a dominant-negative NPM mutant protein in Nf1(-/-) astrocytes rescued actin stress fiber formation and restored cell motility and proliferation to wild-type levels. Together, these data show that neurofibromin regulates actin cytoskeleton dynamics and cell proliferation through a mTOR/Rac1-dependent signaling pathway and identify NPM as a critical mTOR effector mediating these biological properties in Nf1-deficient astrocytes.


A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm.
One of our monoclonal antibodies against the heavy chain of bovine kinesin (H2) also recognized the heavy chain of squid kinesin. The immunofluorescence pattern of H2 in axoplasm was similar to that seen in mammalian cells with antibodies specific for kinesin light and heavy chains, indicating that squid kinesin is also concentrated on membrane-bounded organelles. Although kinesin is assumed to be a motor for translocation of membrane-bounded organelles in fast axonal transport, direct evidence has been lacking. Perfusion of axoplasm with purified H2 at 0.1-0.4 mg/ml resulted in a profound inhibition of both the rates and number of organelles moving in anterograde and retrograde directions in the interior of the axoplasm, and comparable inhibition was noted in bidirectional movement along individual microtubules at the periphery. Maximal inhibition developed over 30-60 min. Perfusion with higher concentrations of H2 (greater than 1 mg of IgG per ml) were less effective, whereas perfusion with 0.04 mg of H2 per ml resulted in minimal inhibition. Movement of membrane-bounded organelles after perfusion with comparable levels of irrelevant mouse IgG (0.04 to greater than 1 mg/ml) were not distinguishable from perfusion with buffer controls. Inhibition of fast axonal transport by an antibody specific for kinesin provides direct evidence that kinesin is involved in the translocation of membrane-bounded organelles in axons. Moreover, the inhibition of bidirectional axonal transport by H2 raises the possibility that kinesin may play some role in both anterograde and retrograde axonal transport.


Determining hazard distances from non-Gaussian lasers.
A simple method is presented for determining the nominal ocular hazard distance from non-Gaussian laser beams. The simplified method currently used for laser safety calculations when determining the laser beam diameter as a function of distance produces errors in the near field. The irradiance or radiant exposure of multimode and non-Gaussian laser beams in the near field may be calculated more precisely by the use of a simple proposed relationship with easily determined parameters.


Comprehensive quantification of herpes simplex virus latency at the single-cell level.
To date, characterization of latently infected tissue with respect to the number of cells in the tissue harboring the viral genome and the number of viral genomes contained within individual latently infected cells has not been possible. This level of cellular quantification is a critical step in determining (i) viral or host cell factors which function in the establishment and maintenance of latency, (ii) the relationship between latency burden and reactivation, and (iii) the effectiveness of vaccines or antivirals in reducing or preventing the establishment of latent infections. Presented here is a novel approach for the quantitative analysis of nucleic acids within the individual cells comprising complex solid tissues. One unique feature is that the analysis reflects the nucleic acids within the individual cells as they were in the context of the intact tissue-hence the name CXA, for contextual analysis. Trigeminal ganglia latently infected with herpes simplex virus (HSV) were analyzed by CXA of viral DNA. Both the type and the number of cells harboring the viral genome as well as the number of viral genomes within the individual latently infected cells were determined. Here it is demonstrated that (i) the long-term repository of HSV-1 DNA in the ganglion is the neuron, (ii) the viral-genome copy number within individual latently infected neurons is variable, ranging over 3 orders of magnitude from <10 to >1,000, (iii) there is a direct correlation between increasing viral input titer and the number of neurons in which latency is established in the ganglion, (iv) increasing viral input titer results in more neurons with greater numbers of viral-genome copies, (v) treatment with acyclovir (ACV) during acute infection reduces the number of latently infected ganglionic neurons 20-fold, and (vi) ACV treatment results in uniformly low (<10)-copy-number latency. This report represents the first comprehensive quantification of HSV latency at the level of single cells. Beyond viral latency, CXA has the potential to advance many studies in which rare cellular events occur in the background of a complex solid tissue mass, including microbial pathogenesis, tumorigenesis, and analysis of gene transfer.


