Severe dystonia, cerebellar atrophy, and cardiomyopathy likely caused by a missense mutation in TOR1AIP1.
Dystonia, cerebellar atrophy, and cardiomyopathy constitute a rare association. We used homozygosity mapping and whole exome sequencing to determine the mutation, western blot and immunolabelling on cultured fibroblasts to demonstrate the lower expression and the mislocalization of the protein. We report on a boy born from consanguineous healthy parents, who presented at three years of age with rapidly progressing dystonia, progressive cerebellar atrophy, and dilated cardiomyopathy. We identified regions of homozygosity and performed whole exome sequencing that revealed a homozygous missense mutation in TOR1AIP1. The mutation, absent in controls, results in a change of a highly conserved glutamic acid to alanine. TOR1AIP1 encodes lamina-associated polypeptide 1 (LAP1), a transmembrane protein ubiquitously expressed in the inner nuclear membrane. LAP1 interacts with torsinA, the protein mutated in DYT1-dystonia. In vitro studies in fibroblasts of the patient revealed reduced expression of LAP1 and its mislocalization and aggregation in the endoplasmic reticulum as underlying pathogenic mechanisms. The pathogenic role of TOR1AIP1 mutation is supported by a) the involvement of a highly conserved amino acid, b) the absence of the mutation in controls, c) the functional interaction of LAP1 with torsinA, and d) mislocalization of LAP1 in patient cells. Of note, cardiomyopathy has been reported in LAP1-null mice and in patients with the TOR1AIP1 nonsense mutation. Other cases will help delineate the clinical spectrum of LAP1-related mutations.


New bioinformatic tools for analysis of nucleotide modifications in eukaryotic rRNA.
This report presents a valuable new bioinformatics package for research on rRNA nucleotide modifications in the ribosome, especially those created by small nucleolar RNA:protein complexes (snoRNPs). The interactive service, which is not available elsewhere, enables a user to visualize the positions of pseudouridines, 2'-O-methylations, and base methylations in three-dimensional space in the ribosome and also in linear and secondary structure formats of ribosomal RNA. Our tools provide additional perspective on where the modifications occur relative to functional regions within the rRNA and relative to other nearby modifications. This package of new tools is presented as a major enhancement of an existing but significantly upgraded yeast snoRNA database available publicly at http://people.biochem.umass.edu/sfournier/fournierlab/snornadb/. The other key features of the enhanced database include details of the base pairing of snoRNAs with target RNAs, genomic organization of the yeast snoRNA genes, and information on corresponding snoRNAs and modifications in other model organisms.


Mass spectrometric characterization of different norandrosterone derivatives by low-cost mass spectrometric detectors using electron ionization and chemical ionization.
The abuse of nortestosterone in sport is an important problem in doping-control analysis. In order to detect the main urinary metabolite of nortestosterone, norandrosterone (NA), sensitive and specific methodology is necessary. In this context the use of a low-cost mass spectrometric detector such as the Finnigan MAT ion-trap detector (ITD) was studied. The electron ionization (EI) and positive-ion chemical ionization (PICI) mass spectra of the methoxime-trimethylsilyl, trimethylsilyl-enol trimethylsilyl ether and pentafluoropropionic ester derivatives of NA are described. The limits of detection of these derivatives are compared with those obtained by the Hewlett-Packard mass selective detector (MSD), another low-cost mass spectrometric detector and operating only in the EI mode. For the derivatives of the reference standard of NA the ITD has in the EI mode the same limit of detection, in the range of 0.5 to 1 ng injected on the column, as the MSD. However, under these conditions the ITD provides more spectrometric information, because it gives full scan data. Moreover, with the same or even improved limits of detection the ITD can operate in the PICI mode. On the other hand, for the analysis of NA isolated from urine samples, the performance of the MSD was better than that of the ITD. The ion trapping technique is probably limited when the chemical background is high.


Indapamide, a thiazide-like diuretic, decreases bone resorption in vitro.
We recently showed that indapamide (IDP), a thiazide-related diuretic, increases bone mass and decreases bone resorption in spontaneously hypertensive rats supplemented with sodium. In the present study, we evaluated the in vitro effects of this diuretic on bone cells, as well as those of hydrochlorothiazide (HCTZ), the reference thiazide, and acetazolamide (AZ), a carbonic anhydrase (CA) inhibitor. We showed that 10(-4) M IDP and 10(-4) M AZ, as well as 10(-5) M pamidronate (APD), decreased bone resorption in organ cultures and in cocultures of osteoblast-like cells and bone marrow cells in the presence of 10(-8) M 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. We investigated the mechanism of this antiresorptive effect of IDP; IDP decreased osteoclast differentiation as the number of osteoclasts developing in coculture of marrow and osteoblast-like cells was decreased markedly. We then investigated whether IDP affected osteoblast-like cells because these cells are involved in the osteoclast differentiation. Indeed, IDP increased osteoblast-like cell proliferation and alkaline phosphatase (ALP) expression. Nevertheless, it did not modify the colony-stimulating factor 1 (CSF-1) production by these cells. In addition, osteoblast-like cells expressed the Na+/Cl- cotransporter that is necessary for the renal action of thiazide diuretics, but IDP inhibited bone resorption in mice lacking this cotransporter, so the inhibition of bone resorption and osteoclast differentiation did not involve this pathway. Thus, we hypothesized that IDP may act directly on cells of the osteoclast lineage. We observed that resorption pits produced by spleen cells cultured in the presence of soluble osteoclast differentiation factor (sODF) and CSF-1 were decreased by 10(-4) M IDP as well as 10(-5) M APD. In conclusion, in vitro IDP increased osteoblast proliferation and decreased bone resorption at least in part by decreasing osteoclast differentiation via a direct effect on hematopoietic precursors.


Association and regulation of heat shock transcription factor 4b with both extracellular signal-regulated kinase mitogen-activated protein kinase and dual-specificity tyrosine phosphatase DUSP26.
The heat shock transcription factors (Hsfs) activate the stress-inducible expression of heat shock proteins (Hsps) and other molecular chaperones in response to stress and, therefore, play an essential role in protein disaggregation and protein folding. In humans, missense mutation in the hsf4 gene causes cataract, and mice bearing a targeted disruption of the hsf4 gene exhibit defects in lens fiber cell differentiation and early cataract formation. Here, we show that Hsf4b is a direct target of the mitogen-activated protein (MAP) kinase extracellular signal-related kinase (ERK) and that phosphorylation of Hsf4b by ERK leads to increased ability of Hsf4b to bind DNA. Surprisingly, Hsf4b also interacts with an ERK-specific dual-specificity tyrosine phosphatase named DUSP26 identified from a yeast two-hybrid screen. While activated ERK phosphorylates Hsf4b, DUSP26 controls the activity of ERK, leading to phosphorylation/dephosphorylation of Hsf4b, altering its ability to bind DNA. Therefore, DUSP26 interaction with Hsf4b places this transcription factor within a regulatory circuit in the MAP kinase signaling pathway.


Single-trait and multi-trait genome-wide association analyses identify novel loci for blood pressure in African-ancestry populations.
Hypertension is a leading cause of global disease, mortality, and disability. While individuals of African descent suffer a disproportionate burden of hypertension and its complications, they have been underrepresented in genetic studies. To identify novel susceptibility loci for blood pressure and hypertension in people of African ancestry, we performed both single and multiple-trait genome-wide association analyses. We analyzed 21 genome-wide association studies comprised of 31,968 individuals of African ancestry, and validated our results with additional 54,395 individuals from multi-ethnic studies. These analyses identified nine loci with eleven independent variants which reached genome-wide significance (P < 1.25×10-8) for either systolic and diastolic blood pressure, hypertension, or for combined traits. Single-trait analyses identified two loci (TARID/TCF21 and LLPH/TMBIM4) and multiple-trait analyses identified one novel locus (FRMD3) for blood pressure. At these three loci, as well as at GRP20/CDH17, associated variants had alleles common only in African-ancestry populations. Functional annotation showed enrichment for genes expressed in immune and kidney cells, as well as in heart and vascular cells/tissues. Experiments driven by these findings and using angiotensin-II induced hypertension in mice showed altered kidney mRNA expression of six genes, suggesting their potential role in hypertension. Our study provides new evidence for genes related to hypertension susceptibility, and the need to study African-ancestry populations in order to identify biologic factors contributing to hypertension.


[Therapeutic strategy for the treatment of non-infectious uveitis proposed by an expert panel].
Conventional immunosuppressive drugs, anti-TNF alpha and other biotherapies used in clinical practice are capable of controlling non-infectious anterior uveitis, posterior uveitis and panuveitis. The present work has been led by a multidisciplinary panel of experts, internists, rheumatologists and ophthalmologists and is based on a review of the literature. In case of corticodependency or sight-threatening disease, conventional immunosuppressive drugs (methotrexate, azathioprine and mycophenolate mofetil) and/or anti-TNF alpha (adalimumab, infliximab) are used to achieve and maintain remission. Interferon is an efficient immunomodulatory treatment, as a second-line therapy, for some therapeutic indications (refractory macular edema, Behçet's vascularitis). Other biologics, especially tocilizumab, are showing promising results. Local treatments (corticosteroids, sirolimus etc.) are adjuvant therapies in case of unilateral inflammatory relapse. Therapeutic response must be evaluated precisely by clinical examination and repeated complementary investigations (laser flare photometry, multimodal imaging, perimetry, electroretinography measures).


Comparison of the effects of deferiprone versus deferoxamine on growth and virulence of Yersinia enterocolitica.
Deferoxamine, a drug used to treat patients with iron overload, has the capacity to promote systemic Y. enterocolitica infections in humans. The aim of this study was to determine whether deferiprone, the only orally active alternative treatment, has the same potential. When Y. enterocolitica IP864 was grown in an iron-poor chemically defined medium, addition of deferoxamine promoted its growth, while various concentrations of deferiprone did not display this activity. Similarly, on iron-poor agar plates, various Y. enterocolitica strains were able to grow around paper disks impregnated with deferoxamine in a dose-dependent manner, while no growth was observed around the deferiprone disks. In a mouse experimental model of infection, the 50% lethal dose (LD(50)) of strain IP864 was decreased by more than 5 log units in mice pretreated with deferoxamine, while a deferiprone pretreatment did not affect it. Therefore, in contrast to deferoxamine, deferiprone does not enhance growth of pathogenic Y. enterocolitica in vitro and does not have the potential to promote Y. enterocolitica septicemia in a mouse model of infection. Deferiprone may thus represent a useful alternative iron-chelation therapy during invasive Y. enterocolitica infections.


