The effects of EBV transformation on gene expression levels and methylation profiles.
Epstein-Barr virus (EBV) transformed lymphoblastoid cell lines (LCLs) provide a conveniently accessible and renewable resource for functional genomic studies in humans. The ability to accumulate multidimensional data pertaining to the same individual cell lines, from complete genomic sequences to detailed gene regulatory profiles, further enhances the utility of LCLs as a model system. A lingering concern, however, is that the changes associated with EBV transformation of B cells reduce the usefulness of LCLs as a surrogate model for primary tissues. To evaluate the validity of this concern, we compared global gene expression and methylation profiles between CD20+ primary B cells sampled from six individuals and six independent replicates of transformed LCLs derived from each sample. These data allowed us to obtain a detailed catalog of the genes and pathways whose regulation is affected by EBV transformation. We found that the expression levels and promoter methylation profiles of more than half of the studied genes were affected by the EBV transformation, including enrichments of genes involved in transcription regulation, cell cycle and immune response. However, we show that most of the differences in gene expression levels between LCLs and B cells are of small magnitude, and that LCLs can often recapitulate the naturally occurring gene expression variation in primary B cells. Thus, our observations suggest that inference of the genetic architecture that underlies regulatory variation in LCLs can typically be generalized to primary B cells. In contrast, inference based on functional studies in LCLs may be more limited to the cell lines.


Evolutionary criteria in thallophytes: a radical alternative.
The classical assumptions, upon which all previous phylogenies for the lower plants (Thallophytes) have been based, are claimed to be erroneous. An alternative view, that the eukaryotic cell arose in the late Precambrian from prokaryotic ancestors by a specific series of symbioses, is referred to here. Mutually consistent phylogenies, one for the prokaryotes, another for the lower eukaryotes, can be constructed on the basis of the symbiotic theory. The resulting prokaryote phylogeny is presented here; it is claimed to be more consistent with cytological data, measured DNA base ratios, and the fossil record than the several classical partial phylogenies for Thallophytes recently published.


Unique kinetics of nicotinic acid-adenine dinucleotide phosphate (NAADP) binding enhance the sensitivity of NAADP receptors for their ligand.
Nicotinic acid-adenine dinucleotide phosphate (NAADP) is a novel and potent Ca(2+)-mobilizing agent in sea urchin eggs and other cell types. Little is known, however, concerning the properties of the putative intracellular NAADP receptor. In the present study we have characterized NAADP binding sites in sea urchin egg homogenates. [(32)P]NAADP bound to a single class of high-affinity sites that were reversibly inhibited by NaCl but insensitive to pH and Ca(2+). Binding of [(32)P]NAADP was lost in preparations that did not mobilize Ca(2+) in response to NAADP, indicating that [(32)P]NAADP probably binds to a receptor mediating Ca(2+) mobilization. Addition of excess unlabelled NAADP, at various times after initiation of [(32)P]NAADP binding, did not result in displacement of bound [(32)P]NAADP. These data show that NAADP becomes irreversibly bound to its receptor immediately upon association. Accordingly, incubation of homogenates with low concentrations of NAADP resulted in maximal labelling of NAADP binding sites. This unique property renders NAADP receptors exquisitely sensitive to their ligand, thereby allowing detection of minute changes in NAADP levels.


An evaluation of the characteristics and outcomes of users of a street triage service: retrospective case note review.
Aims and methodStreet triage services are now common but the population they serve is poorly understood. We aimed to evaluate a local service to determine the characteristics of those using it and their outcomes in the 90 day period following contact. RESULTS: We found that there were high levels of service use and that the vast majority of contacts were via telephone rather than in person. Street triage was used by both existing secondary mental health patients and non-patients. Follow-up rates with secondary services were high in the former and low in the latter case.ImplicationsServices are very busy where they exist and may be replacing traditional crisis services. It is not apparent that they work to increase follow-up among those using them, unless they are already in contact with services. In this service, although there was a joint response model nearly all responses were provided by telephone.Declaration of interestNone.


Polymorphisms in pockets of major histocompatibility complex class I molecules influence peptide preference.
The set of peptides that is bound by a given major histocompatibility complex class I product can be described by one or two properly spaced anchor residues, and two properly spaced peptide termini, approximately 8-10 residues apart. Using radiolabeled peptide libraries, we examined whether mutations in those "pockets" in class I Kb molecules that do not seem critically involved in the interaction with the peptide anchor residues, do exert an effect on the set of preferred peptides. We find that mutations in all the pockets found in the structure of Kb have a significant effect on the peptide preference of the molecule, and their recognition by cytotoxic T cells. Alterations in substrate specificity are also observed for mutations involving residues that interact with main chain atoms in both peptide termini. These findings challenge a static view of the interaction of peptide termini with their respective pockets in the class I molecule, and imply a role for the minor pockets in peptide selectivity.


Evaluating Charge Equilibration Methods To Generate Electrostatic Fields in Nanoporous Materials.
Charge equilibration (Qeq) methods can estimate the electrostatic potential of molecules and periodic frameworks by assigning point charges to each atom, using only a small fraction of the resources needed to compute density functional (DFT)-derived charges. This makes possible, for example, the computational screening of thousands of microporous structures to assess their performance for the adsorption of polar molecules. Recently, different variants of the original Qeq scheme were proposed to improve the quality of the computed point charges. One focus of this research was to improve the gas adsorption predictions in metal-organic frameworks (MOFs), for which many different structures are available. In this work, we review the evolution of the method from the original Qeq scheme, understanding the role of the different modifications on the final output. We evaluated the result of combining different protocols and set of parameters, by comparing the Qeq charges with high quality DFT-derived DDEC charges for 2338 MOF structures. We focused on the systematic errors that are attributable to specific atom types to quantify the final precision that one can expect from Qeq methods in the context of gas adsorption where the electrostatic potential plays a significant role, namely, CO2 and H2S adsorption. In conclusion, both the type of algorithm and the input parameters have a large impact on the resulting charges, and we draw some guidelines to help the user to choose the proper combination of the two for obtaining a meaningful set of charges. We show that, considering this set of MOFs, the accuracy of the original Qeq scheme is often still comparable with the most recent variants, even if it clearly fails in the presence of certain atom types, such as alkali metals.


Activity of Chlorine Dioxide in a Solution of Ions and pH Against Thielaviopsis basicola and Fusarium oxysporum.
Chlorine dioxide (ClO2) is a disinfestant used to control pathogens in water. To determine if interactions between inorganic ions and pH levels effect ClO2 activity in vitro, concentrations of ClO2 (0, 1, 3, 5, 7, 9, 22, 24, 46, 58, and 70 mg/liter) were mixed for 10 min in solutions containing a nitrogen and hard water solution with equal concentrations of ammonium, nitrate, and synthetic hard water (0 and 100 mg/liter) and a divalent metal ion solution with equal concentrations of copper, iron, manganese, and zinc (0, 1, 3, and 5 mg/liter) at pH 5 and 8. Macro- and microconidia of Fusarium oxysporum f. sp. narcissi or conidia and aleuriospores of Thielaviopsis basicola were injected into each suspension for 30 s, captured on filter paper disks that were flushed with water, and plated on 50% potato dextrose agar. Spore germination was quantified after 1 day. ClO2 activity had a similar effect on both fungal species and all types of propagules with interactions among the divalent metal ion solution, nitrogen and hard water solution, and pH treatments. A higher concentration of ClO2 was required at pH 8 than at pH 5 to achieve a lethal dose resulting in 50% mortality of spores (LD50). The addition of the divalent metal ion solution required an increase in ClO2 concentration to maintain a LD50. When combined with the nitrogen and hard water solution, the divalent metal ion solution placed a higher demand on ClO2 at pH 5 and a lower demand on ClO2 at pH 8, thus requiring an increase and decrease in a ClO2 concentration, respectively, to achieve a LD50. Chlorine dioxide doses resulting in 50% mortality ranged from 0.5 to 7.0 mg/liter for conidia of F. oxysporum, 0.5 to 11.9 mg/liter for conidia of T. basicola, and 15.0 to 45.5 mg/liter for aleuriospores of T. basicola.


Hermite Functional Link Neural Network for Solving the Van der Pol-Duffing Oscillator Equation.
Hermite polynomial-based functional link artificial neural network (FLANN) is proposed here to solve the Van der Pol-Duffing oscillator equation. A single-layer hermite neural network (HeNN) model is used, where a hidden layer is replaced by expansion block of input pattern using Hermite orthogonal polynomials. A feedforward neural network model with the unsupervised error backpropagation principle is used for modifying the network parameters and minimizing the computed error function. The Van der Pol-Duffing and Duffing oscillator equations may not be solved exactly. Here, approximate solutions of these types of equations have been obtained by applying the HeNN model for the first time. Three mathematical example problems and two real-life application problems of Van der Pol-Duffing oscillator equation, extracting the features of early mechanical failure signal and weak signal detection problems, are solved using the proposed HeNN method. HeNN approximate solutions have been compared with results obtained by the well known Runge-Kutta method. Computed results are depicted in term of graphs. After training the HeNN model, we may use it as a black box to get numerical results at any arbitrary point in the domain. Thus, the proposed HeNN method is efficient. The results reveal that this method is reliable and can be applied to other nonlinear problems too.


Cell-autonomous and inductive signals can determine the sex of the germ line of drosophila by regulating the gene Sxl.
To investigate the mechanism of sex determination in the germ line, we analyzed the fate of XY germ cells in ovaries, and the fate of XX germ cells in testes. In ovaries, germ cells developed according to their X:A ratio, i.e., XX cells underwent oogenesis, XY cells formed spermatocytes. In testes, however, XY and XX germ cells entered the spermatogenic pathway. Thus, to determine their sex, the germ cells of Drosophila have cell-autonomous genetic information, and XX cells respond to inductive signals of the soma. Results obtained with amorphic and constitutive mutations of Sxl show that both the genetic and the somatic signals act through Sxl to achieve sex determination in germ cells.