Stem cell therapy and the retina.
Retinal degeneration culminating in photoreceptor loss is the leading cause of untreatable blindness in the developed world. In this review, we consider how photoreceptors might be replaced by transplantation and how stem cells might be optimised for use as donor cells in future clinical strategies for retinal repair. We discuss the current advances in human and animal models of retinal cell transplantation, focussing on stem cell and reproductive cloning biology, in relation to the practical issues of retinal transplantation surgery. Stem and progenitor cells can be isolated from a number of sources including embryonic tissue, adult brain and even the retina, prompting many researchers to investigate the potential for using these cells to generate photoreceptors for transplantation. Nevertheless, several obstacles need to be overcome before these techniques can be applied in a clinical setting. Embryonic or stem cells have so far shown little ability to differentiate into retinal phenotypes when transplanted into the adult retina. We have recently noted, however, that donor cells harvested much later, at the photoreceptor precursor developmental stage, can be transplanted successfully and restore visual function. The current challenge is to understand the developmental processes that guide embryonic or adult stem cells towards photoreceptor differentiation, so that large numbers of these cells might be transplanted at the optimal stage. Future advances in reproductive cloning technology could lead to the successful generation of stem cells from adult somatic cells, thereby facilitating auto-transplantation of genetically identical cells in patients requiring photoreceptor replacement.


A Destabilizing Domain Allows for Fast, Noninvasive, Conditional Control of Protein Abundance in the Mouse Eye - Implications for Ocular Gene Therapy.
Temporal and reversible control of protein expression in vivo is a central goal for many gene therapies, especially for strategies involving proteins that are detrimental to physiology if constitutively expressed. Accordingly, we explored whether protein abundance in the mouse retina could be effectively controlled using a destabilizing Escherichia coli dihydrofolate reductase (DHFR) domain whose stability is dependent on the small molecule, trimethoprim (TMP). We intravitreally injected wild-type C57BL6/J mice with an adeno-associated vector (rAAV2/2[MAX]) constitutively expressing separate fluorescent reporters: DHFR fused to yellow fluorescent protein (DHFR.YFP) and mCherry. TMP or vehicle was administered to mice via oral gavage, drinking water, or eye drops. Ocular TMP levels post treatment were quantified by LC-MS/MS. Protein abundance was measured by fundus fluorescence imaging and western blotting. Visual acuity, response to light stimulus, retinal structure, and gene expression were evaluated after long-term (3 months) TMP treatment. Without TMP, DHFR.YFP was efficiently degraded in the retina. TMP achieved ocular concentrations of ∼13.6 μM (oral gavage), ∼331 nM (drinking water), and ∼636 nM (eye drops). Oral gavage and TMP eye drops stabilized DHFR.YFP as quickly as 6 hours, whereas continuous TMP drinking water could stabilize DHFR.YFP for ≥3 months. Stabilization was completely and repeatedly reversible following removal/addition of TMP in all regimens. Long-term TMP treatment had no impact on retina function/structure and had no effect on >99.9% of tested genes. This DHFR-based conditional system is a rapid, efficient, and reversible tool to effectively control protein expression in the retina.


Holding the Nucleosome Together: A Quantitative Description of the DNA-Histone Interface in Solution.
The nucleosome is the fundamental unit of eukaryotic genome packaging in the chromatin. In this complex, the DNA wraps around eight histone proteins to form a superhelical double helix. The resulting bending, stronger than anything observed in free DNA, raises the question of how such a distortion is stabilized by the proteic and solvent environments. In this work, the DNA-histone interface in solution was exhaustively analyzed from nucleosome structures generated by molecular dynamics. An original Voronoi tessellation technique, measuring the topology of interacting elements without any empirical or subjective adjustment, was used to characterize the interface in terms of contact area and occurrence. Our results revealed an interface more robust than previously known, combining extensive, long-lived nonelectrostatic and electrostatic interactions between DNA and both structured and unstructured histone regions. Cation accumulation makes the proximity of juxtaposed DNA gyres in the superhelix possible by shielding the strong electrostatic repulsion of the charged phosphate groups. Overall, this study provides new insights on the nucleosome cohesion, explaining how DNA distortions can be maintained in a nucleoprotein complex.