Adiposity and insulin resistance in humans: the role of the different tissue and cellular lipid depots.
Human adiposity has long been associated with insulin resistance and increased cardiovascular risk, and abdominal adiposity is considered particularly adverse. Intra-abdominal fat is associated with insulin resistance, possibly mediated by greater lipolytic activity, lower adiponectin levels, resistance to leptin, and increased inflammatory cytokines, although the latter contribution is less clear. Liver lipid is also closely associated with, and likely to be an important contributor to, insulin resistance, but it may also be in part the consequence of the lipogenic pathway of insulin action being up-regulated by hyperinsulinemia and unimpaired signaling. Again, intramyocellular triglyceride is associated with muscle insulin resistance, but anomalies include higher intramyocellular triglyceride in insulin-sensitive athletes and women (vs men). Such issues could be explained if the "culprits" were active lipid moieties such as diacylglycerol and ceramide species, dependent more on lipid metabolism and partitioning than triglyceride amount. Subcutaneous fat, especially gluteofemoral, appears metabolically protective, illustrated by insulin resistance and dyslipidemia in patients with lipodystrophy. However, some studies suggest that deep sc abdominal fat may have adverse properties. Pericardial and perivascular fat relate to atheromatous disease, but not clearly to insulin resistance. There has been recent interest in recognizable brown adipose tissue in adult humans and its possible augmentation by a hormone, irisin, from exercising muscle. Brown adipose tissue is metabolically active, oxidizes fatty acids, and generates heat but, because of its small and variable quantities, its metabolic importance in humans under usual living conditions is still unclear. Further understanding of specific roles of different lipid depots may help new approaches to control obesity and its metabolic sequelae.


Improving Medical Communication in Migraine Management: A Modified Delphi Study to Develop a Digital Migraine Tracker.
This study aimed to identify the essential content and amount of information to be collected from people with migraine via a patient-facing smartphone-based migraine tracker for them to share with clinicians during live discussions to assist in optimizing migraine management. The proposed tracker is intended for use in non-interventional research to evaluate disease burden in episodic migraine and chronic migraine patients as assessed by demographic and clinical characteristics and health resource utilization in an integrated delivery network setting. The proposed tracker is not intended for commercial purposes. Epidemiological studies suggest migraine is underdiagnosed and undertreated. Studies of patient-clinician interactions suggest that effective medical communication may help address these issues. Four migraine practice leaders, an epidemiologist with extensive migraine experience, and a measurement expert took part in a modified Delphi panel process to identify data elements that could be collected from people with migraine through a smartphone-based migraine tracker. Importantly, the proposed tracker would not be intended to replace the patient-clinician encounter but to support the encounter through enabling the patient to document migraine symptoms and experiences in a timely and accurate manner for sharing with a clinician as part of a broader face-to-face discussion. The panel reviewed questions derived from the existing migraine diaries in the public domain, those used in clinical trials, and patient-centric surveys assessing the impact of migraine on physical function and other related concepts. Key considerations included identification of the most clinically useful data elements for a shared communication tool for people with migraine under the care of a clinician. The panel also identified numerous functionality requirements for such a tool and provided recommendations on the most effective way to present results to a clinician. The expert panel opined that people with migraine may value the ability to capture a relatively broad range of information for their own migraine-tracking purposes, while clinicians will likely find greater value in a small set of data relevant to the management of migraine. The panel identified the 3 most essential concepts in categories of data for a clinician, for which they coined the term "The 3 Fs": Frequency of days with headache; Frequency of acute medication usage; and Functional impairment. Information on the frequency of days with headache was felt to combine with the information on the frequency of acute medication usage to provide essential insights into current migraine management strategy and its outcomes, and to assist considerations of preventive measures. Functional impairment was treated as an effective surrogate for headache severity and was assessed based on the following: degree of difficulty in performing activities of daily living, impact on absenteeism (taking leave from work or cancelling/avoiding other activities) and presenteeism (performing work or other daily activities, with reduced productivity/capability), and amount of rest required as a result of a migraine attack. The modified Delphi panel process resulted in the selection of 13 questions in 8 categories to elicit sufficient and meaningful data comprising headache occurrence, symptoms, daily/preventive and as-needed/acute medication usage, triggers, ability to concentrate, and functional impairment. The panel also agreed that the tracker should generate 2 distinct reports: one for people with migraine that would include a wider range of data about symptoms and perceived triggers, and a targeted report for the clinician that would place prime emphasis on the 3 Fs for aggregating the results of each headache occurrence and the trend over time. A system that easily captures critical data elements about migraine, with specific feedback displays for patients to share with clinicians during live discussions, may offer some benefit to people with migraine and their clinicians by facilitating more objective communication and optimizing management. The tracker's output may enable people with migraine to track a wide range of data for their own purposes, allowing them to better understand their condition, while a synthesized view of the selected data may support more informed clinical decision-making for the clinician and individualized, evidence-based discussion with the patient. As a result, this shared decision-making tool may enable patients to more accurately convey essential migraine information during live patient-clinician discussions to drive improved management and patient outcomes.


The heterogeneity of cell subtypes from a primary culture of human amniotic fluid.
Heterogeneous human amniotic fluid contains various cell types. Herein, we report on the possibility of simultaneously isolating three subtypes of cells from one primary culture. Using a stainless steel instrument named a colony poculum, two of the three cell subtypes could be efficiently cultured, and these were further characterized. The results indicated that these two cell subtypes had different morphologies and were characterized by different cell marker expression profiles, including the differential expression of CD105, CD117 and EBAF. Furthermore, their gene expression array data revealed their different gene expression profiles. Although both cell types expressed several embryonic stem cell-specific markers, they were non-tumorigenic in vivo. This paper not only provides new insight into the heterogeneity of human amniotic fluid, it also presents a simple yet efficient cell isolation method. These results will contribute to the thorough investigation of the properties and potential future applications of human amniotic fluid-derived cells.


Rhythmic Components in Extracranial Brain Signals Reveal Multifaceted Task Modulation of Overlapping Neuronal Activity.
Oscillatory neuronal activity is implicated in many cognitive functions, and its phase coupling between sensors may reflect networks of communicating neuronal populations. Oscillatory activity is often studied using extracranial recordings and compared between experimental conditions. This is challenging, because there is overlap between sensor-level activity generated by different sources, and this can obscure differential experimental modulations of these sources. Additionally, in extracranial data, sensor-level phase coupling not only reflects communicating populations, but can also be generated by a current dipole, whose sensor-level phase coupling does not reflect source-level interactions. We present a novel method, which is capable of separating and characterizing sources on the basis of their phase coupling patterns as a function of space, frequency and time (trials). Importantly, this method depends on a plausible model of a neurobiological rhythm. We present this model and an accompanying analysis pipeline. Next, we demonstrate our approach, using magnetoencephalographic (MEG) recordings during a cued tactile detection task as a case study. We show that the extracted components have overlapping spatial maps and frequency content, which are difficult to resolve using conventional pairwise measures. Because our decomposition also provides trial loadings, components can be readily contrasted between experimental conditions. Strikingly, we observed heterogeneity in alpha and beta sources with respect to whether their activity was suppressed or enhanced as a function of attention and performance, and this happened both in task relevant and irrelevant regions. This heterogeneity contrasts with the common view that alpha and beta amplitude over sensory areas are always negatively related to attention and performance.


Sequence Identification, Recombinant Production, and Analysis of the Self-Assembly of Egg Stalk Silk Proteins from Lacewing Chrysoperla carnea.
Egg stalk silks of the common green lacewing Chrysoperla carnea likely comprise at least three different silk proteins. Based on the natural spinning process, it was hypothesized that these proteins self-assemble without shear stress, as adult lacewings do not use a spinneret. To examine this, the first sequence identification and determination of the gene expression profile of several silk proteins and various transcript variants thereof was conducted, and then the three major proteins were recombinantly produced in Escherichia coli encoded by their native complementary DNA (cDNA) sequences. Circular dichroism measurements indicated that the silk proteins in aqueous solutions had a mainly intrinsically disordered structure. The largest silk protein, which we named ChryC1, exhibited a lower critical solution temperature (LCST) behavior and self-assembled into fibers or film morphologies, depending on the conditions used. The second silk protein, ChryC2, self-assembled into nanofibrils and subsequently formed hydrogels. Circular dichroism and Fourier transform infrared spectroscopy confirmed conformational changes of both proteins into beta sheet rich structures upon assembly. ChryC3 did not self-assemble into any morphology under the tested conditions. Thereby, through this work, it could be shown that recombinant lacewing silk proteins can be produced and further used for studying the fiber formation of lacewing egg stalks.


A Bayesian perspective on magnitude estimation.
Our representation of the physical world requires judgments of magnitudes, such as loudness, distance, or time. Interestingly, magnitude estimates are often not veridical but subject to characteristic biases. These biases are strikingly similar across different sensory modalities, suggesting common processing mechanisms that are shared by different sensory systems. However, the search for universal neurobiological principles of magnitude judgments requires guidance by formal theories. Here, we discuss a unifying Bayesian framework for understanding biases in magnitude estimation. This Bayesian perspective enables a re-interpretation of a range of established psychophysical findings, reconciles seemingly incompatible classical views on magnitude estimation, and can guide future investigations of magnitude estimation and its neurobiological mechanisms in health and in psychiatric diseases, such as schizophrenia.