Structural and functional abnormalities of retinal ganglion cells measured in vivo at the onset of optic nerve head surface change in experimental glaucoma.
To compare peripapillary retinal nerve fiber layer thickness (RNFLT), RNFL retardance, and retinal function at the onset of optic nerve head (ONH) surface topography change in experimental glaucoma (EG). Thirty-three rhesus macaques had three or more weekly baseline measurements in both eyes of ONH surface topography, peripapillary RNFLT, RNFL retardance, and multifocal electroretinography (mfERG). Laser photocoagulation was then applied to the trabecular meshwork of one eye to induce chronic elevation of IOP and weekly recordings continued alternating between ONH surface topography and RNFLT during one week and RNFL retardance and mfERG the next week. Data were pooled for the group at the onset of ONH surface topography change in each EG eye, which was defined as the first date when either the mean position of the disc (MPD) fell below the 95% confidence limit of each eye's individual baseline range and/or when the topographic change analysis (TCA) map was subjectively judged as having demonstrated change, whichever came first. Analysis of variance with post hoc tests corrected for multiple comparisons were used to assess parameter changes. At onset of ONH surface topography change, there was no significant difference for RNFLT versus baseline or fellow control eyes. RNFL retardance and mfERG were significantly reduced in the recordings just prior (median of 9 days) to ONH onset (P < 0.01) and had progressed significantly (P < 0.001) an average of 17 days later (median of 7 days after ONH onset). RNFLT did not exhibit significant thinning until 15 days after onset of ONH surface topography change (P < 0.001). These results support the hypothesis that during the course of glaucomatous neurodegeneration, axonal cytoskeletal and retinal ganglion cell functional abnormalities exist before thinning of peripapillary RNFL axon bundles begins.


Diabatic free energy curves and coordination fluctuations for the aqueous Ag+Ag2+ redox couple: a biased Born-Oppenheimer molecular dynamics investigation.
Biased Born-Oppenheimer molecular dynamics simulations are performed to compute redox potential and free energy curves for the redox half reaction Ag(+)-->Ag(2+)+e(-) in aqueous solution. The potential energy surfaces of reactant and product state are linearly coupled and the system transferred from the reduced state to the oxidized state by variation of the coupling parameter from 0 to 1. The redox potential is obtained by thermodynamic integration of the average ionization energy of Ag(+). Diabatic free energy curves of reduced (R) and oxidized (O) states are obtained to good statistical accuracy by reweighting and combining the set of biased distributions of the ionization energy. The diabatic free energy curves of Ag(+) and Ag(2+) are parabolic over a wide range of the reaction coordinate in agreement with the linear response assumption that underlies Marcus theory. However, we observe deviations from parabolic behavior in the equilibrium region of Ag(+) and find different values for the reorganization free energy of R (1.4 eV) and O (0.9 eV). The computed reorganization free energy of Ag(2+) is in good agreement with the experimental estimate of 0.9-1.2 eV obtained from photoelectron spectroscopy. As suggested by our calculations, the moderate deviation from linear response behavior found for Ag(+) is likely related to the highly fluxional solvation shell of this ion, which exhibits water exchange reactions on the picosecond time scale of the present molecular dynamics simulation.


Rapid Prediction of Moisture Content in Intact Green Coffee Beans Using Near Infrared Spectroscopy.
Moisture content (MC) is one of the most important quality parameters of green coffee beans. Therefore, its fast and reliable measurement is necessary. This study evaluated the feasibility of near infrared (NIR) spectroscopy and chemometrics for rapid and non-destructive prediction of MC in intact green coffee beans of both Coffeaarabica (Arabica) and Coffeacanephora (Robusta) species. Diffuse reflectance (log 1/R) spectra of intact beans were acquired using a bench top Fourier transform NIR instrument. MC was determined gravimetrically according to The International Organization for Standardization (ISO) 6673. Samples were split into subsets for calibration (n = 64) and independent validation (n = 44). A three-component partial least squares regression (PLSR) model using raw NIR spectra yielded a root mean square error of prediction (RMSEP) of 0.80% MC; a four component PLSR model using scatter corrected spectra yielded a RMSEP of 0.57% MC. A simplified PLS model using seven selected wavelengths (1155, 1212, 1340, 1409, 1724, 1908, and 2249 nm) yielded a similar accuracy (RMSEP: 0.77% MC) which opens the possibility of creating cheaper NIR instruments. In conclusion, NIR diffuse reflectance spectroscopy appears to be suitable for rapid and reliable MC prediction in intact green coffee; no separate model for Arabica and Robusta species is needed.


TU-E-BRB-11: End-To-End Positioning Quality Assurance for Image-Guided Radiosurgery of Multiple Targets Using a Single-Isocenter.
Using a single isocenter significantly reduces delivery times in radiosurgery involving multiple targets. However, because not every target can be placed at isocenter with this type of treatment, a conventional Winston-Lutz test cannot be used. We describe a novel Winston-Lutz like mulitarget test (MTT) for verifying accurate positioning. A target phantom, comprised of an acrylic plate with recesses for three 3/4″ spheres was constructed and a high-resolution (0.5×0.5×0.8 mm) CT scan obtained with PTFE spheres placed in the recesses. The scan was imported into a commercial treatment planning system and multiple beams were prepared, having their isocenter at the centroid of the arrangement of spheres. Every beam incorporated three MLC-defined rectangular apertures that circumscribed the spheres. Custom software selected setup parameters (table, gantry and collimator angle, MLC openings) such that the spheres were centered as precisely as possible within their respective MLC fields, considering the discrete width of collimator leaves. The phantom, with the PTFE replaced by steel spheres, was placed on the treatment couch and imaged using stereoscopic x-ray beams. A 6 degree-of-freedom robotic couch applied translations and rotations to reproduce the CT position. A MV EPID rendered images of the spheres within their respective apertures, allowing identification of sphere and aperture centers. Any error upstream would manifest itself as inaccurate centering of a sphere. Eight beams with table angle 0 and two beams each with table angles 49.7, 89.8, 272.3, and 310.1 were selected. The maximum calculated distance between any sphere and the respective aperture center was 0.07 mm. The median difference measured from the MV images ranged from 0.1 mm to 1.4 mm with a median of 0.8 mm. The MTT is a practical end-to-end test for quality assurance of the entire positioning process in multitarget radiosurgery, from CT scanning to beam delivery.


An experimental study of intraspecific variation, developmental timing, and heterochrony in fishes.
Heterochrony is widely regarded as an important evolutionary mechanism, one that may underlie most, if not all, morphological evolution, yet relatively few studies have examined variation in the sequence of development. Even fewer studies have been designed so that intraspecific variation in the relative sequence of developmental events can be assessed, although this variation must be the basis for evolutionary change. Intraspecific variation in developmental ossification sequences was documented from the zebrafish (Danio rerio) by Cubbage and Mabee (1996) and from the Siamese fighting fish (Betta splendens) by Mabee and Trendler (1996), but a quantitative analysis of the patterns within this variation was not made. Here, we quantify the effect of rearing temperature on the sequence of ossification and characterize the levels and patterns of intraspecific variation in these fishes. For Danio, there were no temperature effects on the sequence of bone development across the cranium, cranial region development, cartilage versus dermal bones, or lateral line bone versus nonassociated bones. Likewise the level of variation in relative sequence (position) of ossification was low, about two ranks, across temperatures. At higher temperatures, we found higher levels of variation in iterated cranial bones and less in bones forming early in the sequence. No temperature effects on variation were found among regions, between lateral line-associated bones and nonassociated bones, between median and paired bones, or across the entire sequence, indicating concordant variability among the three temperatures. Individual bones with the highest levels of variability were not consistent among temperatures. Baseline patterns of intraspecific variation in Danio were compared to those of Betta. For both species, the level of intraspecific variation in sequence position was low and the variability of cranial bones was concordant. Individual bones with the highest levels of variability were not consistent between species. In both species, variation was widespread (distributed evenly across the sequence). We used comparisons (among regions, between dermal and cartilage bones, between lateral line-associated and other bones, between median and paired bones, between iterated and noniterated bones, between feeding-associated bones and others) to see which subsets were most variable and thus potentially useful in predicting high levels of evolutionary change. The only subset of bones that was significantly more variable than others was cartilage bones. If interspecific patterns are parallel to these intraspecific differences, cartilage bones would be expected to show higher levels of heterochrony. Although concordance across the cranial ossification sequence and among regions in Danio, Betta, and two other teleosts, Oryzias and Barbus, suggests an evolutionarily conserved pattern of ossification, identity in sequence position across taxa was not observed for any bone. Thus, variation existed in sequence position across temperatures and species. Intraspecific variation of this sort may influence the morphological outcome and evolutionary trajectories of species.


A global logrank test for adaptive treatment strategies based on observational studies.
In studying adaptive treatment strategies, a natural question that is of paramount interest is whether there is any significant difference among all possible treatment strategies. When the outcome variable of interest is time-to-event, we propose an inverse probability weighted logrank test for testing the equivalence of a fixed set of pre-specified adaptive treatment strategies based on data from an observational study. The weights take into account both the possible selection bias in an observational study and the fact that the same subject may be consistent with more than one treatment strategy. The asymptotic distribution of the weighted logrank statistic under the null hypothesis is obtained. We show that, in an observational study where the treatment selection probabilities need to be estimated, the estimation of these probabilities does not have an effect on the asymptotic distribution of the weighted logrank statistic, as long as the estimation of the parameters in the models for these probabilities is n-consistent. Finite sample performance of the test is assessed via a simulation study. We also show in the simulation that the test can be pretty robust to misspecification of the models for the probabilities of treatment selection. The method is applied to analyze data on antidepressant adherence time from an observational database maintained at the Department of Veterans Affairs' Serious Mental Illness Treatment Research and Evaluation Center.