Effects of a physiological insulin concentration on the endothelin-sensitive Ca2+ store in porcine coronary artery smooth muscle.
The effect of insulin to attenuate the Ca2+ and contractile response of vascular smooth muscle to a number of agonists has been described previously, but the Ca2+ regulatory mechanisms of insulin action remain unclear. We determined the effect of a physiological insulin concentration (300 pmol/l) on the Ca2+ response of vascular smooth muscle cells of the porcine right coronary artery to endothelin 1 (ET-1); furthermore, we examined the cellular Ca2+ stores affected by insulin (i.e., Ca2+ stores releasable by inositol 1,4,5-trisphosphate, caffeine, and ionomycin). We measured the Ca2+ responses of acutely isolated single smooth muscle cells with the fluorescent Ca2+ indicator Fura-2. Acute insulin exposure (20 min) significantly attenuated the Ca2+ response of single smooth muscle cells to 10 nmol/l ET-1. This inhibitory effect of insulin was observed both in the presence and absence of extracellular Ca2+. In contrast with the effects on ET-1-induced Ca2+ responses, insulin did not inhibit the Ca2+ response to 5 mmol/l caffeine, an agent that directly releases sarcoplasmic reticulum Ca2+ stores. Insulin was also without effect on the total cellular Ca2+ store released by 1 micromol/l ionomycin, a Ca2+-transporting ionophore. When ET-1 and caffeine were given in succession, a sizable caffeine-sensitive Ca2+ store could be released from insulin-treated cells but not control cells, indicating that the sarcoplasmic reticulum Ca2+ store of insulin-treated cells was not depleted by ET-1. Generalized depletion of the sarcoplasmic reticulum Ca2+ store is not one of the cellular mechanisms involved in the effect of insulin on coronary smooth muscle; instead, the effect may be due to an inhibitory influence on transmembrane signal transduction, such as diminished ET-1-induced inositol 1,4,5-trisphosphate production or reduced ability of this phosphoinositol to release stored Ca2+.


Assessment of Cr(VI)-induced cytotoxicity and genotoxicity using high content analysis.
Oral exposure to high concentrations of hexavalent chromium [Cr(VI)] induces intestinal redox changes, villus cytotoxicity, crypt hyperplasia, and intestinal tumors in mice. To assess the effects of Cr(VI) in a cell model relevant to the intestine, undifferentiated (proliferating) and differentiated (confluent) Caco-2 cells were treated with Cr(VI), hydrogen peroxide or rotenone for 2-24 hours. DNA damage was then assessed by nuclear staining intensity of 8-hydroxydeoxyguanosine (8-OHdG) and phosphorylated histone variant H2AX (γ-H2AX) measured by high content analysis methods. In undifferentiated Caco-2, all three chemicals increased 8-OHdG and γ-H2AX staining at cytotoxic concentrations, whereas only 8-OHdG was elevated at non-cytotoxic concentrations at 24 hr. Differentiated Caco-2 were more resistant to cytotoxicity and DNA damage than undifferentiated cells, and there were no changes in apoptotic markers p53 or annexin-V. However, Cr(VI) induced a dose-dependent translocation of the unfolded protein response transcription factor ATF6 into the nucleus. Micronucleus (MN) formation was assessed in CHO-K1 and A549 cell lines. Cr(VI) increased MN frequency in CHO-K1 only at highly cytotoxic concentrations. Relative to the positive control Mitomycin-C, Cr(VI) only slightly increased MN frequency in A549 at mildly cytotoxic concentrations. The results demonstrate that Cr(VI) genotoxicity correlates with cytotoxic concentrations, and that H2AX phosphorylation occurs at higher concentrations than oxidative DNA damage in proliferating Caco-2 cells. The findings suggest that in vitro genotoxicity of Cr(VI) is primarily oxidative in nature at low concentrations. Implications for in vivo intestinal toxicity of Cr(VI) will be discussed.


