Transcriptional regulation of main metabolic pathways of cyoA, cydB, fnr, and fur gene knockout Escherichia coli in C-limited and N-limited aerobic continuous cultures.
It is important to understand the cellular responses emanating from environmental perturbations to redesign the networks for practical applications. In particular, the carbon (C) metabolism, nitrogen (N) assimilation, and energy generation are by far important, where those are interconnected and integrated to maintain cellular integrity. In our previous study, we investigated the effect of C/N ratio on the metabolic regulation of gdhA, glnL, glt B,D mutants as well as wild type Escherichia coli (Kumar and Shimizu, MCF, 1-17, 9:8,2010), where it was shown that the transcript levels of cyoA and cydB which encode the terminal oxidases, fnr and fur which encode global regulators were significantly up-regulated under N-limited condition as compared to C-limited condition. In the present study, therefore, the effects of such single-gene knockout on the metabolic regulation were investigated to clarify the roles of those genes in the aerobic continuous culture at the dilution rate of 0.2 h(-1). The specific glucose consumption rates and the specific CO2 production rates of cyoA, cydB, fnr, and fur mutants were all increased as compared to the wild type under both C-limited and N-limited conditions. The former phenomenon was consistent with the up-regulations of the transcript levels of ptsG and ptsH, which are consistent with down-regulations of crp and mlc genes. Moreover, the increase in the specific glucose consumption rate was also caused by up-regulations of the transcript levels of pfkA, pykF and possibly zwf, where those are consistent with the down regulations of cra, crp and mlc genes. Moreover, the transcript levels of rpoN together with glnK, glnB, glnE were up-regulated, and thus the transcript levels of glnA,L,G, and gltB,D as well as nac were up-regulated, while gdhA was down-regulated. This implies the interconnection between cAMP-Crp and PII-Ntr systems. Moreover, cyoA, cydB, fnr and fur gene deletions up-regulated the transcript levels of respiration (nuoA, ndh, cyoA, cydB, and atpA) and the oxidative stress related genes such as soxR, S and sodA, where this was further enhanced under N-limitation. In the cases of cyoA and cydB mutants, arcA, fnr, fur, cydB (for cyoA mutant), and cyoA (for cydB mutant) genes were up-regulated, which may be due to incomplete oxidation of quinol. It was also shown that fur gene transcript level was up-regulated in accordance with the activation of respiratory chain genes. It was shown that the deletion of fur gene activated the enterobactin pathway. The present result demonstrated how the fermentation characteristics could be explained by the transcript levels of metabolic pathway genes as well as global regulators in relation to the knockout of such single genes as cyoA, cydB, fnr, and fur, and clarified the complex gene network regulation in relation to glycolysis, TCA cycle, respiration, and N-regulated pathways. The present result is quite important in understanding the metabolic regulation for metabolic engineering. Moreover, the present result may be useful in improving the specific glucose consumption rate and activation of the TCA cycle by modulating the respiratory chain genes and the related global regulators. The result obtained under N-limited condition may be useful for the heterologous protein production under N-limitation.


Differential metabolic effects of saturated versus polyunsaturated fats in ketogenic diets.
Ketogenic diets (KDs) are used for treatment of refractory epilepsy and metabolic disorders. The classic saturated fatty acid-enriched (SAT) KD has a fat:carbohydrate plus protein ratio of 4:1, in which the predominant fats are saturated. We hypothesized that a polyunsaturated fat-enriched (POLY) KD would induce a similar degree of ketosis with less detrimental effects on carbohydrate and lipid metabolism. Twenty healthy adults were randomized to two different weight-maintaining KDs for 5 d. Diets were 70% fat, 15% carbohydrate, and 15% protein. The fat contents were 60 or 15% saturated, 15 or 60% polyunsaturated, and 25% monounsaturated for SAT and POLY, respectively. Changes in serum beta-hydroxybutyrate, insulin sensitivity (S(I)), and lipid profiles were measured. Mean circulating beta-hydroxybutyrate levels increased 8.4 mg/dl in the POLY group (P = 0.0004), compared with 3.1 mg/dl in the SAT group (P = 0.07). S(I) increased significantly in the POLY group (P = 0.02), whereas total and low-density lipoprotein cholesterol increased significantly in the SAT group (both P = 0.002). These data demonstrate that a short-term POLY KD induces a greater level of ketosis and improves S(I), without adversely affecting total and low-density lipoprotein cholesterol, compared with a traditional SAT KD. Thus, a POLY KD may be superior to a classical SAT KD for chronic administration.


Effect of long-term treatment with an inhaled corticosteroid (budesonide) on airway hyperresponsiveness and clinical asthma in nonsteroid-dependent asthmatics.
Several short-term studies have shown that inhaled steroids can reduce airway hyper-responsiveness in asthma. To evaluate whether prolonged treatment can bring about full recovery, this double-blind, randomized, controlled trial examined the effect of budesonide, 400 micrograms daily for 1 yr, on airway hyperresponsiveness. The time course and characteristics of improvements and associated changes in clinical asthma severity were also evaluated. Thirty-two stable adult asthmatics, requiring bronchodilators alone, were selected. Before and monthly throughout the study, airway responsiveness to methacholine was measured and clinical asthma severity assessed by questionnaire, daily bronchodilator use, and number of asthma exacerbations. Patients receiving budesonide showed a fourfold mean improvement in airway responsiveness compared with those receiving placebo (p less than 0.0005), whose responsiveness remained very stable. Fifteen of the 16 budesonide subjects improved and 5 returned to the normal range. Largest improvements occurred during the first 3 months but, in some, were still progressing slowly at 1 yr. Improvements in responsiveness were accompanied by significant improvements in asthma symptoms, bronchodilator use, and number of asthma exacerbations. The results show that regular, prolonged use of inhaled steroid can produce marked improvements in airway hyperresponsiveness, sometimes with full resolution, and these improvements are accompanied by clinically significant improvements in clinical asthma.


Rapid processing of letters, digits and symbols: what purely visual-attentional deficit in developmental dyslexia?
Visual-attentional theories of dyslexia predict deficits for dyslexic children not only for the perception of letter strings but also for non-alphanumeric symbol strings. This prediction was tested in a two-alternative forced-choice paradigm with letters, digits, and symbols. Children with dyslexia showed significant deficits for letter and digit strings but not for symbol strings. This finding is difficult to explain for visual-attentional theories of dyslexia which postulate identical deficits for letters, digits and symbols. Moreover, dyslexics showed normal W-shaped serial position functions for letter and digit strings, which suggests that their deficit is not due to an abnormally small attentional window. Finally, the size of the deficit was identical for letters and digits, which suggests that poor letter perception is not just a consequence of the lack of reading. Together then, our results show that symbols that map onto phonological codes are impaired (i.e. letters and digits), whereas symbols that do not map onto phonological codes are not impaired. This dissociation suggests that impaired symbol-sound mapping rather than impaired visual-attentional processing is the key to understanding dyslexia.


Energetics of zinc-mediated interactions in the allosteric pathways of metal sensor proteins.
A metal-mediated interprotomer hydrogen bond has been implicated in the allosteric mechanism of DNA operator binding in several metal-sensing proteins. Using computational methods, we investigate the energetics of such zinc-mediated interactions in members of the ArsR/SmtB family of proteins (CzrA, SmtB, CadC, and NmtR) and the MarR family zinc-uptake repressor AdcR, which feature similar interactions, but in sites that differ widely in their allosteric responsiveness. We provide novel structural insight into previously uncharacterized allosteric forms of these proteins using computational methodologies. We find this metal-mediated interaction to be significantly stronger (∼8 kcal/mol) at functional allosteric metal binding sites compared to a nonresponsive site (CadC) and the apo-proteins. Simulations of the apo-proteins further reveal that the high interaction energy works to overcome the considerable disorder at these hydrogen-bonding sites and functions as a "switch" to lock in a weak DNA-binding conformation once metal is bound. These findings suggest a conserved functional role of metal-mediated second coordination shell hydrogen bonds at allosterically responsive sites in zinc-sensing transcription regulators.


Effect of time and frequency manipulation on syllable perception in developmental dyslexics.
Many people with developmental dyslexia have difficulty perceiving stop consonant contrasts as effectively as other people and it has been suggested that this may be due to perceptual limitations of a temporal nature. Accordingly, we predicted that perception of such stimuli by listeners with dyslexia might be improved by stretching them in time-equivalent to speaking slowly. Conversely, their perception of the same stimuli ought to be made even worse by compressing them in time-equivalent to speaking quickly. We tested 15 children with dyslexia on their ability to identify correctly consonant-vowel-consonant (CVC) stimuli that had been stretched or compressed in the time domain. We also tested their perception of the same CVC stimuli after the formant transitions had been stretched or compressed in the frequency domain. Contrary to our predictions, we failed to find any systematic improvement in their performance with either manipulation. We conclude that simple manipulations in the time and frequency domains are unlikely to benefit the ability of people with dyslexia to discriminate between CVCs containing stop consonants.


Effects of CapG overexpression on agonist-induced motility and second messenger generation.
Actin modulating proteins that bind polyphosphoinositides, such as phosphatidylinositol 4, 5-bisphosphate (PIP2), can potentially participate in receptor signaling by restructuring the membrane cytoskeleton and modulating second messenger generation through the phosphoinositide cycle. We examined these possibilities by overexpressing CapG, an actin filament end capping, Ca(2+)- and polyphosphoinositide-binding protein of the gelsolin family. High level transient overexpression decreased actin filament staining in the center of the cells but not in the cell periphery. Moderate overexpression in clonally selected cell lines did not have a detectible effect on actin filament content or organization. Nevertheless, it promoted a dose-dependent increase in rates of wound healing and chemotaxis. The motile phenotype was similar to that observed with gelsolin overexpression, which in addition to capping, also severs and nucleates actin filaments. CapG overexpressing clones are more responsive to platelet-derived growth factor than control-transfected clones. They form more circular dorsal membrane ruffles, have higher phosphoinositide turnover, inositol 1,4,5-trisphosphate generation and Ca2+ signaling. These responses are consistent with enhanced PLC gamma activity. Direct measurements of PIP2 mass showed that the CapG effect on PLC gamma was not due primarily to an increase in the PIP2 substrate concentration. The observed changes in cell motility and membrane signaling are consistent with the hypothesis that PIP(2)-binding actin regulatory proteins modulate phosphoinositide turnover and second messenger generation in vivo. We infer that CapG and related proteins are poised to coordinate membrane signaling with actin filament dynamics following cell stimulation.


