Computer Simulation of Ion-Selective Membrane Electrodes and Related Systems by Finite-Element Procedures.
A simple but powerful numerical simulation for analyzing the electrochemical behavior of ion-selective membranes and liquid junctions is presented. The computer modeling makes use of a finite-element procedure in the space and time domains, which can be easily processed (e. g., with MS Excel software) without the need for complex mathematical evaluations. It leads to convincing results on the dynamic evolution of concentration profiles, potentials, and fluxes in the studied systems. The treatment accounts for influences of convection, flow, or stirring in the sample solution that act on the boundary diffusion layer and it is even capable of including the effects of an electrolyte flow through the whole system. To minimize the number of arbitrary parameters, interfacial reactions are assumed to be near local equilibrium, and space-charge influences are considered via phase-boundary potential differences. The applicability of the computer simulation is demonstrated for different ion-selective membranes as well as for liquid junctions. The numerical results are in excellent agreement with experimental data.


Circulating antibodies to cow's milk proteins in ulcerative colitis.
Sera from patients with ulcerative colitis (51), Crohn's disease (30), hypolactasia (13), untreated adult coeliac disease (11), irritable colon syndrome (24), and sera from 38 healthy control subjects were tested for antibodies to the principal cow's milk proteins-casein, alpha-lactalbumin, and beta-lactoglobulin. The red-cell-linked antigen-antiglobulin reaction was used to determine the titres of direct agglutinating antibodies and IgA and IgG incomplete antibodies. Apart from patients with coeliac disease, direct agglutinating antibodies were found infrequently and then in low titres. Approximately 50% of subjects had low titres of IgA and IgG antibodies. However, the titres found in sera from patients with ulcerative colitis did not differ from those found in the control subjects or in patients with Crohn's disease, hypolactasia, or irritable colon syndrome. Patients with untreated coeliac disease frequently had high antibody titres to the milk proteins. In all subjects tested, incomplete antibodies of IgA or IgG immunoglobulin class occurred with equal frequency. The frequent occurrence in adults of low titres of antibodies to the milk proteins may be due to continued absorption of minute amounts of protein. Absorption of allergens may be facilitated by mucosal damage, such as that of coeliac disease, with stimulation of antibody production. At the present time, however, there is little evidence to suggest that milk allergy is a factor in the aetiology of ulcerative colitis.


Contrasting neuropathology and functional recovery after spinal cord injury in developing and adult rats.
Conflicting findings exist regarding the link between functional recovery and the regrowth of spinal tracts across the lesion leading to the restoration of functional contacts. In the present study, we investigated whether functional locomotor recovery was attributable to anatomical regeneration at postnatal day 1 (PN1), PN7, PN14 and in adult rats two months after transection injury at the tenth thoracic segment of the spinal cord. The Basso, Beattie, and Bresnahan scores showed that transection led to a failure of hindlimb locomotor function in PN14 and adult rats. However, PN1 and PN7 rats showed a significant level of stepping function after complete spinal cord transection. Unexpectedly, unlike the transected PN14 and adult rats in which the spinal cord underwent limited secondary degeneration and showed a scar at the lesion site, the rats transected at PN1 and PN7 showed massive secondary degeneration both anterograde and retrograde, leaving a >5-mm gap between the two stumps. Furthermore, retrograde tracing with fluorogold (FG) also showed that FG did not cross the transection site in PN1 and PN7 rats as in PN14 and adult rats, and re-transection of the cord caused no apparent loss in locomotor performance in the rats transected at PN1. Thus, these three lines of evidence strongly indicated that the functional recovery after transection in neonatal rats is independent of regrowth of spinal tracts across the lesion site. Our results support the notion that the recovery of locomotor function in developing rats may be due to intrinsic adaptations in the spinal circuitry below the lesion that control hindlimb locomotor activity rather than the regrowth of spinal tracts across the lesion. The difference in secondary degeneration between neonatal and adult rats remains to be explored.


Phenotypic profile of expanded NK cells in chronic lymphoproliferative disorders: a surrogate marker for NK-cell clonality.
Currently, the lack of a universal and specific marker of clonality hampers the diagnosis and classification of chronic expansions of natural killer (NK) cells. Here we investigated the utility of flow cytometric detection of aberrant/altered NK-cell phenotypes as a surrogate marker for clonality, in the diagnostic work-up of chronic lymphoproliferative disorders of NK cells (CLPD-NK). For this purpose, a large panel of markers was evaluated by multiparametric flow cytometry on peripheral blood (PB) CD56(low) NK cells from 60 patients, including 23 subjects with predefined clonal (n = 9) and polyclonal (n = 14) CD56(low) NK-cell expansions, and 37 with CLPD-NK of undetermined clonality; also, PB samples from 10 healthy adults were included. Clonality was established using the human androgen receptor (HUMARA) assay. Clonal NK cells were found to show decreased expression of CD7, CD11b and CD38, and higher CD2, CD94 and HLADR levels vs. normal NK cells, together with a restricted repertoire of expression of the CD158a, CD158b and CD161 killer-associated receptors. In turn, NK cells from both clonal and polyclonal CLPD-NK showed similar/overlapping phenotypic profiles, except for high and more homogeneous expression of CD94 and HLADR, which was restricted to clonal CLPD-NK. We conclude that the CD94(hi)/HLADR+ phenotypic profile proved to be a useful surrogate marker for NK-cell clonality.


Diurnal changes in electrocorticogram sleep slow-wave activity during development in rats.
According to the homeostatic regulation of sleep, sleep pressure accumulates during wakefulness, further increases during sleep deprivation and dissipates during subsequent sleep. Sleep pressure is electrophysiologically reflected by electroencephalogram slow-wave activity during non-rapid eye movement sleep, and is thought to be stable across time. During childhood and adolescence the brain undergoes massive reorganization processes. Slow-wave activity during these developmental periods has been shown in humans to follow an inverted U-shaped trajectory, which recently was replicated in rats. The goal of this study was to investigate in rats the diurnal changes of slow-wave activity during the inverted U-shaped developmental trajectory of slow-wave activity. To do so, we performed longitudinal electrocorticogram recordings, and compared the level of slow-wave activity at the beginning with the slow-wave activity level at the end of 24-h baselines in two sets of Sprague-Dawley rats. In younger animals (n = 17) we investigated specific postnatal days when overall slow-wave activity increases (postnatal day 26), peaks (postnatal day 28) and decreases (>postnatal day 28). The same analysis was performed in older animals (postnatal day 48, n = 6). Our results show a gain of slow-wave activity across 24 h on postnatal day 26, followed by no net changes on postnatal day 28, which was then followed by a loss of slow-wave activity during subsequent days (>postnatal day 28). Older animals did not show any net changes in slow-wave activity across 24 h. These results cannot be explained by differences in vigilance states. Thus, slow-wave activity during this developmental period may not only reflect the trajectory of sleep pressure but may additionally reflect maturational processes.


Latent Supervised Learning.
A new machine learning task is introduced, called latent supervised learning, where the goal is to learn a binary classifier from continuous training labels which serve as surrogates for the unobserved class labels. A specific model is investigated where the surrogate variable arises from a two-component Gaussian mixture with unknown means and variances, and the component membership is determined by a hyperplane in the covariate space. The estimation of the separating hyperplane and the Gaussian mixture parameters forms what shall be referred to as the change-line classification problem. A data-driven sieve maximum likelihood estimator for the hyperplane is proposed, which in turn can be used to estimate the parameters of the Gaussian mixture. The estimator is shown to be consistent. Simulations as well as empirical data show the estimator has high classification accuracy.


3-Methylglutaconic aciduria--lessons from 50 genes and 977 patients.
Elevated urinary excretion of 3-methylglutaconic acid is considered rare in patients suspected of a metabolic disorder. In 3-methylglutaconyl-CoA hydratase deficiency (mutations in AUH), it derives from leucine degradation. In all other disorders with 3-methylglutaconic aciduria the origin is unknown, yet mitochondrial dysfunction is thought to be the common denominator. We investigate the biochemical, clinical and genetic data of 388 patients referred to our centre under suspicion of a metabolic disorder showing 3-methylglutaconic aciduria in routine metabolic screening. Furthermore, we investigate 591 patients with 50 different, genetically proven, mitochondrial disorders for the presence of 3-methylglutaconic aciduria. Three percent of all urine samples of the patients referred showed 3-methylglutaconic aciduria, often in correlation with disorders not reported earlier in association with 3-methylglutaconic aciduria (e.g. organic acidurias, urea cycle disorders, haematological and neuromuscular disorders). In the patient cohort with genetically proven mitochondrial disorders 11% presented 3-methylglutaconic aciduria. It was more frequently seen in ATPase related disorders, with mitochondrial DNA depletion or deletion, but not in patients with single respiratory chain complex deficiencies. Besides, it was a consistent feature of patients with mutations in TAZ, SERAC1, OPA3, DNAJC19 and TMEM70 accounting for mitochondrial membrane related pathology. 3-methylglutaconic aciduria is found quite frequently in patients suspected of a metabolic disorder, and mitochondrial dysfunction is indeed a common denominator. It is only a discriminative feature of patients with mutations in AUH, TAZ, SERAC1, OPA3, DNAJC19 TMEM70. These conditions should therefore be referred to as inborn errors of metabolism with 3-methylglutaconic aciduria as discriminative feature.