Andrographolide Ameliorates Inflammation and Fibrogenesis and Attenuates Inflammasome Activation in Experimental Non-Alcoholic Steatohepatitis.
Therapy for nonalcoholic steatohepatitis (NASH) is limited. Andrographolide (ANDRO), a botanical compound, has a potent anti-inflammatory activity due to its ability to inhibit NF-κB. ANDRO has been also shown to inhibit the NLRP3 inflammasome, a relevant pathway in NASH. Our aim was to evaluate the effects of ANDRO in NASH and its influence on inflammasome activation in this setting. Thus, mice were fed a choline-deficient-amino-acid-defined (CDAA) diet with/without concomitant ANDRO administration (1 mg/kg, 3-times/week). Also, we assessed serum levels of alanine-aminotransferase (ALT), liver histology, hepatic triglyceride content (HTC) and hepatic expression of pro-inflammatory, pro-fibrotic and inflammasome genes. Inflammasome activation was also evaluated in fat-laden HepG2 cells. Our results showed that ANDRO administration decreased HTC and attenuated hepatic inflammation and fibrosis in CDAA-fed mice. ANDRO treatment determined a strong reduction in hepatic macrophage infiltration and reduced hepatic mRNA levels of both pro-inflammatory and pro-fibrotic genes. In addition, mice treated with ANDRO showed reduced expression of inflammasome genes. Finally, ANDRO inhibited LPS-induced interleukin-1β expression through NF-κB inhibition in fat-laden HepG2 cells and inflammasome disassembly. In conclusion, ANDRO administration reduces inflammation and fibrosis in experimental NASH. Inflammasome modulation by a NF-κB-dependent mechanism may be involved in the therapeutic effects of ANDRO.


All common bipedal gaits emerge from a single passive model.
In this paper, we systematically investigate passive gaits that emerge from the natural mechanical dynamics of a bipedal system. We use an energetically conservative model of a simple spring-leg biped that exhibits well-defined swing leg dynamics. Through a targeted continuation of periodic motions of this model, we systematically identify different gaits that emerge from simple bouncing in place. We show that these gaits arise along one-dimensional manifolds that bifurcate into different branches with distinctly different motions. The branching is associated with repeated breaks in symmetry of the motion. Among others, the resulting passive dynamic gaits include walking, running, hopping, skipping and galloping. Our work establishes that the most common bipedal gaits can be obtained as different oscillatory motions (or nonlinear modes) of a single mechanical system with a single set of parameter values. For each of these gaits, the timing of swing leg motion and vertical motion is matched. This work thus supports the notion that different gaits are primarily a manifestation of the underlying natural mechanical dynamics of a legged system. Our results might explain the prevalence of certain gaits in nature, and may provide a blueprint for the design and control of energetically economical legged robots.


Current status on plasma biomarkers for acute mesenteric ischemia.
Clinical diagnosis of acute mesenteric ischemia is difficult. The aim of this review is to provide current status on the search for an accurate plasma biomarker for acute mesenteric ischemia. A search using the medical subject heading terms marker and mesenteric ischemia or intestinal ischemia or superior mesenteric artery occlusion or mesenteric venous thrombosis in the Medline and Embase databases from 1980 to 2011. Studies without a control group or a control group consisted of healthy individuals (human studies), or studies on intestinal reperfusion were excluded. Twenty animal and twelve human studies were identified. In human studies, the studied series of patients had a control group that had a need of laparotomy (n = 2), suspected acute mesenteric ischemia (n = 7), acute abdomen (n = 2) or systemic inflammatory response syndrome (n = 1). D: -dimer has been found to be the most consistent highly sensitive early marker, but specificity was low. The follow-up study on α-glutathione S-transferase yielded inferior sensitivity and accuracy than the preliminary study, clearly questioning the value of this marker. Intestinal fatty acid binding globulin (I-FABP) and D: -lactate are both interesting markers, but the results were conflicting. Different cut-off levels have been used in the studies on I-FABP. The encouraging preliminary result of cobalt-albumin and urinary FABP as an accurate marker needs to be addressed in other study populations. The early clinical and laboratory diagnosis of intestinal ischemia remains a challenge. None of the proposed plasma-derived tests for acute mesenteric ischemia has as yet entered routine clinical practice. The proposed biomarkers need to be evaluated in a prospective clinical research project in patients with acute abdomen.


Hand before foot? Cortical somatotopy suggests manual dexterity is primitive and evolved independently of bipedalism.
People have long speculated whether the evolution of bipedalism in early hominins triggered tool use (by freeing their hands) or whether the necessity of making and using tools encouraged the shift to upright gait. Either way, it is commonly thought that one led to the other. In this study, we sought to shed new light on the origins of manual dexterity and bipedalism by mapping the neural representations in the brain of the fingers and toes of living people and monkeys. Contrary to the 'hand-in-glove' notion outlined above, our results suggest that adaptations underlying tool use evolved independently of those required for human bipedality. In both humans and monkeys, we found that each finger was represented separately in the primary sensorimotor cortex just as they are physically separated in the hand. This reflects the ability to use each digit independently, as required for the complex manipulation involved in tool use. The neural mapping of the subjects' toes differed, however. In the monkeys, the somatotopic representation of the toes was fused, showing that the digits function predominantly as a unit in general grasping. Humans, by contrast, had an independent neurological representation of the big toe (hallux), suggesting association with bipedal locomotion. These observations suggest that the brain circuits for the hand had advanced beyond simple grasping, whereas our primate ancestors were still general arboreal quadrupeds. This early adaptation laid the foundation for the evolution of manual dexterity, which was preserved and enhanced in hominins. In hominins, a separate adaptation, involving the neural separation of the big toe, apparently occurred with bipedality. This accords with the known fossil evidence, including the recently reported hominin fossils which have been dated to 4.4 million years ago.


In vitro activity of tedizolid and linezolid against Staphylococcus epidermidis isolated from prosthetic joint infections.
Prosthetic joint infections (PJIs) are rare but long-lasting and are serious complications without any spontaneous resolution, requiring additional surgery and long-term treatment with antibiotics. Staphylococci are the most important aetiological agents of PJIs, and among the coagulase-negative staphylococci Staphylococcus epidermidis is the most common. However, S. epidermidis often displays multidrug resistance (MDR), demanding additional treatment options. The objective was to examine the effectiveness of tedizolid and linezolid against S. epidermidis isolated from PJIs. The standard antibiotic susceptibility pattern of S. epidermidis (n = 183) obtained from PJIs was determined by disc diffusion test, and MIC was determined by Etest for tedizolid, linezolid, and vancomycin. Tedizolid displayed MIC values ranging from 0.094 to 0.5 mg/L (MIC50: 0.19 mg/L, MIC90: 0.38 mg/L), linezolid MIC values ranging from 0.25 to 2 mg/L (MIC50: 0.75 mg/L, MIC90: 1 mg/L), and vancomycin MIC values ranging from 0.5 to 3 mg/L (MIC50 and MIC90 both 2 mg/L). According to the disc diffusion test, 153/183 (84%) isolates were resistant to ≥3 antibiotic groups, indicating MDR. In conclusion, S. epidermidis isolates from PJIs were fully susceptible, and the MIC50 and MIC90 values for tedizolid were two- to four-fold dilution steps lower compared with linezolid. Tedizolid is not approved, and there are no reports of long-term treatment, but it may display better tolerability and fewer adverse effects than linezolid; it thus could be a possible treatment option for PJIs, alone or in combination with rifampicin.


Electromagnetic field of extremely low frequency has an impact on selected chemical components of the honeybee.
The electromagnetic field (EMF) is an environmental factor affecting living organisms. The aim of this study was to demonstrate the effect of an extremely low frequency electromagnetic field (ELF-EMF) on selected chemical components of the honeybee (Apis mellifera L.) using Fourier Transform Infrared (FTIR) spectroscopy. The FTIR method provides information on the chemical structure of compounds through identification and analysis of functional groups. The honeybees were treated with EMF at a frequency of 50 Hz and magnetic induction of 1.6 mT for 2, 6, 12, 24 and 48 hours. Analysis of FTIR spectra showed that EMF exposure longer than 2 hours induced changes in the structure of chemical compounds, especially in the IR region corresponding to DNA, RNA, phospholipids and protein vibrations, compared to control samples (bees not EMF treated). The results confirm the effect of EMF on bees depending on the duration of exposure.


Collective antiskyrmion-mediated phase transition and defect-induced melting in chiral magnetic films.
Magnetic phase transitions are a manifestation of competing interactions whose behavior is critically modified by defects and becomes even more complex when topological constraints are involved. In particular, the investigation of skyrmions and skyrmion lattices offers insight into fundamental processes of topological-charge creation and annihilation upon changing the magnetic state. Nonetheless, the exact physical mechanisms behind these phase transitions remain unresolved. Here, we show numerically that it is possible to collectively reverse the polarity of a skyrmion lattice in a field-induced first-order phase transition via a transient antiskyrmion-lattice state. We thus propose a new type of phase transformation where a skyrmion lattice inverts to another one due to topological constraints. In the presence of even a single defect, the process becomes a second-order phase transition with gradual topological-charge melting. This radical change in the system's behavior from a first-order to a second-order phase transition demonstrates that defects in real materials could prevent us from observing collective topological phenomena. We have systematically compared ultra-thin films with isotropic and anisotropic Dzyaloshinskii-Moriya interactions (DMIs), and demonstrated a nearly identical behavior for such technologically relevant interfacial systems.


Simultaneous therapy with antiplatelet and anticoagulant drugs in symptomatic cardiovascular disease.
Twenty of approximately 1000 patients attending the arteriosclerosis clinic at MIT during a 13 year period were treated simultaneously with aspirin and warfarin for symptomatic atherosclerotic (19) or rheumatic (1) heart or vascular disease. The average duration of therapy was 5.8 years. Thirteen patients suffered from familial hyperlipoproteinemia; only one patient had none of the major risk factors for arteriosclerosis. Refractory symptoms were related to the central nervous system in 13, peripheral vascular system in 5 and the heart in 2. All twenty patients became asymptomatic or showed marked clinical improvement on aspirin plus warfarin therapy. While on this therapy, complications, both thrombotic and hemorrhagic, occurred in 7 of the 20 patients (graft embolus in 1, and bleeding in 6; with one death as a result of intracranial bleeding) and sudden death, probably from acute myocardial ischemia, in a further 2 patients. We conclude that when alternative therapies are impossible or have proven to be of no avail in patients suffering from the complications of advanced atherosclerosis, the simultaneous administration of aspirin and warfarin may be a therapeutic alternative, although very close and careful followup of the patients' prothrombin times and clinical status is essential.