Predictors of long-term adherence to continuous positive airway pressure therapy in patients with obstructive sleep apnea and cardiovascular disease in the SAVE study.
To determine the clinical variables that best predict long- term continuous positive airway pressure (CPAP) adherence among patients with cardiovascular disease who have obstructive sleep apnea (OSA). 12-mo prospective within-trial observational study. Centers in China, Australia, and New Zealand participating in the Sleep Apnea cardioVascular Endpoints (SAVE) study. There were 275 patients age 45-70 y with cardiovascular disease (i.e., previously documented transient ischemic attack, stroke, or coronary artery disease) and OSA (4% oxygen desaturation index (ODI) > 12) who were randomized into the CPAP arm of the SAVE trial prior to July 1, 2010. Age, sex, country of residence, type of cardiovascular disease, baseline ODI, severity of sleepiness, and Hospital Anxiety and Depression Scale (HADS) scores plus CPAP side effects and adherence at 1 mo were entered in univariate analyses in an attempt to identify factors predictive of CPAP adherence at 12 mo. Variables with P < 0.2 were then included in a multivariate analysis using a linear mixed model with sites as a random effect and 12-mo CPAP use as the dependent outcome variable. CPAP adherence at 1, 6, and 12 mo was (mean ± standard deviation) 4.4 ± 2.0, 3.8 ± 2.3, and 3.3 ± 2.4 h/night, respectively. CPAP use at 1 mo (effect estimate ± standard error, 0.65 ± 0.07 per h increase, P < 0.001) and side effects at 1 mo (-0.24 ± 0.092 per additional side effect, P = 0.009) were the only independent predictors of 12- mo CPAP adherence. Continuous positive airway pressure use in patients with coexisting cardiovascular disease and moderate to severe obstructive sleep apnea decreases significantly over 12 months. This decline can be predicted by early patient experiences with continuous positive airway pressure (i.e., adherence and side effects at 1 month), raising the possibility that intensive early interventions could improve long-term continuous positive airway pressure compliance in this patient population. Clinical Trials, http://www.clinicaltrials.gov, NCT00738179.


Effects of poly aluminum chloride dosing positions on the performance of a pilot scale anoxic/oxic-membrane bioreactor (A/O-MBR).
The effects of poly aluminum chloride (PACl) dosing positions on the performance of a pilot scale anoxic/oxic membrane bioreactor were investigated. PACl dosage was optimized at 19.5 mg Al2O3/L by jar test. Nutrients removal efficiencies and sludge properties were systematically investigated during periods with no PACl dosing (phase I), with PACl dosing in oxic tank (phase II) and then in anoxic tank (phase III). The results showed that total phosphorus removal efficiency increased from 18 to 88% in phase II and 85% in phase III with less than 0.5 mg P/L in effluent. Ammonia nitrogen removal efficiencies reached 99% in all phases and chemical oxygen demand removal efficiencies reached 92%, 91% and 90% in the three phases, respectively. Total nitrogen removal efficiency decreased from 59% in phase I to 49% in phases II and III. Dosing PACl in the oxic tank resulted in smaller sludge particle size, higher zeta potential, better sludge settleability and lower membrane fouling rate in comparison with dosing PACl in the anoxic tank.


Enhanced detection limit by dark mode perturbation in 2D photonic crystal slab refractive index sensors.
We demonstrate for the first time a 300nm thick, 300μm × 300μm 2D dielectric photonic crystal slab membrane with a quality factor of 10,600 by coupling light to slightly perturbed dark modes through alternating nano-hole sizes. The newly created fundamental guided resonances greatly reduce nano-fabrication accuracy requirements. Moreover, we created a new layer architecture resulting in electric field enhancement at the interface between the slab and sensing regions, and spectral sensitivity of >800 nm/RIU, that is, >0.8 of the single-mode theoretical upper limit of spectral sensitivity.


Clinician knowledge, clinician barriers, and perceived parental barriers regarding human papillomavirus vaccination: Association with initiation and completion rates.
We tested the hypothesis that clinician knowledge, clinician barriers, and perceived parental barriers relevant to the human papillomavirus (HPV) vaccination account for the variation in vaccine delivery at the practice-site level. We conducted a survey from October 2015 through January 2016 among primary care clinicians (n=280) in a 27-county geographic region to assess clinician knowledge, clinician barriers, and perceived parental barriers regarding HPV vaccination. Primary care clinicians included family medicine physicians, general pediatricians, and family and pediatric nurse-practitioners. We also used the Rochester Epidemiology Project to measure HPV vaccination delivery. Specifically we used administrative data to measure receipt of at least one valid HPV vaccine dose (initiation) and receipt of three valid HPV vaccine doses (completion) among 9-18year old patients residing in the same 27-county geographic region. We assessed associations of clinician survey data with variation in vaccine delivery at the clinical site using administrative data on patients aged 9-18years (n=68,272). Consistent with our hypothesis, we found that greater knowledge of HPV and the HPV vaccination was associated with higher rates of HPV vaccination initiation (Incidence rate ratio [IRR]=1.05) and completion of three doses (IRR=1.28). We also found support for the hypothesis that greater perceived parental barriers to the HPV vaccination were associated with lower rates of initiation (IRR=0.94) and completion (IRR=0.90). These IRRs were statistically significant even after adjustment for site-level characteristics including percent white, percent female, percent ages 9-13, and percent with government insurance or self-pay at each site. Clinician knowledge and their report of the frequency of experiencing parental barriers are associated with HPV vaccine delivery rates-initiation and completion. Higher measures of knowledge correlated with higher rates. Fewer perceived occurrences of parental barriers correlated with lower rates. These data can guide efforts to improve HPV vaccine delivery in clinical settings.


Identifying Individual-Cancer-Related Genes by Rebalancing the Training Samples.
The identification of individual-cancer-related genes typically is an imbalanced classification issue. The number of known cancer-related genes is far less than the number of all unknown genes, which makes it very hard to detect novel predictions from such imbalanced training samples. A regular machine learning method can either only detect genes related to all cancers or add clinical knowledge to circumvent this issue. In this study, we introduce a training sample rebalancing strategy to overcome this issue by using a two-step logistic regression and a random resampling method. The two-step logistic regression is to select a set of genes that related to all cancers. While the random resampling method is performed to further classify those genes associated with individual cancers. The issue of imbalanced classification is circumvented by randomly adding positive instances related to other cancers at first, and then excluding those unrelated predictions according to the overall performance at the following step. Numerical experiments show that the proposed resampling method is able to identify cancer-related genes even when the number of known genes related to it is small. The final predictions for all individual cancers achieve AUC values around 0.93 by using the leave-one-out cross validation method, which is very promising, compared with existing methods.


Flow around fishlike shapes studied using multiparticle collision dynamics.
Empirical measurements of hydrodynamics of swimming fish are very difficult. Therefore, modeling studies may be of great benefit. Here, we investigate the suitability for such a study of a recently developed mesoscale method, namely, multiparticle collision dynamics. As a first step, we confine ourselves to investigations at intermediate Reynolds numbers of objects that are stiff. Due to the lack of empirical data on the hydrodynamics of stiff fishlike shapes we use a previously published numerical simulation of the shapes of a fish and a tadpole for comparison. Because the shape of a tadpole resembles that of a circle with an attached splitter plate, we exploit the knowledge on hydrodynamic consequences of such an attachment to test the model further and study the effects of splitter plates for objects of several shapes at several Reynolds numbers. Further, we measure the angles of separation of flow around a circular cylinder and make small adjustments to the boundary condition and the method to drive the flow. Our results correspond with empirical data and with results from other models.


Changes in electroencephalographic activity associated with learning a novel motor task.
This study was designed to examine changes in EEG activity associated with the learning of a novel task. Right-handed adults (N = 61) were randomly assigned to experimental and control groups. Subjects'EEG was recorded at 10 sites. Subjects' performance was assessed using 8-s trials on a mirror star trace. On the acquisition day, the experimental subjects performed 175 trials while the control subjects performed 10 trials, sat quietly for the amount of time needed to perform 155 trials, and then performed 10 more trials. On the retention day, all subjects performed 20 trials. There was a significant Group x Day x Trial interaction that showed that performance improved across trial blocks and across days; however, after the first 10 acquisition trials, the experimental subjects were always significantly better than the control subjects. Analysis of the EEG data showed a significant four-way interaction that showed that following the the first 10 acquisition trials, the experimental subjects had more alpha activity than the control subjects. It is concluded that there are consistent EEG changes in the alpha band that are associated with learning a motor task.


Myosin isoform switching during assembly of the Drosophila flight muscle thick filament lattice.
During muscle development myosin molecules form symmetrical thick filaments, which integrate with the thin filaments to produce the regular sarcomeric lattice. In Drosophila indirect flight muscles (IFMs) the details of this process can be studied using genetic approaches. The weeP26 transgenic line has a GFP-encoding exon inserted into the single Drosophila muscle myosin heavy chain gene, Mhc. The weeP26 IFM sarcomeres have a unique MHC-GFP-labelling pattern restricted to the sarcomere core, explained by non-translation of the GFP exon following alternative splicing. Characterisation of wild-type IFM MHC mRNA confirmed the presence of an alternately spliced isoform, expressed earlier than the major IFM-specific isoform. The two wild-type IFM-specific MHC isoforms differ by the presence of a C-terminal 'tailpiece' in the minor isoform. The sequential expression and assembly of these two MHCs into developing thick filaments suggest a role for the tailpiece in initiating A-band formation. The restriction of the MHC-GFP sarcomeric pattern in weeP26 is lifted when the IFM lack the IFM-specific myosin binding protein flightin, suggesting that it limits myosin dissociation from thick filaments. Studies of flightin binding to developing thick filaments reveal a progressive binding at the growing thick filament tips and in a retrograde direction to earlier assembled, proximal filament regions. We propose that this flightin binding restricts myosin molecule incorporation/dissociation during thick filament assembly and explains the location of the early MHC isoform pattern in the IFM A-band.