Proteolytic cleavage of covalently linked cell wall proteins by Candida albicans Sap9 and Sap10.
The cell wall of the human-pathogenic fungus Candida albicans is a robust but also dynamic structure which mediates adaptation to changing environmental conditions during infection. Sap9 and Sap10 are cell surface-associated proteases which function in C. albicans cell wall integrity and interaction with human epithelial cells and neutrophils. In this study, we have analyzed the enzymatic properties of Sap9 and Sap10 and investigated whether these proteases cleave proteins on the fungal cell surface. We show that Sap9 and Sap10, in contrast to other aspartic proteases, exhibit a near-neutral pH optimum of proteolytic activity and prefer the processing of peptides containing basic or dibasic residues. However, both proteases also cleaved at nonbasic sites, and not all tested peptides with dibasic residues were processed. By digesting isolated cell walls with Sap9 or Sap10, we identified the covalently linked cell wall proteins (CWPs) Cht2, Ywp1, Als2, Rhd3, Rbt5, Ecm33, and Pga4 as in vitro protease substrates. Proteolytic cleavage of the chitinase Cht2 and the glucan-cross-linking protein Pir1 by Sap9 was verified using hemagglutinin (HA) epitope-tagged versions of both proteins. Deletion of the SAP9 and SAP10 genes resulted in a reduction of cell-associated chitinase activity similar to that upon deletion of CHT2, suggesting a direct influence of Sap9 and Sap10 on Cht2 function. In contrast, cell surface changes elicited by SAP9 and SAP10 deletion had no major impact on the phagocytosis and killing of C. albicans by human macrophages. We propose that Sap9 and Sap10 influence distinct cell wall functions by proteolytic cleavage of covalently linked cell wall proteins.


Piezo-optic tensor of crystals from quantum-mechanical calculations.
An automated computational strategy is devised for the ab initio determination of the full fourth-rank piezo-optic tensor of crystals belonging to any space group of symmetry. Elastic stiffness and compliance constants are obtained as numerical first derivatives of analytical energy gradients with respect to the strain and photo-elastic constants as numerical derivatives of analytical dielectric tensor components, which are in turn computed through a Coupled-Perturbed-Hartree-Fock/Kohn-Sham approach, with respect to the strain. Both point and translation symmetries are exploited at all steps of the calculation, within the framework of periodic boundary conditions. The scheme is applied to the determination of the full set of ten symmetry-independent piezo-optic constants of calcium tungstate CaWO4, which have recently been experimentally reconstructed. Present calculations unambiguously determine the absolute sign (positive) of the π61 constant, confirm the reliability of 6 out of 10 experimentally determined constants and provide new, more accurate values for the remaining 4 constants.


Recovery of plasmid pEMB1, whose toxin-antitoxin system stabilizes an ampicillin resistance-conferring β-lactamase gene in Escherichia coli, from natural environments.
Non-culture-based procedures were used to investigate plasmids showing ampicillin resistance properties in two different environments: remote mountain soil (Mt. Jeombong) and sludge (Tancheon wastewater treatment plant). Total DNA extracted from the environmental samples was directly transformed into Escherichia coli TOP10, and a single and three different plasmids were obtained from the mountain soil and sludge samples, respectively. Interestingly, the restriction fragment length polymorphism pattern of the plasmid from the mountain soil sample, designated pEMB1, was identical to the pattern of one of the three plasmids from the sludge sample. Complete DNA sequencing of plasmid pEMB1 (8,744 bp) showed the presence of six open reading frames, including a β-lactamase gene. Using BLASTX, the orf5 and orf6 genes were suggested to encode a CopG family transcriptional regulator and a plasmid stabilization system, respectively. Functional characterization of these genes using a knockout orf5 plasmid (pEMB1ΔparD) and the cloning and expression of orf6 (pET21bparE) indicated that these genes were antitoxin (parD) and toxin (parE) genes. Plasmid stability tests using pEMB1 and pEMB1ΔparDE in E. coli revealed that the orf5 and orf6 genes enhanced plasmid maintenance in the absence of ampicillin. Using a PCR-based survey, pEMB1-like plasmids were additionally detected in samples from other human-impacted sites (sludge samples) and two other remote mountain soil samples, suggesting that plasmids harboring a β-lactamase gene with a ParD-ParE toxin-antitoxin system occurs broadly in the environment. This study extends knowledge about the dissemination and persistence of antibiotic resistance genes in naturally occurring microbial populations.


Increasing cellulose accessibility is more important than removing lignin: a comparison of cellulose solvent-based lignocellulose fractionation and soaking in aqueous ammonia.
While many pretreatments attempt to improve the enzymatic digestibility of biomass by removing lignin, this study shows that improving the surface area accessible to cellulase is a more important factor for achieving a high sugar yield. Here we compared the pretreatment of switchgrass by two methods, cellulose solvent- and organic solvent-based lignocellulose fractionation (COSLIF) and soaking in aqueous ammonia (SAA). Following pretreatment, enzymatic hydrolysis was conducted at two cellulase loadings, 15 filter paper units (FPU)/g glucan and 3 FPU/g glucan, with and without BSA blocking of lignin absorption sites. The hydrolysis results showed that the lignin remaining after SAA had a significant negative effect on cellulase performance, despite the high level of delignification achieved with this pretreatment. No negative effect due to lignin was detected for COSLIF-treated substrate. SEM micrographs, XRD crystallinity measurements, and cellulose accessibility to cellulase (CAC) determinations confirmed that COSLIF fully disrupted the cell wall structure, resulting in a 16-fold increase in CAC, while SAA caused a 1.4-fold CAC increase. A surface plot relating the lignin removal, CAC, and digestibility of numerous samples (both pure cellulosic substrates and lignocellulosic materials pretreated by several methods) was also developed to better understand the relative impacts of delignification and CAC on glucan digestibility.


Energy-dependent fitness: a quantitative model for the evolution of yeast transcription factor binding sites.
We present a genomewide cross-species analysis of regulation for broad-acting transcription factors in yeast. Our model for binding site evolution is founded on biophysics: the binding energy between transcription factor and site is a quantitative phenotype of regulatory function, and selection is given by a fitness landscape that depends on this phenotype. The model quantifies conservation, as well as loss and gain, of functional binding sites in a coherent way. Its predictions are supported by direct cross-species comparison between four yeast species. We find ubiquitous compensatory mutations within functional sites, such that the energy phenotype and the function of a site evolve in a significantly more constrained way than does its sequence. We also find evidence for substantial evolution of regulatory function involving point mutations as well as sequence insertions and deletions within binding sites. Genes lose their regulatory link to a given transcription factor at a rate similar to the neutral point mutation rate, from which we infer a moderate average fitness advantage of functional over nonfunctional sites. In a wider context, this study provides an example of inference of selection acting on a quantitative molecular trait.


Characterization of a fourth adaptor-related protein complex.
Adaptor protein complexes (APs) function as vesicle coat components in different membrane traffic pathways; however, there are a number of pathways for which there is still no candidate coat. To find novel coat components related to AP complexes, we have searched the expressed sequence tag database and have identified, cloned, and sequenced a new member of each of the four AP subunit families. We have shown by a combination of coimmunoprecipitation and yeast two-hybrid analysis that these four proteins (epsilon, beta4, mu4, and sigma4) are components of a novel adaptor-like heterotetrameric complex, which we are calling AP-4. Immunofluorescence reveals that AP-4 is localized to approximately 10-20 discrete dots in the perinuclear region of the cell. This pattern is disrupted by treating the cells with brefeldin A, indicating that, like other coat proteins, the association of AP-4 with membranes is regulated by the small GTPase ARF. Immunogold electron microscopy indicates that AP-4 is associated with nonclathrin-coated vesicles in the region of the trans-Golgi network. The mu4 subunit of the complex specifically interacts with a tyrosine-based sorting signal, indicating that, like the other three AP complexes, AP-4 is involved in the recognition and sorting of cargo proteins with tyrosine-based motifs. AP-4 is of relatively low abundance, but it is expressed ubiquitously, suggesting that it participates in a specialized trafficking pathway but one that is required in all cell types.


Promoting resident wellness: evaluation of a time-off policy to increase residents' utilization of health care services.
To evaluate awareness and utilization of a new institutional policy to grant residents time off to access personal and family health care. In 2012, two years after policy implementation, an electronic survey was sent to all 546 residents and fellows at a tertiary care academic medical center in the United States. Residents were asked questions regarding awareness of the time-off policy, use of the policy, health care status, reasons for policy use, and barriers to use. A total of 490 (90%) residents responded. Eighty-nine percent of those surveyed were aware of the policy. Of those who were aware, 49.7% used the policy to access health care. Top reasons for policy use were for personal routine or preventive health care, dental care, and urgent health care needs. The most commonly reported barrier to policy use was concern about the impact the resident's absence would have on colleagues. Implementation of policies to prospectively schedule residents' time off during business hours to address health care needs is an important means to promote resident wellness. Such policies remove one commonly cited barrier to residents' access to health care. However, residents still reported concerns about impact on peers and patients as the main reason they were reluctant to take the time off to address their health care needs. More work is needed on both wellness policy implementation practices and on refining the systems that will allow seamless and guiltless transitions of care.


Polymorphisms in Th1-type cell-mediated response genes and risk of gastric cancer.
Helicobacter pylori infection, the dominant risk factor for gastric cancers, has been shown to elicit T helper type 1 (Th1) polarized immunological responses. We conducted a population-based study of 305 gastric cancer cases and 427 age- and gender-matched controls in Warsaw, Poland, to evaluate the association with several variants in genes responsible for Th1-cell-mediated response. Genotyping was performed on genomic DNA by TaqMan(TM) assays to determine TNFA (-308 G>A, -417 G>A, -555 G>A, -1036 C>T, -1042 C>A, -1210 T>C), IL1A (-889 C>T), IFNGR2 (Ex7-128 T>C, Ex2-34 C>G and Ex2-16 A>G) and IL12A (IVS2-798 T>A, IVS2-701 C>A and Ex7+277 G>A) polymorphisms. We used unconditional logistic regression to estimate odds ratios (ORs) and 95% confidence intervals (CIs), adjusting for sex, age, education and smoking status. Out of six single nucleotide polymorphisms (SNPs) tested in TNFA, gastric cancer risk was significantly associated with the TNFA (-308 G>A) polymorphism, with ORs of 1.4 (95% CI: 1.0-2.0) for the G/A and 2.5 (95% CI: 1.3-4.9) for the A/A genotype carriers, when compared with the more frequent genotype (G/G) (P-trend < 0.001). Among the three tested SNPs in the IFNGR2 gene, only the Ex7-128C>T polymorphism was associated with increased risk, with ORs of 1.5 (95% CI: 1.0-2.3) for T/C and 1.7 (95% CI: 1.1-2.7) for C/C carriers when compared with T/T carriers (P-trend = 0.01). Subjects carrying both IFNGR2 Ex7-128 C/C and TNFA -308 A/A genotypes had the highest risk (OR = 5.5, 95% CI: 1.5-19.4), although the interaction was not statistically significant. IL1A (-889 C>T) and the three examined IL12A variants were unrelated to gastric cancer risk. Our findings suggest that two Th1-related polymorphisms (TNFA -308 A>G and IFNGR2 Ex7-128 C>T) may increase the risk of gastric cancer.