Infection by a Giant Virus (AaV) Induces Widespread Physiological Reprogramming in Aureococcus anophagefferens CCMP1984 - A Harmful Bloom Algae.
While viruses with distinct phylogenetic origins and different nucleic acid types can infect and lyse eukaryotic phytoplankton, "giant" dsDNA viruses have been found to be associated with important ecological processes, including the collapse of algal blooms. However, the molecular aspects of giant virus-host interactions remain largely unknown. Aureococcus anophagefferens virus (AaV), a giant virus in the Mimiviridae clade, is known to play a critical role in regulating the fate of brown tide blooms caused by the pelagophyte Aureococcus anophagefferens. To understand the physiological response of A. anophagefferens CCMP1984 upon AaV infection, we studied the transcriptomic landscape of this host-virus pair over an entire infection cycle using a RNA-sequencing approach. A massive transcriptional response of the host was evident as early as 5 min post-infection, with modulation of specific processes likely related to both host defense mechanism(s) and viral takeover of the cell. Infected Aureococcus showed a relative suppression of host-cell transcripts associated with photosynthesis, cytoskeleton formation, fatty acid, and carbohydrate biosynthesis. In contrast, host cell processes related to protein synthesis, polyamine biosynthesis, cellular respiration, transcription, and RNA processing were overrepresented compared to the healthy cultures at different stages of the infection cycle. A large number of redox active host-selenoproteins were overexpressed, which suggested that viral replication and assembly progresses in a highly oxidative environment. The majority (99.2%) of annotated AaV genes were expressed at some point during the infection cycle and demonstrated a clear temporal-expression pattern and an increasing relative expression for the majority of the genes through the time course. We detected a putative early promoter motif for AaV, which was highly similar to the early promoter elements of two other Mimiviridae members, indicating some degree of evolutionary conservation of gene regulation within this clade. This large-scale transcriptome study provides insights into the Aureococcus cells infected by a giant virus and establishes a foundation to test hypotheses regarding metabolic and regulatory processes critical for AaV and other Mimiviridae members.


Extraction of phenolics and changes in antioxidant activity of red wines during vinification.
The moderate consumption of alcoholic beverages has been associated with protection against the development of coronary heart disease. Although alcohol itself can help prevent coronary heart disease through a number of mechanisms, red wine appears to offer protection above and beyond that attributable to alcohol alone. Red wine is a complex fluid containing grape, yeast, and wood-derived phenolic compounds, the majority of which have been recognized as potent antioxidants. The aim of this study was to investigate the major phenolic contributors to the antioxidant activity of wine. To this end, four wines were followed during the first 7-9 days of vinification. Individual phenolic compounds were quantified by HPLC, and antioxidant activity was determined by electron spin resonance spectroscopy. The extraction of the phenolics was found to be influenced by vinification procedure, grape quality, and grape variety. Although fermenting wines reached a total phenolic content comparable to that of a bottled wine after 9 days of vinification, the antioxidant activity was significantly lower than that of a finished wine. This suggests that the larger polyphenolic complexes and condensation products that appear during aging make a sizable contribution to the overall antioxidant activity of red wines.


MHCII-independent CD4+ T cells protect injured CNS neurons via IL-4.
A body of experimental evidence suggests that T cells mediate neuroprotection following CNS injury; however, the antigen specificity of these T cells and how they mediate neuroprotection are unknown. Here, we have provided evidence that T cell-mediated neuroprotection after CNS injury can occur independently of major histocompatibility class II (MHCII) signaling to T cell receptors (TCRs). Using two murine models of CNS injury, we determined that damage-associated molecular mediators that originate from injured CNS tissue induce a population of neuroprotective, IL-4-producing T cells in an antigen-independent fashion. Compared with wild-type mice, IL-4-deficient animals had decreased functional recovery following CNS injury; however, transfer of CD4+ T cells from wild-type mice, but not from IL-4-deficient mice, enhanced neuronal survival. Using a culture-based system, we determined that T cell-derived IL-4 protects and induces recovery of injured neurons by activation of neuronal IL-4 receptors, which potentiated neurotrophin signaling via the AKT and MAPK pathways. Together, these findings demonstrate that damage-associated molecules from the injured CNS induce a neuroprotective T cell response that is independent of MHCII/TCR interactions and is MyD88 dependent. Moreover, our results indicate that IL-4 mediates neuroprotection and recovery of the injured CNS and suggest that strategies to enhance IL-4-producing CD4+ T cells have potential to attenuate axonal damage in the course of CNS injury in trauma, inflammation, or neurodegeneration.


Respiratory function during pressure support ventilation.
Pressure support ventilation (PSV) is a pressure assist form of mechanical ventilatory support that augments the patient's spontaneous inspiratory efforts with a clinician selected level of positive airway pressure. To understand the effects of PSV on respiratory function, experiments were performed on 15 stable patients requiring synchronized intermittent mandatory ventilation (SIMV), as well as on a mechanical model simulating these patients' ventilatory systems. In the clinical study, gas exchange, airway pressures, blood pressure and heart rate were measured while SIMV was replaced by enough PSV to approximate the baseline SIMV tidal volume (VT). Measurements were repeated while this PSV level was then reduced in three 5 cm H2O steps every 10 to 15 minutes. It was found that PSV was a reasonable form of mechanical ventilatory support in patients with spontaneous ventilatory drives. It improves patient comfort, reduces the patient's ventilatory work, and provides a more balanced pressure and volume change form of muscle work to the patient. The clinical significance of these properties during the weaning process remain to be determined.


Incorporating telemedicine into the integrated care of the COPD patient a summary of an interdisciplinary workshop held in Stresa, Italy, 7-8 September 2017.
This report is a summary of a workshop focusing on using telemedicine to facilitate the integrated care of chronic obstructive pulmonary disease (COPD). Twenty-five invited participants from 8 countries met for one and one-half days in Stresa, Italy on 7-8 September 2017, to discuss this topic. Participants included physiotherapists, nurses, a nurse practitioner, and physicians. While evidence-based data are always at the center of sound inference and recommendations, at this point in time the science behind telemedicine in COPD remains under-developed; therefore, this document reflects expert opinion and consensus. While telemedicine has great potential to expand and improve the care of our COPD patients, its application is still in its infancy. While studies have demonstrated its effectiveness in some patient-centered outcomes, the results are by no means consistently positive. Whereas this tool may potentially reduce health care costs by moving some medical interventions from centralized locations in to patient's home, its cost-effectiveness has had mixed results and telemonitoring has yet to prove its worth in the COPD population. These discordant results should not be unexpected in view of patient complexity and the heterogeneity of telemedicine. This is reflected in the very limited support offered by the National Health Services to a wider application of telemedicine in the integrated care of COPD patients. However, this situation should challenge us to develop the necessary science to clarify the role of telemedicine in the medical management of our patients, providing a better and definitive scientific basis to this approach.


Fragman: an R package for fragment analysis.
Determination of microsatellite lengths or other DNA fragment types is an important initial component of many genetic studies such as mutation detection, linkage and quantitative trait loci (QTL) mapping, genetic diversity, pedigree analysis, and detection of heterozygosity. A handful of commercial and freely available software programs exist for fragment analysis; however, most of them are platform dependent and lack high-throughput applicability. We present the R package Fragman to serve as a freely available and platform independent resource for automatic scoring of DNA fragment lengths diversity panels and biparental populations. The program analyzes DNA fragment lengths generated in Applied Biosystems® (ABI) either manually or automatically by providing panels or bins. The package contains additional tools for converting the allele calls to GenAlEx, JoinMap® and OneMap software formats mainly used for genetic diversity and generating linkage maps in plant and animal populations. Easy plotting functions and multiplexing friendly capabilities are some of the strengths of this R package. Fragment analysis using a unique set of cranberry (Vaccinium macrocarpon) genotypes based on microsatellite markers is used to highlight the capabilities of Fragman. Fragman is a valuable new tool for genetic analysis. The package produces equivalent results to other popular software for fragment analysis while possessing unique advantages and the possibility of automation for high-throughput experiments by exploiting the power of R.


Occupational risk of lung cancer among lifetime non-smoking women in Shanghai, China.
Occupational lung carcinogens have been primarily studied in men. The aim of this study was to investigate occupational lung cancer risk in a cohort of Chinese non-smoking women. In 1996-2000, 71 067 non-smoking women who had held a job outside the home were interviewed for the prospective Shanghai Women's Health Study in China. Exposure to lung carcinogens was assessed by matching occupation and industry titles from lifetime occupational histories with lists of jobs identified by the International Agency for Research on Cancer to have potential exposure to: (1) known (A-list); or (2) suspected (B-list) carcinogens. In addition, similar occupational titles were grouped independent of the a priori defined lists. Relative risks (RRs) were calculated using Cox proportional hazards regression. During follow-up through 2005, 219 incident lung cancer cases were diagnosed. Jobs on the A-list and B-list were held by 0.8-6.7% and 2.7-9.4% of the cohort, respectively. Overall, ever holding any job on the A-list or B-list was not associated with lung cancer incidence. Indications of excess risk were found for two subgroups: painters (A-list) and rubber workers (B-list) (RR = 2.0 and 1.7, respectively, p<or=0.1). An exploratory analysis of 35 occupational categories independent of the lists showed significantly increased risks for leather products/shoes, wood/paper products and miscellaneous production/transportation. The former two of these categories were similar to subgroups of the B-list, but broader than the specific a priori defined jobs. Significantly elevated lung cancer risk was associated with employment in some broad occupational categories that also included jobs with potential exposure to suspected carcinogens (B-list). The results suggest that although similar exposures to those described on the B-list may play a role in this cohort of Chinese women, carcinogenic exposure may not be restricted only to the jobs on the B-list.