Head and neck malignancies in Croatian renal transplant recipients.
Renal transplantation is associated with increased incidence of cancer. We reviewed a large series of renal transplant recipients to determine the incidence and outcome of patients with malignant changes located at the head and neck. A total of 1232 renal transplant recipients have been followed at Department of Dialysis University Hospital Centre Zagreb from 1972 to 2009. Demographic data, localization and disease outcome were evaluated in patients who developed cancer. Twenty one patients (1.7%) developed 27 head and neck malignancies. The average time from transplantation to development of cancer was 56.8 months. The mean length of follow-up was 9.4+/-4.8 years. Eighteen malignancies were cutaneous in origin and 9 were noncutaneous. Of cutaneous malignancies, 88.9% were basal cell carcinoma; one patient had Merkell-cell carcinoma and one patient developed squamous cell carcinoma. Six cases of basocellular skin cancer were recorded in one fair-skin patient. Noncutaneous malignancies involved the oral cavity (2 cases of Kaposi's sarcoma and one pharyngeal cancer) and the thyroid gland in 3 patients each. Two patients had post-transplant lymphoproliferative disorder occurring at the head and neck. One patient had brain tumor. Radical surgery, radiation, and/or chemotherapy were necessary in 33.3% of patients. Immunosuppression was reduced in all patients, and 12 patients were switched from the calcineurin-based immunosuppression to sirolimus. They all have stable graft function. None of the patients died from cancer. Immunosuppression was ceased in one patient with Kaposi's sarcoma who returned to dialysis and died 10 years later from heart failure. An increased incidence of cancer occurring in the head and neck was recorded. Careful skin examination and oral examination is mandatory for discovering cancer before dissemination. Sirolimus is safe alternative to calcineurin-based immunosuppression in patients who developed head and neck malignancies.


Utilization of Facial Image Analysis Technology for Blink Detection: A Validation Study.
The assessment of anterior eye diseases and the understanding of psychological functions of blinking can benefit greatly from a validated blinking detection technology. In this work, we proposed an algorithm based on facial recognition built on current video processing technologies to automatically filter and analyze blinking movements. We compared electrooculography (EOG), the gold standard of blinking measurement, with manual video tape recording counting (mVTRc) and our proposed automated video tape recording analysis (aVTRa) in both static and dynamic conditions to validate our aVTRa method. We measured blinking in both static condition, where the subject was sitting still with chin fixed on the table, and dynamic condition, where the subject's face was not fixed and natural communication was taking place between the subject and interviewer. We defined concordance of blinks between measurement methods as having less than 50 ms difference between eyes opening and closing. The subjects consisted of seven healthy Japanese volunteers (3 male, four female) without significant eye disease with average age of 31.4±7.2. The concordance of EOG vs. aVTRa, EOG vs. mVTRc, and aVTRa vs. mVTRc (average±SD) were found to be 92.2±10.8%, 85.0±16.5%, and 99.6±1.0% in static conditions and 32.6±31.0%, 28.0±24.2%, and 98.5±2.7% in dynamic conditions, respectively. In static conditions, we have found a high blink concordance rate between the proposed aVTRa versus EOG, and confirmed the validity of aVTRa in both static and dynamic conditions.


Committee Opinion No. 637: Marijuana Use During Pregnancy and Lactation.
Cannabis sativa (marijuana) is the illicit drug most commonly used during pregnancy. The self-reported prevalence of marijuana use during pregnancy ranges from 2% to 5% in most studies. A growing number of states are legalizing marijuana for medicinal or recreational purposes, and its use by pregnant women could increase even further as a result. Because of concerns regarding impaired neurodevelopment, as well as maternal and fetal exposure to the adverse effects of smoking, women who are pregnant or contemplating pregnancy should be encouraged to discontinue marijuana use. Obstetrician-gynecologists should be discouraged from prescribing or suggesting the use of marijuana for medicinal purposes during preconception, pregnancy, and lactation. Pregnant women or women contemplating pregnancy should be encouraged to discontinue use of marijuana for medicinal purposes in favor of an alternative therapy for which there are better pregnancy-specific safety data. There are insufficient data to evaluate the effects of marijuana use on infants during lactation and breastfeeding, and in the absence of such data, marijuana use is discouraged.


Efficacy and safety of stereotactic radiosurgery for glomus jugulare tumors.
Since the mid-1990s the use of radiosurgery for glomus jugulare tumors has grown in popularity. Despite its increased use, follow-up periods for radiosurgery are short and the numbers of patients reported are small. To add to the available information, the authors report their experience with the application of linear accelerator (LINAC) or CyberKnife modalities in 13 patients with 16 tumors. All patients were treated with frame-based LINAC or CyberKnife radiosurgery, with doses ranging from 1400 to 2700 cGy. Patients were retrospectively assessed for posttreatment side effects, which included hearing loss, tongue weakness, and vocal hoarseness. The patients' most recent magnetic resonance (MR) images were also assessed for changes in tumor size. The median follow-up duration was 41 months and the mean follow-up period was 60 months. All tumors remained stable or decreased in size on follow-up MR images. All patients had stable neurological symptoms, and one experienced transient ipsilateral tongue weakness and hearing loss, both of which subsequently resolved. One patient experienced transient ipsilateral vocal cord paresis; however, this individual had received previous external-beam radiation therapy. The authors' findings continue to support radiosurgery as an effective and safe method of treatment for glomus jugulare tumors that results in low rates of morbidity.


Reflection and refraction of an arbitrary electromagnetic wave at a plane interface separating an isotropic and a biaxial medium.
Exact solutions are obtained for the reflected and transmitted fields resulting when an arbitrary electromagnetic field is incident on a plane interface separating an isotropic medium and a biaxially anisotropic medium in which one of the principal axes is along the interface normal. From our exact solutions for the reflected fields resulting when a plane TE or TM wave is incident on the plane interface, it can be inferred that the reflected field contains both a TE and a TM component. This gives a change in polarization that can be utilized to determine the properties of the biaxial medium. The time-harmonic solution for the reflected field is in the form of two quadruple integrals, one of which is a superposition of plane waves polarized perpendicular to the plane of incidence and the other a superposition of plane waves polarized parallel to the plane of incidence. The time-harmonic solution for the transmitted field is also in the form of two quadruple integrals. Each of these is a superposition of extraordinary plane waves with displacement vectors that are perpendicular to the direction of phase propagation.


A novel multilayer immunoisolating encapsulation system overcoming protrusion of cells.
Application of alginate-microencapsulated therapeutic cells is a promising approach for diseases that require a local and constant supply of therapeutic molecules. However most conventional alginate microencapsulation systems are associated with low mechanical stability and protrusion of cells which is associated with higher surface roughness and limits their clinical application. Here we have developed a novel multilayer encapsulation system that prevents cells from protruding from capsules. The system was tested using a therapeutic protein with anti-tumor activity overexpressed in mammalian cells. The cell containing core of the multilayer capsule was formed by flexible alginate, creating a cell sustaining environment. Surrounded by a poly-L-lysine layer the flexible core was enveloped in a high-G alginate matrix that is less flexible and has higher mechanical stability, which does not support cell survival. The cells in the core of the multilayer capsule did not show growth impairment and protein production was normal for periods up to 70 days in vitro. The additional alginate layer also lowered the surface roughness compared to conventional cell containing alginate-PLL capsules. Our system provides a solution for two important, often overlooked phenomena in cell encapsulation: preventing cell protrusion and improving surface roughness.


Idiopathic IgA nephropathy: pathogenesis, histopathology, and therapeutic options.
IgA nephropathy is one of the most common causes of glomerulonephritis in the world. Proliferative and crescentic forms of IgA are found in up to 30% of cases and are associated with nephrotic-range proteinuria, accelerated hypertension, and accelerated decline toward ESRD. Despite its prevalence and clinical importance, there is no unifying nomenclature or consensus for the treatment of specific histologic subgroups. As a consequence, the development of clinically effective treatment regimens for IgA nephropathy have lagged behind other, less common forms of glomerulonephritis. Herein is reviewed the pathogenesis and histologic subtypes of IgA nephropathy and how conventional and immunosuppressive therapies have an impact on renal survival and recurrence rates. The use of known clinical risk factors for disease progression in conjunction with specific histologic features can be a guide to both induction and consolidation therapies for individual patients with IgA nephropathy.


Tinker-HP: a massively parallel molecular dynamics package for multiscale simulations of large complex systems with advanced point dipole polarizable force fields.
We present Tinker-HP, a massively MPI parallel package dedicated to classical molecular dynamics (MD) and to multiscale simulations, using advanced polarizable force fields (PFF) encompassing distributed multipoles electrostatics. Tinker-HP is an evolution of the popular Tinker package code that conserves its simplicity of use and its reference double precision implementation for CPUs. Grounded on interdisciplinary efforts with applied mathematics, Tinker-HP allows for long polarizable MD simulations on large systems up to millions of atoms. We detail in the paper the newly developed extension of massively parallel 3D spatial decomposition to point dipole polarizable models as well as their coupling to efficient Krylov iterative and non-iterative polarization solvers. The design of the code allows the use of various computer systems ranging from laboratory workstations to modern petascale supercomputers with thousands of cores. Tinker-HP proposes therefore the first high-performance scalable CPU computing environment for the development of next generation point dipole PFFs and for production simulations. Strategies linking Tinker-HP to Quantum Mechanics (QM) in the framework of multiscale polarizable self-consistent QM/MD simulations are also provided. The possibilities, performances and scalability of the software are demonstrated via benchmarks calculations using the polarizable AMOEBA force field on systems ranging from large water boxes of increasing size and ionic liquids to (very) large biosystems encompassing several proteins as well as the complete satellite tobacco mosaic virus and ribosome structures. For small systems, Tinker-HP appears to be competitive with the Tinker-OpenMM GPU implementation of Tinker. As the system size grows, Tinker-HP remains operational thanks to its access to distributed memory and takes advantage of its new algorithmic enabling for stable long timescale polarizable simulations. Overall, a several thousand-fold acceleration over a single-core computation is observed for the largest systems. The extension of the present CPU implementation of Tinker-HP to other computational platforms is discussed.