5' tRNA halves are highly expressed in the primate hippocampus and might sequence-specifically regulate gene expression.
Fragments of mature tRNAs have long been considered as mere degradation products without physiological function. However, recent reports show that tRNA-derived small RNAs (tsRNAs) play prominent roles in diverse cellular processes across a wide spectrum of species. Contrasting the situation in other small RNA pathways the mechanisms behind these effects appear more diverse, more complex, and are generally less well understood. In addition, surprisingly little is known about the expression profiles of tsRNAs across different tissues and species. Here, we provide an initial overview of tsRNA expression in different species and tissues, revealing very high levels of 5' tRNA halves (5' tRHs) particularly in the primate hippocampus. We further modulated the regulation capacity of selected 5' tRHs in human cells by transfecting synthetic tsRNA mimics ("overexpression") or antisense-RNAs ("inhibition") and identified differentially expressed transcripts based on RNA-seq. We then used a novel k-mer mapping approach to dissect the underlying targeting rules, suggesting that 5' tRHs silence genes in a sequence-specific manner, while the most efficient target sites align to the mid-region of the 5' tRH and are located within the CDS or 3' UTR of the target. This amends previous observations that tsRNAs guide Argonaute proteins to silence their targets via a miRNA-like 5' seed match and suggests a yet unknown mechanism of regulation. Finally, our data suggest that some 5' tRHs that are also able to sequence-specifically stabilize mRNAs as up-regulated mRNAs are also significantly enriched for 5' tRH target sites.


Expression of glutamate receptor subunit/subtype messenger RNAS for NMDAR1, GLuR1, GLuR2 and mGLuR5 by accumbal projection neurons.
Nucleus accumbens neurons are the targets of glutamatergic inputs. By coupling in situ hybridization for glutamate receptor mRNAs with retrograde transport of Fluoro-Gold, the present study examined the relationship between the distribution patterns of glutamate receptor subtypes/subunits and the output pathways of the nucleus accumbens to the ventral pallidum and ventral tegmental area. Following iontophoretic deposits of Fluoro-Gold into the ventral pallidum, neurons in both the nucleus accumbens shell and core were retrogradely labeled. A high percentage of accumbens neurons retrogradely labeled from the ventral pallidum were double-labeled for mRNAs encoding for mGluR5 (82+/-4.1%), NMDAR1 (71+/-3.5%), GluR1 (70+/-6.1%) and GluR2 (76+/-3.6%). No significant difference in the proportion of double-labeled neurons between the core and shell was observed. Following the deposit of Fluoro-Gold into the ventral tegmental area, only the accumbens shell neurons were retrogradely labeled. The proportion of neurons expressing NMDAR1, GluR1 and GluR2 were somewhat less in the projection to the ventral tegmental area compared to the ventral pallidum since approximately 60% of the neurons retrogradely-labeled from the ventral tegmental area expressed these transcripts. In contrast to the high proportion of mGluR5-containing neurons in the nucleus accumbens innervating the ventral pallidum, only half of the neurons projecting to the ventral tegmental area expressed mGluR5. These data show that accumbens neurons innervating the ventral pallidum and ventral tegmental area differ in the relative proportion of expressed mRNA encoding mGluR5, implying differential postsynaptic impact by glutamate transmission on neurons contributing to the two major efferent pathways of the nucleus accumbens.


An Integrated Proteomic and Transcriptomic Analysis Reveals the Venom Complexity of the Bullet Ant Paraponera clavata.
A critical hurdle in ant venom proteomic investigations is the lack of databases to comprehensively and specifically identify the sequence and function of venom proteins and peptides. To resolve this, we used venom gland transcriptomics to generate a sequence database that was used to assign the tandem mass spectrometry (MS) fragmentation spectra of venom peptides and proteins to specific transcripts. This was performed alongside a shotgun liquid chromatography-mass spectrometry (LC-MS/MS) analysis of the venom to confirm that these assigned transcripts were expressed as proteins. Through the combined transcriptomic and proteomic investigation of Paraponera clavata venom, we identified four times the number of proteins previously identified using 2D-PAGE alone. In addition to this, by mining the transcriptomic data, we identified several novel peptide sequences for future pharmacological investigations, some of which conform with inhibitor cysteine knot motifs. These types of peptides have the potential to be developed into pharmaceutical or bioinsecticide peptides.


Physical activity volume in relation to risk of atrial fibrillation. A non-linear meta-regression analysis.
Background Light physical activity is known to reduce atrial fibrillation risk, whereas moderate to vigorous physical activity may result in an increased risk. However, the question of what volume of physical activity can be considered beneficial remains poorly understood. The scope of the present work was to examine the relation between physical activity volume and atrial fibrillation risk. Design A comprehensive systematic review was performed following the PRISMA guidelines. Methods A non-linear meta-regression considering the amount of energy spent in physical activity was carried out. The first derivative of the non-linear relation between physical activity and atrial fibrillation risk was evaluated to determine the volume of physical activity that carried the minimum atrial fibrillation risk. Results The dose-response analysis of the relation between physical activity and atrial fibrillation risk showed that physical activity at volumes of 5-20 metabolic equivalents per week (MET-h/week) was associated with significant reduction in atrial fibrillation risk (relative risk for 19 MET-h/week = 0.92 (0.87, 0.98). By comparison, physical activity volumes exceeding 20 MET-h/week were unrelated to atrial fibrillation risk (relative risk for 21 MET-h/week = 0.95 (0.88, 1.02). Conclusion These data show a J-shaped relation between physical activity volume and atrial fibrillation risk. Physical activity at volumes of up to 20 MET-h/week is associated with reduced atrial fibrillation risk, whereas volumes exceeding 20 MET-h/week show no relation with risk.


Initial assessment in the management of adult epistaxis: systematic review.
The initial assessment of epistaxis patients commonly includes: first aid measures, observations, focused history taking, and clinical examinations and investigations. This systematic review aimed to identify evidence that informs how the initial assessment of these patients should be conducted. A systematic review of the literature was performed using a standardised methodology and search strategy. Seventeen articles were included. Factors identified were: co-morbidity, intrinsic patient factors, coagulation screening and ice pack use. Hypertension and anticoagulant use were demonstrated to adversely affect outcomes. Coagulation screening is useful in patients on anticoagulant medication. Four studies could not be accessed. Retrospective methodology and insufficient statistical analysis limit several studies. Sustained ambulatory hypertension, anticoagulant therapy and posterior bleeding may be associated with recurrent epistaxis, and should be recorded. Oral ice pack use may decrease severity and can be considered as first aid. Coagulation studies are appropriate for patients with a history of anticoagulant use or bleeding diatheses.


The Polycomb-group gene eed is required for normal morphogenetic movements during gastrulation in the mouse embryo.
We have characterized an induced mutation, called embryonic ectoderm development or eed, that disrupts A-P patterning of the mouse embryo during gastrulation. Positional cloning of this gene revealed it to be the highly conserved homologue of the Drosophila gene extra sex combs, which is required for maintenance of long-term transcriptional repression of homeotic gene expression. Mouse embryos homozygous for loss-of-function alleles of eed initiate gastrulation but display abnormal mesoderm production. Very little embryonic mesoderm is produced; in contrast, extraembryonic mesoderm is relatively abundant. These observations, along with mRNA in situ hybridization analyses, suggested a defect in the anterior primitive streak, from which much of the embryonic mesoderm of the wild-type embryo is derived. To analyse this defect, we initiated clonal analysis of the pre-streak epiblast in eed mutant embryos, using the lineage tracer horseradish peroxidase (HRP). The results of these studies indicate that epiblast cells ingress through the anterior streak, but the newly formed mesoderm does not migrate anteriorly and is mislocalized to the extraembryonic compartment. Abnormal localization of mesoderm to the extraembryonic region did not appear to be due to a restriction and alteration of distal epiblast cell fate, since the majority of clones produced from regions fated to ingress through the anterior streak were mixed, displaying descendants in both embryonic and extraembryonic derivatives. eed mutant embryos also fail to display proper epiblast expansion, particularly with respect to the A-P axis. Based on patterns of clonal spread and calculated clone doubling times for the epiblast, this does not appear to be due to decreased epiblast growth. Rather, epiblast, which is normally fated to make a substantial contribution to the axial midline, appears to make mesoderm preferentially. The data are discussed in terms of global morphogenetic movements in the mouse gastrula and a disruption of signalling activity in the anterior primitive streak.


Fabrication and laser patterning of polystyrene optical oxygen sensor films for lab-on-a-chip applications.
We present a novel and simple method for patterning oxygen-sensitive polystyrene thin films and demonstrate its potential for integration with microfluidic lab-on-a-chip devices. Optical oxygen sensing films composed of polystyrene with an embedded luminescent oxygen-sensitive dye present a convenient option for the measurement of oxygen levels in microfluidic and lab-on-a-chip devices; however, patterning and integrating the films with poly(dimethylsiloxane) (PDMS) microfluidic devices has proven difficult due to a residue after dry etch patterning that inhibits subsequent PDMS bonding. Our new method uses mask-less laser ablation by a commercial laser ablation system to define the outline of the structures and subsequent bulk film removal by aqueous lift-off. Because the bulk film is peeled or lifted off of the substrate rather than etched, the process is compatible with standard PDMS plasma bonding. We used ToF-SIMS analysis to investigate how laser ablation facilitates this fabrication process as well as why dry etching polystyrene inhibits PDMS plasma bonding. The results of this analysis showed evidence of chemical species formed during the laser ablation and dry etching processes that can produce these effects. Our new method's mask-less nature, simplicity, speed, and compatibility with PDMS bonding make it ideally suited for single-use lab-on-a-chip applications. To demonstrate the method's compatibility with PDMS microfluidics, we also present a demonstration of the sensors' integration into a microfluidic oxygen gradient generator device.