University of North Carolina/Emory Center for Innovative Technology (iTech) for Addressing the HIV Epidemic Among Adolescents and Young Adults in the United States: Protocol and Rationale for Center Development.
Over a fifth of all new HIV infections in the United States occur among persons aged 13 24 years, with most of these diagnoses occurring among gay and bisexual males (81%). While the epidemic of HIV in the United States has leveled off for many age groups, the annual number of new HIV diagnoses among young men who have sex with men (YMSM; 13-24 years old) remains high. Traditional approaches to continuum improvement for youth have been insufficient, and targeted interventions are urgently needed for young people at risk for or infected with HIV. Interventions delivered through mobile health technology represent a promising approach for improving outcomes in this population. Mobile phones have nearly reached saturation among youth, making mobile technology a particularly promising tool for reaching this population. The University of North Carolina/Emory Center for Innovative Technology (iTech) is a National Institutes of Health cooperative agreement as part of the Adolescent Medicine Trials Network for HIV/AIDS Interventions. iTech aims to impact the HIV epidemic by conducting innovative, interdisciplinary research on technology-based interventions across the HIV prevention and care continuum for adolescents and young adults in the United States, particularly YMSM, by providing the following: (1) evaluation of novel approaches to identifying youth with undiagnosed HIV infections; (2) evaluation of multilevel, combination prevention approaches, particularly relevant to gender- and sexual-minority youth facing co-occurring health risks; (3) evaluation of uptake of and adherence to biomedical prevention modalities; and 4) evaluation of interventions designed to promote or optimize engagement in care and antiretroviral therapy adherence in HIV-positive youth, to optimize viral load suppression. iTech brings together multidisciplinary experts in the fields of adolescent HIV treatment and prevention, development and evaluation of technology-based interventions, HIV surveillance and epidemiology, and intervention design and evaluation. This initiative will support 8 efficacy trials and 2 exploratory projects, each led by 2 principal investigators. Taken together, the studies address all of the key steps of the HIV prevention and care continuum for youth in the United States. Each proposal uses technology in a scientifically rigorous and innovative way to access, engage, and impact at-risk or infected youth. Nine iTech subject recruitment venues are spread across 8 US cities. Three cores (management, analytic, and technology) support all iTech activities and form the research network's infrastructure, facilitating all aspects of study implementation and evaluation. Formative work has already begun on many of the above-mentioned iTech trials. We expect the first randomized controlled trials to begin in mid-2018. Additional details can be found in the individual intervention protocol papers in this issue. Through its comprehensive research portfolio, iTech aims to effectively advance HIV prevention and care for youth through technology-based, youth-relevant interventions that maximize adaptability and sustainability. RR1-10.2196/10365.


Short photoperiod-induced decrease of histamine H3 receptors facilitates activation of hypothalamic neurons in the Siberian hamster.
Nonhibernating seasonal mammals have adapted to temporal changes in food availability through behavioral and physiological mechanisms to store food and energy during times of predictable plenty and conserve energy during predicted shortage. Little is known, however, of the hypothalamic neuronal events that lead to a change in behavior or physiology. Here we show for the first time that a shift from long summer-like to short winter-like photoperiod, which induces physiological adaptation to winter in the Siberian hamster, including a body weight decrease of up to 30%, increases neuronal activity in the dorsomedial region of the arcuate nucleus (dmpARC) assessed by electrophysiological patch-clamping recording. Increased neuronal activity in short days is dependent on a photoperiod-driven down-regulation of H3 receptor expression and can be mimicked in long-day dmpARC neurons by the application of the H3 receptor antagonist, clobenproprit. Short-day activation of dmpARC neurons results in increased c-Fos expression. Tract tracing with the trans-synaptic retrograde tracer, pseudorabies virus, delivered into adipose tissue reveals a multisynaptic neuronal sympathetic outflow from dmpARC to white adipose tissue. These data strongly suggest that increased activity of dmpARC neurons, as a consequence of down-regulation of the histamine H3 receptor, contributes to the physiological adaptation of body weight regulation in seasonal photoperiod.


Lymphangiogenesis: fuel, smoke, or extinguisher of inflammation's fire?
Lymphangiogenesis is a recognized hallmark of inflammatory processes in tissues and organs as diverse as the skin, heart, bowel, and airways. In clinical and animal models wherein the signaling processes of lymphangiogenesis are manipulated, most studies demonstrate that an expanded lymphatic vasculature is necessary for the resolution of inflammation. The fundamental roles that lymphatics play in fluid clearance and immune cell trafficking from the periphery make these results seemingly obvious as a mechanism of alleviating locally inflamed environments: the lymphatics are simply providing a drain. Depending on the tissue site, lymphangiogenic mechanism, or induction timeframe, however, evidence shows that inflammation-associated lymphangiogenesis (IAL) may worsen the pathology. Recent studies have identified lymphatic endothelial cells themselves to be local regulators of immune cell activity and its consequential phenotypes - a more active role in inflammation regulation than previously thought. Indeed, results focusing on the immunocentric roles of peripheral lymphatic function have revealed that the basic drainage task of lymphatic vessels is a complex balance of locally processed and transported antigens as well as interstitial cytokine and immune cell signaling: an interplay that likely defines the function of IAL. This review will summarize the latest findings on how IAL impacts a series of disease states in various tissues in both preclinical models and clinical studies. This discussion will serve to highlight some emerging areas of lymphatic research in an attempt to answer the question relevant to an array of scientists and clinicians of whether IAL helps to fuel or extinguish inflammation. Impact statement Inflammatory progression is present in acute and chronic tissue pathologies throughout the body. Lymphatic vessels play physiological roles relevant to all medical fields as important regulators of fluid balance, immune cell trafficking, and immune identity. Lymphangiogenesis is often concurrent with inflammation and can potentially aide or worsen disease progression. How new lymphatic vessels impact inflammation and by which mechanism is an important consideration in current and future clinical therapies targeting inflammation and/or vasculogenesis. This review identifies, across a range of tissue-specific pathologies, the current understanding of inflammation-associated lymphangiogenesis in the progression or resolution of inflammation.


Intrahepatic cholestasis of pregnancy: correlation of preterm delivery with bile acids.
The aim of this study was to determine the incidence, obstetrical and fetal complication rates of intrahepatic cholestasis of pregnancy in patients managed actively around 38 weeks and evaluate the correlation of these results with liver function tests and bile acids. In this cohort study 3710 women were booked for delivery, of which 32 pregnant women were diagnosed as intrahepatic cholestasis of pregnancy. All data concerning obstetric- medical history, laboratory results, symptom onset time, pruritus degree, treatment response, and delivery time and infants information were recorded in the study protocol. Statistical analyses were conducted with SPSS 12.0 version and correlations were assessed by Spearman Rank correlation analysis. The incidence of intrahepatic cholestasis of pregnancy was 0.86%. The symptoms appeared around 32 weeks. 16.6% multiparas had a previously affected pregnancy and 21.8% of intrahepatic cholestasis of pregnancy patients had family history of intrahepatic cholestasis of pregnancy. Symptom onset varied according to season (p<0.05). Most patients (69.5%) were diagnosed in winter and the beginning of spring. There were no reported cases of clinical maternal jaundice, bleeding tendency or stillbirth. Pruritus was decreased by ursodeoxycholic acid treatment. Total bile acids tended to be higher in patients with preterm delivery (r=0.409, p=0.038). Total bile acids are correlated with preterm delivery. An attempt to deliver at around 38 weeks may improve perinatal outcome.


Deprivation and health in one general practice.
A detailed comparison made in late 1984 of 587 matched pairs of patients from neighbouring deprived and more endowed urban communities, both served by the same experienced primary health care team, showed much worse morbidity, with almost three times as much mental illness, in the deprived group. This group also had 60% more hospital admissions and 75% more casualty attendances. In contrast, they had a much lower uptake of preventive health care, especially childhood immunisations and cervical cytology in older women. A much higher birthrate within the deprived group, contributing to ill health by worsening overcrowding and deprivation, indicates that family planning is ineffective where it is most needed. Practical measures have now been instituted to improve the preventive care of the deprived community.


Analysis of ultrasonic frequency response of surface attached fiber Bragg grating.
In recent years, fiber Bragg grating (FBG), for the well-known advantages over other fiber optic sensors, has attracted more attention in ultrasonic inspection for structure health monitoring (SHM). Spectrum shift of FBG to ultrasonic wave is caused by the refractive index profile changing along the FBG, which can be attributed to nonuniform perturbation caused by strain-optic and geometric effects of ultrasonic wave. Response of FBG to the above two effects was analyzed firstly by the V-I transmission matrix model, showing high computing efficiency. Based on this model, spectra response of FBG under changing ultrasonic frequencies was simulated and discussed. In experiment, the system was able to detect a wideband ultrasonic wave ranging from 15 to 1380 kHz. These results would provide a guideline for an FBG-based acoustic detection system design in a specific ultrasonic frequency.


Male and female brain evolution is subject to contrasting selection pressures in primates.
The claim that differences in brain size across primate species has mainly been driven by the demands of sociality (the "social brain" hypothesis) is now widely accepted. Some of the evidence to support this comes from the fact that species that live in large social groups have larger brains, and in particular larger neocortices. Lindenfors and colleagues (BMC Biology 5:20) add significantly to our appreciation of this process by showing that there are striking differences between the two sexes in the social mechanisms and brain units involved. Female sociality (which is more affiliative) is related most closely to neocortex volume, but male sociality (which is more competitive and combative) is more closely related to subcortical units (notably those associated with emotional responses). Thus different brain units have responded to different selection pressures.


Mathematical modeling of an aqueous film coating process in a Bohle Lab-Coater, part 1: development of the model.
The purpose of this study was to develop a model to predict (1) air and product temperatures, (2) product moisture, and (3) air humidity during an aqueous coating process using a Bohle Lab-Coater. Because of the geometrical properties and the airflow, the drum of the Bohle Lab-Coater can in principle be divided into 2 zones of equal size-the drying and the spraying zones. For each zone, 4 balance equations could be set up describing the change of the air humidity, the product moisture, the enthalpy of the air, and the enthalpy of the product in each zone. For this purpose, knowledge regarding heat and mass transfer and also the motion of the tablets in drums was used. Based on the considerations of the heat and mass transfer, a set of first-order coupled ordinary differential equations (ODEs) was developed. This set of ODEs can be solved numerically. In this part, the development of the model is described in detail, whereas the application of the model can be found in part 2.