Increased sequence coverage through combined targeting of variant and conserved epitopes correlates with control of HIV replication.
A major challenge in the development of an HIV vaccine is that of contending with the extensive sequence variability found in circulating viruses. Induction of HIV-specific T-cell responses targeting conserved regions and induction of HIV-specific T-cell responses recognizing a high number of epitope variants have both been proposed as strategies to overcome this challenge. We addressed the ability of cytotoxic T lymphocytes from 30 untreated HIV-infected subjects with and without control of virus replication to recognize all clade B Gag sequence variants encoded by at least 5% of the sequences in the Los Alamos National Laboratory HIV database (1,300 peptides) using gamma interferon and interleukin-2 (IFN-γ/IL-2) FluoroSpot analysis. While targeting of conserved regions was similar in the two groups (P = 0.47), we found that subjects with control of virus replication demonstrated marginally lower recognition of Gag epitope variants than subjects with normal progression (P = 0.05). In viremic controllers and progressors, we found variant recognition to be associated with viral load (r = 0.62, P = 0.001). Interestingly, we show that increased overall sequence coverage, defined as the overall proportion of HIV database sequences targeted through the Gag-specific repertoire, is inversely associated with viral load (r = -0.38, P = 0.03). Furthermore, we found that sequence coverage, but not variant recognition, correlated with increased recognition of a panel of clade B HIV founder viruses (r = 0.50, P = 0.004). We propose sequence coverage by HIV Gag-specific immune responses as a possible correlate of protection that may contribute to control of virus replication. Additionally, sequence coverage serves as a valuable measure by which to evaluate the protective potential of future vaccination strategies.


Single-cycle influenza virus vaccine generates lung CD8+ Trm that cross-react against viral variants and subvert virus escape mutants.
Influenza virus-specific tissue-resident memory (Trm) CD8+ T cells located along the respiratory tract provide cross-strain protection against a breadth of influenza viruses. We show that immunization with a single-cycle influenza virus vaccine candidate (S-FLU) results in the deposition of influenza virus nucleoprotein (NP)-specific CD8+ Trm along the respiratory tract that were more cross-reactive against viral variants and less likely to drive the development of cytotoxic T lymphocyte (CTL) escape mutants, as compared to the lung memory NP-specific CD8+ T cell pool established following influenza infection. This immune profile was linked to the limited inflammatory response evoked by S-FLU vaccination, which increased TCR repertoire diversity within the memory CD8+ T cell compartment. Cumulatively, this work shows that S-FLU vaccination evokes a clonally diverse, cross-reactive memory CD8+ T cell pool, which protects against severe disease without driving the virus to rapidly evolve and escape, and thus represents an attractive vaccine for use against rapidly mutating influenza viruses.


Tropical Livestock Units: Re-evaluating a Methodology.
The dynamic between humans, livestock, and wildlife is evolving owing to growth in populations, a finite global landmass, and shifting climatic conditions. This change comes with certain benefits in terms of food security, nutrition, and livelihoods as livestock populations increase, but is not without risk. The role of livestock in infectious disease emergence, environmental degradation, and the development of antimicrobial resistance is becoming more apparent. An understanding of these risks and development of mitigation tactics, especially in low- and middle-income countries where the pace of change is most rapid, is increasingly based on comprehensive models and tools built to map livestock populations at the global, regional or national level. Translation of model estimates into evidence is often underpinned by a quantification of livestock biomass to support policy development and implementation. This paper discusses the application of the Tropical Livestock Unit in the context of measuring biomass. It examines the established method of calculation, designating all cattle a standard weight of 175 kg, and compares it to two proposed alternatives. In doing so, the potential to refine estimates of biomass in low and middle-income countries is explored, though this concept could be extrapolated to higher income economies as well. Publicly available data from six countries in sub-Saharan Africa was utilized to demonstrate how breed liveweight, herd structures, and growth rates have the potential to dramatically alter the estimates of cattle biomass in each country. Establishing standardized data collection procedures to capture this information on a regular basis would grant a better understanding of the true nature of livestock populations, aid in the development of superior disease prevention and response measures, bolster food security initiatives through improving livestock production, and inform the intelligent management of shared ecosystems to improve conservation and biodiversity.


Abrin poisoning in an 18-month-old child.
When discussing known poisons and their clinical consequences, few physicians are aware of the deadly poison abrin. The common symptoms of abrin toxicity include nausea, vomiting, abdominal pain, and diarrhea with or without bloody stool. However, with an estimated fatal human dose of less than 1 microgram/kg, death due to complications such as liver failure, renal failure, and cerebral edema are possible. An 18-month-old male presented to the emergency department with an abrupt onset of fever, vomiting, diarrhea, and dehydration. The parents had assumed the child was suffering from a severe gastroenteritis until they noticed 3 consecutive diapers containing colorful seeds that were identified by poison control as belonging to the Abrus precatorius plant. The child's gastrointestinal symptoms were consistent with reported cases of abrin poisoning, but the patient also had an isolated and significantly elevated alkaline phosphatase, with testing unable to reveal the responsible pathological process. To reduce morbidity and possible mortality, parents and pediatricians alike should be conscious of the danger the Abrus precatorius seed poses to the pediatric population. Children are at a greater risk of ingesting these seeds due to their colorful appearance, and the consequences could be fatal. Through this case report we hope to raise public awareness regarding this toxin. This includes the management of known cases, as well as the possibility of encountering an isolated elevated alkaline phosphatase level as a laboratory finding if ingestion occurs.


Anti-Trypanosoma cruzi activity of green tea (Camellia sinensis) catechins.
The trypanocidal action of green tea catechins against two different developmental stages of Trypanosoma cruzi is reported for the first time. This activity was assayed with the nonproliferative bloodstream trypomastigote and with the intracellular replicative amastigote parasite forms. An ethyl acetate fraction from Camellia sinensis green tea leaves, which contains most of the polyphenolic compounds and the maximal trypanocidal activity, was obtained by fractionation of the aqueous extract with organic solvents. The active compounds present in this extract were further purified by LH-20 column chromatography and were identified by high-performance liquid chromatography analysis with a photo diode array detector and gas chromatography coupled to mass spectroscopy. The following flavan-3-ols derivatives, known as catechins, were identified: catechin, epicatechin, gallocatechin, epigallocatechin, catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate. The purified compounds lysed more than 50% of the parasites present in the blood of infected BALB/c mice at concentrations as low as 0.12 to 85 pM. The most active compounds were gallocatechin gallate and epigallocatechin gallate, with minimal bactericidal concentrations that inhibited 50% of isolates tested of 0.12 and 0.53 pM, respectively. The number of amastigotes in infected Vero cells decreased by 50% in the presence of each of these compounds at 100 nM. The effects of the catechins on the recombinant T. cruzi arginine kinase, a key enzyme in the energy metabolism of the parasite, were assayed. The activity of this enzyme was inhibited by about 50% by nanomolar concentrations of catechin gallate or gallocatechin gallate, whereas the other members of the group were less effective. On the basis of these results, we suggest that these compounds could be used to sterilize blood and, eventually, as therapeutic agents for Chagas' disease.


[Parenting experiences of parents of children with ADHD: approaching the normal].
The purpose of this study was to explore the experiences of parenting from the parents of children with ADHD and to develop a grounded theory about their experiences. This study was a qualitative research study that used a grounded theory to understand and discuss the parenting experiences of parents of children with ADHD in a greater depth. Grounded data was collected through an in-depth interview from twelve participants with ADHD children. The data was analyzed using the grounded theory method. "Approaching the normal" emerged as the core phenomenone. Parenting experiences for parents of children with ADHD referred to a process of bringing the children, back on track of normal growth development, and their functioning in the family at a normal level and establishing a new normalcy. The process of approaching the normal involved a basic social-psychological process, such as 'living a day in hell', 'accepting', 'confronting the conflicts', 'lowering expectations', and 'making a new normalcy'. The outcomes of this study, which observed the parenting process of parents of children with ADHD, could enhance nurses' understanding of ADHD and help nurses become major mental health service providers for the mental health of children with ADHD and their families.


Selective inhibition of protein arginine methyltransferase 5 blocks initiation and maintenance of B-cell transformation.
Epigenetic events that are essential drivers of lymphocyte transformation remain incompletely characterized. We used models of Epstein-Barr virus (EBV)-induced B-cell transformation to document the relevance of protein arginine methyltransferase 5 (PRMT5) to regulation of epigenetic-repressive marks during lymphomagenesis. EBV(+) lymphomas and transformed cell lines exhibited abundant expression of PRMT5, a type II PRMT enzyme that promotes transcriptional silencing of target genes by methylating arginine residues on histone tails. PRMT5 expression was limited to EBV-transformed cells, not resting or activated B lymphocytes, validating it as an ideal therapeutic target. We developed a first-in-class, small-molecule PRMT5 inhibitor that blocked EBV-driven B-lymphocyte transformation and survival while leaving normal B cells unaffected. Inhibition of PRMT5 led to lost recruitment of a PRMT5/p65/HDAC3-repressive complex on the miR96 promoter, restored miR96 expression, and PRMT5 downregulation. RNA-sequencing and chromatin immunoprecipitation experiments identified several tumor suppressor genes, including the protein tyrosine phosphatase gene PTPROt, which became silenced during EBV-driven B-cell transformation. Enhanced PTPROt expression following PRMT5 inhibition led to dephosphorylation of kinases that regulate B-cell receptor signaling. We conclude that PRMT5 is critical to EBV-driven B-cell transformation and maintenance of the malignant phenotype, and that PRMT5 inhibition shows promise as a novel therapeutic approach for B-cell lymphomas.