Localization of primary language areas by arcuate fascicle fiber tracking.
To reduce the risk of disabling postoperative functional deficit in patients with lesions in the dominant hemisphere, information about the localization of eloquent language areas is mandatory. To demonstrate the feasibility of arcuate fascicle (AF) tractography for proper localization of eloquent language areas in the superior temporal (STG) and inferior frontal gyrus (IFG). Between January and June 2010, we performed surgery in 13 patients with highly eloquent lesions with close spatial relationship to the primary language areas. All of them received preoperative diffusion tensor imaging for AF tractography. The STG and IFG were delineated at the ends of the AF. Five patients underwent functional magnetic resonance imaging of the primary language areas. The results were compared with tractography. Tractography of the AF without prior knowledge of the localization of the STG and IFG was feasible in all cases. In the cases with functional magnetic resonance imaging, the activation maps matched the tractography results. In all but 1 patient, preservation of the primary language areas was possible, proven by the good neurological outcome. One patient suffered from a language dysfunction caused by a lesion in the medial and inferior temporal gyrus along the surgical pathway. Tractography of the AF is a useful tool for identification of parts of the main primary language areas. Using tractography as a localization procedure to determine the primary language areas aids in the delineation of the STG and IFG and thus may help reduce the risk of postoperative permanent neurological deficit.


[Insulin resistance and atherosclerosis].
The western way of life favours the development of a state of insulin resistance, in genetically predisposed subjects. In this state, greater levels of insulin are necessary so that an answer can be obtained and, consequently, hyperinsulinism occurs. Insulin has several target tissues, thus insulin resistance is associated with the dysfunction of a multiplicity of tissues, organs and systems in the body (Syndrome X). All of those dysfunctions together with hyperinsulinism can greatly enhance the risk of atherosclerotic vascular disease. In this article we review the dysfunction at several levels, including blood pressure, endothelium, lipid metabolism and fibrinolytic system and the way they can, together with hyperinsulinism, induce atherogenesis. We review some of the therapeutic options that can reduce this state of insulin resistance as well as the morbidity and mortality associated with atherosclerosis.


Ideal and real treatment planning processes for people with serious mental illness in public mental health care.
Treatment planning processes are a fundamental component of evidence-based practice in mental health for people with serious mental illness (SMI), who often present with complex concerns and require an interdisciplinary treatment team. It is unclear how well treatment planning practices in usual care settings for SMI adhere to best practices guidelines. In this study, we used qualitative methods to increase understanding of typical treatment planning practices. Twelve mental health providers completed a participatory dialogue focused on discussing perceptions of ideal and real treatment planning processes. Content analysis of the transcription from the dialogue was used to identify major themes and subthemes. Analysis revealed 6 primary themes with 23 subthemes. Providers described the ideal treatment planning process as dynamic and collaborative, including thorough assessment and inclusion of all stakeholders including the consumer, providers, and family members. Real treatment planning was described as directed by institutional and regulatory needs, resulting in treatment plans that were not personalized and not communicated to frontline staff or the consumer. These results indicate that providers have a strong understanding of evidence-based principles of treatment decision-making. However, actual treatment planning processes rarely live up to those principles. Providers identified several obstacles to enacting best practices. Although many obstacles were system-level, providers themselves also contributed to the gap between ideal and real treatment planning. Additional training and education may help to close this gap. Consumer self-advocacy is also important, given that providers often see themselves as lacking agency to make changes. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Mishearing as a Side Effect of Rational Language Comprehension in Noise.
Language comprehension in noise can sometimes lead to mishearing, due to the noise disrupting the speech signal. Some of the difficulties in dealing with the noisy signal can be alleviated by drawing on the context - indeed, top-down predictability has shown to facilitate speech comprehension in noise. Previous studies have furthermore shown that strong reliance on the top-down predictions can lead to increased rates of mishearing, especially in older adults, which are attributed to general deficits in cognitive control in older adults. We here propose that the observed mishearing may be a simple consequence of rational language processing in noise. It should not be related to failure on the side of the older comprehenders, but instead would be predicted by rational processing accounts. To test this hypothesis, we extend earlier studies by running an online listening experiment with younger and older adults, carefully controlling the target and direct competitor in our stimuli. We show that mishearing is directly related to the perceptibility of the signal. We furthermore add an analysis of wrong responses, which shows that results are at odds with the idea that participants overly strongly rely on context in this task, as most false answers are indeed close to the speech signal, and not to the semantics of the context.


Automatic Detection of Hard Exudates in Color Retinal Images Using Dynamic Threshold and SVM Classification: Algorithm Development and Evaluation.
Diabetic retinopathy (DR) is one of the most common causes of visual impairment. Automatic detection of hard exudates (HE) from retinal photographs is an important step for detection of DR. However, most of existing algorithms for HE detection are complex and inefficient. We have developed and evaluated an automatic retinal image processing algorithm for HE detection using dynamic threshold and fuzzy C-means clustering (FCM) followed by support vector machine (SVM) for classification. The proposed algorithm consisted of four main stages: (i) imaging preprocessing; (ii) localization of optic disc (OD); (iii) determination of candidate HE using dynamic threshold in combination with global threshold based on FCM; and (iv) extraction of eight texture features from the candidate HE region, which were then fed into an SVM classifier for automatic HE classification. The proposed algorithm was trained and cross-validated (10 fold) on a publicly available e-ophtha EX database (47 images) on pixel-level, achieving the overall average sensitivity, PPV, and F-score of 76.5%, 82.7%, and 76.7%. It was tested on another independent DIARETDB1 database (89 images) with the overall average sensitivity, specificity, and accuracy of 97.5%, 97.8%, and 97.7%, respectively. In summary, the satisfactory evaluation results on both retinal imaging databases demonstrated the effectiveness of our proposed algorithm for automatic HE detection, by using dynamic threshold and FCM followed by an SVM for classification.


Smoking reduction interventions for smoking cessation.
The standard way most people are advised to stop smoking is by quitting abruptly on a designated quit day. However, many people who smoke have tried to quit many times and may like to try an alternative method. Reducing smoking behaviour before quitting could be an alternative approach to cessation. However, before this method can be recommended it is important to ensure that abrupt quitting is not more effective than reducing to quit, and to determine whether there are ways to optimise reduction methods to increase the chances of cessation. To assess the effect of reduction-to-quit interventions on long-term smoking cessation. We searched the Cochrane Tobacco Addiction Group Specialised Register, MEDLINE, Embase and PsycINFO for studies, using the terms: cold turkey, schedul*, cut* down, cut-down, gradual*, abrupt*, fading, reduc*, taper*, controlled smoking and smoking reduction. We also searched trial registries to identify unpublished studies. Date of the most recent search: 29 October 2018. Randomised controlled trials in which people who smoked were advised to reduce their smoking consumption before quitting smoking altogether in at least one trial arm. This advice could be delivered using self-help materials or behavioural support, and provided alongside smoking cessation pharmacotherapies or not. We excluded trials that did not assess cessation as an outcome, with follow-up of less than six months, where participants spontaneously reduced without being advised to do so, where the goal of reduction was not to quit altogether, or where participants were advised to switch to cigarettes with lower nicotine levels without reducing the amount of cigarettes smoked or the length of time spent smoking. We also excluded trials carried out in pregnant women. We followed standard Cochrane methods. Smoking cessation was measured after at least six months, using the most rigorous definition available, on an intention-to-treat basis. We calculated risk ratios (RRs) and 95% confidence intervals (CIs) for smoking cessation for each study, where possible. We grouped eligible studies according to the type of comparison (no smoking cessation treatment, abrupt quitting interventions, and other reduction-to-quit interventions) and carried out meta-analyses where appropriate, using a Mantel-Haenszel random-effects model. We also extracted data on quit attempts, pre-quit smoking reduction, adverse events (AEs), serious adverse events (SAEs) and nicotine withdrawal symptoms, and meta-analysed these where sufficient data were available. We identified 51 trials with 22,509 participants. Most recruited adults from the community using media or local advertising. People enrolled in the studies typically smoked an average of 23 cigarettes a day. We judged 18 of the studies to be at high risk of bias, but restricting the analysis only to the five studies at low or to the 28 studies at unclear risk of bias did not significantly alter results.We identified very low-certainty evidence, limited by risk of bias, inconsistency and imprecision, comparing the effect of reduction-to-quit interventions with no treatment on cessation rates (RR 1.74, 95% CI 0.90 to 3.38; I2 = 45%; 6 studies, 1599 participants). However, when comparing reduction-to-quit interventions with abrupt quitting (standard care) we found evidence that neither approach resulted in superior quit rates (RR 1. 01, 95% CI 0.87 to 1.17; I2 = 29%; 22 studies, 9219 participants). We judged this estimate to be of moderate certainty, due to imprecision. Subgroup analysis provided some evidence (P = 0.01, I2 = 77%) that reduction-to-quit interventions may result in more favourable quit rates than abrupt quitting if varenicline is used as a reduction aid. Our analysis comparing reduction using pharmacotherapy with reduction alone found low-certainty evidence, limited by inconsistency and imprecision, that reduction aided by pharmacotherapy resulted in higher quit rates (RR 1. 68, 95% CI 1.09 to 2.58; I2 = 78%; 11 studies, 8636 participants). However, a significant subgroup analysis (P < 0.001, I2 = 80% for subgroup differences) suggests that this may only be true when fast-acting NRT or varenicline are used (both moderate-certainty evidence) and not when nicotine patch, combination NRT or bupropion are used as an aid (all low- or very low-quality evidence). More evidence is likely to change the interpretation of the latter effects.Although there was some evidence from within-study comparisons that behavioural support for reduction to quit resulted in higher quit rates than self-help resources alone, the relative efficacy of various other characteristics of reduction-to-quit interventions investigated through within- and between-study comparisons did not provide any evidence that they enhanced the success of reduction-to-quit interventions. Pre-quit AEs, SAEs and nicotine withdrawal symptoms were measured variably and infrequently across studies. There was some evidence that AEs occurred more frequently in studies that compared reduction using pharmacotherapy versus no pharmacotherapy; however, the AEs reported were mild and usual symptoms associated with NRT use. There was no clear evidence that the number of people reporting SAEs, or changes in withdrawal symptoms, differed between trial arms. There is moderate-certainty evidence that neither reduction-to-quit nor abrupt quitting interventions result in superior long-term quit rates when compared with one another. Evidence comparing the efficacy of reduction-to-quit interventions with no treatment was inconclusive and of low certainty. There is also low-certainty evidence to suggest that reduction-to-quit interventions may be more effective when pharmacotherapy is used as an aid, particularly fast-acting NRT or varenicline (moderate-certainty evidence). Evidence for any adverse effects of reduction-to-quit interventions was sparse, but available data suggested no excess of pre-quit SAEs or withdrawal symptoms. We downgraded the evidence across comparisons due to risk of bias, inconsistency and imprecision. Future research should aim to match any additional components of multicomponent reduction-to-quit interventions across study arms, so that the effect of reduction can be isolated. In particular, well-conducted, adequately-powered studies should focus on investigating the most effective features of reduction-to-quit interventions to maximise cessation rates.