Bisphenol A may cause testosterone reduction by adversely affecting both testis and pituitary systems similar to estradiol.
Bisphenol A (BPA) causes reproductive toxicities, but the mechanisms are still unclear. In the present study, we sought to clarify these mechanisms in comparison with those of 17beta-estradiol (E2). Prepubertal Wistar/ST male rats (4 weeks old) were subcutaneously administered BPA (0, 20, 100 and 200 mg/kg/day) or E2 (10 and 100 microg/kg/day) for 6 weeks. Both BPA and E2 treatments decreased plasma and testicular testosterone levels, and plasma luteinizing hormone (LH), but not E2 and follicle-stimulating hormone levels, though E2 treatment increased its plasma level. In relation to the decreased testosterone levels, BPA and E2 decreased expressions of steroidogenic enzymes and cholesterol carrier protein in Leydig cells. Thus, decreased testosterone levels in plasma might have resulted from decreased expressions of these enzymes and protein as well as from decreased plasma LH levels. Interestingly, the changes in steroidogenic enzymes and carrier protein were observed at lower levels of exposure to BPA or E2 than those inhibiting plasma LH levels. Microscopically, 200 mg/kg BPA and 100 microg/kg E2 significantly decreased Leydig cell numbers in the testis. In addition, BPA and E2 also decreased expression of estrogen receptor alpha-mRNA, which might be related to the decreased numbers of Leydig cells. Thus, BPA directly affects not only the Leydig cells but also the pituitary gland, but the former may be impaired at lower exposure concentrations than the latter.


Engineered commensal microbes for diet-mediated colorectal-cancer chemoprevention.
Chemoprevention-the use of medication to prevent cancer-can be augmented by the consumption of produce enriched with natural metabolites. However, chemopreventive metabolites are typically inactive and have low bioavailability and poor host absorption. Here, we show that engineered commensal microbes can prevent carcinogenesis and promote the regression of colorectal cancer through a cruciferous vegetable diet. The engineered commensal Escherichia coli bound specifically to the heparan sulphate proteoglycan on colorectal cancer cells and secreted the enzyme myrosinase to transform host-ingested glucosinolates-natural components of cruciferous vegetables-to sulphoraphane, an organic small molecule with known anticancer activity. The engineered microbes coupled with glucosinolates resulted in >95% proliferation inhibition of murine, human and colorectal adenocarcinoma cell lines in vitro. We also show that murine models of colorectal carcinoma fed with the engineered microbes and the cruciferous vegetable diet displayed significant tumour regression and reduced tumour occurrence.


PTPRD (protein tyrosine phosphatase receptor type delta) is associated with restless legs syndrome.
We identified association of restless legs syndrome (RLS) with PTPRD at 9p23-24 in 2,458 affected individuals and 4,749 controls from Germany, Austria, Czechia and Canada. Two independent SNPs in the 5' UTR of splice variants expressed predominantly in the central nervous system showed highly significant P values (rs4626664, P(nominal/lambda corrected) = 5.91 x 10(-10), odds ratio (OR) = 1.44; rs1975197, P(nominal/lambda corrected) = 5.81 x 10(-9), OR = 1.31). This work identifies PTPRD as the fourth genome-wide significant locus for RLS.


Wolf spiders show graded antipredator behavior in the presence of chemical cues from different sized predators.
The wolf spider, Pardosa milvina, displays effective antipredator behavior (reduced activity) in the presence of silk and excreta cues from adults of another cooccurring wolf spider, Hogna helluo. However, Pardosa and Hogna engage in size-structured intraguild predation, where Pardosa may be either the prey or predator of Hogna. We tested the ability of adult female Pardosa to vary antipredator responses toward kairomones produced by Hogna that vary in size. Hogna were maintained on filter paper for 24 hr. We then presented the paper to adult female Pardosa simultaneously paired with a blank sheet of paper. One treatment had two sheets of blank paper to serve as a control. The Hogna stimulus treatments were as follows (N = 15/treatment): (1) 1 Hogna half the mass of Pardosa; (2) 1 Hogna of equal mass of a Pardosa; (3) 1 adult Hogna, 30 times the mass of Pardosa; and (4) 8 Hogna each 0.25 the mass of Pardosa. Pardosa decreased activity in the presence of kairomones from Hogna of equal or larger size, but showed no change in activity in the presence of a blank control or from a single Hogna smaller than itself. Pardosa showed a reduction in activity in the presence of cues from eight small Hogna. Pardosa avoided substrates with adult Hogna cues, but showed no avoidance response to any other treatment. These results suggest that Pardosa is showing graded antipredator behavior relative to the quantity of predator kairomones present rather than directly discriminating among the different sizes of the predator.


Self-advanced fast light propagation in an optical fiber based on Brillouin scattering.
We experimentally demonstrate an extremely simple technique to achieve pulse advancements in optical fibers by using both spontaneous amplified and stimulated Brillouin scattering. It is shown that the group velocity of a light signal is all-optically controlled by its average power while it propagates through an optical fiber. The signal generates an intense back-propagating Stokes emission that causes a loss on the signal through depletion. This narrowband loss gives rise to a fast light propagation at the exact signal frequency. The Stokes emission self-adapts in real time to the Brillouin properties of the fiber and to a wide extent to the signal bandwidth.


A Comparison of Internal Jugular Vein Cannulation by Ultrasound-Guided and Anatomical Landmark Technique in Resource-Limited Emergency Department Setting.
Central venous catheter placement is a frequently performed procedure in emergency medicine as well as critical care unit. We aimed to compare real-time ultrasonography (USG)-guided and the traditional anatomical landmark (AL) technique for the insertion of internal jugular vein (IJV) catheters in an emergency department (ED) setting. Patients requiring IJV catheterization were prospectively recruited over a period of 1 year at a single center. Cannulation was done either by the AL or USG technique, according to ED physician's discretion. A preset pro forma was completed for each central line placed. Variables were compared using the independent t-test, Fisher's exact test, and the nonparametric Mann-Whitney U-test. Seventy patients were enrolled, of which 35 patients underwent IJV cannulation by USG-guided technique (USG group) and 35 patients by the AL technique (AL group). There were a 100% success rate (35/35) for cannulation in the USG group and a 91.4% success rate (32/35) in the AL group. The catheter was placed on the first attempt in 17 (48.6%) patients in the AL group and 32 (91.4%) patients in the USG group. In th AL group, there were three failed cannulation attempts in comparison to the USG group. The mean start to flash time for the AL technique was 16.59 s (±10.67) and 4.86 s (±2.18) in the USG group. The mean cannulation time was 305.88 s (±66.84) in the AL group and 293.03 s (±71.15) in the USG group. A total of seven acute complications were noted, of which 2 (5.7%) in the USG group and 5 (14.3%) in the AL group. The real-time USG guided technique significantly reduces the number of attempts to cannulate, has a higher first-pass success rate, a quicker flash time, and fewer complications when compared to the AL technique. In EDs equipped with USG, insertion of IJV catheters under real-time USG guidance should become the standard of care.


High-risk case identification for use in comprehensive complex care management.
The current study aimed to develop a patient selection process for muiltimorbid care management that balances the needs to accurately identify patients who are at risk for future high costs and assures that those selected can clinically benefit from proactive care management. Six physicians were surveyed on characteristics of their current (2012) patients to elicit clinical considerations for high-risk patient identification. Data from 2010-2011 were extracted from Clalit Health Services' (Israel's largest managed care organization) comprehensive database to derive the Adjusted Clinical Groups (ACG) predictive model risk scores for risk of future high costs. Model discriminatory power was assessed using the c-statistic and positive predictive value (PPV), before and after application of the clinical exclusion criteria. Inclusion criteria were refined based on physician input from a survey on 375 patients. Recommended reasons for exclusion: active cancer, schizophrenia, dialysis, residence in nursing homes or long-term care facilities, and age 95 years or older. The high-risk group included 5341 patients (mean 50 patients per physician). The c-statistic of the ACG model before and after exclusions applied was 0.80 and 0.75, respectively. After exclusion, the PPV for the 6% highest risk patients was 40%. High-risk patients' age, number of chronic conditions, and utilization were substantially higher than those of all other patients. This study shows that a validated predictive modeling tool provides acceptable discriminatory power for selecting multimorbid patients for participation in proactive care management, even after some of the highest risk patients are excluded because of priori clinical considerations.


Alveolar nerve unfolding technique for synoptic analysis: visualization and quantification of inferior alveolar nerve tracings in three-dimensional cone-beam computed tomography.
The aim of the technique presented here is to visualize the anatomical context of the inferior alveolar nerve (IAN) canal. For 2 cases, cone-beam computed tomography images of the mandible were obtained from patient files together with the manual preoperative IAN canal tracings. For both cases, similar to simulated panoramic images, a two-dimensional image is extracted from a three-dimensional cone-beam computed tomography image. Unlike panoramic images, the unfolding does not follow the general curvature of the mandible but follows the nerve tracing closely and places the traced nerve track on a horizontal central line. Because of the centering of the nerve tracing together with the nerve canal and its surroundings in a two-dimensional representation, the technique (ANUTSA [Alveolar Nerve Unfolding Technique for Synoptic Analysis]) allowed the first case to evidence the adjacency of root tips along the IAN, whereas in the second case the degree of penetration of the IAN by an implant is revealed. The global aspect of the representation through unfolding allowed for the detection of the anomalies and the IAN-penetrating lesion along the IAN canal at a glance.