Muscle activation and the slow component rise in oxygen uptake during cycling.
During constant-rate high-intensity exercise, a steady state for oxygen uptake (VO2) is not achieved and, after the initial rapid increase, VO2 continues to increase slowly. The mechanism underlying the slow-component rise in VO2 during high-intensity exercise is unknown. It has been hypothesized that increased muscle use may be a contributing factor, but only limited electromyograph (EMG) data are available supporting this hypothesis. The purpose of this study was to determine whether there is an association between the VO2 slow component and muscle use assessed by contrast shifts in magnetic resonance images (magnetic resonance imaging (MRI)). The VO2 slow component was measured in 16 subjects during two 15-min bouts of cycling performed at high and low intensities. EMG and MRI transverse relaxation times (T2) were obtained after 3 and 15 min to determine muscle activity at each intensity. Low-intensity cycling produced no VO2 slow component, and no increases in muscle activity, except for a small increase (P < 0.05) in the T2 of the vastus lateralis. During high-intensity cycling, VO2, T2 of the vastus lateralis, rectus femoris and whole leg, and EMG activity and median power frequency of the vastus lateralis rose significantly (P < 0.05) from 3 to 15 min. Percent increases in VO2 and muscle T2 were related during high-intensity cycling (r = 0.63), but not during low-intensity cycling (r = 0.00). We conclude that increased muscle use is in part responsible for the slow component rise in oxygen uptake. The results support the hypothesis that during constant-rate exercise at intensities above lactate threshold, progressively greater use of fast-twitch motor units increases energy demand and causes concomitant progressive increases in VO2 and lactate.


A case report of clonal EBV-like memory CD4+ T cell activation in fatal checkpoint inhibitor-induced encephalitis.
Checkpoint inhibitors produce durable responses in numerous metastatic cancers, but immune-related adverse events (irAEs) complicate and limit their benefit. IrAEs can affect organ systems idiosyncratically; presentations range from mild and self-limited to fulminant and fatal. The molecular mechanisms underlying irAEs are poorly understood. Here, we report a fatal case of encephalitis arising during anti-programmed cell death receptor 1 therapy in a patient with metastatic melanoma. Histologic analyses revealed robust T cell infiltration and prominent programmed death ligand 1 expression. We identified 209 reported cases in global pharmacovigilance databases (across multiple cancer types) of encephalitis associated with checkpoint inhibitor regimens, with a 19% fatality rate. We performed further analyses from the index case and two additional cases to shed light on this recurrent and fulminant irAE. Spatial and multi-omic analyses pinpointed activated memory CD4+ T cells as highly enriched in the inflamed, affected region. We identified a highly oligoclonal T cell receptor repertoire, which we localized to activated memory cytotoxic (CD45RO+GZMB+Ki67+) CD4 cells. We also identified Epstein-Barr virus-specific T cell receptors and EBV+ lymphocytes in the affected region, which we speculate contributed to neural inflammation in the index case. Collectively, the three cases studied here identify CD4+ and CD8+ T cells as culprits of checkpoint inhibitor-associated immune encephalitis.


International Working Group consensus response evaluation criteria in lymphoma (RECIL 2017).
In recent years, the number of approved and investigational agents that can be safely administered for the treatment of lymphoma patients for a prolonged period of time has substantially increased. Many of these novel agents are evaluated in early-phase clinical trials in patients with a wide range of malignancies, including solid tumors and lymphoma. Furthermore, with the advances in genome sequencing, new "basket" clinical trial designs have emerged that select patients based on the presence of specific genetic alterations across different types of solid tumors and lymphoma. The standard response criteria currently in use for lymphoma are the Lugano Criteria which are based on [18F]2-fluoro-2-deoxy-D-glucose positron emission tomography or bidimensional tumor measurements on computerized tomography scans. These differ from the RECIST criteria used in solid tumors, which use unidimensional measurements. The RECIL group hypothesized that single-dimension measurement could be used to assess response to therapy in lymphoma patients, producing results similar to the standard criteria. We tested this hypothesis by analyzing 47 828 imaging measurements from 2983 individual adult and pediatric lymphoma patients enrolled on 10 multicenter clinical trials and developed new lymphoma response criteria (RECIL 2017). We demonstrate that assessment of tumor burden in lymphoma clinical trials can use the sum of longest diameters of a maximum of three target lesions. Furthermore, we introduced a new provisional category of a minor response. We also clarified response assessment in patients receiving novel immune therapy and targeted agents that generate unique imaging situations.


Delayed feedback during sensorimotor learning selectively disrupts adaptation but not strategy use.
In sensorimotor adaptation tasks, feedback delays can cause significant reductions in the rate of learning. This constraint is puzzling given that many skilled behaviors have inherently long delays (e.g., hitting a golf ball). One difference in these task domains is that adaptation is primarily driven by error-based feedback, whereas skilled performance may also rely to a large extent on outcome-based feedback. This difference suggests that error- and outcome-based feedback may engage different learning processes, and these processes may be associated with different temporal constraints. We tested this hypothesis in a visuomotor adaptation task. Error feedback was indicated by the terminal position of a cursor, while outcome feedback was indicated by points. In separate groups of participants, the two feedback signals were presented immediately at the end of the movement, after a delay, or with just the error feedback delayed. Participants learned to counter the rotation in a similar manner regardless of feedback delay. However, the aftereffect, an indicator of implicit motor adaptation, was attenuated with delayed error feedback, consistent with the hypothesis that a different learning process supports performance under delay. We tested this by employing a task that dissociates the contribution of explicit strategies and implicit adaptation. We find that explicit aiming strategies contribute to the majority of the learning curve, regardless of delay; however, implicit learning, measured over the course of learning and by aftereffects, was significantly attenuated with delayed error-based feedback. These experiments offer new insight into the temporal constraints associated with different motor learning processes.


Establishing the transcriptional programme for blood: the SCL stem cell enhancer is regulated by a multiprotein complex containing Ets and GATA factors.
Stem cells are a central feature of metazoan biology. Haematopoietic stem cells (HSCs) represent the best-characterized example of this phenomenon, but the molecular mechanisms responsible for their formation remain obscure. The stem cell leukaemia (SCL) gene encodes a basic helix-loop-helix (bHLH) transcription factor with an essential role in specifying HSCs. Here we have addressed the transcriptional hierarchy responsible for HSC formation by characterizing an SCL 3' enhancer that targets expression to HSCs and endothelium and their bipotential precursors, the haemangioblast. We have identified three critical motifs, which are essential for enhancer function and bind GATA-2, Fli-1 and Elf-1 in vivo. Our results suggest that these transcription factors are key components of an enhanceosome responsible for activating SCL transcription and establishing the transcriptional programme required for HSC formation.


Poly(GP) proteins are a useful pharmacodynamic marker for C9ORF72-associated amyotrophic lateral sclerosis.
There is no effective treatment for amyotrophic lateral sclerosis (ALS), a devastating motor neuron disease. However, discovery of a G4C2 repeat expansion in the C9ORF72 gene as the most common genetic cause of ALS has opened up new avenues for therapeutic intervention for this form of ALS. G4C2 repeat expansion RNAs and proteins of repeating dipeptides synthesized from these transcripts are believed to play a key role in C9ORF72-associated ALS (c9ALS). Therapeutics that target G4C2 RNA, such as antisense oligonucleotides (ASOs) and small molecules, are thus being actively investigated. A limitation in moving such treatments from bench to bedside is a lack of pharmacodynamic markers for use in clinical trials. We explored whether poly(GP) proteins translated from G4C2 RNA could serve such a purpose. Poly(GP) proteins were detected in cerebrospinal fluid (CSF) and in peripheral blood mononuclear cells from c9ALS patients and, notably, from asymptomatic C9ORF72 mutation carriers. Moreover, CSF poly(GP) proteins remained relatively constant over time, boding well for their use in gauging biochemical responses to potential treatments. Treating c9ALS patient cells or a mouse model of c9ALS with ASOs that target G4C2 RNA resulted in decreased intracellular and extracellular poly(GP) proteins. This decrease paralleled reductions in G4C2 RNA and downstream G4C2 RNA-mediated events. These findings indicate that tracking poly(GP) proteins in CSF could provide a means to assess target engagement of G4C2 RNA-based therapies in symptomatic C9ORF72 repeat expansion carriers and presymptomatic individuals who are expected to benefit from early therapeutic intervention.


The vaccinia-related kinases phosphorylate the N' terminus of BAF, regulating its interaction with DNA and its retention in the nucleus.
The vaccinia-related kinases (VRKs) comprise a branch of the casein kinase family whose members are characterized by homology to the vaccinia virus B1 kinase. The VRK orthologues encoded by Caenorhabditis elegans and Drosophila melanogaster play an essential role in cell division; however, substrates that mediate this role have yet to be elucidated. VRK1 can complement the temperature sensitivity of a vaccinia B1 mutant, implying that VRK1 and B1 have overlapping substrate specificity. Herein, we demonstrate that B1, VRK1, and VRK2 efficiently phosphorylate the extreme N' terminus of the BAF protein (Barrier to Autointegration Factor). BAF binds to both DNA and LEM domain-containing proteins of the inner nuclear membrane; in lower eukaryotes, BAF has been shown to play an important role during the reassembly of the nuclear envelope at the end of mitosis. We demonstrate that phosphorylation of ser4 and/or thr2/thr3 abrogates the interaction of BAF with DNA and reduces its interaction with the LEM domain. Coexpression of VRK1 and GFP-BAF greatly diminishes the association of BAF with the nuclear chromatin/matrix and leads to its dispersal throughout the cell. Cumulatively, our data suggest that the VRKs may modulate the association of BAF with nuclear components and hence play a role in maintaining appropriate nuclear architecture.


Effect of catecholamines on the ventricular myocyte action potential in raised extracellular potassium.
We describe the relationship between catecholamines and raised extracellular potassium ([K+]o) on action potential parameters and calcium currents in isolated ventricular myocytes of the guinea-pig and relate these findings to the problem of understanding how the heart is protected from exercise-induced hyperkalaemia ([K+]a up to 8.5 mM). Action potential duration (APD90), amplitude and upstroke velocity were recorded in stimulated (2Hz) guinea-pig ventricular myocytes using whole-cell patch electrode recordings (37 degrees C). Cells were superfused with normal K+ Tyrode and with raised K+ Tyrode in the presence of either noradrenaline, adrenaline or raised calcium. Inward calcium current was measured using voltage clamp. Raised K+ (8, 12, 16 mM K+ Tyrode) caused a significant (P < 0.01) depolarisation, shortened the APD90 and decreased the action potential amplitude and upstroke velocity. In raised K+ Tyrode addition of noradrenaline (0.08-0.1 microM) or adrenaline (0.1-0.2 microM) increased action potential amplitude (P < 0.01), APD90 (P < 0.01) and upstroke velocity (P < 0.01) (measured only in 16 mM K+ Tyrode). In 12 mM K+ Tyrode raised Ca2+ (5-6 mM) increased action potential amplitude (P < 0.05) and shortened APD90 (P < 0.05). Addition of NA (0.08-0.1 microM) increased the inward Ca2+ current. All effects were fully reversible. In raised [K+]o increases in catecholamines and [Ca2+]o cause changes in action potential parameters that would be expected to maintain propagation of the cardiac action potential in the whole heart. Thus, in the ventricular myocyte the increase in conductance to Ca2+ caused by catecholamines may be one factor that is important in minimising the potentially adverse effects of exercise-induced hyperkalaemia.