PRMT4-mediated arginine methylation negatively regulates retinoblastoma tumor suppressor protein and promotes E2F-1 dissociation.
The retinoblastoma protein (pRb/p105) tumor suppressor plays a pivotal role in cell cycle regulation by blockage of the G1-to-S-phase transition. pRb tumor suppressor activity is governed by a variety of posttranslational modifications, most notably phosphorylation by cyclin-dependent kinase (Cdk) complexes. Here we report a novel regulation of pRb through protein arginine methyltransferase 4 (PRMT4)-mediated arginine methylation, which parallels phosphorylation. PRMT4 specifically methylates pRb at the pRb C-terminal domain (pRb C(term)) on arginine (R) residues R775, R787, and R798 in vitro and R787 in vivo. Arginine methylation is important for efficient pRb C(term) phosphorylation, as manifested by the reduced phosphorylation of a methylation-impaired mutant, pRb (R3K). A methylmimetic form of pRb, pRb (R3F), disrupts the formation of the E2F-1/DP1-pRb complex in cells as well as in an isolated system. Finally, studies using a Gal4-E2F-1 reporter system show that pRb (R3F) expression reduces the ability of pRb to repress E2F-1 transcriptional activation, while pRb (R3K) expression further represses E2F-1 transcriptional activation relative to that for cells expressing wild-type pRb. Together, our results suggest that arginine methylation negatively regulates the tumor suppressor function of pRb during cell cycle control, in part by creating a better substrate for Cdk complex phosphorylation and disrupting the interaction of pRb with E2F-1.


A Computational Fluid-Structure Interaction Study for Carotids With Different Atherosclerotic Plaques.
Atherosclerosis is a systemic disease that leads to accumulation of deposits, known as atherosclerotic plaques, within the walls of the carotids. In particular, three types of plaque can be distinguished: soft, fibrous, and calcific. Most of the computational studies who investigated the interplay between the plaque and the blood flow on patient-specific geometries used nonstandard medical images to directly delineate and segment the plaque and its components. However, these techniques are not so widely available in the clinical practice. In this context, the aim of our work was twofold: (i) to propose a new geometric tool that allowed to reconstruct a plausible plaque in the carotids from standard images and (ii) to perform three-dimensional (3D) fluid-structure interaction (FSI) simulations where we compared some fluid-dynamic and structural quantities among 15 patients characterized by different typologies of plaque. Our results highlighted that both the morphology and the mechanical properties of different plaque components play a crucial role in determining the vulnerability of the plaque.


Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters.
Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA) and TOTO-1 [1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3dihydro(benzo-1,3-oxazole)-2-methylidene]-1-(3'-trimethylammoniumpropyl)-pyridinium tetraiodide] for (stressed) lactic acid bacteria (C. J. Bunthof, K. Bloemen, P. Breeuwer, F. M. Rombouts, and T. Abee, Appl. Environ. Microbiol. 67:2326-2335, 2001). cFDA stains intact cells with enzymatic activity, and TOTO-1 stains membrane-permeabilized cells. Here we used this assay to study the viability of bacterial suspensions in milk, dairy fermentation starters, and probiotic products. To facilitate FCM analysis of bacteria in milk, a commercially available milk-clearing solution was used. The procedure was optimized to increase the signal-to-noise ratio. FCM enumerations were accurate down to a concentration of 10(5) cells ml(-1). The level of retrieval of Lactobacillus plantarum WCFS 1 suspended in milk was high, and viability was not affected by the procedure. The plate counts for cleared samples of untreated cell suspensions were nearly as high as the total FCM counts, and the correlation was strong (r > 0.99). In dairy fermentation starters and in probiotic products the FCM total cell counts were substantially higher than the numbers of CFU. Three functional populations could be distinguished: culturable cells, cells that are intact and metabolically active but not culturable, and permeabilized cells. The proportions of the populations differed in the products tested. This FCM method provides tools to assess the functionality of different populations in fermentation starters and probiotic products.


Susceptibility locus for Alzheimer's disease on chromosome 10.
The apolipoprotein E (APOE) gene is the only genetic risk factor that has so far been linked to risk for late-onset Alzheimer's disease (LOAD). However, 50 percent of Alzheimer's disease cases do not carry an APOE4 allele, suggesting that other risk factors must exist. We performed a two-stage genome-wide screen in sibling pairs with LOAD to detect other susceptibility loci. Here we report evidence for an Alzheimer's disease locus on chromosome 10. Our stage one multipoint lod score (logarithm of the odds ratio for linkage/no linkage) of 2.48 (266 sibling pairs) increased to 3.83 in stage 2 (429 sibling pairs) close to D10S1225 (79 centimorgans). This locus modifies risk for Alzheimer's disease independent of APOE genotype.


Inactivation of Escherichia coli K-12 in Liquid Egg White By a Flow-through Pulsed Uv Light Treatment System.
Unpasteurized liquid egg can be contaminated with pathogenic microorganisms and may cause foodborne outbreaks. Thus, it is essential to decontaminate the liquid egg to ensure food safety. Pulsed UV light is one of the emerging technologies for food decontamination in recent years. This static treatment system has been studied previously in our laboratory. However, continuous processing using a flow-through treatment system needs to be evaluated for potential commercial applications. Therefore, in this study, a flow-through treatment system of pulsed UV light was evaluated and optimized for inactivation of Escherichia coli K12NSR for liquid egg white decontamination. Treatment factors including flow rate (40 to 80 mL/min), number of passes (one to three passes), and distance from the sample to the pulsed UV light strobe (5 to 13 cm) were optimized using response surface methodology. This methodology suggested three passes with 40 mL/min flow rate and a 5-cm distance as the optimum conditions. The model was then validated for the maximum reduction of E. coli K12NSR, which was measured as 1.57 log CFU/mL at the optimal conditions. The energy doses of the pulsed UV light and temperature changes of the liquid egg white during the treatment were measured. Furthermore, several quality parameters were assessed at the optimum treatment conditions to determine the impact of the flow-through pulsed UV processing on the quality of liquid egg white. The results showed significant differences in pH, lipid oxidation, turbidity, and color between control and pulsed UV light-treated samples (P < 0.05). However, there was no significant difference in foaming ability or foam stability between pulsed UV light-treated samples and the control. Overall, this study demonstrated the potential of flow-through pulsed UV light to decontaminate liquid egg white, but further research is needed for optimal enhancement.


Electronic imitation of behavioral and psychological synaptic activities using TiOx/Al2O3-based memristor devices.
Seeking an effective electronic synapse to emulate biological synaptic behavior is fundamental for building brain-inspired computers. An emerging two-terminal memristor, in which the conductance can be gradually modulated by external electrical stimuli, is widely considered as the strongest competitor of the electronic synapse. Here, we show the capability of TiOx/Al2O3-based memristor devices to imitate synaptic behaviors. Along with analog resistive switching performances, the devices replicate the bio-synapse behaviors of potentiation/depression, short-term-plasticity, and long-term-potentiation, which show that TiOx/Al2O3-based memristors may be useful as electronic synapses. The essential memorizing capabilities of the brain are dependent on the connection strength between neurons, and the memory types change from short-term memory to long-term memory. In the TiOx/Al2O3-based electronic synaptic junction, the memorizing levels can change their state via a standard rehearsal process and also via newly introduced process called "impact of event" i.e. the impact of pulse amplitude, and the width of the input pulse. The devices show a short-term to long-term memory effect with the introduction of intermediate mezzanine memory. The experimental achievements using the TiOx/Al2O3 electronic synapses are finally psychologically modeled by considering the mezzanine level. It is highly recommended that similar phenomena should be investigated for other memristor systems to check the authenticity of this model.


Impact of early surgery on survival of patients with severe mitral regurgitation.
Optimal timing of surgery in degenerative mitral regurgitation (MR) remains a controversial topic. The impact of current ACC/AHA guideline recommendations about optimal timing of surgery on outcomes is untested and contemporary data are lacking. To assess the association between the timing of surgery and long-term survival in patients with severe MR. A cohort of 481 patients with severe, degenerative mitral regurgitation (1995-2007) from the Duke Cardiovascular Disease Databank who fulfilled at least one ACC/AHA guideline indication for surgery was identified. Exclusion criteria were rheumatic disease, congenital mitral valve (MV) disease, hypertrophic cardiomyopathy, coronary disease in more than one vessel, endocarditis, other severe valve disease, h/o valve repair/replacement. Patients were grouped into early surgery (in ≤ 2 months of presenting with surgical indications) and late surgery (>2 months) groups. An adjusted Cox regression model was constructed for time to death after 2 months with a time-dependent covariate term for late surgery. 168 patients had early surgery (median time to surgery 0.42 months) with 153 followed up after 2 months, 94 had late surgery (median time to surgery 8.75 months) and 219 medically managed. 127/168 in the early surgery group and 84/94 in the late surgery group received MV repair (p=0.02). Over 5.6 years' (median) follow-up there were 35 deaths (21%) in the early surgery group, with two occurring before 2 months and 20 (21%) in the late group. In the multivariable model, those undergoing early surgery had a lower hazard for death than those who underwent late surgery (HR=0.54 (95% CI 0.30 to 0.97), p=0.039). MV repair was independently associated with survival (HR=0.45 (95% CI 0.25 to 0.83), p=0.01). In patients with severe MR who presented with guideline indications for surgery, those selected for earlier surgery had improved survival. These data support the current guidelines for early referral to surgery in patients with severe MR for enlarged left ventricular dimensions, reduced ejection fraction and symptoms rather than delaying surgery. Larger randomised trials are needed to definitively answer the question of optimal timing of surgery in patients with severe degenerative MR.


Tunable reflector with active magnetic metamaterials.
We placed active magnetic metamaterials on metallic surface to implement a tunable reflector with excellent agile performance. By incorporating active elements into the unit cells of the magnetic metamaterial, this active magnetic metamaterial can be tuned to switch function of the reflector among a perfect absorber, a perfect reflector and a gain reflector. This brings about DC control lines to electrically tune the active magnetic metamaterial with positive loss, zero loss and even negative loss. The design, analytical and numerical simulation methods, and experimental results of the tunable reflector are presented.