What is the value of regional cerebral saturation in post-cardiac arrest patients? A prospective observational study.
The aim of this study was to elucidate the possible role of cerebral saturation monitoring in the post-cardiac arrest setting. Cerebral tissue saturation (SctO2) was measured in 107 successfully resuscitated out-of-hospital cardiac arrest patients for 48 hours between 2011 and 2015. All patients were treated with targeted temperature management, 24 hours at 33 °C and rewarming at 0.3 °C per hour. A threshold analysis was performed as well as a linear mixed models analysis for continuous SctO2 data to compare the relation between SctO2 and favorable (cerebral performance category (CPC) 1-2) and unfavorable outcome (CPC 3-4-5) at 180 days post-cardiac arrest in OHCA patients. Of the 107 patients, 50 (47 %) had a favorable neurological outcome at 180 days post-cardiac arrest. Mean SctO2 over 48 hours was 68 % ± 4 in patients with a favorable outcome compared to 66 % ± 5 for patients with an unfavorable outcome (p = 0.035). No reliable SctO2 threshold was able to predict favorable neurological outcome. A significant different course of SctO2 was observed, represented by a logarithmic and linear course of SctO2 in patients with favorable outcome and unfavorable outcome, respectively (p < 0.001). During the rewarming phase, significant higher SctO2 values were observed in patients with a favorable neurological outcome (p = 0.046). This study represents the largest post-resuscitation cohort evaluated using NIRS technology, including a sizeable cohort of balloon-assisted patients. Although a significant difference was observed in the overall course of SctO2 between OHCA patients with a favorable and unfavorable outcome, the margin was too small to likely represent functional outcome differentiation based on SctO2 alone. As such, these results given such methodology as performed in this study suggest that NIRS is insufficient by itself to serve in outcome prognostication, but there may remain benefit when incorporated into a multi-neuromonitoring bedside assessment algorithm.


Efficient plasmonic scattering of colloidal silver particles through annealing-induced changes.
We present an experimental study on the influence of annealing temperature on morphological changes of colloidal silver nanoparticles (Ag NPs) and their optical response. Monodisperse colloidal Ag NPs with diameter of 164 nm ± 15 nm have been fabricated by a facile two-step synthesis approach. The annealing effects on the Ag NPs have been investigated by means of optical measurements, scanning electron microscopy, x-ray photoelectron spectroscopy and x-ray diffraction. By annealing up to 440 °C morphology and chemical compositions of the Ag NPs changed. These changes affect the particle size and distribution, surface morphology, crystallinity and, most importantly, the oxidation state of the surface layer. The removal of an oxide layer leads to stronger light scattering from the nanoparticles and decreases parasitic light absorption at wavelengths above 400 nm. Strong light trapping has been observed for a silicon thin film with embedded annealed Ag NPs. This study demonstrates that colloidal Ag NPs for plasmonic solar cells need to be carefully processed and that they can potentially achieve high scatter efficiencies.


Pediatric skull base reconstruction: case report of a tunneled temporoparietal fascia flap.
The authors of this report present a pediatric case involving the use of a tunneled temporoparietal fascia flap to reconstruct a skull base defect for a multiply recurrent clival chordoma and cerebrospinal fluid leak, demonstrate the surgical technique through illustrations and intraoperative photos, and review the pertinent literature. A 9-year-old female patient underwent extensive clival chordoma resection via both the endoscopic and open approaches, which ultimately exhausted the bilateral nasoseptal flaps and other intranasal reconstructive options. Following proton beam radiation and initiation of chemotherapy, tumor recurrence was managed with further endoscopic resection, which was complicated by a recalcitrant cerebrospinal fluid leak. A tunneled temporoparietal fascia flap was used to provide vascular tissue to augment an endoscopic repair of the leak and reconstruction of the skull base. While the nasoseptal flap remains the workhorse for many pediatric and adult endoscopic skull base reconstructions, the tunneled temporoparietal fascia flap has a demonstrated efficacy in adults when the nasoseptal flap and other intranasal flaps are unavailable. This report documents a pediatric case, serving as a step toward establishing this technique in the pediatric population.


The role of forward osmosis and microfiltration in an integrated osmotic-microfiltration membrane bioreactor system.
This study investigates the performance of an integrated osmotic and microfiltration membrane bioreactor (O/MF-MBR) system for wastewater treatment and reclamation. The O/MF-MBR system simultaneously used microfiltration (MF) and forward osmosis (FO) membranes to extract water from the mixed liquor of an aerobic bioreactor. The MF membrane facilitated the bleeding of dissolved inorganic salts and thus prevented the build-up of salinity in the bioreactor. As a result, sludge production and microbial activity were relatively stable over 60 days of operation. Compared to MF, the FO process produced a better permeate quality in terms of nutrients, total organic carbon, as well as hydrophilic and biologically persistent trace organic chemicals (TrOCs). The high rejection by the FO membrane also led to accumulation of hydrophilic and biologically persistent TrOCs in the bioreactor, consequently increasing their concentration in the MF permeate. On the other hand, hydrophobic and readily biodegradable TrOCs were minimally detected in both MF and FO permeates, with no clear difference in the removal efficiencies between two processes.


A supra-cellular model for coupling of bone resorption to formation during remodeling: lessons from two bone resorption inhibitors affecting bone formation differently.
The bone matrix is maintained functional through the combined action of bone resorbing osteoclasts and bone forming osteoblasts, in so-called bone remodeling units. The coupling of these two activities is critical for securing bone replenishment and involves osteogenic factors released by the osteoclasts. However, the osteoclasts are separated from the mature bone forming osteoblasts in time and space. Therefore the target cell of these osteoclastic factors has remained unknown. Recent explorations of the physical microenvironment of osteoclasts revealed a cell layer lining the bone marrow and forming a canopy over the whole remodeling surface, spanning from the osteoclasts to the bone forming osteoblasts. Several observations show that these canopy cells are a source of osteoblast progenitors, and we hypothesized therefore that they are the likely cells targeted by the osteogenic factors of the osteoclasts. Here we provide evidence supporting this hypothesis, by comparing the osteoclast-canopy interface in response to two types of bone resorption inhibitors in rabbit lumbar vertebrae. The bisphosphonate alendronate, an inhibitor leading to low bone formation levels, reduces the extent of canopy coverage above osteoclasts. This effect is in accordance with its toxic action on periosteoclastic cells. In contrast, odanacatib, an inhibitor preserving bone formation, increases the extent of the osteoclast-canopy interface. Interestingly, these distinct effects correlate with how fast bone formation follows resorption during these respective treatments. Furthermore, canopy cells exhibit uPARAP/Endo180, a receptor able to bind the collagen made available by osteoclasts, and reported to mediate osteoblast recruitment. Overall these observations support a mechanism where the recruitment of bone forming osteoblasts from the canopy is induced by osteoclastic factors, thereby favoring initiation of bone formation. They lead to a model where the osteoclast-canopy interface is the physical site where coupling of bone resorption to bone formation occurs.


Patient survival and tumor characteristics associated with CHEK2:p.I157T - findings from the Breast Cancer Association Consortium.
P.I157T is a CHEK2 missense mutation associated with a modest increase in breast cancer risk. Previously, another CHEK2 mutation, the protein truncating c.1100delC has been associated with poor prognosis of breast cancer patients. Here, we have investigated patient survival and characteristics of breast tumors of germ line p.I157T carriers. We included in the analyses 26,801 European female breast cancer patients from 15 studies participating in the Breast Cancer Association Consortium. We analyzed the association between p.I157T and the clinico-pathological breast cancer characteristics by comparing the p.I157T carrier tumors to non-carrier and c.1100delC carrier tumors. Similarly, we investigated the p.I157T associated risk of early death, breast cancer-associated death, distant metastasis, locoregional relapse and second breast cancer using Cox proportional hazards models. Additionally, we explored the p.I157T-associated genomic gene expression profile using data from breast tumors of 183 Finnish female breast cancer patients (ten p.I157T carriers) (GEO: GSE24450). Differential gene expression analysis was performed using a moderated t test. Functional enrichment was investigated using the DAVID functional annotation tool and gene set enrichment analysis (GSEA). The tumors were classified into molecular subtypes according to the St Gallen 2013 criteria and the PAM50 gene expression signature. P.I157T was not associated with increased risk of early death, breast cancer-associated death or distant metastasis relapse, and there was a significant difference in prognosis associated with the two CHEK2 mutations, p.I157T and c.1100delC. Furthermore, p.I157T was associated with lobular histological type and clinico-pathological markers of good prognosis, such as ER and PR expression, low TP53 expression and low grade. Gene expression analysis suggested luminal A to be the most common subtype for p.I157T carriers and CDH1 (cadherin 1) target genes to be significantly enriched among genes, whose expression differed between p.I157T and non-carrier tumors. Our analyses suggest that there are fundamental differences in breast tumors of CHEK2:p.I157T and c.1100delC carriers. The poor prognosis associated with c.1100delC cannot be generalized to other CHEK2 mutations.


[Forcing patients with bipolar disorder to make a financial contribution to secondary psychiatric care may make them forsake care completely].
In 2012 patients were required to make a personal financial contribution for secondary mental health care over and above their standard insurance fee. According to current guidelines, the majority of patients with bipolar disorder must be treated as outpatients at psychiatric clinics. It was to be expected that some patients would decide to discontinue their outpatient treatment on account of the newly imposed personal financial contribution. To obtain insight into the size and characteristics of the group of patients with bipolar disorder who were thinking about giving up treatment or had already decided to give it up because of the imposition of the personal financial contribution; also to find out which factors influenced patients' decisions and to discover how patients perceived the role of the GP as the provider of subsequent psychiatric care. We conducted an exploratory study by sending a survey to all outpatients receiving treatment at three clinics specialising in the treatment of bipolar disorder. 640 patients responded to the survey (55% response); 15% of these patients were thinking about giving up treatment or had already decided to stop their treatment. They were influenced primarily by financial considerations. Two-thirds of the respondents did not consider that the GP was as a suitable alternative to outpatient care at a clinic. Even patients with moderate to serious symptoms were considering leaving secondary care. The obligatory financial contribution for secondary mental health care could have considerable consequences for a small number of patients with severe mood disorder who are currently treated as outpatients in secondary facilities. The increase in the compulsory & squo;own risk' insurance fee as from 2013 could have similar consequences.