Lifestyle and Chronic Pain across the Lifespan: An Inconvenient Truth?
Chronic pain has a tremendous personal and socioeconomic impact and remains difficult to treat. Therefore, it is important to provide an update on the current understanding regarding lifestyle factors in people with chronic pain across the lifespan. Lifestyle factors such as physical (in)activity, sedentary behavior, stress, poor sleep, unhealthy diet, and smoking are associated with chronic pain severity and sustainment. This applies to all age categories, that is, chronic pain across the lifespan. Yet current treatment options often do not or only partly address the many lifestyle factors associated with chronic pain or attempt to address them in a standard format rather than providing an individually tailored multimodal lifestyle intervention. The evidence regarding lifestyle factors is available in adults, but limited in children and older adults having chronic pain, providing important avenues for future research. In conclusion, it is proposed that treatment approaches for people with chronic pain should address all relevant lifestyle factors concomitantly in an individually-tailored multimodal intervention. Ultimately, this should lead to improved outcomes and decrease the psychological and socioeconomic burden of chronic pain. Level of Evidence: IV.


Sex in America online: an exploration of sex, marital status, and sexual identity in internet sex seeking and its impacts.
This was an exploratory study of sex and relationship seeking on the Internet, based on a survey of 15,246 respondents in the United States Seventy-five percent of men and 41% of women had intentionally viewed or downloaded porn. Men and gays/lesbians were more likely to access porn or engage in other sex-seeking behaviors online compared with straights or women. A symmetrical relationship was revealed between men and women as a result of viewing pornography, with women reporting more negative consequences, including lowered body image, partner critical of their body, increased pressure to perform acts seen in pornographic films, and less actual sex, while men reported being more critical of their partners' body and less interested in actual sex. Married and divorced were more likely than singles to go online seeking a serious relationship. Only 2% of users met the threshold of compulsive use established by previous studies.


Genomic landscape of colorectal carcinogenesis.
The molecular pathogenesis of solid tumour was first assessed in colorectal cancer (CRC). To date, ≤ 100 genes with somatic alterations have been found to inter-connectively promote neoplastic transformation through specific pathways. The process of colorectal carcinogenesis via genome landscape is reviewed on the basis of an adenoma-to-carcinoma sequence, as shown by serial histological and epidemiological observations. The relevant literatures from PubMed (1980-2021) have been reviewed for this article. The major routes of CRC development, chromosomal instability (CIN), microsatellite instability (MSI), and the serrated route either via CIN or MSI, proceed through the respective molecular pathway of colorectal carcinogenesis. Particular aspects of CRC carcinogenesis can also be determined by evaluating familial CRCs (FCRC) and genotype-phenotype correlations. Specific causative gene alterations still leave to be identified in several FCRCs. Otherwise, recently verified FCRC can be particularly notable, for example, EPCAM-associated Lynch syndrome, polymerase proofreading-associated polyposis, RNF43-associated polyposis syndrome or NTHL1 tumour syndrome, and hereditary mixed polyposis syndrome. The oncogenic landscape is described, including representative pathway genes, the three routes of carcinogenesis, familial CRCs, genotype-phenotype correlations, the identification of causative genes, and consensus molecular subtypes (CMS). Whole genome research using multi-gene panels (MGPs) has facilitated high through-put detection of previously unidentified genes involved in colorectal carcinogenesis. New approaches designed to identify rare variants are recommended to consider their alterations implicated in the molecular pathogenesis.


Conversational language use as a predictor of early reading development: language history as a moderating variable.
The present study examined the nature of concurrent and predictive associations between conversational language use and reading development during early school-age years. Language and reading data from 380 twins in the Western Reserve Reading Project were examined via phenotypic correlations and multilevel modeling on exploratory latent factors. In the concurrent prediction of children's early reading abilities, a significant interaction emerged between children's conversational language abilities and their history of reported language difficulties. Specifically, conversational language concurrently predicted reading development above and beyond variance accounted for by formal vocabulary scores, but only in children with a history of reported language difficulties. A similar trend was noted in predicting reading skills 1 year later, but the interaction was not statistically significant. Findings suggest a more nuanced view of the association between spoken language and early reading than is commonly proposed. One possibility is that children with and without a history of reported language difficulties rely on different skills, or the same skills to differing degrees, when completing early reading-related tasks. Future studies should examine the causal link between conversational language and early reading specifically in children with a history of reported language difficulties.


Dual-frequency heterodyne ellipsometer for characterizing generalized elliptically birefringent media.
This research proposed a dual-frequency heterodyne ellipsometer (DHE) in which a dual-frequency collinearly polarized laser beam with equal amplitude and zero phase difference between p- and s-polarizations is setup. It is based on the polarizer-sample-analyzer, PSA configuration of the conventional ellipsometer. DHE enables to characterize a generalized elliptical phase retarder by treating it as the combination of a linear phase retarder and a polarization rotator. The method for measuring elliptical birefringence of an elliptical phase retarder based on the equivalence theorem of an unitary optical system was derived and the experimental verification by use of DHE was demonstrated too. The experimental results show the capability of DHE on characterization of a generalized phase retardation plate accurately.


Effect of Detarium microcarpum (Dm) and Mucuna flagellipes (Mf) gums on the quality of white bread.
Incorporation of Detarium Microcarpum (Dm) and Mucuna flagellipes (Mf) water soluble polysaccharides (gums) at 0.0 to 0.5% levels in wheat flour was studied to evaluate their effect on the rheological properties of wheat flour dough and white bread quality. At all levels of incorporation, there were increases (p < or = 0.05) in water absorption of the dough. Doughs containing gums had higher (p < or = 0.05) mixing tolerance index than the control. Set back viscosities decreased by 4.0 RVU and 9.0 RVU with increased levels of Dm and Mf gum incorporation, respectively. Significantly (p < or = 0.05) higher oven spring occurred in all the gum substituted white bread when compared to the control. The 0.5% gum substituted breads had a significantly (p < or = 0.05) higher sensory score for crumb grain, texture but lower (p < 0.05) crumb firmness than the control as determined instrumentally. Textural analysis after 5 days storage revealed that Dm and Mf gums improved moisture retention properties of the bread and reduced crumb firming tendency.


The health effects of chrysotile: current perspective based upon recent data.
This review substantiates kinetically and pathologically the differences between chrysotile and amphiboles. The serpentine chrysotile is a thin walled sheet silicate while the amphiboles are double-chain silicates. These different chemistries result in chrysotile clearing very rapidly from the lung (T(1/2)=0.3 to 11 days) while amphiboles are among the slowest clearing fibers known (T(1/2)=500 days to infinity). Across the range of mineral fiber solubilities chrysotile lies towards the soluble end of the scale. Chronic inhalation toxicity studies with chrysotile in animals have unfortunately been performed at very high exposure concentrations resulting in lung overload. Consequently their relevance to human exposures is extremely limited. Chrysotile following subchronic inhalation at a mean exposure of 76 fibers L>20 microm/cm(3) (3413 total fibers/cm(3)) resulted in no fibrosis (Wagner score 1.8-2.6), at any time point and no difference with controls in BrdU response or biochemical and cellular parameters. The long chrysotile fibers were observed to break apart into small particles and smaller fibers. Toxicologically, chrysotile which rapidly falls apart in the lung behaves more like non-fibrous mineral dusts while response to amphibole asbestos reflects its insoluble fibrous structure. Recent quantitative reviews of epidemiological studies of mineral fibers have determined the potency of chrysotile and amphibole asbestos for causing lung cancer and mesothelioma in relation to fiber type have also differentiated between these two minerals. The most recent analyses also concluded that it is the longer, thinner fibers that have the greatest potency as has been reported in animal inhalation toxicology studies. However, one of the major difficulties in interpreting these studies is that the original exposure estimates rarely differentiated between chrysotile and amphiboles. Not unlike some other respirable particulates, to which humans are, or have been heavily occupationally exposed, there is evidence that heavy and prolonged exposure to chrysotile can produce lung cancer. The value of the present and other similar studies is that they show that low exposures to pure chrysotile do not present a detectable risk to health. Since total dose over time decides the likelihood of disease occurrence and progression, they also suggest that the risk of an adverse outcome may be low if even any high exposures experienced were of short duration.


The precision of video and photocell tracking systems and the elimination of tracking errors with infrared backlighting.
Automated tracking offers a number of advantages over both manual and photocell tracking methodologies, including increased reliability, validity, and flexibility of application. Despite the advantages that video offers, our experience has been that video systems cannot track a mouse consistently when its coat color is in low contrast with the background. Furthermore, the local lab lighting can influence how well results are quantified. To test the effect of lighting, we built devices that provide a known path length for any given trial duration, at a velocity close to the average speed of a mouse in the open-field and the circular water maze. We found that the validity of results from two commercial video tracking systems (ANY-maze and EthoVision XT) depends greatly on the level of contrast and the quality of the lighting. A photocell detection system was immune to lighting problems but yielded a path length that deviated from the true length. Excellent precision was achieved consistently, however, with video tracking using infrared backlighting in both the open field and water maze. A high correlation (r=0.98) between the two software systems was observed when infrared backlighting was used with live mice.