Myeloid Differentiation Factor 88 Signaling in Bone Marrow-Derived Cells Promotes Gastric Tumorigenesis by Generation of Inflammatory Microenvironment.
It has been established that COX-2 and downstream signaling by prostaglandin E2 (PGE2) play a key role in tumorigenesis through generation of inflammatory microenvironment. Toll-like receptor (TLR) signaling through myeloid differentiation factor 88 (MyD88) also regulates inflammatory responses in tumors. However, the relationship between these distinct pathways in tumorigenesis is not yet fully understood. We herein investigated the role of MyD88 in gastric tumorigenesis using Gan mice, which develop inflammation-associated gastric tumors due to the simultaneous activation of the COX-2/PGE2 pathway and Wnt signaling. Notably, the disruption of Myd88 in Gan mice resulted in the significant suppression of gastric tumorigenesis with the inhibition of inflammatory responses, even though COX-2/PGE2 pathway is constitutively activated. Moreover, Myd88 disruption in bone marrow-derived cells (BMDCs) in Gan mice also suppressed inflammation and tumorigenesis, indicating that MyD88 signaling in BMDCs regulates the inflammatory microenvironment. We also found that expression of Tlr2 and its coreceptor Cd14 was induced in tumor epithelial cells in Gan mice, which was suppressed by the disruption of Myd88. It has already been shown that TLR2/CD14 signaling is important for stemness of intestinal epithelial cells. These results indicate that MyD88 in BMDCs, together with COX-2/PGE2 pathway, plays an essential role in the generation of the inflammatory microenvironment, which may promote tumorigenesis through induction of TLR2/CD14 pathway in tumor epithelial cells. These results suggest that inhibition of TLR/MyD88 signaling together with COX-2/PGE2 pathway will be an effective preventive strategy for gastric cancer.


Promotion and prevention within a decentralized framework: changing health care in Brazil and Chile.
The study aims to define configurations of factors from local, regional and national levels that, within a context of decentralized health systems, enable or hinder change towards a health care model of promotion and disease prevention as advocated in current Latin American public health discourse. The project made in-depth case studies of prevention and promotion activities in eight local health systems in rural and urban Brazil and Chile allowing three levels of comparison: national, regional and local. The data are based on interviews, secondary sources, policy documentation and observations. The results are summarized as pathways through configurations of factors leading to what are termed active and basic degrees of activity related to disease prevention and health promotion at the local level. The results raise considerations for policy-makers at each of the three levels of health systems regarding vertical and horizontal system structures, relationships to local government, management options and human resource incentives. The findings from this study can serve as a guide or menu of issues to consider in the implementation of new models of health care provision, not only in Brazil and Chile, but also elsewhere in Latin America and more widely in the world.


Bending dynamics of fluctuating biopolymers probed by automated high-resolution filament tracking.
Microscope images of fluctuating biopolymers contain a wealth of information about their underlying mechanics and dynamics. However, successful extraction of this information requires precise localization of filament position and shape from thousands of noisy images. Here, we present careful measurements of the bending dynamics of filamentous (F-)actin and microtubules at thermal equilibrium with high spatial and temporal resolution using a new, simple but robust, automated image analysis algorithm with subpixel accuracy. We find that slender actin filaments have a persistence length of approximately 17 microm, and display a q(-4)-dependent relaxation spectrum, as expected from viscous drag. Microtubules have a persistence length of several millimeters; interestingly, there is a small correlation between total microtubule length and rigidity, with shorter filaments appearing softer. However, we show that this correlation can arise, in principle, from intrinsic measurement noise that must be carefully considered. The dynamic behavior of the bending of microtubules also appears more complex than that of F-actin, reflecting their higher-order structure. These results emphasize both the power and limitations of light microscopy techniques for studying the mechanics and dynamics of biopolymers.


CaMKII phosphorylation of neuroligin-1 regulates excitatory synapses.
Neuroligins are postsynaptic cell adhesion molecules that are important for synaptic function through their trans-synaptic interaction with neurexins (NRXNs). The localization and synaptic effects of neuroligin-1 (NL-1, also called NLGN1) are specific to excitatory synapses with the capacity to enhance excitatory synapses dependent on synaptic activity or Ca(2+)/calmodulin kinase II (CaMKII). Here we report that CaMKII robustly phosphorylates the intracellular domain of NL-1. We show that T739 is the dominant CaMKII site on NL-1 and is phosphorylated in response to synaptic activity in cultured rodent neurons and sensory experience in vivo. Furthermore, a phosphodeficient mutant (NL-1 T739A) reduces the basal and activity-driven surface expression of NL-1, leading to a reduction in neuroligin-mediated excitatory synaptic potentiation. To the best of our knowledge, our results are the first to demonstrate a direct functional interaction between CaMKII and NL-1, two primary components of excitatory synapses.


Loss-of-function mutations of ILDR1 cause autosomal-recessive hearing impairment DFNB42.
By using homozygosity mapping in a consanguineous Pakistani family, we detected linkage of nonsyndromic hearing loss to a 7.6 Mb region on chromosome 3q13.31-q21.1 within the previously reported DFNB42 locus. Subsequent candidate gene sequencing identified a homozygous nonsense mutation (c.1135G>T [p.Glu379X]) in ILDR1 as the cause of hearing impairment. By analyzing additional consanguineous families with homozygosity at this locus, we detected ILDR1 mutations in the affected individuals of 10 more families from Pakistan and Iran. The identified ILDR1 variants include missense, nonsense, frameshift, and splice-site mutations as well as a start codon mutation in the family that originally defined the DFNB42 locus. ILDR1 encodes the evolutionarily conserved immunoglobulin-like domain containing receptor 1, a putative transmembrane receptor of unknown function. In situ hybridization detected expression of Ildr1, the murine ortholog, early in development in the vestibule and in hair cells and supporting cells of the cochlea. Expression in hair cell- and supporting cell-containing neurosensory organs is conserved in the zebrafish, in which the ildr1 ortholog is prominently expressed in the developing ear and neuromasts of the lateral line. These data identify loss-of-function mutations of ILDR1, a gene with a conserved expression pattern pointing to a conserved function in hearing in vertebrates, as underlying nonsyndromic prelingual sensorineural hearing impairment.


Neuroinflammatory signals in Alzheimer disease and APP/PS1 transgenic mice: correlations with plaques, tangles, and oligomeric species.
To understand neuroinflammation-related gene regulation during normal aging and in sporadic Alzheimer disease (sAD), we performed functional genomics analysis and analyzed messenger RNA (mRNA) expression by quantitative reverse transcription-polymerase chain reaction of 22 genes involved in neuroinflammation-like responses in the cerebral cortex of wild-type and APP/PS1 transgenic mice. For direct comparisons, mRNA expression of 18 of the same genes was then analyzed in the entorhinal cortex, orbitofrontal cortex, and frontal cortex area 8 of middle-aged human subjects lacking Alzheimer disease-related pathology and in older subjects with sAD pathology covering Stages I-II/0(A), III-IV/A-B, and V-VI/C of Braak and Braak classification. Modifications of cytokine and immune mediator mRNA expression were found with normal aging in wild-type mice and in middle-aged individuals and patients with early stages of sAD-related pathology; these were accompanied by increased protein expression of certain mediators in ramified microglia. In APP/PS1 mice, inflammatory changes coincided with β-amyloid (Aβ) deposition; increased levels of soluble oligomers paralleled the modified mRNA expression of cytokines and mediators in wild-type mice. In patients with sAD, regulation was stage- and region-dependent and not merely acceleration and exacerbation of mRNA regulation with aging. Gene regulation at first stages of AD was not related to hyperphosphorylated tau deposition in neurofibrillary tangles, Aβ plaque burden, concentration of Aβ1-40 (Aβ40) and Aβ1-42 (Aβ42), or fibrillar Aβ linked to membranes but rather to increased levels of soluble oligomers. Thus, species differences and region- and stage-dependent inflammatory responses in sAD, particularly at the initial stages, indicate the need to identify new anti-inflammatory compounds with specific molecular therapeutic targets.