Exogenous low-dose hydrogen peroxide enhances drought tolerance of soybean (Glycine max L.) through inducing antioxidant system.
Hydrogen peroxide (H(2)O(2)) functions as a signal molecule in plants under abiotic and biotic stress. In this study, the role of exogenous H(2)O(2) in improving drought tolerance in two soybean cultivars (Glycine max L. Merrill) differing in their tolerance to drought was evaluated. Plants were grown in plastic pots with normal irrigation in a phytotron. Four weeks after radicle emergence, either 1 mM H(2)O(2) or distilled water was sprayed as foliar onto the leaves of each plant, after drought stress was applied. Leaf samples were harvested on the 4(th) and 7(th) days of the drought. Antioxidant-related enzyme activity, such as the superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), hydrogen peroxide (H(2)O(2)) and malondialdehyde (MDA) content was measured during the drought period. Drought stress decreased leaf water potential, relative water content and photosynthetic pigment content but enhanced lipid peroxidation and endogenous H(2)O(2) concentration. By contrast, exogenous low dose H(2)O(2) improved water status, pigment content and lipid peroxidation under drought stress. Endogenous H(2)O(2) concentration was reduced by exogenous H(2)O(2) as compared to drought treatment alone. H(2)O(2) pre-treatment induced all the antioxidant enzyme activities, to a greater extent than the control leaves, during drought. H(2)O(2) pretreatment further enhanced the activities of antioxidant enzymes in the tolerant cultivar compared to the sensitive cultivar. Results suggested that low dose H(2)O(2) pre-treatment alleviated water loss and H(2)O(2) content and increased drought stress tolerance by inducing the antioxidant system.


Crystal structure of a monomeric retroviral protease solved by protein folding game players.
Following the failure of a wide range of attempts to solve the crystal structure of M-PMV retroviral protease by molecular replacement, we challenged players of the protein folding game Foldit to produce accurate models of the protein. Remarkably, Foldit players were able to generate models of sufficient quality for successful molecular replacement and subsequent structure determination. The refined structure provides new insights for the design of antiretroviral drugs.


Dietary fructose accelerates the development of diabetes in UCD-T2DM rats: amelioration by the antioxidant, alpha-lipoic acid.
Sustained fructose consumption has been shown to induce insulin resistance and glucose intolerance, in part, by promoting oxidative stress. Alpha-lipoic acid (LA) is an antioxidant with insulin-sensitizing activity. The effect of sustained fructose consumption (20% of energy) on the development of T2DM and the effects of daily LA supplementation in fructose-fed University of California, Davis-Type 2 diabetes mellitus (UCD-T2DM) rats, a model of polygenic obese T2DM, was investigated. At 2 mo of age, animals were divided into three groups: control, fructose, and fructose + LA (80 mg LA.kg body wt(-1).day(-1)). One subset was followed until diabetes onset, while another subset was euthanized at 4 mo of age for tissue collection. Monthly fasted blood samples were collected, and an intravenous glucose tolerance test (IVGTT) was performed. Fructose feeding accelerated diabetes onset by 2.6 +/- 0.5 mo compared with control (P < 0.01), without affecting body weight. LA supplementation delayed diabetes onset in fructose-fed animals by 1.0 +/- 0.7 mo (P < 0.05). Fructose consumption lowered the GSH/GSSG ratio, while LA attenuated the fructose-induced decrease of oxidative capacity. Insulin sensitivity, as assessed by IVGTT, decreased in both fructose-fed and fructose + LA-supplemented rats. However, glucose excursions in fructose-fed LA-supplemented animals were normalized to those of control via increased glucose-stimulated insulin secretion. Fasting plasma triglycerides were twofold higher in fructose-fed compared with control animals at 4 mo, and triglyceride exposure during IVGTT was increased in both the fructose and fructose + LA groups compared with control. In conclusion, dietary fructose accelerates the onset of T2DM in UCD-T2DM rats, and LA ameliorates the effects of fructose by improving glucose homeostasis, possibly by preserving beta-cell function.