Organizing Principles of Human Cortical Development--Thickness and Area from 4 to 30 Years: Insights from Comparative Primate Neuroanatomy.
The human cerebral cortex undergoes a protracted, regionally heterogeneous development well into young adulthood. Cortical areas that expand the most during human development correspond to those that differ most markedly when the brains of macaque monkeys and humans are compared. However, it remains unclear to what extent this relationship derives from allometric scaling laws that apply to primate brains in general, or represents unique evolutionary adaptations. Furthermore, it is unknown whether the relationship only applies to surface area (SA), or also holds for cortical thickness (CT). In 331 participants aged 4 to 30, we calculated age functions of SA and CT, and examined the correspondence of human cortical development with macaque to human expansion, and with expansion across nonhuman primates. CT followed a linear negative age function from 4 to 30 years, while SA showed positive age functions until 12 years with little further development. Differential cortical expansion across primates was related to regional maturation of SA and CT, with age trajectories differing between high- and low-expanding cortical regions. This relationship adhered to allometric scaling laws rather than representing uniquely macaque-human differences: regional correspondence with human development was as large for expansion across nonhuman primates as between humans and macaque.


Content disputes in Wikipedia reflect geopolitical instability.
Indicators that rank countries according socioeconomic measurements are important tools for regional development and political reform. Those currently in widespread use are sometimes criticized for a lack of reproducibility or the inability to compare values over time, necessitating simple, fast and systematic measures. Here, we applied the 'guilt by association' principle often used in biological networks to the information network within the online encyclopedia Wikipedia to create an indicator quantifying the degree to which pages linked to a country are disputed by contributors. The indicator correlates with metrics of governance, political or economic stability about as well as they correlate with each other, and though faster and simpler, it is remarkably stable over time despite constant changes in the underlying disputes. For some countries, changes over a four year period appear to correlate with world events related to conflicts or economic problems.


Developing intuition: neural correlates of cognitive-skill learning in caudate nucleus.
The superior capability of cognitive experts largely depends on automatic, quick information processing, which is often referred to as intuition. Intuition develops following extensive long-term training. There are many cognitive models on intuition development, but its neural basis is not known. Here we trained novices for 15 weeks to learn a simple board game and measured their brain activities in early and end phases of the training while they quickly generated the best next-move to a given board pattern. We found that the activation in the head of caudate nucleus developed over the course of training, in parallel to the development of the capability to quickly generate the best next-move, and the magnitude of the caudate activity was correlated with the subject's performance. In contrast, cortical activations, which already appeared in the early phase of training, did not further change. Thus, neural activation in the caudate head, but not those in cortical areas, tracked the development of capability to quickly generate the best next-move, indicating that circuitries including the caudate head may automate cognitive computations.


Modelling corneal epithelial wound closure in the presence of physiological electric fields via a moving boundary formalism.
A new framework for the modelling of corneal epithelial wound healing is presented, which can include the presence of a physiological electric field. The difficulty inherent in the inclusion of this biological phenomenon motivates our use of a moving boundary formalism. A key conclusion is that the model predicts a linear relation between the wound healing speed and the physiological electric field strengths over a physiologically large range of electric field strength. Another key point is that this linear relationship between electric field strength and wound healing speed is robust to variations in critical parameters that are difficult to estimate. The linearity is also robust to different realizations of the modelling framework presented.


Alkylresorcinol synthases expressed in Sorghum bicolor root hairs play an essential role in the biosynthesis of the allelopathic benzoquinone sorgoleone.
Sorghum bicolor is considered to be an allelopathic crop species, producing phytotoxins such as the lipid benzoquinone sorgoleone, which likely accounts for many of the allelopathic properties of Sorghum spp. Current evidence suggests that sorgoleone biosynthesis occurs exclusively in root hair cells and involves the production of an alkylresorcinolic intermediate (5-[(Z,Z)-8',11',14'-pentadecatrienyl]resorcinol) derived from an unusual 16:3Delta(9,12,15) fatty acyl-CoA starter unit. This led to the suggestion of the involvement of one or more alkylresorcinol synthases (ARSs), type III polyketide synthases (PKSs) that produce 5-alkylresorcinols using medium to long-chain fatty acyl-CoA starter units via iterative condensations with malonyl-CoA. In an effort to characterize the enzymes responsible for the biosynthesis of the pentadecyl resorcinol intermediate, a previously described expressed sequence tag database prepared from isolated S. bicolor (genotype BTx623) root hairs was first mined for all PKS-like sequences. Quantitative real-time RT-PCR analyses revealed that three of these sequences were preferentially expressed in root hairs, two of which (designated ARS1 and ARS2) were found to encode ARS enzymes capable of accepting a variety of fatty acyl-CoA starter units in recombinant enzyme studies. Furthermore, RNA interference experiments directed against ARS1 and ARS2 resulted in the generation of multiple independent transformant events exhibiting dramatically reduced sorgoleone levels. Thus, both ARS1 and ARS2 are likely to participate in the biosynthesis of sorgoleone in planta. The sequences of ARS1 and ARS2 were also used to identify several rice (Oryza sativa) genes encoding ARSs, which are likely involved in the production of defense-related alkylresorcinols.


Hematologic and immunomodulatory effects of an interleukin-1 receptor antagonist coinfusion during low-dose endotoxemia in healthy humans.
Endotoxin is a component of gram-negative bacteria that causes hematologic and immunologic changes through its induction of cytokines. Interleukin-1 receptor antagonist (IL-1Ra) is a naturally occurring inhibitor of IL-1 that competes with IL-1 for occupancy of cell-surface receptors but possesses no agonist activity. We investigated the ability of human recombinant IL-1Ra to block the effects of low-dose endotoxin. Fourteen healthy male volunteers between 18 and 30 years old were injected intravenously with 3 ng/kg Escherichia coli endotoxin. Concurrent with the injections, nine volunteers received a 3-hour continuous intravenous infusion of IL-1Ra. The other five subjects were given a 3-hour infusion of saline. Volunteers injected with endotoxin experienced a threefold increase in circulating neutrophils over baseline. This neutrophilia was significantly reduced by 48% in subjects administered endotoxin plus IL-1Ra (P = .0253). Ex vivo mitogen-induced peripheral blood mononuclear cell proliferation decreased by greater than 60% at 3 and 6 hours after endotoxin injection (P = .0053). This endotoxin-induced reduction in mitogen response was reversed in subjects coinjected with IL-1Ra (P = .0253). Endotoxin-induced symptoms, fever, and tachycardia were unaffected by IL-1Ra. IL-1 appears to be an important mediator in endotoxemia because some of its hematologic and immunomodulatory effects can be blocked by IL-1Ra.


Association between Working Time and Burnout Syndrome in Peruvian Military during the Second Epidemic Wave of COVID-19.
There is scant evidence on the impact of the COVID-19 pandemic on burnout in front-line military personnel and how working time may influence on this condition. We aimed to determine the association between working time and Burnout syndrome in military personnel. A cross-sectional study was conducted using secondary data among 576 military personnel from Lambayeque, Peru during the second wave of COVID-19 in 2021. We used the Maslach Burnout Inventory instrument to measure Burnout Syndrome. We evaluated its association with work time, measured as the number of months that the military member worked during the pandemic. The prevalence of burnout syndrome was 9%. Of the total sample, 39.1% and 10.3% presented depersonalization and emotional exhaustion, respectively. Military personnel working for more than 18 months had a 104% higher prevalence of Burnout syndrome (PR: 2.04, 95%CI: 1.02-4.10). Exposure to a prolonged work time during the pandemic increased the prevalence of Burnout syndrome in military personnel. This information helps to understand the potential effects of the pandemic on this population and provides insight into the time the military members would need rest to prevent Burnout syndrome.


Transcriptional networks in the human epididymis.
The epithelial lining of the human epididymis is critical for sperm maturation. This process requires distinct specialized functions in the head, body, and tail of the duct. These region-specific properties are maintained by distinct gene expression profiles which are governed by transcription factor networks, non-coding RNAs, and other factors. We used genome-wide protocols including DNase-seq, RNA-seq and ChIP-seq to characterize open (active) chromatin, the transcriptome and occupancy of specific transcription factors (TFs) respectively, in caput, corpus, and cauda segments of adult human epididymis tissue and primary human epididymis epithelial (HEE) cell cultures derived from them. RNA-seq following TF depletion or activation, combined with gene ontology analysis also determined TF targets. Among regional differentially expressed transcripts were epithelial-selective transcription factors (TFs), microRNAs, and antiviral response genes. Caput-enriched TFs included hepatocyte nuclear factor 1 (HNF1) and the androgen receptor (AR), both of which were also predicted to occupy cis-regulatory elements identified as open chromatin in HEE cells. HNF1 targets were identified genome-wide using ChIP-seq, in HEE cells. Next, siRNA-mediated depletion of HNF1 revealed a pivotal role for this TF in coordinating epithelial water and solute transport in caput epithelium. The importance of AR in HEE cells was shown by AR ChIP-seq, and by RNA-seq after synthetic androgen (R1881) treatment. AR has a distinct transcriptional program in the HEE cells and likely recruits different co-factors (RUNX1 and CEBPβ) in comparison to those used in prostate epithelium. Our data identify many transcription factors that regulate the development and differentiation of HEE cells. Moreover, a comparison between immature and adult HEE cells showed key TFs in the transition to fully differentiated function of this epithelium. These data may help identify new targets to treat male infertility and have the potential to open new avenues for male contraception.


Specialist health care services use in a European cohort of infants born very preterm.
Children born very preterm require additional specialist care because of the health and developmental risks associated with preterm birth, but information on their health service use is sparse. We sought to describe the use of specialist services by children born very preterm in Europe. We analysed data from the multi-regional, population-based Effective Perinatal Intensive Care in Europe (EPICE) cohort of births before 32 weeks' gestation in 11 European countries. Perinatal data were abstracted from medical records and parents completed a questionnaire at 2 years corrected age (4322 children; 2026 females, 2296 males; median gestational age 29wks, interquartile range [IQR] 27-31wks; median birthweight 1230g, IQR 970-1511g). We compared parent-reported use of specialist services by country, perinatal risk (based on gestational age, small for gestational age, and neonatal morbidities), maternal education, and birthplace. Seventy-six per cent of the children had consulted at least one specialist, ranging across countries from 53.7% to 100%. Ophthalmologists (53.4%) and physiotherapists (48.0%) were most frequently consulted, but individual specialists varied greatly by country. Perinatal risk was associated with specialist use, but the gradient differed across countries. Children with more educated mothers had higher proportions of specialist use in three countries. Large variations in the use of specialist services across Europe were not explained by perinatal risk and raise questions about the strengths and limits of existing models of care. Use of specialist services by children born very preterm varied across Europe. This variation was observed for types and number of specialists consulted. Perinatal risk was associated with specialist care, but did not explain country-level differences. In some countries, mothers' educational level affected use of specialist services.