Managing Constipation in Adults With Cancer.
Constipation is common in individuals with cancer, occurring in almost 60% of patients overall. The incidence increases in patients with advanced disease, particularly in those receiving opioid analgesics or medications with anticholinergic properties. Constipation is not uniformly assessed and therefore not recognized and appropriately managed in many instances. This can increase patients' physical and psychological distress. Furthermore, there is scant research to support current management strategies for constipation. The objectives of this review are to explore the incidence of and risk factors for constipation in patients with cancer, to discuss the extent of the problem, to explore the nonpharmacologic and pharmacologic measures for constipation and fecal impaction, and to synthesize a laxative management. An extensive review of medical, pharmacy, and nursing literature was done to explore the physiology and pathogenesis of constipation; detail the mechanisms of action, onset of effect, approximate costs, and adverse effects of drugs for constipation; and condense clinical expert consensus recommendations for constipation, particularly in patients with cancer. Advanced practitioners (APs) and other clinicians play crucial roles in identifying individuals at risk for and experiencing constipation to help them use effective regimens, including over-the-counter laxatives, and perhaps adjunctive nondrug measures. Clinicians and patients must develop an agreed-upon language for identifying the severity and effects of constipation. In addition, both should understand which laxatives are most appropriate and which should be avoided for particular patients. Two prescription agents are also available, and understanding when they should be used is important for APs.


Diversity of Transmission Outcomes Following Co-Infection of Sheep with Strains of Bluetongue Virus Serotype 1 and 8.
Bluetongue virus (BTV) causes an economically important disease, bluetongue (BT), in susceptible ruminants and is transmitted primarily by species of Culicoides biting midges (Diptera: Ceratopogonidae). Since 2006, northern Europe has experienced multiple incursions of BTV through a variety of routes of entry, including major outbreaks of strains of BTV serotype 8 (BTV-8) and BTV serotype 1 (BTV-1), which overlapped in distribution within southern Europe. In this paper, we examined the variation in response to coinfection with strains of BTV-1 and BTV-8 using an in vivo transmission model involving Culicoides sonorensis, low passage virus strains, and sheep sourced in the United Kingdom. In the study, four sheep were simultaneously infected using BTV-8 and BTV-1 intrathoracically inoculated C. sonorensis and co-infections of all sheep with both strains were established. However, there were significant variations in both the initiation and peak levels of virus RNA detected throughout the experiment, as well as in the infection rates in the C. sonorensis that were blood-fed on experimentally infected sheep at peak viremia. This is discussed in relation to the potential for reassortment between these strains in the field and the policy implications for detection of BTV strains.


Characterization of TMPRSS2-ERG fusion high-grade prostatic intraepithelial neoplasia and potential clinical implications.
More than 1,300,000 prostate needle biopsies are done annually in the United States with up to 16% incidence of isolated high-grade prostatic intraepithelial neoplasia (HGPIN). HGPIN has low predictive value for identifying prostate cancer on subsequent needle biopsies in prostate-specific antigen-screened populations. In contemporary series, prostate cancer is detected in approximately 20% of repeat biopsies following a diagnosis of HGPIN. Further, discrete histologic subtypes of HGPIN with clinical implication in management have not been characterized. The TMPRSS2-ERG gene fusion that has recently been described in prostate cancer has also been shown to occur in a subset of HGPIN. This may have significant clinical implications given that TMPRSS2-ERG fusion prostate cancer is associated with a more aggressive clinical course. In this study, we assessed a series of HGPIN lesions and paired prostate cancer for the presence of TMPRSS2-ERG gene fusion. Fusion-positive HGPIN was observed in 16% of the 143 number of lesions, and in all instances, the matching cancer shared the same fusion pattern. Sixty percent of TMPRSS2-ERG fusion prostate cancer had fusion-negative HGPIN. Given the more aggressive nature of TMPRSS2-ERG prostate cancer, the findings of this study raise the possibility that gene fusion-positive HGPIN lesions are harbingers of more aggressive disease. To date, pathologic, molecular, and clinical variables do not help stratify which men with HGPIN are at increased risk for a cancer diagnosis. Our results suggest that the detection of isolated TMPRSS2-ERG fusion HGPIN would improve the positive predictive value of finding TMPRSS2-ERG fusion prostate cancer in subsequent biopsies.


Triple uptake inhibitors: therapeutic potential in depression and beyond.
Drugs that interfere with the uptake and/or metabolism of biogenic amines have been used to treat depression for > 4 decades. Early medications such as tricyclic antidepressants and monoamine oxidase inhibitors are effective but possess many side effects that limit their usefulness. Selective serotonin reuptake inhibitors (SSRIs) or selective noradrenaline reuptake inhibitors (SNRIs) are the results of rational design to find drugs that are as effective as the tricyclic antidepressants, but with more selectivity towards a single monoamine transporter. The SSRI class of drugs, which includes fluoxetine, paroxetine and sertraline, were previously viewed as the agents of choice for treating major depression. Recently, inhibitors of both serotonin and noradrenaline uptake ('dual uptake inhibitors'; SSRI/SNRI such as venlafaxine, duloxetine and milnacipran) have gained acceptance in the market. However, neither the SSRIs nor the SSRI/SNRI are fully satisfactory due to a delayed onset of action, low rate of response and side effect that can affect compliance. An important recent development has been the emergence of the triple uptake inhibitors (SSRI/SNRI/selective dopamine reuptake inhibitor), which inhibit the uptake of all three neurotransmitters that are most closely linked to depression: serotonin, noradrenaline and dopamine. Preclinical studies and clinical trials indicate that a drug inhibiting the uptake of all three of these neurotransmitters could produce a more rapid onset of action and possess greater efficacy than traditional antidepressants. This review discusses the evolution of biogenic amine-based therapies, the emerging strategies involved in the design and synthesis of novel triple uptake inhibitors as antidepressants and the therapeutic potential of triple uptake inhibitors.


Future orientation and suicide ideation and attempts in depressed adults ages 50 and over.
The objective of this study was to test the hypothesis that future orientation is associated with lower levels of suicide ideation and lower likelihood of suicide attempt in a sample of patients in treatment for major depression. Two hundred two participants (116 female, 57%) ages 50-88 years were recruited from inpatient and outpatient settings. All were diagnosed with major depression using a structured diagnostic interview. Suicide ideation was assessed with the Scale for Suicide Ideation (both current and worst point ratings), and a measure of future orientation was created to assess future expectancies. The authors predicted that greater future orientation would be associated with less current and worst point suicide ideation, and would distinguish current and lifetime suicide attempters from nonattempters. Hypotheses were tested using multivariate logistic regression and linear regression analyses that accounted for age, gender, hopelessness, and depression. As hypothesized, higher future orientation scores were associated with lower current suicidal ideation, less intense suicidal ideation at its worst point, and lower probability of a history of attempted suicide after accounting for covariates. Future orientation was not associated with current attempt status. Future orientation holds promise as a cognitive variable associated with decreased suicide risk; a better understanding of its putative protective role is needed. Treatments designed to enhance future orientation might decrease suicide risk.


Living in two homes-a Swedish national survey of wellbeing in 12 and 15 year olds with joint physical custody.
The practice of joint physical custody, where children spend equal time in each parent's home after they separate, is increasing in many countries. It is particularly common in Sweden, where this custody arrangement applies to 30 per cent of children with separated parents. The aim of this study was to examine children's health-related quality of life after parental separation, by comparing children living with both parents in nuclear families to those living in joint physical custody and other forms of domestic arrangements. Data from a national Swedish classroom study of 164,580 children aged 12 and 15-years-old were analysed by two-level linear regression modelling. Z-scores were used to equalise scales for ten dimensions of wellbeing from the KIDSCREEN-52 and the KIDSCREEN-10 Index and analysed for children in joint physical custody in comparison with children living in nuclear families and mostly or only with one parent. Living in a nuclear family was positively associated with almost all aspects of wellbeing in comparison to children with separated parents. Children in joint physical custody experienced more positive outcomes, in terms of subjective wellbeing, family life and peer relations, than children living mostly or only with one parent. For the 12-year-olds, beta coefficients for moods and emotions ranged from -0.20 to -0.33 and peer relations from -0.11 to -0.20 for children in joint physical custody and living mostly or only with one parent. The corresponding estimates for the 15-year-olds varied from -0.08 to -0.28 and from -0.03 to -0.13 on these subscales. The 15-year-olds in joint physical custody were more likely than the 12-year-olds to report similar wellbeing levels on most outcomes to the children in nuclear families. Children who spent equal time living with both parents after a separation reported better wellbeing than children in predominantly single parent care. This was particularly true for the 15-year-olds, while the reported wellbeing of 12-years-olds was less satisfactory. There is a need for further studies that can account for the pre and post separation context of individual families and the wellbeing of younger age groups in joint physical custody.


Ligation of 4-1BB (CDw137) regulates graft-versus-host disease, graft-versus-leukemia, and graft rejection in allogeneic bone marrow transplant recipients.
4-1BB is expressed on activated CD4(+) and CD8(+) T cells; its ligand, 4-1BB ligand is expressed on APCs. Despite expression on both T cell subpopulations, 4-1BB has been reported to predominantly affect CD8(+) T cell responses. By quantifying graft-vs-host disease alloresponses in vivo, we demonstrate that both CD4(+) and CD8(+) T cell-mediated alloresponses are regulated by 4-1BB/4-1BB ligand interactions to approximately the same extent. 4-1BB receptor-facilitated CD4(+) T cell-mediated alloresponses were partly CD28 independent. In two distinct marrow graft rejection systems, host CD8(+) and CD4(+) T cells each separately contributed to host anti-donor T cell-mediated allograft rejection. alpha 4-1BB mAb increased the graft-vs-leukemia effect of a suboptimal number of donor splenocytes given later post bone marrow transplantation by bolstering allogeneic responses resulting in leukemia elimination. In summary, 4-1BB ligation is a potent regulator of CD4(+) and CD8(+) T cell-mediated allogeneic responses in vivo. Modifying the ligation of 4-1BB represents a new approach to altering the graft-vs-host disease and graft-vs-leukemia effects of allogeneic T cells post bone marrow transplantation.