Incidence of behavior problems in toddlers and preschool children from families living in poverty.
Few studies have examined the incidence of behavior problems in toddlers and preschool children from families living in poverty. The available research suggests behavior problems occur at higher rates in children living in poverty and may have long-term negative outcomes if not identified and properly treated. This study included an ethnically representative sample of 357 children, five years of age and younger, from a diverse, low-income, urban area. All families' incomes met the federal threshold for living in poverty. Behavior problems were assessed by parent report through a questionnaire specifically designed for low-income families. Boys and younger children were reported as demonstrating a higher rate of externalizing behaviors than girls and older children. The overall rate of children scoring at least one standard deviation above the sample's mean for challenging behaviors was 17.4% and was not related to the child's gender, age or ethnicity. This study also sampled children's positive behaviors, which is unique in studies of behavior problems. Gender and age were not related to the frequency of reported positive behaviors. Ethnicity did influence scores on the positive scale. African American children appeared to present their parents more difficulty on items reflecting cooperative behaviors than Caucasian or Latino children. The implications of the study are discussed based on the recognized need for universal screening of behavior problems in young children and the small number professional training programs targeting the identification and treatment of early childhood behavior problems, despite the availability of evidence-based treatment programs tailored to young children in low-income families.


Safety and pharmacokinetics of multiple doses of intravenous ofloxacin in healthy volunteers.
The safety and pharmacokinetics of ofloxacin in 48 healthy male volunteers were studied in a two-center, randomized, double-blind, placebo-controlled study. Ofloxacin (200 or 400 mg) or placebo was administered as 1-h infusions every 12 h for 7 days. Plasma ofloxacin concentrations were measured by high-performance liquid chromatography. Mean harmonic half-lives ranged from 4.28 to 4.98 h in the 200-mg dosing group and from 5.06 to 6.67 h in the 400-mg dosing group. Intragroup comparisons of trough plasma concentration-versus-time data from study days 2 through 7 revealed that steady state was achieved by day 2 of both multiple-dose regimens. Intergroup comparisons of mean harmonic half-lives, the areas under the concentration-time curve from 0 to 12 and 0 to 60 h, clearance, and apparent volume of distribution (area method) revealed that the pharmacokinetics of ofloxacin are dose independent. Both ofloxacin dosage regimens appeared to be reasonably well tolerated. The two dosage regimens of ofloxacin, 200 or 400 mg every 12 h, appear to be safe and provide serum drug concentrations in excess of the MICs for most susceptible pathogens over the entire dosing interval.


Adenovirus-mediated expression of aquaporin-5 in epithelial cells.
A recombinant adenovirus coding for rat aquaporin-5 was constructed and plaque purified. The recombinant adenovirus (AdrAQP5) mediated the expression of aquaporin-5 in rat and human salivary cell lines and in dog kidney cells in vitro as demonstrated by Northern blot and Western blot analyses, and by confocal microscopy after immunofluorescent labeling. In kidney cells, expression of the transgene was optimal if cells were infected at their basolateral surface, a phenomenon associated with the distribution of integrin receptors on these cells. The expressed aquaporin-5 protein was functionally active because viral-mediated gene transfer resulted in a significant increase in the osmotically directed net fluid secretion rate across monolayers of kidney cells. AdrAQP5 should provide an efficient and useful means to impart facilitated water permeability to cells lacking such a pathway.


Imagery Rescripting for Obsessive Compulsive Disorder: A single case experimental design in 12 cases.
Some individuals with Obsessive Compulsive Disorder (OCD) may experience recurrent intrusive distressing images, which may be emotionally linked to past aversive memories. Our aim was to investigate whether Imagery Rescripting was an effective intervention for such individuals with OCD. Twelve cases who experienced intrusive distressing images are presented in a A1BA2CA3 single case experimental design. After a baseline of symptom monitoring (A1), participants had a control intervention of talking about the memory related image (B), followed by symptom monitoring (A2), a single session of Imagery Rescripting (C) and further monitoring for up to 3 months (A3). Minimal change was seen following the control intervention. However, at 3 months following ImRs, there was a drop in the Yale-Brown Obsessive Compulsive Scale, with a decrease from a mean of 24.1 to 10.7. Reliable improvement was achieved in 9 out of the 12 cases and clinically significant change in 7 out of 12 at 3-month follow up. The limitations are that all cases were selected on the basis that they had an aversive memory linked to their imagery. Imagery Rescripting is a promising therapeutic technique for OCD as an adjunct to CBT where intrusive images are linked to aversive memories.