New highly sensitive rodent and human tests for soluble amyloid precursor protein alpha quantification: preclinical and clinical applications in Alzheimer's disease.
Amyloid precursor protein (APP), a key molecule in Alzheimer's disease (AD), is metabolized in two alternative cleavages, generating either the amyloidogenic peptides involved in AD pathology or the soluble form of APP (sAPPα). The level of amyloidogenic peptides in human cerebrospinal fluid (CSF) is considered to be a biomarker of AD, whereas the level of sAPPα in CSF as a biomarker has not been clearly established. sAPPα has neurotrophic and neuroprotective properties. Stimulating its formation and secretion is a promising therapeutic target in AD research. To this end, very sensitive tests for preclinical and clinical research are required. The tests are based on homogenous time-resolved fluorescence and require no washing steps. We describe two new rapid and sensitive tests for quantifying mouse and human sAPPα. These 20 μl-volume tests quantify the levels of: i) endogenous mouse sAPPα in the conditioned medium of mouse neuron primary cultures, as well as in the CSF of wild-type mice, ii) human sAPPα in the CSF of AD mouse models, and iii) human sAPPα in the CSF of AD and non-AD patients. These tests require only 5 μl of conditioned medium from 5 × 10(4) mouse primary neurons, 1 μl of CSF from wild-type and transgenic mice, and 0.5 μl of human CSF. The high sensitivity of the mouse sAPPα test will allow high-throughput investigations of molecules capable of increasing the secretion of endogenous sAPPα in primary neurons, as well as the in vivo validation of molecules of interest through the quantification of sAPPα in the CSF of treated wild-type mice. Active molecules could then be tested in the AD mouse models by quantifying human sAPPα in the CSF through the progression of the disease. Finally, the human sAPPα test could strengthen the biological diagnosis of AD in large clinical investigations. Taken together, these new tests have a wide field of applications in preclinical and clinical studies.


A finite element study of short dental implants in the posterior maxilla.
Elevated bite forces and reduced bone densities and dimensions associated with posterior regions of the maxilla cause relatively high failure rates when short dental implants are placed to substitute missing teeth. This study uses the finite element method to evaluate four distinctly different short implant designs (Bicon, Neodent, Nobel Biocare, and Straumann) for their influences on the von Mises stress characteristics within the posterior maxilla. Finite element models of the supporting bone and tooth crowns are developed based on computed tomography data, and implant geometries are obtained from manufacturers' catalogs. The finite element models are meshed using three-dimensional hexahedral and wedge-shaped brick elements. Assumptions made in the analyses are: linear elastic material properties for bone, 50% osseointegration between bone and implant, and crown height-implant length ratio of 2:1. Bicon's neck indentation produced reduced stress in the cortical bone when compared with the Nobel Biocare and Straumann systems. The increased taper of the Neodent design decreased the stress level in cancellous bone. Nobel Biocare's rounded thread crest and reduced thread pitch produced a smoother stress profile. Straumann's increased thread pitch produced elevated stress in the cancellous bone. Generally, stresses were concentrated in the crestal bone region around the implant neck, attributable to the inclined nature of the masticatory force. Nobel Biocare and Bicon systems are recommended for use in type 4 cancellous and cortical bone, respectively.


Triatominae as a model of morphological plasticity under ecological pressure.
The use of biochemical and genetic characters to explore species or population relationships has been applied to taxonomic questions since the 60s. In responding to the central question of the evolutionary history of Triatominae, i.e. their monophyletic or polyphyletic origin, two important questions arise (i) to what extent is the morphologically-based classification valid for assessing phylogenetic relationships? and (ii) what are the main mechanisms underlying speciation in Triatominae? Phenetic and genetic studies so far developed suggest that speciation in Triatominae may be a rapid process mainly driven by ecological factors.


Analyses of the mouthpart kinematics in Periplaneta americana (Blattodea, Blattidae) using synchrotron-based X-ray cineradiography.
The kinematics of the biting and chewing mouthparts of insects is a complex interaction of various components forming multiple jointed chains. The non-invasive technique of in vivo cineradiography by means of synchrotron radiation was employed to elucidate the motion cycles of the mouthparts in the cockroach Periplaneta americana. Digital X-ray footage sequences were used in order to calculate pre-defined angles and distances, each representing characteristic aspects of the movement pattern. We were able to analyze the interactions of the mouthpart components and to generate a functional model of maxillary movement by integrating kinematic results, morphological dissections and fluorescence microscopy. During the opening and closing cycles, which take about 450-500 ms on average, we found strong correlations between the measured maxillary and mandibular angles, indicating a strong neural coordination of these movements. This is manifested by strong antiphasic courses of the maxillae and the mandibles, antiphasic patterns of the rotation of the cardo about its basic articulation at the head and by the deflection between the cardo and stipes. In our functional model of the maxilla, its movement pattern is explained by the antagonistic activity of four adductor-promotor muscles and two abductor-remotor muscles. However, beyond the observed intersegmental and bilateral stereotypy, certain amounts of variation across subsequent cycles within a sequence were observed with respect to the degree of correlation between the various mouthparts, the maximum, minimum and time course of the angular movements. Although generally correlated with the movement pattern of the mandibles and the maxillary cardo-stipes complex, such plastic behaviour was especially observed in the maxillary palpi and the labium.


In vivo evaluation of carbon fullerene toxicity using embryonic zebrafish.
There is a pressing need to develop rapid whole animal-based testing assays to assess the potential toxicity of engineered nanomaterials. To meet this challenge, the embryonic zebrafish model was employed to determine the toxicity of fullerenes. Embryonic zebrafish were exposed to graded concentrations of fullerenes [C(60), C(70), and C(60)(OH)(24)] during early embryogenesis and the resulting morphological and cellular responses were defined. Exposure to 200 μg/L C(60) and C(70) induced a significant increased in malformations, pericardial edema, and mortality; while the response to C(60)(OH)(24) exposure was less pronounced at concentrations an order of magnitude higher. Exposure to C(60) induced both necrotic and apoptotic cellular death throughout the embryo. While C(60)(OH)(24) induced an increase in embryonic cellular death, it did not induce apoptosis. Our findings concur with results obtained in other models indicating that C(60)(OH)(24) is significantly less toxic than C(60). These studies also suggest that that the embryonic zebrafish model is well-suited for the rapid assessment of nanomaterial toxicity.


Hand and sex differences in the isthmus and genu of the human corpus callosum. A postmortem morphological study.
The midsagittal area of the corpus callosum was measured in its entirety and in seven subdivisions in a sample of 50 brains consecutively obtained from autopsies of individuals who had neuropsychological testing before death. A 12-item test of hand preference was used as an index of the pattern of interhemispheric functional asymmetry. Callosal size was analysed for two factors: hand preference, classified as consistent-right-hand preference (CRH) versus non consistent-right-hand preference, and sex. The group of nCRH (n = 18) was found to have a larger overall callosal area, with the greatest difference occurring in the posterior body segments, especially the isthmus. The isthmus probably includes interhemispheric fibres from posterior parietal and superior temporal cortex which involves cortical regions related to functional asymmetry. The results of variation in callosal morphology are discussed as part of a possible substrate of functional asymmetry and due to variation in axonal elimination in early brain development. Sex differences were found in several aspects of callosal anatomy. (1) The difference between hand groups in the posterior body occurred in interaction with sex:handedness was a factor in callosal size in males (n = 15), but not in females (n = 35). This result is consistent with the general hypothesis of females having less clear lateralization than males. (2) Females did not have a larger overall callosum or a larger splenium, either in absolute size or size proportional to brain weight. The latter measure was considered since callosal area correlated with cerebrum weight (r = 0.48). In contrast, female of both hand groups were found to have a larger proportional isthmus compared to CRH males. (3) Of all callosal regions, only the genu and a part of the anterior body were found to be larger in absolute size in males than females. (4) Callosal size decreased with chronological age in males, but not in females.


Amphiphilic dynamic NDI and PDI probes: imaging microdomains in giant unilamellar vesicles.
Dynamic amphiphiles provide access to transmembrane ion transport, differential sensing and cellular uptake. In this report, we introduce dynamic amphiphiles with fluorescent tails. Core-substituted naphthalenediimides (cNDIs) and perylenediimides (cPDIs) are tested. Whereas the latter suffer from poor partitioning, dynamic cNDI amphiphiles are found to be purifiable by RP-HPLC, to partition selectively into liquid-disordered (Ld) microdomains of mixed lipid bilayers and to activate DNA as transporters. Importantly, fluorescence properties, partitioning and activity can be modulated by changes in the structure of mixed amphiphiles. These results confirm the potential of dynamic fluorescent amphiphiles to selectively label extra- and intracellular membrane domains and visualize biological function.


Matrix stiffness regulates migration of human lung fibroblasts.
In patients with pulmonary diseases such as idiopathic pulmonary fibrosis and severe acute respiratory distress syndrome, progressive pulmonary fibrosis is caused by dysregulated wound healing via activation of fibroblasts after lung inflammation or severe damage. Migration of fibroblasts toward the fibrotic lesions plays an important role in pulmonary fibrosis. Fibrotic tissue in the lung is much stiffer than normal lung tissue. Emerging evidence supports the hypothesis that the stiffness of the matrix is not only a consequence of fibrosis, but also can induce fibroblast activation. Nevertheless, the effects of substrate rigidity on migration of lung fibroblasts have not been fully elucidated. We evaluated the effects of substrate stiffness on the morphology, α-smooth muscle actin (α-SMA) expression, and cell migration of primary human lung fibroblasts by using polyacrylamide hydrogels with stiffnesses ranging from 1 to 50 kPa. Cell motility was assessed by platelet-derived growth factor (PDGF)-induced chemotaxis and random walk migration assays. As the stiffness of substrates increased, fibroblasts became spindle-shaped and spread. Expression of α-SMA proteins was higher on the stiffer substrates (25 kPa gel and plastic dishes) than on the soft 2 kPa gel. Both PDGF-induced chemotaxis and random walk migration of fibroblasts precultured on stiff substrates (25 kPa gel and plastic dishes) were significantly higher than those of cells precultured on 2 kPa gel. Transfection of the fibroblasts with short interfering RNA for α-SMA inhibited cell migration. These findings suggest that fibroblast activation induced by a stiff matrix is involved in mechanisms of the pathophysiology of pulmonary fibrosis.