Evaluation of physical activity counseling in primary care using direct observation of the 5As.
The 5As (ask, advise, assess, assist, arrange) are recommended as a strategy for brief physical activity counseling in primary care. There is no reference standard for measurement, however, and patient participation is not well understood. This study's objectives were to (1) develop a coding scheme to measure the 5As using audio-recordings of primary care visits and (2) describe the degree to which patients and physicians accomplish the 5As. We developed a coding scheme using previously published definitions of the 5As, direct-observation measures, and evaluation of audio-recorded discussions of physical activity. We applied the coding scheme to 361 audio-recorded visits by patients reporting low levels of physical activity and 28 physicians in northeast Ohio. The coding scheme achieved good inter-rater agreement for each of the 5As (κ = 0.62-1.0). A total of 135 visits included discussion of physical activity. Although ask tasks occurred in 91% of visits, it infrequently elicited sufficient detail about current activity. Patient readiness to change physical activity (assess) was infrequently directly elicited by the physician (24%), but readiness was commonly expressed by the patient in response to an assessment of current level of physical activity (53%). Ambivalence was infrequently followed by physician assistance (49%). Our newly developed measure showed that (1) physicians infrequently assess patient readiness to change, (2) patient expressions of ambivalence are common, and (3) specific mention of recommended guidelines for exercise is nearly absent. Future work should increase clinician skills in exploring ambivalence and readiness to change, as well as improve explicit mention of recommended guidelines for physical activity.


Automatic segmentation of thalamic nuclei from diffusion tensor magnetic resonance imaging.
The nuclei of the thalamus have traditionally been delineated by their distinct cyto/myeloarchitectural appearance on histology. Here, we show that diffusion tensor magnetic resonance imaging (DTI) can noninvasively resolve the major thalamic nuclei based on the characteristic fiber orientation of the corticothalamic/thalamocortical striations within each nucleus. Using an automatic clustering algorithm, we extracted the Talairach coordinates for the individual thalamic nuclei. The center-of-mass coordinates for the segmented nuclei were found to agree strongly with those obtained from a histological atlas. The ability to resolve thalamic nuclei with DTI will allow for morphometric analysis of specific nuclei and improved anatomical localization of functional activation in the thalamus.


Three-dimensional in vitro tumor models for cancer research and drug evaluation.
Cancer occurs when cells acquire genomic instability and inflammation, produce abnormal levels of epigenetic factors/proteins and tumor suppressors, reprogram the energy metabolism and evade immune destruction, leading to the disruption of cell cycle/normal growth. An early event in carcinogenesis is loss of polarity and detachment from the natural basement membrane, allowing cells to form distinct three-dimensional (3D) structures that interact with each other and with the surrounding microenvironment. Although valuable information has been accumulated from traditional in vitro studies in which cells are grown on flat and hard plastic surfaces (2D culture), this culture condition does not reflect the essential features of tumor tissues. Further, fundamental understanding of cancer metastasis cannot be obtained readily from 2D studies because they lack the complex and dynamic cell-cell communications and cell-matrix interactions that occur during cancer metastasis. These shortcomings, along with lack of spatial depth and cell connectivity, limit the applicability of 2D cultures to accurate testing of pharmacologically active compounds, free or sequestered in nanoparticles. To recapitulate features of native tumor microenvironments, various biomimetic 3D tumor models have been developed to incorporate cancer and stromal cells, relevant matrix components, and biochemical and biophysical cues, into one spatially and temporally integrated system. In this article, we review recent advances in creating 3D tumor models employing tissue engineering principles. We then evaluate the utilities of these novel models for the testing of anticancer drugs and their delivery systems. We highlight the profound differences in responses from 3D in vitro tumors and conventional monolayer cultures. Overall, strategic integration of biological principles and engineering approaches will both improve understanding of tumor progression and invasion and support discovery of more personalized first line treatments for cancer patients.


The induction of growth arrest DNA damage-inducible gene 45 beta in human hepatoma cell lines by S-adenosylmethionine.
Down-regulation of GADD45beta, which is known to influence cell growth control, apoptosis, and cellular response to DNA damage, has been verified to be specific in hepatocellular carcinoma and consistent with the degree of malignancy. Here, we identified promoter elements for several transcriptional factors in the proximal promoter of GADD45beta using the luciferase assay. As a methyl donor for biological transmethylation reactions, S-adenosylmethionine (SAMe) could restore GADD45beta expression in HepG2 in Northern blot analyses and quantitative real-time polymerase chain reaction. Activity and binding capacity of nuclear factor (NF)-kappaB were confirmed to be specifically induced by SAMe, as evidenced by electrophoretic mobility shift assay, enzyme-linked immunosorbent assay, and a decrease of IkappaBalpha in Western blot analyses. The most upstream NF-kappaB-binding site was crucial for transcriptional activation. In contrast to NF-kappaB, although there is an E2F-1-binding site adjacent to the NF-kappaB sites, treatment with SAMe could not induce E2F-1-binding activity. Despite showing a similar GADD45beta promoter regulatory pattern as HepG2 (p53 wild type), Hep3B (p53-null) did not exhibit GADD45beta induction by SAMe, and the induction could be partially recovered on reconstituting p53 in Hep3B. Thus, our results suggest that GADD45beta induction by SAMe via NF-kappaB may represent a novel mechanism of SAMe-mediated hepatoprotection, with p53 playing an important role.


Pulmonary outcome in former preterm, very low birth weight children with bronchopulmonary dysplasia: a case-control follow-up at school age.
To assess and compare long-term pulmonary outcomes in former preterm-born, very low birth weight (VLBW) children with and without bronchopulmonary dysplasia (BPD) born in the surfactant era. Pulmonary function tests (ie, spirometry, body plethysmography, and gas transfer testing) were performed in children with a history of VLBW and BPD (n = 28) and compared with a matched preterm-born VLBW control group (n = 28). Medical history was evaluated by questionnaire. At time of follow-up (mean age, 9.5 years), respiratory symptoms (36% vs 8%) and receipt of asthma medication (21% vs 0%) were significantly more frequent in the preterm-born children with previous BPD than in those with no history of BPD. The children with a history of BPD had significantly lower values for forced expiratory volume in 1 second (z-score -1.27 vs -0.4; P = .008), forced vital capacity (z-score -1.39 vs -0.71 z-score; P = .022), and forced expiratory flow rate at 50% of forced vital capacity (z-score -2.21 vs -1.04; P = .048) compared with the preterm control group. Preterm-born children with a history of BPD are significantly more likely to have lung function abnormalities, such as airway obstruction and respiratory symptoms, at school age compared with preterm-born children without BPD.


Masked-Volume-Wise PCA and "reference Logan" illustrate similar regional differences in kinetic behavior in human brain PET study using [11C]-PIB.
Kinetic modeling using reference Logan is commonly used to analyze data obtained from dynamic Positron Emission Tomography (PET) studies on patients with Alzheimer's disease (AD) and healthy volunteers (HVs) using amyloid imaging agent N-methyl [11C]2-(4'-methylaminophenyl)-6-hydroxy-benzothiazole, [11C]-PIB. The aim of the present study was to explore whether results obtained using the newly introduced method, Masked Volume Wise Principal Component Analysis, MVW-PCA, were similar to the results obtained using reference Logan. MVW-PCA and reference Logan were performed on dynamic PET images obtained from four Alzheimer's disease (AD) patients on two occasions (baseline and follow-up) and on four healthy volunteers (HVs). Regions of interest (ROIs) of similar sizes were positioned in different parts of the brain in both AD patients and HVs where the difference between AD patients and HVs is largest. Signal-to-noise ratio (SNR) and discrimination power (DP) were calculated for images generated by the different methods and the results were compared both qualitatively and quantitatively. MVW-PCA generated images that illustrated similar regional binding patterns compared to reference Logan images and with slightly higher quality, enhanced contrast, improved SNR and DP, without being based on modeling assumptions. MVW-PCA also generated additional MVW-PC images by using the whole dataset, which illustrated regions with different and uncorrelated kinetic behaviors of the administered tracer. This additional information might improve the understanding of kinetic behavior of the administered tracer. MVW-PCA is a potential multivariate method that without modeling assumptions generates high quality images, which illustrated similar regional changes compared to modeling methods such as reference Logan. In addition, MVW-PCA could be used as a new technique, applicable not only on dynamic human brain studies but also on dynamic cardiac studies when using PET.


Construction of a public CHO cell line transcript database using versatile bioinformatics analysis pipelines.
Chinese hamster ovary (CHO) cell lines represent the most commonly used mammalian expression system for the production of therapeutic proteins. In this context, detailed knowledge of the CHO cell transcriptome might help to improve biotechnological processes conducted by specific cell lines. Nevertheless, very few assembled cDNA sequences of CHO cells were publicly released until recently, which puts a severe limitation on biotechnological research. Two extended annotation systems and web-based tools, one for browsing eukaryotic genomes (GenDBE) and one for viewing eukaryotic transcriptomes (SAMS), were established as the first step towards a publicly usable CHO cell genome/transcriptome analysis platform. This is complemented by the development of a new strategy to assemble the ca. 100 million reads, sequenced from a broad range of diverse transcripts, to a high quality CHO cell transcript set. The cDNA libraries were constructed from different CHO cell lines grown under various culture conditions and sequenced using Roche/454 and Illumina sequencing technologies in addition to sequencing reads from a previous study. Two pipelines to extend and improve the CHO cell line transcripts were established. First, de novo assemblies were carried out with the Trinity and Oases assemblers, using varying k-mer sizes. The resulting contigs were screened for potential CDS using ESTScan. Redundant contigs were filtered out using cd-hit-est. The remaining CDS contigs were re-assembled with CAP3. Second, a reference-based assembly with the TopHat/Cufflinks pipeline was performed, using the recently published draft genome sequence of CHO-K1 as reference. Additionally, the de novo contigs were mapped to the reference genome using GMAP and merged with the Cufflinks assembly using the cuffmerge software. With this approach 28,874 transcripts located on 16,492 gene loci could be assembled. Combining the results of both approaches, 65,561 transcripts were identified for CHO cell lines, which could be clustered by sequence identity into 17,598 gene clusters.