GRHL3 activates FSCN1 to relax cell-cell adhesions between migrating keratinocytes during wound reepithelialization.
The migrating keratinocyte wound front is required for skin wound closure. Despite significant advances in wound healing research, we do not fully understand the molecular mechanisms that orchestrate collective keratinocyte migration. Here, we show that, in the wound front, the epidermal transcription factor Grainyhead like-3 (GRHL3) mediates decreased expression of the adherens junction protein E-cadherin; this results in relaxed adhesions between suprabasal keratinocytes, thus promoting collective cell migration and wound closure. Wound fronts from mice lacking GRHL3 in epithelial cells (Grhl3-cKO) have lower expression of Fascin-1 (FSCN1), a known negative regulator of E-cadherin. Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) on wounded keratinocytes shows decreased wound-induced chromatin accessibility near the Fscn1 gene in Grhl3-cKO mice, a region enriched for GRHL3 motifs. These data reveal a wound-induced GRHL3/FSCN1/E-cadherin pathway that regulates keratinocyte-keratinocyte adhesion during wound-front migration; this pathway is activated in acute human wounds and is altered in diabetic wounds in mice, suggesting translational relevance.


Biomarkers of protein oxidation in human disease.
Oxidative stress is caused by an imbalance between the production of reactive species of oxygen and nitrogen (RS) and the ability to either detoxify the reactive intermediates produced or repair the resulting damage. Ultimately, oxidative stress conveys the alteration in cellular function caused by the reaction of RS with cellular constituents. Oxidative stress has been extensively reported to participate in the progression of a variety of human diseases including cancer, neurodegenerative disorders and diabetes. Oxidation of proteins is thought to be one of the major mechanisms by which oxidative stress is integrated into cellular signal transduction pathways. Thus, recent research efforts have been aimed to identify the role of specific oxidative protein modifications in the signal transduction events mediating the etiology of human diseases progression. The identification of these oxidative modifications has also raised the possibility of using this knowledge to develop new methods to diagnose diseases before they are clinically evident. In this work, we summarize the mechanisms by which RS generate distinct oxidative modifications. Furthermore, we also review the potential of these oxidative modifications to be used as early biomarkers of human disease.


EEG indices of reward motivation and target detectability in a rapid visual detection task.
A large corpus of data has demonstrated the sensitivity of behavioral and neural measures to variation in the availability of reward. The present study aimed to extend this work by exploring reward motivation in an RSVP task using complex satellite imagery. We found that reward motivation significantly influenced neural activity both in the preparatory period and in response to target images. Pre-stimulus alpha activity and, to a lesser degree, P3 and CNV amplitude were found to be significantly predictive of reward condition on single trials. Target-locked P3 amplitude was modulated both by reward condition and by variation in target detectability inherent to our task. We further quantified this exogenous influence, showing that P3 differences reflected single-trial variation in P3 amplitude for different targets. These findings provide theoretical insight into the neural indices of reward in an RSVP task, and have important applications in the field of satellite imagery analysis.