Widespread omnivory and low temporal and spatial variation in the diet of fishes in a hydrologically variable northern Australian river.
This article examines the trophic ecology of freshwater fishes (22 species in 15 families) in a wet and dry tropical Australian river of high intra-annual and interannual hydrological variability. Seven major trophic groups were identified by cluster analysis; however, four food items (filamentous algae, chironomid larvae, Trichoptera larvae and Ephemeroptera nymphs) comprised almost half of the average diet of all species. The influence of species, fish size, spatial effects and temporal effects on food use was investigated using redundancy analysis. Size, time and space accounted for little of the perceived variation. Ontogenetic changes in diet were minor and limited to a few large species. Spatial variation in trophic composition of the fish assemblages reflected the effects of the Burdekin Falls and dam, a major geographic barrier, on species distributions. Little spatial variation in diet was detected after accounting for this biogeographical effect. Temporal variations in flow, although marked, had little effect on variations in fish diet composition due to the low temporal diversity of food resources in physically monotonous sand and gravel channels. Species identity accounted for<50% of the observed variation in food choice; omnivory and generalism were pronounced. The aquatic food web of the Burdekin River appears simple, supported largely by autochthonous production (filamentous and benthic microalgae, and to some extent, aquatic macrophytes). Allochthonous food resources appear to be unimportant. The generalist feeding strategies, widespread omnivory and absence of pronounced trophic segregation reported here for Burdekin River fishes may be common to variable and intermittent rivers of subtropical and tropical northern Australia with similar fish communities and may be a general feature of rivers of low habitat diversity and characterized by flow regimes that vary greatly both within and between years.


Factors affecting pregnancy outcomes in young women treated with fertility-preserving therapy for well-differentiated endometrial cancer or atypical endometrial hyperplasia.
Patients hoping to preserve their fertility receive conservative treatment with high-dose medroxyprogesterone acetate (MPA) for well-differentiated endometrioid adenocarcinoma (EC) or atypical endometrial hyperplasia (AEH) . Such treatment generally involves frequent intrauterine operations, including dilation and curettage (D&C) and endometrial biopsy (EMB), which could result in endometritis, endometrial thinning, or intrauterine adhesion. In turn, any of these outcomes could adversely affect implantation and pregnancy development. The current study thus aimed to identify factors that might affect pregnancy following conservative treatment by MPA. We compared a pregnancy group (45 patients) with a non-pregnancy group (53 patients) of MPA-treated patients to evaluate the factors affecting clinical pregnancy establishment. We undertook a multivariate logistic regression analysis based on factors shown by univariate analysis to be significantly different between the groups. Univariate analysis identified number of D&C, endometrial thickness, duration of MPA administration, age of pregnancy permission (the age at which a patient was first allowed to attempt pregnancy after disappearance of the lesion), period of disappearance of lesions, and recurrence as independent variables. The odds ratios (95 % confidence interval) of multivariate analysis for disease recurrence, endometrial thickness during ovulation, and age of pregnancy permission were 0.283 (0.102-0.785), 1.677 (1.251-2.248), and 0.889 (0.792-0.998), respectively. There was no significant difference in the other independent variables between groups. We identified three factors considered to affect pregnancy establishment following conservative treatment with MPA: recurrence, endometrial thickness during ovulation, and the age of the pregnancy permission. Introduction of infertility treatment including assisted reproductive technology (ART) soon after achieving tumor disappearance by MPA would therefore be beneficial for patients with disease recurrence, thin endometrium, or a higher age of pregnancy permission.


Shiga toxin-producing Escherichia coli.
In the United States, it is estimated that non-O157 Shiga toxin-producing Escherichia coli (STEC) cause more illnesses than STEC O157:H7, and the majority of cases of non-O157 STEC infections are due to serogroups O26, O45, O103, O111, O121, and O145, referred to as the top six non-O157 STEC. The diseases caused by non-O157 STEC are generally milder than those induced by O157 STEC; nonetheless, non-O157 STEC strains have also been associated with serious illnesses such as hemorrhagic colitis and hemolytic uremic syndrome, as well as death. Ruminants, particularly cattle, are reservoirs for both O157 and non-O157 STEC, which are transmitted to humans by person-to-person or animal contact and by ingestion of food or water contaminated with animal feces. Improved strategies to control STEC colonization and shedding in cattle and contamination of meat and produce are needed. In general, non-O157 STEC respond to stresses such as acid, heat, and other stresses induced during food preparation similar to O157 STEC. Similar to O157:H7, the top six non-O157 STEC are classified as adulterants in beef by the USDA Food Safety and Inspection Service, and regulatory testing for these pathogens began in June 2012. Due to the genetic and phenotypic variability of non-O157 STEC strains, the development of accurate and reliable methods for detection and isolation of these pathogens has been challenging. Since the non-O157 STEC are responsible for a large portion of STEC-related illnesses, more extensive studies on their physiology, genetics, pathogenicity, and evolution are needed in order to develop more effective control strategies.


"Not a university type": focus group study of social class, ethnic, and sex differences in school pupils' perceptions about medical school.
To investigate what going to medical school means to academically able 14-16 year olds from different ethnic and socioeconomic backgrounds in order to understand the wide socioeconomic variation in applications to medical school. Focus group study. Six London secondary schools. 68 academically able and scientifically oriented pupils aged 14-16 years from a wide range of social and ethnic backgrounds. Pupils' perceptions of medical school, motivation to apply, confidence in ability to stay the course, expectations of medicine as a career, and perceived sources of information and support. There were few differences by sex or ethnicity, but striking differences by socioeconomic status. Pupils from lower socioeconomic groups held stereotyped and superficial perceptions of doctors, saw medical school as culturally alien and geared towards "posh" students, and greatly underestimated their own chances of gaining a place and staying the course. They saw medicine as having extrinsic rewards (money) but requiring prohibitive personal sacrifices. Pupils from affluent backgrounds saw medicine as one of a menu of challenging career options with intrinsic rewards (fulfillment, achievement). All pupils had concerns about the costs of study, but only those from poor backgrounds saw costs as constraining their choices. Underachievement by able pupils from poor backgrounds may be more to do with identity, motivation, and the cultural framing of career choices than with low levels of factual knowledge. Policies to widen participation in medical education must go beyond a knowledge deficit model and address the complex social and cultural environment within which individual life choices are embedded.


The effect of real-time gait retraining on hip kinematics, pain and function in subjects with patellofemoral pain syndrome.
Patellofemoral pain syndrome (PFPS) is the most common overuse injury in runners. Recent research suggests that hip mechanics play a role in the development of this syndrome. Currently, there are no treatments that directly address the atypical mechanics associated with this injury. The purpose of this study was to determine whether gait retraining using real-time feedback improves hip mechanics and reduces pain in subjects with PFPS. Ten runners with PFPS participated in this study. Real-time kinematic feedback of hip adduction (HADD) during stance was provided to the subjects as they ran on a treadmill. Subjects completed a total of eight training sessions. Feedback was gradually removed over the last four sessions. Variables of interest included peak HADD, hip internal rotation (HIR), contralateral pelvic drop, as well as pain on a verbal analogue scale and the lower-extremity function index. We also assessed HADD, HIR and contralateral pelvic drop during a single leg squat. Comparisons of variables of interest were made between the initial, final and 1-month follow-up visit. Following the gait retraining, there was a significant reduction in HADD and contralateral pelvic drop while running. Although not statistically significant, HIR decreased by 23% following gait retraining. The 18% reduction in HADD during a single leg squat was very close to significant. There were also significant improvements in pain and function. Subjects were able to maintain their improvements in running mechanics, pain and function at a 1-month follow-up. An unexpected benefit of the retraining was an 18% and 20% reduction in instantaneous and average vertical load rates, respectively. Gait retraining in individuals with PFPS resulted in a significant improvement of hip mechanics that was associated with a reduction in pain and improvements in function. These results suggest that interventions for PFPS should focus on addressing the underlying mechanics associated with this injury. The reduction in vertical load rates may be protective for the knee and reduce the risk for other running-related injuries.


Voluntary wheel running decreases adipose tissue mass and expression of leptin mRNA in Osborne-Mendel rats.
The purpose of this study was to assess the effects of voluntary wheel running on the expression of leptin mRNA in rats that are either sensitive (OM) or resistant (S5B/Pl) to diet-induced obesity. Male OM and S5B/Pl rats had ad libitum access to standard rodent diet and water. At 3-5 weeks of age, animals of both strains were randomly assigned to either an exercise or sedentary control group. The exercise groups had 24-h access to a running wheel, and they trained for 7 weeks. During weeks 1-4, animals in both OM and S5B/Pl exercise groups progressively increased their running. During weeks 5-7, S5B/Pl exercisers tended to run more than did OM (approximately 60 vs. 45 km/week), but by the end of the study both groups had an equally greater heart weight (mg/g body weight) and planteris citrate synthase activity than their sedentary controls. Oral glucose tolerance tests performed during the last week of training revealed that compared with their appropriate controls, insulin sensitivity was enhanced (P < 0.05) in OM but not in the S5B/Pl wheel-running groups. Inguinal, epididymal, and retroperitoneal fat pads weighed less in the running than in the nonrunning groups of both strains (P < 0.01). Additionally, exercised animals had an increased percentage of smaller cells (40-60 microm; P < 0.05) and a decreased percentage of larger cells (120-160 microm; P < 0.05) in the epididymal fat depot. Epididymal leptin mRNA measured by Northern blot analysis was reduced in the exercise-trained rats of both strains (P < 0.05). Furthermore, serum leptin was reduced in exercise-trained compared with the control animals of both strains. In comparison to S5B/Pl, control OM animals exhibited both a higher expression and higher circulating levels of leptin (P < 0.05). While serum leptin levels were decreased and food intake was increased in the exercise-trained animals of both strains (P < 0.05), the exact relationship between exercise, leptin, and food intake in this rat model of dietary obesity remains to be determined. Nonetheless, these results suggest that the expression and secretion of leptin can be influenced by exercise training and that these changes (i.e., reduced expression and secretion of protein) can occur independently of changes in whole-body insulin sensitivity and susceptibility to diet-induced obesity.