Male hormonal contraception: effects of injections of testosterone undecanoate and depot medroxyprogesterone acetate at eight-week intervals in chinese men.
Surveys indicate that one form of acceptable male hormonal contraception would consist of injections given at 2- to 3-month intervals. This report describes a study of depot medroxyprogesterone acetate (DMPA) and testosterone undecanoate (TU) injected at 8-wk intervals for suppression of spermatogenesis in healthy Chinese men. After screening, 30 healthy volunteers were enrolled and randomly assigned to one of three dose groups (n = 10/group): 1000 mg TU (group A); 1000 mg TU plus 150 mg DMPA (group B); 1000 mg TU plus 300 mg DMPA (group C). All doses were given as im injections at 8-wk intervals. The study consisted of an 8-wk control (baseline) period, a 24-wk treatment period, and a 24-wk recovery period. Consistent azoospermia or severe oligozoospermia was achieved and maintained in all volunteers during the treatment period, except for two men in the TU-alone group who experienced a rebound in sperm concentrations. An 8-wk regimen of TU plus DMPA at both tested combination doses effectively suppressed spermatogenesis to azoospermia in Chinese men. All volunteers tolerated the injections; no serious adverse effects were reported. The lower-dose combination is recommended for further testing in an expanded clinical trial or contraceptive efficacy study.


Epithelial mesenchymal transition (EMT) in prostate growth and tumor progression.
Epithelial-mesenchymal transition (EMT) and its reversal, mesenchymal-epithelial transition (MET), are essential morphological processes during development and in the regulation of stem cell pluripotency, yet these processes are also activated in pathological contexts, such as in fibrosis and cancer progression. Multi-component signaling pathways cooperate in initiation of EMT and MET programs, via transcriptional, post-transcriptional, translational, and post-translational regulation. EMT is required for tissue regeneration and normal embryonic development as it enables epithelial cells to acquire the mesenchymal phenotype, conferring them migratory and dynamic properties towards forming three-dimensional structures during gastrulation and organ formation. Uncontrolled activation of such phenomenon and the pathways signaling EMT events in adult life, leads to cancer growth and orchestrated by signaling interactions from the microenvironment, epithelial tumor cells with enhanced polarity, become invasive and rapidly metastasize to distant sites. Loss of epithelial markers (E-cadherin) and gain of mesenchymal markers (N-cadherin), at the leading edge of solid tumors is associated with progression to metastasis. This review will explore the contribution of EMT to embryonic development of GU organs and the functional consequences of EMT-MET cycles in prostate tumorigenesis. Recent insights identifying key players driving EMT and its reversal to MET during prostate cancer progression to metastatic castration-resistant disease will be discussed, with specific focus on androgen receptor (AR) and transforming growth factor-β (TGF-β) signaling in the context of their predictive and targeting value in prostate cancer progression.


A rare case of the upper extremity diffuse large B-cell lymphoma mimicking soft tissue sarcoma in an elderly patient.
Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma, with about 30% of new cases presenting with extranodal disease. Lesions originating from soft tissues of the upper extremities are extremely rare and may mimic other malignancies like sarcoma. We present a case of an elderly patient with right upper extremity (RUE) mass which was proven to be DLBCL instead of sarcoma. We emphasize the increasing need for investigating new therapeutic options for patients of extreme age and/or with underlying heart disease.


Does Robotic-Assisted Computer Navigation Affect Acetabular Cup Positioning in Total Hip Arthroplasty in the Obese Patient? A Comparison Study.
Obese populations present challenges for acetabular cup placement during total hip arthroplasty (THA). This study examines the accuracy of acetabular cup inclination and version in the obese patient with robotic-assisted computer navigation. A total of 105 patients underwent robotic-assisted computer navigation THA with a posterior approach. Groups were divided on body mass index (BMI, kg/m(2)) of <30, 30-35, and >35. There was no statistical difference between the BMI <30 (n=59), BMI 30-35 (n=34) and BMI >35 (n=12) groups for acetabular inclination (P=0.43) or version (P=0.95). Robotic-assisted computer navigation provided accurate and reproducible placement of the acetabular cup within safe zones for inclination and version in the obese patient.


Effects of Mutations on Structure-Function Relationships of Matrix Metalloproteinase-1.
Matrix metalloproteinase-1 (MMP-1) is one of the most widely studied enzymes involved in collagen degradation. Mutations of specific residues in the MMP-1 hemopexin-like (HPX) domain have been shown to modulate activity of the MMP-1 catalytic (CAT) domain. In order to reveal the structural and conformational effects of such mutations, a molecular dynamics (MD) study was performed of in silico mutated residues in the X-ray crystallographic structure of MMP-1 complexed with a collagen-model triple-helical peptide (THP). The results indicate an important role of the mutated residues in MMP-1 interactions with the THP and communication between the CAT and the HPX domains. Each mutation has a distinct impact on the correlated motions in the MMP-1•THP. An increased collagenase activity corresponded to the appearance of a unique anti-correlated motion and decreased correlated motions, while decreased collagenase activity corresponded both to increased and decreased anti-correlated motions.


Confirmation of TACO1 as a Leigh Syndrome Disease Gene in Two Additional Families.
In 2009, we identified TACO1 as a novel mitochondrial disease gene in a single family, however no second family has been described to confirm the role of TACO1 in mitochondrial disease. In this report, we describe two independent consanguineous families carrying pathogenic variants in TACO1, confirming the phenotype. Detailed clinical investigations and whole exome sequencing with haplotype analysis have been performed in several members of the two reported families. Clinical phenotype of the patients confirms the originally reported phenotype of a childhood-onset progressive cerebellar and pyramidal syndrome with optic atrophy and learning difficulties. Brain MRI showed periventricular white matter lesions with multiple cystic defects, suggesting leukoencephalopathy in both patients. One patient carried the previously described homozygous TACO1 variant (p.His158ProfsTer8) and haplotype analysis suggested that this variant is a rare founder mutation. The second patient from another family carried a homozygous novel frame shift variant (p.Cys85PhefsTer15). The identification of two Turkish families with similar characteristic clinical presentation and an additional homozygous nonsense mutation confirms that TACO1 is a human mitochondrial disease gene. Although most patients with this clinical presentation undergo next generation sequencing analysis, screening for selected founder mutations in the Turkish population based on the precise clinical presentation may reduce time and cost of finding the genetic diagnosis even in the era of massively parallel sequencing.


Orexin A modulates prolactin production by regulating BMP-4 activity in rat pituitary lactotorope cells.
The impact of orexins on anterior pituitary function has yet to be clarified. We studied the effects of orexin A and its interaction with the bone morphogenetic protein (BMP) system on the regulatory role of prolactin synthesis using rat lactotrope GH3 cells expressing BMP-4. Orexin type 1 receptor (OX1R), but not type 2 receptor (OX2R), was predominantly expressed in GH3 cells. Orexin A suppressed forskolin-induced, but not basal, prolactin mRNA expression without reducing cAMP levels. Of note, orexin A suppressed BMP-4-induced prolactin mRNA and cAMP synthesis. Impairment of the effects of orexin by chemical inhibitors suggested involvement of the P38 pathway in the OX1R activity that suppresses BMP-4-induced PRL expression. Given that inhibition of BMP-receptor signaling reduced prolactin mRNA levels, endogenous BMP action is likely to be linked to the activation of prolactin synthesis by GH3 cells. Orexin A was revealed to suppress Smad1/5/9 phosphorylation and Id-1 transcription induced by BMP-4, which was restored in the presence of orexin-receptor antagonists, suggesting that the inhibitory effect of orexin A occurred via OX1R. Orexin A also reduced ALK-3 expression but increased inhibitory Smad6/7 expression, while BMP-4 treatment downregulated OX1R expression. These results indicated that orexin A plays an inhibitory role in prolactin production through suppression of endogenous BMP activity in GH3 cells, suggesting that a new functional role of the interaction between orexin and BMP-4 is modulation of prolactin levels in lactotrope cells.


Involuntary attention with uncertainty: peripheral cues improve perception of masked letters, but may impair perception of low-contrast letters.
Improvements of perceptual performance following the presentation of peripheral cues have been ascribed to accelerated accrual of information, enhanced contrast perception, and decision bias. We investigated effects of peripheral cues on the perception of Gabor and letter stimuli. Non-predictive, peripheral cues improved perceptual accuracy when the stimuli were masked. In contrast, peripheral cues degraded perception of low-contrast letters and did not affect the perception of low-contrast Gabors. The results suggest that involuntary attention accelerates accrual of information but are not entirely consistent with the idea that involuntary attention enhances subjective contrast. Rather, peripheral cues may cause crowding with single letter targets of low contrast. Further, we investigated the effect of the amount of uncertainty on involuntary attention. Cueing effects were (initially) larger when there were more possible target locations. In addition, cueing effects were larger when error feedback was absent and observers had no knowledge of results. Despite these strategic factors, location uncertainty was not sufficient to produce cueing effects, showing that location uncertainty paired with non-predictive cues reveals perceptual and not (only) decisional processes.


Axonal transport of rubidium and thallium in the olfactory nerve of mice.
Following intranasal administration of radioactive (86)Rb(+) and (201)Tl(+) in mice, we observed this direct transport via the olfactory nerve pathway. The (86)RbCl and (201)TlCl solutions were administered to two groups of mice, the unilateral intranasal and intravenous administration groups. After sacrifice, their heads were divided into the right and left side, which were then subdivided into seven parts; the nasal mucosa and brain regions were separated. Following the unilateral intranasal administration, uptake after 6 h by the olfactory bulb was significantly higher on the ipsilateral side ((86)Rb, 0.7 %dose; (201)Tl, 0.5 %dose) than on the contralateral side ((86)Rb, 0.08 %dose; (201)Tl, 0.15 %dose). Moreover, the (86)Rb and (201)Tl that accumulated in the olfactory bulb were gradually transported to other brain regions of the olfactory tract, the telencephalon and the diencephalon on the side corresponding to the nostril used for administration. Significant differences were observed between the right and left side of the brain regions 6 and 12 h after administration. Further, (201)Tl autoradiography clearly showed striped patterns of dense accumulation, localized in the region around the glomerular layer and granule cell layer of the olfactory bulb and around the olfactory cortex. These results provide clear evidence of axonal transport via the olfactory nerve pathway, from nasal cavity to the olfactory bulb, as well as to the olfactory cortex through the synaptic junctions. The olfactory transport of the (86)Rb(+) and (201)Tl(+) is thought to represent the behavior of K(+) in the olfactory system.