Dairy products and inflammation: A review of the clinical evidence.
Inflammation is a major biological process regulating the interaction between organisms and the environment, including the diet. Because of the increase in chronic inflammatory diseases, and in light of the immune-regulatory properties of breastfeeding, the ability of dairy products to modulate inflammatory processes in humans is an important but unresolved issue. Here, we report a systematic review of 52 clinical trials investigating inflammatory markers in relation to the consumption of dairy products. An inflammatory score (IS) was defined to quantitatively evaluate this interaction. The IS was significantly positive for the entire data set, indicating an anti-inflammatory activity in humans. When the subjects were stratified according to their health status, the IS was strongly indicative of an anti-inflammatory activity in subjects with metabolic disorders and of a pro-inflammatory activity in subjects allergic to bovine milk. Stratifying the data by product categories associated both low-fat and high-fat products, as well as fermented products, with an anti-inflammatory activity. Remarkably, the literature is characterized by a large gap in knowledge on bioavailability of bioactive nutrients. Future research should thus better combine food and nutritional sciences to adequately follow the fate of these nutrients along the gastrointestinal and metabolic axes.


Towards policies that capture the expected value of biomolecular diversity for drug discovery, human health, and well-being.
This paper aims to help policy makers with a characterization of the intrinsic value of biodiversity and its role as a critical foundation for sustainable development, human health, and well-being. Our objective is to highlight the urgent need to overcome economic, disciplinary, national, cultural, and regional barriers, in order to work out innovative measures to create a sustainable future and prevent the mutual extinction of humans and other species. We emphasize the pervasive neglect paid to the cross-dependency of planetary health, the health of individual human beings and other species. It is critical that social and natural sciences are taken into account as key contributors to forming policies related to biodiversity, conservation, and health management. We are reaching the target date of Nagoya treaty signatories to have accomplished measures to prevent biodiversity loss, providing a unique opportunity for policy makers to make necessary adjustments and refocus targets for the next decade. We propose recommendations for policy makers to explore novel avenues to halt the accelerated global loss of biodiversity. Beyond the critical ecological functions biodiversity performs, its enormous untapped the repertoire of natural molecular diversity is needed for solving accelerating global healthcare challenges.


Aerobic fitness effects in fibromyalgia.
To compare 2 exercise modalities, aerobic fitness training and stretching exercises, in patients with fibromyalgia (FM) in relation to function, pain, quality of life, depression, and anxiety, and to correlate the cardiorespiratory fitness gain with symptom improvement. Seventy-six women with FM between 18 and 60 years old were randomized to either an aerobic program or stretching program, for 20 weeks. They were evaluated at the beginning of the program and after 10 and 20 weeks in relation to the improvement of aerobic fitness, flexibility, function, Fibromyalgia Impact Questionnaire (FIQ), Short-form Health Survey (SF-36), and depression and anxiety levels. Ventilatory anaerobic threshold (VT) and maximum oxygen uptake (VO2max) were determined by expired gas analyses. Aerobic exercise was superior to stretching in relation to VO2 max, VT, function, depression, pain, and the emotional aspects and mental health domains of SF-36. Patients in the stretching group showed no improvement in depression, "role emotional," and "mental health." No association was noted between improvement in aerobic fitness as measured by VT and the improvement of pain, function, or scores in FIQ and SF-36. Our results confirm that aerobic exercise is beneficial to patients with FM, but the cardiorespiratory fitness gain is not related to improvement of FM symptoms.