SCAPER, a novel cyclin A-interacting protein that regulates cell cycle progression.
Cyclin A/Cdk2 plays an important role during S and G2/M phases of the eukaryotic cell cycle, but the mechanisms by which it regulates cell cycle events are not fully understood. We have biochemically purified and identified SCAPER, a novel protein that specifically interacts with cyclin A/Cdk2 in vivo. Its expression is cell cycle independent, and it associates with cyclin A/Cdk2 at multiple phases of the cell cycle. SCAPER localizes primarily to the endoplasmic reticulum. Ectopic expression of SCAPER sequesters cyclin A from the nucleus and results specifically in an accumulation of cells in M phase of the cell cycle. RNAi-mediated depletion of SCAPER decreases the cytoplasmic pool of cyclin A and delays the G1/S phase transition upon cell cycle re-entry from quiescence. We propose that SCAPER represents a novel cyclin A/Cdk2 regulatory protein that transiently maintains this kinase in the cytoplasm. SCAPER could play a role in distinguishing S phase- from M phase-specific functions of cyclin A/Cdk2.


A Delphi study to identify healthcare users' priorities for cancer care in Greece.
User involvement is widely promoted within the European healthcare sector as an effective means of developing patient-centred health services. In addition, user involvement is recommended as an important aspect of developing effective cancer services. Despite the acknowledged benefits of user involvement, this concept has not been fully endorsed in Greece. Greek healthcare users may be increasingly involved in the decisions made for their health, but their involvement in cancer services planning remains very limited. The aim of this study was to identify healthcare users' key areas of cancer care and services that needed to be developed or improved in Greece and prioritise them through a consensus building study. A Delphi technique was used to collect data from a sample of 30 healthcare users in two rounds. The response rate for each round was over 80%. The main consensual issues prioritised by the healthcare users were the improvement of education for health professionals and the development of effective treatments. Other areas that were also rated highly included the organisation of cancer services, the need for more resources and improvements in the attitude of healthcare professionals. Healthcare users' priorities were focused on timely diagnosis and treatment by well-educated healthcare professionals, demonstrating a strong will for survival. Based on the findings from this study and the literature recommendations, it is expected that the re-organisation and fair distribution of the existing resources could improve the experience of healthcare users. In addition, the number of employed nurses should increase and improvements in healthcare professionals' education should be implemented in order to develop and improve cancer services. Public health education needs to be emphasised and prioritised especially in areas such as prevention and screening. The Greek government needs to address and encourage user involvement in planning healthcare services with the introduction of relevant legislation.


Spectral-reflectance linear models for optical color-pattern recognition.
We propose a new method of color-pattern recognition by optical correlation that uses a linear description of spectral reflectance functions and the spectral power distribution of illuminants that contains few parameters. We report on a method of preprocessing color input scenes in which the spectral functions are derived from linear models based on principal-component analysis. This multichannel algorithm transforms the red-green-blue (RGB) components into a new set of components that permit a generalization of the matched filter operations that are usually applied in optical pattern recognition with more-stable results under changes in illumination in the source images. The correlation is made in the subspace spanned by the coefficients that describe all reflectances according to a suitable basis for linear representation. First we illustrate the method in a control experiment in which the scenes are captured under known conditions of illumination. The discrimination capability of the algorithm improves upon the conventional RGB multichannel decomposition used in optical correlators when scenes are captured under different illuminant conditions and is slightly better than color recognition based on uniform color spaces (e.g., the CIELab system). Then we test the coefficient method in situations in which the target is captured under a reference illuminant and the scene that contains the target under an unknown spectrally different illuminant. We show that the method prevents false alarms caused by changes in the illuminant and that only two coefficients suffice to discriminate polychromatic objects.


Real-Time Neural Signals Decoding onto Off-the-Shelf DSP Processors for Neuroprosthetic Applications.
The control of upper limb neuroprostheses through the peripheral nervous system (PNS) can allow restoring motor functions in amputees. At present, the important aspect of the real-time implementation of neural decoding algorithms on embedded systems has been often overlooked, notwithstanding the impact that limited hardware resources have on the efficiency/effectiveness of any given algorithm. Present study is addressing the optimization of a template matching based algorithm for PNS signals decoding that is a milestone for its real-time, full implementation onto a floating-point digital signal processor (DSP). The proposed optimized real-time algorithm achieves up to 96% of correct classification on real PNS signals acquired through LIFE electrodes on animals, and can correctly sort spikes of a synthetic cortical dataset with sufficiently uncorrelated spike morphologies (93% average correct classification) comparably to the results obtained with top spike sorter (94% on average on the same dataset). The power consumption enables more than 24 h processing at the maximum load, and latency model has been derived to enable a fair performance assessment. The final embodiment demonstrates the real-time performance onto a low-power off-the-shelf DSP, opening to experiments exploiting the efferent signals to control a motor neuroprosthesis.


Influence of the type of post and core on in vitro marginal continuity, fracture resistance, and fracture mode of lithia disilicate-based all-ceramic crowns.
The influence of different crown foundations on marginal seal and fracture resistance of ceramic crowns placed on endodontically treated teeth has not been clearly established. The purpose of this study was to evaluate the marginal continuity and fracture behavior of high-strength all-ceramic crowns with different substructures in endodontically treated premolars. Forty-eight human mandibular premolars were assigned to 6 groups, including a no-treatment group (UNTREATED) and a group for which the access cavity was restored with composite resin (Tetric Ceram) (COMP). In the remaining 4 groups, teeth were prepared to receive all-ceramic crowns with 0.8-mm-wide shoulders and axial dentin heights of 2 mm. No posts were used in the ENDOCROWN group. Glass fiber posts (FRC Postec) were used in group FRC-POST. Group ZRO-POST received zirconia ceramic posts (CosmoPost), and group GOLD-POST received cast gold posts (CM). Experimental lithia disilicate ceramic crowns were made and adhesively cemented (Variolink). All teeth were subjected to thermal cycling and mechanical loading (TCML) in a masticatory simulator (1,200,000 loads, 49 N, 1.7 Hz, 3000 temperature cycles of 5 degrees C-50 degrees C-5 degrees C). Marginal continuity was evaluated with scanning electron microscopy at x200. All specimens were loaded to failure in a universal testing machine at 0.5 mm/min after TCML. Data were analyzed using 1-way ANOVA and post hoc t tests with Bonferroni correction (alpha=.05). Initially, mean values (SD) between 72.4 (15.8)% (ENDOCROWN) and 94.8 (3)% (FRC-POST) for continuous margins were found. With TCML, marginal continuity decreased significantly only in FRC-POST, to 75.5 (8.4)%, and in ENDOCROWN, to 44.7 (14.5)%. Mechanical load testing measured mean loads to failure between 1092.4 (307.8) N (FRC-POST) and 1253.7 (226.5) N (ZRO-POST) without significant differences between groups. Deep root fractures were observed in half of the specimens, irrespective of their groups. Marginal continuity of the crowns studied was better and more stress resistant when posts and cores were included in the restoration of endodontically treated teeth with complete ceramic crowns. The placement of a post-and-core foundation did not influence the pattern of failure.


Glucose levels and risk of dementia.
Diabetes is a risk factor for dementia. It is unknown whether higher glucose levels increase the risk of dementia in people without diabetes. We used 35,264 clinical measurements of glucose levels and 10,208 measurements of glycated hemoglobin levels from 2067 participants without dementia to examine the relationship between glucose levels and the risk of dementia. Participants were from the Adult Changes in Thought study and included 839 men and 1228 women whose mean age at baseline was 76 years; 232 participants had diabetes, and 1835 did not. We fit Cox regression models, stratified according to diabetes status and adjusted for age, sex, study cohort, educational level, level of exercise, blood pressure, and status with respect to coronary and cerebrovascular diseases, atrial fibrillation, smoking, and treatment for hypertension. During a median follow-up of 6.8 years, dementia developed in 524 participants (74 with diabetes and 450 without). Among participants without diabetes, higher average glucose levels within the preceding 5 years were related to an increased risk of dementia (P=0.01); with a glucose level of 115 mg per deciliter (6.4 mmol per liter) as compared with 100 mg per deciliter (5.5 mmol per liter), the adjusted hazard ratio for dementia was 1.18 (95% confidence interval [CI], 1.04 to 1.33). Among participants with diabetes, higher average glucose levels were also related to an increased risk of dementia (P=0.002); with a glucose level of 190 mg per deciliter (10.5 mmol per liter) as compared with 160 mg per deciliter (8.9 mmol per liter), the adjusted hazard ratio was 1.40 (95% CI, 1.12 to 1.76). Our results suggest that higher glucose levels may be a risk factor for dementia, even among persons without diabetes. (Funded by the National Institutes of Health.)


Concomitant Disruption of CD4 and CD8 Genes Facilitates the Development of Double Negative αβ TCR+ Peripheral T Cells That Respond Robustly to Staphylococcal Superantigen.
Mature peripheral double negative T (DNT) cells expressing αβ TCR but lacking CD4/CD8 coreceptors play protective as well as pathogenic roles. To better understand their development and functioning in vivo, we concomitantly inactivated CD4 and CD8 genes in mice with intact MHC class I and class II molecules with the hypothesis that this would enable the development of DNT cells. We also envisaged that these DNT cells could be activated by bacterial superantigens in vivo as activation of T cells by superantigens does not require CD4 and CD8 coreceptors. Because HLA class II molecules present superantigens more efficiently than murine MHC class II molecules, CD4 CD8 double knockout (DKO) mice transgenically expressing HLA-DR3 or HLA-DQ8 molecules were generated. Although thymic cellularity was comparable between wild type (WT) and DKO mice, CD3+ αβ TCR+ thymocytes were significantly reduced in DKO mice, implying defects in thymic-positive selection. Splenic CD3+ αβ TCR+ cells and Foxp3+ T regulatory cells were present in DKO mice but significantly reduced. However, the in vivo inflammatory responses and immunopathology elicited by acute challenge with the staphylococcal superantigen enterotoxin B were comparable between WT and DKO mice. Choric exposure to staphylococcal enterotoxin B precipitated a lupus-like inflammatory disease with characteristic lympho-monocytic infiltration in lungs, livers, and kidneys, along with production of anti-nuclear Abs in DKO mice as in WT mice. Overall, our results suggest that DNT cells can develop efficiently in vivo and chronic exposure to bacterial superantigens may precipitate a lupus-like autoimmune disease through activation of DNT cells.