Evolutionary testing using an extended Chaining Approach.
Fitness functions derived from certain types of white-box test goals can be inadequate for evolutionary software test data generation (Evolutionary Testing), due to a lack of search guidance to the required test data. Often this is because the fitness function does not take into account data dependencies within the program under test, and the fact that certain program statements may need to have been executed prior to the target structure in order for it to be feasible. This paper proposes a solution to this problem by hybridizing Evolutionary Testing with an extended Chaining Approach. The Chaining Approach is a method which identifies statements on which the target structure is data dependent, and incrementally develops chains of dependencies in an event sequence. By incorporating this facility into Evolutionary Testing, and by performing a test data search for each generated event sequence, the search can be directed into potentially promising, unexplored areas of the test object's input domain. Results presented in the paper show that test data can be found for a number of test goals with this hybrid approach that could not be found by using the original Evolutionary Testing approach alone. One such test goal is drawn from code found in the publicly available libpng library.


Artificial Intelligence-Assisted Loop Mediated Isothermal Amplification (AI-LAMP) for Rapid Detection of SARS-CoV-2.
Until vaccines and effective therapeutics become available, the practical solution to transit safely out of the current coronavirus disease 19 (CoVID-19) lockdown may include the implementation of an effective testing, tracing and tracking system. However, this requires a reliable and clinically validated diagnostic platform for the sensitive and specific identification of SARS-CoV-2. Here, we report on the development of a de novo, high-resolution and comparative genomics guided reverse-transcribed loop-mediated isothermal amplification (LAMP) assay. To further enhance the assay performance and to remove any subjectivity associated with operator interpretation of results, we engineered a novel hand-held smart diagnostic device. The robust diagnostic device was further furnished with automated image acquisition and processing algorithms and the collated data was processed through artificial intelligence (AI) pipelines to further reduce the assay run time and the subjectivity of the colorimetric LAMP detection. This advanced AI algorithm-implemented LAMP (ai-LAMP) assay, targeting the RNA-dependent RNA polymerase gene, showed high analytical sensitivity and specificity for SARS-CoV-2. A total of ~200 coronavirus disease (CoVID-19)-suspected NHS patient samples were tested using the platform and it was shown to be reliable, highly specific and significantly more sensitive than the current gold standard qRT-PCR. Therefore, this system could provide an efficient and cost-effective platform to detect SARS-CoV-2 in resource-limited laboratories.


Making psychological theory useful for implementing evidence based practice: a consensus approach.
Evidence-based guidelines are often not implemented effectively with the result that best health outcomes are not achieved. This may be due to a lack of theoretical understanding of the processes involved in changing the behaviour of healthcare professionals. This paper reports the development of a consensus on a theoretical framework that could be used in implementation research. The objectives were to identify an agreed set of key theoretical constructs for use in (1) studying the implementation of evidence based practice and (2) developing strategies for effective implementation, and to communicate these constructs to an interdisciplinary audience. Six phases of work were conducted to develop a consensus: (1) identifying theoretical constructs; (2) simplifying into construct domains; (3) evaluating the importance of the construct domains; (4) interdisciplinary evaluation; (5) validating the domain list; and (6) piloting interview questions. The contributors were a "psychological theory" group (n = 18), a "health services research" group (n = 13), and a "health psychology" group (n = 30). Twelve domains were identified to explain behaviour change: (1) knowledge, (2) skills, (3) social/professional role and identity, (4) beliefs about capabilities, (5) beliefs about consequences, (6) motivation and goals, (7) memory, attention and decision processes, (8) environmental context and resources, (9) social influences, (10) emotion regulation, (11) behavioural regulation, and (12) nature of the behaviour. A set of behaviour change domains agreed by a consensus of experts is available for use in implementation research. Applications of this domain list will enhance understanding of the behaviour change processes inherent in implementation of evidence-based practice and will also test the validity of these proposed domains.


Process intensification for production of Streptococcus pneumoniae whole-cell vaccine.
The available pneumococcal conjugate vaccines provide protection against only those serotypes that are included in the vaccine, which leads to a selective pressure and serotype replacement in the population. An alternative low-cost, safe and serotype-independent vaccine was developed based on a nonencapsulated pneumococcus strain. This study evaluates process intensification to improve biomass production and shows for the first time the use of perfusion-batch with cell recycling for bacterial vaccine production. Batch, fed-batch, and perfusion-batch were performed at 10 L scale using a complex animal component-free culture medium. Cells were harvested at the highest optical density, concentrated and washed using microfiltration or centrifugation to compare cell separation methods. Higher biomass was achieved using perfusion-batch, which removes lactate while retaining cells. The biomass produced in perfusion-batch would represent at least a fourfold greater number of doses per cultivation than in the previously described batch process. Each strategy yielded similar vaccines in terms of quality as evaluated by western blot and animal immunization assays, indicating that so far, perfusion-batch is the best strategy for the intensification of pneumococcal whole-cell vaccine production, as it can be integrated to the cell separation process keeping the same vaccine quality.


Tunable plasmon resonances and two-dimensional anisotropy of angular optical response of overlapped nanoshells.
Symmetry breaking of metallic nanoparticles results in many unique optical properties. We use the discrete dipole approximation method to study the optical properties of overlapped nanoshells which further break the rotational symmetry compared with the semishells. The optical properties of the nanoparticles can be tuned from the visible to near infrared regime by varying the geometry parameters and the hybrid components of nanoparticles. The calculated extinction spectra show the two-dimensional anisotropy of the angular optical response of the nanoparticles. The plasmon hybridization model provides a way to interpret the resonance modes of the nanoparticles. The tunable plasmon resonances, the enhanced local fields and the anisotropic optical properties suggest that the overlapped nanoshells have potential applications in surface-enhanced spectroscopy and "smart" coating in windows or display devices.


Local changes in the wake electroencephalogram precedes lane departures.
The objective of this exploratory study is to investigate if lane departures are associated with local sleep, measured via source-localized electroencephalography (EEG) theta power in the 5-9 Hz frequency range. Thirty participants drove in an advanced driving simulator, resulting in 135 lane departures at high levels of self-reported sleepiness. These lane departures were compared to matching non-departures at the same sleepiness level within the same individual. There was no correspondence between lane departures and global theta activity. However, at the local level an increased risk for lane departures was associated with increased theta content in brain regions related to motor function.


Physiologically based boundary conditions in finite element modelling.
Finite element analysis has been used extensively in the study of bone loading and implant performance, such as in the femur. The boundary conditions applied vary widely, generally producing excessive femoral deformation, and although it has been shown that the muscle forces influence femoral deflections and loading, little consideration has been given to the displacement constraints. It is hypothesised that careful application of physiologically based constraints can produce physiological deformation, and therefore straining, of the femur. Joint contact forces and a complete set of muscle forces were calculated based on the geometry of the Standardised Femur using previously validated musculoskeletal models. Five boundary condition cases were applied to a finite element model of the Standardised Femur: (A) diaphyseally constrained with hip contact and abductor forces; (B) case A plus vasti forces; (C) case A with complete set of muscle forces; (D) distally constrained with all muscle forces; (E) physiological constraints with all muscle forces. It was seen that only the physiological boundary conditions, case E, produced physiological deflections (< 2.0mm) of the femoral head in both the coronal and sagittal planes, which resulted in minimal reaction forces at the constrained nodes. Strains in the mid-diaphysis varied by up to 600 micro-strain under walking loads and 1000 micro-strain under stair climbing loads. The mode of loading, as indicated by the strain profiles on the cortex also varied substantially under these boundary conditions, which has important consequences for studies that examine localised bone loading such as fracture or bone remodelling simulations.


[Pulsed electromagnetic fields (PEMF)--results in evidence based medicine].
Therapy with electromagnetic fields has a very old tradition in medicine. The indications are widespread, whereas little is known about the effects. Controlled randomizied studies with positive results for pulsed electromagnetic fields (PEMF) are available for osteotomies, the healing of skin wounds, and osteoarthritis. Comparison of the studies is difficult because of the different doses applied and intervals of therapy. Therefore recommendations regarding an optimal dosis and interval are, depending on the disease, quite variable.


Phylogenetic and molecular insights into the evolution of multidrug-resistant porcine enterotoxigenic Escherichia coli in Australia.
This study investigated the phylogeny and molecular epidemiology of Australian porcine enterotoxigenic Escherichia coli (ETEC) isolates (n=70) by performing multilocus sequence typing (MLST), random amplified polymorphic DNA (RAPD) analysis, virulence gene analysis, plasmid, bacteriocin, integron and antimicrobial resistance gene typing, and antimicrobial susceptibility phenotyping. Isolates of the most commonly observed O serogroup (O149) were highly clonal with a lower frequency of antimicrobial resistance compared with the less common O141 serogroup isolates, which were more genetically diverse and resistant to a greater array of antimicrobials. The O149 and O141 isolates belonged to sequence types (STs) ST100 and ST1260, respectively. A small number of new STs were identified for the least common serogroups, including O157 (ST4245), O138 (ST4244), O139 (ST4246) and O8 (ST4247). A high frequency of plasmid replicons was observed among all ETEC isolates. However, O149 isolates predominantly carried IncFIB, I1, HI1 and FIC, whereas O141 isolates carried a more varied array, including IncI1, FIB, FIC, HI1, I1, Y and, most significantly, A/C. O141 isolates also possessed a greater diversity of bacteriocins, with almost one-half of the isolates carrying colicin E3 (44.4%; 12/27) and E7 (48.1%; 13/27). This study shows that Australian porcine ETEC are distinct from isolates obtained in other parts of the world with respect to the MLST profile and the absence of resistance to critically important antimicrobials, including third-generation cephalosporins and fluoroquinolones.