First Report of Raffaelea canadensis Causing Laurel Wilt Disease Symptoms on Avocado in California.
Laurel wilt disease is a newly described vascular disease of redbay (Persea borbonia (L.) Spreng.) and other members of the Lauraceae family in the southeastern United States. The disease, caused by the fungus Raffaelea lauricola and vectored by a nonnative redbay ambrosia beetle (Xyleborus glabratus Eichhoff), was first detected in Georgia in 2003 (1). Laurel wilt has caused extensive mortality of native redbay in Georgia, Florida, South Carolina, and recently, Mississippi. The avocado, Persea americana, is in the Lauraceae family and has been shown to be susceptible to the laurel wilt pathogen in Florida (3). The potential spread of this pathogen into California is of concern to the commercial avocado industry. During a survey in 2010 in a Temecula, CA avocado orchard with a history of root rot, an avocado (cv. Hass) tree with a diameter at breast height (DBH) of 45 cm was found to be showing typical laurel wilt disease symptoms. The crown was approximately 80% declined and exhibited dead branches without leaves. Black-to-brown discolored sapwood under the bark and many ambrosia beetle exit holes within 1 to 1.5 m up the bole were also observed. A Raffaelea sp. was consistently isolated from symptomatic branch tissue (from two different branches) plated onto cycloheximide-streptomycin malt agar (2) and incubated at room temperature for 2 weeks. Small subunit (18S) sequences of rDNA (approximately 1,150 bp) of three Raffaelea isolates were amplified using primers NS1 and NS4 (4) and deposited into GenBank under Accession Nos. JF327799, JF327800, and JF327801. A BLASTn search of all three sequences revealed high homology (98, 99, and 98% respectively) to an accession of R. canadensis associated with a species of ambrosia beetle (GenBank Accession No. AY858665). Pathogenicity testing was conducted by pipetting 50 μl of a 105 conidia per ml suspension of each of two isolates (UCR1080 and UCR1081) into five 2-mm-diameter holes on each of two avocado (cv. Hass) trees (10 to 15 cm DBH). Isolate UCR1080 was inoculated into three holes on Tree 1 and two holes on Tree 2. Isolate UCR1081 was inoculated into two holes on Tree 1 and three holes on Tree 2. Sterile water was used as a control in five 2-mm-diameter holes on each tree. Holes were drilled to the cambium within 1 to 2 m up the bole using a 0.157-cm electric drill. Four months later, phloem tissue was peeled back, lesion lengths were measured, and pieces of necrotic tissue were cultured for completion of Koch's postulates. R. canadensis was consistently reisolated from necrotic tissue but not from control treatments. To our knowledge, this is the first report of R. canadensis associated with wilt on avocado in California. R. canadensis is closely related to R. lauricola, however, its impact on the California avocado industry is unknown at this time. References: (1) S. W. Fraedrich et al. Plant Dis. 92:215, 2008. (2) T. C. Harrington et al. Mycotaxon 111:337, 2010. (3) A. E. Mayfield et al. Plant Dis. 92:976, 2008. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.


Molecular dynamics simulations of melting and sintering of Si nanoparticles: a comparison of different force fields and computational models.
Melting and sintering of silicon nanoparticles are investigated by means of classical molecular dynamics simulations to disclose the dependence of modelling on the system type, the simulation procedure and interaction potential. The capability of our parametrization of a reactive force field ReaxFF to describe such processes is assessed through a comparison with formally simpler Stillinger-Weber and Tersoff potentials, which are frequently used for simulating silicon-based materials. A substantial dependence of both the predicted melting point and its variation as a function of the nanoparticle size on the simulation model is also highlighted. The outcomes of the molecular dynamics simulations suggest that the trend of the nanoparticulate sintering/coalescence time vs. temperature could provide a valid tool to determine the melting points of nanoparticles theoretically/experimentally.


Direct Recording of Trans-Plasma Membrane Electron Currents Mediated by a Member of the Cytochrome b561 Family of Soybean.
Trans-plasma membrane electron transfer is achieved by b-type cytochromes of different families, and plays a fundamental role in diverse cellular processes involving two interacting redox couples that are physically separated by a phospholipid bilayer, such as iron uptake and redox signaling. Despite their importance, no direct recordings of trans-plasma membrane electron currents have been described in plants. In this work, we provide robust electrophysiological evidence of trans-plasma membrane electron flow mediated by a soybean (Glycine max) cytochrome b561 associated with a dopamine β-monooxygenase redox domain (CYBDOM), which localizes to the plasma membrane in transgenic Arabidopsis (Arabidopsis thaliana) plants and CYBDOM complementary RNA-injected Xenopus laevis oocytes. In oocytes, two-electrode voltage clamp experiments showed that CYBDOM-mediated currents were activated by extracellular electron acceptors in a concentration- and type-specific manner. Current amplitudes were voltage dependent, strongly potentiated in oocytes preinjected with ascorbate (the canonical electron donor for cytochrome b561), and abolished by mutating a highly conserved His residue (H292L) predicted to coordinate the cytoplasmic heme b group. We believe that this unique approach opens new perspectives in plant transmembrane electron transport and beyond.