Sequence-controlled RNA self-processing: computational design, biochemical analysis, and visualization by AFM.
Reversible chemistry allowing for assembly and disassembly of molecular entities is important for biological self-organization. Thus, ribozymes that support both cleavage and formation of phosphodiester bonds may have contributed to the emergence of functional diversity and increasing complexity of regulatory RNAs in early life. We have previously engineered a variant of the hairpin ribozyme that shows how ribozymes may have circularized or extended their own length by forming concatemers. Using the Vienna RNA package, we now optimized this hairpin ribozyme variant and selected four different RNA sequences that were expected to circularize more efficiently or form longer concatemers upon transcription. (Two-dimensional) PAGE analysis confirms that (i) all four selected ribozymes are catalytically active and (ii) high yields of cyclic species are obtained. AFM imaging in combination with RNA structure prediction enabled us to calculate the distributions of monomers and self-concatenated dimers and trimers. Our results show that computationally optimized molecules do form reasonable amounts of trimers, which has not been observed for the original system so far, and we demonstrate that the combination of theoretical prediction, biochemical and physical analysis is a promising approach toward accurate prediction of ribozyme behavior and design of ribozymes with predefined functions.


Not just passengers: pigeons, Columba livia, can learn homing routes while flying with a more experienced conspecific.
For animals that travel in groups, the directional choices of conspecifics are potentially a rich source of information for spatial learning. In this study, we investigate how the opportunity to follow a locally experienced demonstrator affects route learning by pigeons over repeated homing flights. This test of social influences on navigation takes advantage of the individually distinctive routes that pigeons establish when trained alone. We found that pigeons learn routes just as effectively while flying with a partner as control pigeons do while flying alone. However, rather than learning the exact route of the demonstrator, the paired routes shifted over repeated flights, which suggests that the birds with less local experience also took an active role in the navigational task. The efficiency of the original routes was a key factor in how far they shifted, with less efficient routes undergoing the greatest changes. In this context, inefficient routes are unlikely to be maintained through repeated rounds of social transmission, and instead more efficient routes are achieved because of the interaction between social learning and information pooling.


What determines future smoking intentions of 12- to 13-year-old UK African-Caribbean, Indian, Pakistani and white young people?
It is important to know when designing adolescent smoking interventions how ethnicity and gender influence intention. This paper reports an investigation into how ethnicity influences the smoking intentions of disadvantaged UK African-Caribbean (n = 275), Indian (n = 397), Pakistani (n = 687) and white (n = 1792) 12-13 year olds. The Attitudes-Social influences-Efficacy (ASE) model underpinned the study. It states that ASE determinants (advantages, disadvantages, social acceptance, social norms, modelling, perceived pressure and self-efficacy) directly influence behavioural intention. External factors (country, ethnicity and gender) indirectly influence intention by influencing ASE determinants. ASE determinant scores and future smoking intentions were measured. Linear regression analyses showed that smoking intention varied by ethnicity and gender. Differences in ASE scores largely explained these variations. Ethnicity and gender did not modify the predictive effects of equivalent ASE determinant scores on intention. Being a white boy had a small independent direct influence on intention, which was ascribed to affective beliefs underpinning fitness and sporting prowess. Otherwise, ethnicity had no independent direct effects on intention. Culturally appropriate interventions that aim to change cognitions underpinning ASE determinants and, thus, ASE scores would, consequently, be expected to be equally effective amongst disadvantaged UK African-Caribbean, Indian, Pakistani and white adolescents.


The joint structure of normal and pathological personality: further evidence for a dimensional model.
The literature proposes a joint structure of normal and pathological personality with higher-order factors mainly based on the five-factor model of personality (FFM). The purpose of the present study was to examine the joint structure of the FFM and the DSM-IV personality disorders (PDs) and to discuss this structure with regard to higher-order domains commonly reported in the literature. We applied a canonical correlation analysis, a series of principal component analyses with oblique Promax rotation and a bi-factor analysis with Geomin rotation on 511 subjects of the general population of Zurich, Switzerland, using data from the ZInEP Epidemiology Survey. The 5 FFM traits and the 10 DSM-IV PD dimensions shared 77% of total variance. Component extraction tests pointed towards a two- and three-component solution. The two-component solution comprised a first component with strong positive loadings on neuroticism and all 10 PD dimensions and a second component with strong negative loadings on extraversion and openness and positive loadings on schizoid and avoidant PDs. The three-component solution added a third component with strong positive loadings on conscientiousness and agreeableness and a negative loading on antisocial PD. The bi-factor model provided evidence for 1 general personality dysfunction factor related to neuroticism and 5 group factors, although the interpretability of the latter was limited. Normal and pathological personality domains are not isomorphic or superposable, although they share a substantial proportion of variance. The two and three higher-order domains extracted in the present study correspond well to equivalent factor-solutions reported in the literature. Moreover, these superordinate factors can consistently be integrated within a hierarchical structure of alternative four- and five-factor models. The top of the hierarchy presumably constitutes a general personality dysfunction factor which is closely related to neuroticism.


Cervical cancer outcome by type of health care facilities: National Cancer Database, 2004-2015.
The National Cancer Database from 2004 to 2015 was analyzed to identify cervical cancer outcomes associated with demographic and clinical characteristics measured by types of facility. Chi-Square tests were used to compare proportions and logistic regression to determine factors associated with cervical cancer outcomes. Women treated at Academic/Research Programs (ARPs) were younger at diagnosis, more likely black, less educated and more in Stage 2, lived further away from treatment facilities, had less comorbidities and better 5-year survival, and were more likely to be alive at 30 and 90 days after surgery compared to other programs. Women treated at Community Cancer Programs were more likely 75 and older at diagnosis, more likely to receive radiation treatment and more in Stage 4, more living in rural areas and less than 10 miles from the facility, and had more comorbidities, and lower 5-year survival compared to other programs. Women treated at Comprehensive Community Cancer Programs were more likely white and educated, had more private insurance, and underwent surgery. Women treated at Integrated Network Cancer Programs were more likely to live in urban, south region, and in Stage 1B2, had more surgery and one comorbidity, and died fewer than 30 days after surgery. The type of facility and treatment had varied effects on mortality and 5-year survival. Considering the different cervical cancer outcomes from different health care facilities, further research is needed to identify what factors influence women to choose a health care facility for their treatment and how this choice can affect different health outcomes.


Motion discrimination in cortically blind patients.
Some patients with brain damage affecting the striate cortex, though clinically blind in their field defects, can still discriminate visual stimuli when forced choice procedures are used. Such patients seem particularly sensitive to moving stimuli in their scotomata, though there are conflicting reports as to whether they can discriminate the direction of motion. We tested three patients with areas of cortical blindness for their ability to detect and discriminate the direction of motion of a variety of first-order motion stimuli, namely bars, gratings, plaids and random dot kinematograms depicting translation and motion in depth, during forced choice tasks. The patients could detect the presence of movement in any kind of stimulus, and could discriminate the direction of single bars, but none could discriminate the direction of motion of the more complex stimuli (gratings, plaids and random dot kinematograms) or discriminate between 0 and 100% coherent random dot kinematograms at any speed tested (from 4 to 64 degrees /s). Similar results were obtained from one of the patients who was additionally tested with second-order versions of the translated bar and random dot kinematograms, eliminating light scatter as an explanation. Overall, the results suggest that motion processing in the scotoma is severely impaired, and that the puzzling discrepancies between previous studies can be accounted for by the type of stimulus used. The motion discrimination impairment caused by brain damage affecting the primary visual cortex is inconsistent with the proposed existence of a subcortical pathway to extrastriate cortical motion areas (such as areas MT and MST) which bypasses the striate cortex and is specialized for analysing 'fast' motion.


Pancreatic resections for advanced M1-pancreatic carcinoma: the value of synchronous metastasectomy.
For M1 pancreatic adenocarcinomas pancreatic resection is usually not indicated. However, in highly selected patients synchronous metastasectomy may be appropriate together with pancreatic resection when operative morbidity is low. From January 1, 2004 to December, 2007 a total of 20 patients with pancreatic malignancies were retrospectively evaluated who underwent pancreatic surgery with synchronous resection of hepatic, adjacent organ, or peritoneal metastases for proven UICC stage IV periampullary cancer of the pancreas. Perioperative as well as clinicopathological parameters were evaluated. There were 20 patients (9 men, 11 women; mean age 58 years) identified. The primary tumor was located in the pancreatic head (n = 9, 45%), in pancreatic tail (n = 9, 45%), and in the papilla Vateri (n = 2, 10%). Metastases were located in the liver (n = 14, 70%), peritoneum (n = 5, 25%), and omentum majus (n = 2, 10%). Lymphnode metastases were present in 16 patients (80%). All patients received resection of their tumors together with metastasectomy. Pylorus preserving duodenopancreatectomy was performed in 8 patients, distal pancreatectomy in 8, duodenopancreatectomy in 2, and total pancreatectomy in 2. Morbidity was 45% and there was no perioperative mortality. Median postoperative survival was 10.7 months (2.6-37.7 months) which was not significantly different from a matched-pair group of patients who underwent pancreatic resection for UICC adenocarcinoma of the pancreas (median survival 15.6 months; P = .1). Pancreatic resection for M1 periampullary cancer of the pancreas can be performed safely in well-selected patients. However, indication for surgery has to be made on an individual basis.