Metatranscriptomic reconstruction reveals RNA viruses with the potential to shape carbon cycling in soil.
Viruses impact nearly all organisms on Earth, with ripples of influence in agriculture, health, and biogeochemical processes. However, very little is known about RNA viruses in an environmental context, and even less is known about their diversity and ecology in soil, 1 of the most complex microbial systems. Here, we assembled 48 individual metatranscriptomes from 4 habitats within a planted soil sampled over a 22-d time series: Rhizosphere alone, detritosphere alone, rhizosphere with added root detritus, and unamended soil (4 time points and 3 biological replicates). We resolved the RNA viral community, uncovering a high diversity of viral sequences. We also investigated possible host organisms by analyzing metatranscriptome marker genes. Based on viral phylogeny, much of the diversity was Narnaviridae that may parasitize fungi or Leviviridae, which may infect Proteobacteria. Both host and viral communities appear to be highly dynamic, and rapidly diverged depending on experimental conditions. The viral and host communities were structured based on the presence of root litter. Clear temporal dynamics by Leviviridae and their hosts indicated that viruses were replicating. With this time-resolved analysis, we show that RNA viruses are diverse, abundant, and active in soil. When viral infection causes host cell death, it may mobilize cell carbon in a process that may represent an overlooked component of soil carbon cycling.


Tactile Object Familiarity in the Blind Brain Reveals the Supramodal Perceptual-Mnemonic Nature of the Perirhinal Cortex.
This study is the first to investigate the neural underpinnings of tactile object familiarity in the blind during both perception and memory. In the sighted, the perirhinal cortex (PRC) has been implicated in the assessment of visual object familiarity-a crucial everyday task-as evidenced by reduced activation when an object becomes familiar. Here, to examine the PRC's role in tactile object familiarity in the absence of vision, we trained blind participants on a unique memory-guided drawing technique and measured brain activity while they perceptually explored raised-line drawings, drew them from tactile memory, and scribbled (control). Functional magnetic resonance imaging (fMRI) before and after a week of training revealed a significant decrease in PRC activation from pre- to post-training (i.e., from unfamiliar to familiar) during perceptual exploration as well as memory-guided drawing, but not scribbling. This familiarity-based reduction is the first evidence that the PRC represents tactile object familiarity in the blind. Furthermore, the finding of this effect during both tactile perception and tactile memory provides the critical link in establishing the PRC as a structure whose representations are supramodal for both perception and memory.


Commodity risk assessment of Quercus petraea plants from the UK.
The European Commission requested the EFSA Panel on Plant Health to prepare and deliver risk assessments for commodities listed in Commission Implementing Regulation (EU) 2018/2019 as 'High-risk plants, plant products and other objects'. This Scientific Opinion covers plant health risks posed by plants of Quercus petraea imported from the UK as: (a) bundles of 1- to 2-year-old whips and seedlings, (b) 1- to 7-year-old bare root plants for planting and (c) less than 1- to 15-year-old plants in pots, taking into account the available scientific information, including the technical information provided by the UK. All pests associated with the commodity were evaluated against specific criteria for their relevance for this opinion. Two EU quarantine pests, Cronartium quercuum and Phytophthora ramorum (non-EU isolates), two protected zone quarantine pests, Cryphonectria parasitica and Thaumetopoea processionea, and four pests not regulated in the EU, Coniella castaneicola, Meloidogyne mali, Phytophthora kernoviae and Trinophylum cribratum, fulfilled all relevant criteria and were selected for further evaluation. For the selected pests, the risk mitigation measures included in the technical dossier from the UK were evaluated taking into account the possible limiting factors. For these pests an expert judgement is given on the likelihood of pest freedom taking into consideration the risk mitigation measures acting on the pest, including uncertainties associated with the assessment. In the assessment of risk, the age of the plants was considered, reasoning that older trees are more likely to be infested mainly due to longer exposure time and larger size. The degree of pest freedom varies among the pests evaluated, with C. castaneicola being the pests most frequently expected on the imported plants. The Expert knowledge elicitation indicated with 95% certainty that between 9,711 and 10,000 per 10,000 less than 1- to 15-year-old plants in pots will be free from C. castaneicola.


Analysis of binding site hot spots on the surface of Ras GTPase.
We have recently discovered an allosteric switch in Ras, bringing an additional level of complexity to this GTPase whose mutants are involved in nearly 30% of cancers. Upon activation of the allosteric switch, there is a shift in helix 3/loop 7 associated with a disorder to order transition in the active site. Here, we use a combination of multiple solvent crystal structures and computational solvent mapping (FTMap) to determine binding site hot spots in the "off" and "on" allosteric states of the GTP-bound form of H-Ras. Thirteen sites are revealed, expanding possible target sites for ligand binding well beyond the active site. Comparison of FTMaps for the H and K isoforms reveals essentially identical hot spots. Furthermore, using NMR measurements of spin relaxation, we determined that K-Ras exhibits global conformational dynamics very similar to those we previously reported for H-Ras. We thus hypothesize that the global conformational rearrangement serves as a mechanism for allosteric coupling between the effector interface and remote hot spots in all Ras isoforms. At least with respect to the binding sites involving the G domain, H-Ras is an excellent model for K-Ras and probably N-Ras as well. Ras has so far been elusive as a target for drug design. The present work identifies various unexplored hot spots throughout the entire surface of Ras, extending the focus from the disordered active site to well-ordered locations that should be easier to target.


Predictors of dental rehabilitation in children aged 3-12 years.
The aim of this study was to evaluate the proportion of completed treatments and to study the factors affecting the full mouth dental rehabilitation in pediatric patients treated by undergraduate students at the College of Dental Medicine Teaching Clinics, University of Sharjah. A retrospective study was conducted on 270 children aged less than 12 years (mean age 7.6, SD 2.04). Comprehensive dental rehabilitation reports of child patients that were completed by final year dental undergraduate students from the year 2009 to 2011 were reviewed. Data on complete history, oral examination, dental charting, and treatment plan were collected from pediatric dentistry case sheet. Dental caries was charted using WHO 1997 criteria. Dental treatment needs and completion of dental care delivered to children involved in this study were assessed using DMFT/deft scores. Percentages of treatment provided included completed restorations (94%) and space management (84%) in primary dentition, whereas 98% of restoration and 94% of required sealants were completed in permanent dentition. The percentage of completed dental treatment including sealant placement was 61%. Age of the child and the number of decayed teeth present before the start of the treatment significantly correlated with the children in the incomplete treatment category (P < 0.05). Therefore, a worthy dental care was provided in a holistic approach to the children attending College of Dental Medicine training clinics. Age of the child and the number of decayed teeth were the factors affecting dental rehabilitation in children aged 3-12 years.


Chemical and technological properties of bologna-type sausages with added black quinoa wet-milling coproducts as binder replacer.
The objective of this study was to evaluate different strategies for adding 3% black quinoa (either as whole seeds or as a fiber-rich fraction of quinoa from its wet-milling process) to bologna-type sausage. This addition was evaluated in terms of its influence on nutritional composition and technological properties (emulsion stability, pH, water activity, color changes, textural properties, residual nitrite level and lipid oxidation). Both strategies resulted in commercially feasible sausages with increased nutritive properties (dietary fiber) and with some modifications in their technological properties. Compared with control sausages, they showed better emulsion stability, lower water activity and lipid oxidation values (interesting properties for sausages shelf-life). Color changes were more evident when the fiber-rich fraction was added. The residual nitrite level increased with the addition of quinoa so that it would be necessary to incorporate less nitrites, or it might even be unnecessary, contributing to the production of more natural products.


Fixation identification in centroid versus start-point modes using eye-tracking data.
Fixation-identification algorithms, needed for analyses of eye movements, may typically be separated into three categories, viz. (i) velocity-based algorithms, (ii) area-based algorithms, and (iii) dispersion-based algorithms. Dispersion-based algorithms are commonly used but this application introduces some difficulties, one being optimization. Basically, there are two modes to reach this goal of optimization, viz., the start-point mode and the centroid mode. The aim of the present study was to compare and evaluate these two dispersion-based algorithms. Manual inspections were made of 1,400 fixations in each mode. Odds ratios showed that by using the centroid mode for fixation detection, a valid fixation is 2.86 times more likely to be identified than by using the start-point mode. Moreover, the algorithm based on centroid mode dispersion showed a good interpretation speed, accuracy, robustness, and ease of implementation, as well as adequate parameter settings.


Mass spectrometry reveals synergistic effects of nucleotides, lipids, and drugs binding to a multidrug resistance efflux pump.
Multidrug resistance is a serious barrier to successful treatment of many human diseases, including cancer, wherein chemotherapeutics are exported from target cells by membrane-embedded pumps. The most prevalent of these pumps, the ATP-Binding Cassette transporter P-glycoprotein (P-gp), consists of two homologous halves each comprising one nucleotide-binding domain and six transmembrane helices. The transmembrane region encapsulates a hydrophobic cavity, accessed by portals in the membrane, that binds cytotoxic compounds as well as lipids and peptides. Here we use mass spectrometry (MS) to probe the intact P-gp small molecule-bound complex in a detergent micelle. Activation in the gas phase leads to formation of ions, largely devoid of detergent, yet retaining drug molecules as well as charged or zwitterionic lipids. Measuring the rates of lipid binding and calculating apparent KD values shows that up to six negatively charged diacylglycerides bind more favorably than zwitterionic lipids. Similar experiments confirm binding of cardiolipins and show that prior binding of the immunosuppressant and antifungal antibiotic cyclosporin A enhances subsequent binding of cardiolipin. Ion mobility MS reveals that P-gp exists in an equilibrium between different states, readily interconverted by ligand binding. Overall these MS results show how concerted small molecule binding leads to synergistic effects on binding affinities and conformations of a multidrug efflux pump.