SMCHD1 mutations associated with a rare muscular dystrophy can also cause isolated arhinia and Bosma arhinia microphthalmia syndrome.
Arhinia, or absence of the nose, is a rare malformation of unknown etiology that is often accompanied by ocular and reproductive defects. Sequencing of 40 people with arhinia revealed that 84% of probands harbor a missense mutation localized to a constrained region of SMCHD1 encompassing the ATPase domain. SMCHD1 mutations cause facioscapulohumeral muscular dystrophy type 2 (FSHD2) via a trans-acting loss-of-function epigenetic mechanism. We discovered shared mutations and comparable DNA hypomethylation patterning between these distinct disorders. CRISPR/Cas9-mediated alteration of smchd1 in zebrafish yielded arhinia-relevant phenotypes. Transcriptome and protein analyses in arhinia probands and controls showed no differences in SMCHD1 mRNA or protein abundance but revealed regulatory changes in genes and pathways associated with craniofacial patterning. Mutations in SMCHD1 thus contribute to distinct phenotypic spectra, from craniofacial malformation and reproductive disorders to muscular dystrophy, which we speculate to be consistent with oligogenic mechanisms resulting in pleiotropic outcomes.


The HoxD cluster is a dynamic and resilient TAD boundary controlling the segregation of antagonistic regulatory landscapes.
The mammalian HoxD cluster lies between two topologically associating domains (TADs) matching distinct enhancer-rich regulatory landscapes. During limb development, the telomeric TAD controls the early transcription of Hoxd genes in forearm cells, whereas the centromeric TAD subsequently regulates more posterior Hoxd genes in digit cells. Therefore, the TAD boundary prevents the terminal Hoxd13 gene from responding to forearm enhancers, thereby allowing proper limb patterning. To assess the nature and function of this CTCF-rich DNA region in embryos, we compared chromatin interaction profiles between proximal and distal limb bud cells isolated from mutant stocks where various parts of this boundary region were removed. The resulting progressive release in boundary effect triggered inter-TAD contacts, favored by the activity of the newly accessed enhancers. However, the boundary was highly resilient, and only a 400-kb deletion, including the whole-gene cluster, was eventually able to merge the neighboring TADs into a single structure. In this unified TAD, both proximal and distal limb enhancers nevertheless continued to work independently over a targeted transgenic reporter construct. We propose that the whole HoxD cluster is a dynamic TAD border and that the exact boundary position varies depending on both the transcriptional status and the developmental context.


Detecting effects of herbicide runoff: The use of Cassiopea maremetens as a biomonitor to hexazinone.
Herbicides are an integral part of global agricultural activity but can be advected into local drainages that can discharge to coastal marine systems. Herbicide runoff can impact coastal marine organisms, including those associated with coral reefs and coastal mangrove forests. In this study, the symbiotic sedentary jellyfish Cassiopea maremetens were exposed to analytical grade hexazinone to determine their sensitivity and potential for recovery after exposure to a press herbicide event of 14 days followed by a recovery period of matching duration. Bell surface area, photosynthetic yield (i.e. effective quantum yield, EQY), statolith count and zooxanthellae density were analyzed. Most metrics demonstrated significant decreases when exposed to higher concentrations of hexazinone, while EQY was significantly decreased at exposure concentrations from 31 μg/L hexazinone and above. In contrast, zooxanthellae density (cells/mm2) increased in the highest concentrations compared to control animals. At the end of the exposure period the EC50 for bell surface area, EQY, and statolith count were 176 μg/L, 81.96 μg/L, and 304.3 μg/L, respectively. Jellyfish were able to recover to similar start values for all measured metrics at the end of the 14-day recovery period, with EQY showing recovery by Day 7 of the recovery period. This study demonstrated that statolith counts as an estimate of age were not affected by herbicides. We conclude that the depressed metrics from herbicide related impacts of C. maremetens are effective indicators of a relatively recent herbicide perturbation in that the recovery timeframe for these jellyfish is relatively short.


Infantile hemangioma-like vascular lesion in a 26-year-old woman after abortion.
A 26-year-old woman (G2P1A1) presented with a 5-week history of multiple red marks on her body after a therapeutic abortion. A physical examination found 15 palpable red marks on her head, neck, chest, arms and legs. Proliferating endothelial cells, which expressed CD31, CD34, von Willebrand factor, but not Glut-1 and merosin, were observed in the lesional area by histopathological analyses. Histocompatibility antigen typing of 2 lesions was identical to a sample from peripheral blood. Accelerated regression was observed in 2 lesions treated by intralesional injection of betamethasone, while spontaneous regression was observed within 9 months in the remaining lesions without any treatment. Rapid growth, spontaneous regression and histological analyses in this case support the diagnosis of 'infantile hemangioma-like vascular lesion'.


The anesthetic effect of air at atmospheric pressure.
Nitrogen has recognized narcotic potential at hyperbaric pressures. No narcotic effect of helium has been demonstrated at any pressure. We evaluated the effect of nitrogen in air at one atmosphere on human performance by comparing it with helium-oxygen using a four-alternative divided-attention task that requires rapid response to auditory and visual signal changes. There was a 9.3 per cent decrease in response time when subjects breathed helium-oxygen, a signigicant change (P less than 0.001). This change could not be ascribed to practice since the order of presentation of gases did not have a significant effect. It concluded that the nitrogen in ambient air slightly but measurable impairs human performance compared with a non-anesthetic gas such as helium.


Reducing disorder-induced losses for slow light photonic crystal waveguides through Bloch mode engineering.
We present theory and measurements of disorder-induced losses for low loss 1.5 mm long slow light photonic crystal waveguides. A recent class of dispersion engineered waveguides increases the bandwidth of slow light and shows lower propagation losses for the same group index. Our theory and experiments explain how Bloch mode engineering can substantially reduce scattering losses for the same slow light group velocity regime.


Clemizole hydrochloride is a novel and potent inhibitor of transient receptor potential channel TRPC5.
Canonical transient receptor potential channel 5 (TRPC5) is a nonselective, Ca(2+)-permeable cation channel that belongs to the large family of transient receptor potential channels. It is predominantly found in the central nervous system with a high expression density in the hippocampus, the amygdala, and the frontal cortex. Several studies confirm that TRPC5 channels are implicated in the regulation of neurite length and growth cone morphology. We identified clemizole as a novel inhibitor of TRPC5 channels. Clemizole efficiently blocks TRPC5 currents and Ca(2+) entry in the low micromolar range (IC50 = 1.0-1.3 µM), as determined by fluorometric intracellular free Ca(2+) concentration ([Ca(2+)]i) measurements and patch-clamp recordings. Clemizole blocks TRPC5 currents irrespectively of the mode of activation, for example, stimulation of G protein-coupled receptors, hypo-osmotic buffer conditions, or by the direct activator riluzole. Electrophysiological whole-cell recordings revealed that the block was mostly reversible. Moreover, clemizole was still effective in blocking TRPC5 single channels in excised inside-out membrane patches, hinting to a direct block of TRPC5 by clemizole. Based on fluorometric [Ca(2+)]i measurements, clemizole exhibits a sixfold selectivity for TRPC5 over TRPC4β (IC50 = 6.4 µM), the closest structural relative of TRPC5, and an almost 10-fold selectivity over TRPC3 (IC50 = 9.1 µM) and TRPC6 (IC50 = 11.3 µM). TRPM3 and M8 as well as TRPV1, V2, V3, and V4 channels were only weakly affected by markedly higher clemizole concentrations. Clemizole was not only effective in blocking heterologously expressed TRPC5 homomers but also TRPC1:TRPC5 heteromers as well as native TRPC5-like currents in the U-87 glioblastoma cell line.


Graphene on Ru(0001): a 25 x 25 supercell.
The structure of a single layer of graphene on Ru(0001) has been studied using surface x-ray diffraction. A surprising superstructure containing 1250 carbon atoms has been determined, whereby 25 x 25 graphene unit cells lie on 23 x 23 unit cells of Ru. Each supercell contains 2 x 2 crystallographically inequivalent subcells caused by corrugation. Strong intensity oscillations in the superstructure rods demonstrate that the Ru substrate is also significantly corrugated down to several monolayers and that the bonding between graphene and Ru is strong and cannot be caused by van der Waals bonds. Charge transfer from the Ru substrate to the graphene expands and weakens the C-C bonds, which helps accommodate the in-plane tensile stress. The elucidation of this superstructure provides important information in the potential application of graphene as a template for nanocluster arrays.


Exercise is associated with younger methylome and transcriptome profiles in human skeletal muscle.
Exercise training prevents age-related decline in muscle function. Targeting epigenetic aging is a promising actionable mechanism and late-life exercise mitigates epigenetic aging in rodent muscle. Whether exercise training can decelerate, or reverse epigenetic aging in humans is unknown. Here, we performed a powerful meta-analysis of the methylome and transcriptome of an unprecedented number of human skeletal muscle samples (n = 3176). We show that: (1) individuals with higher baseline aerobic fitness have younger epigenetic and transcriptomic profiles, (2) exercise training leads to significant shifts of epigenetic and transcriptomic patterns toward a younger profile, and (3) muscle disuse "ages" the transcriptome. Higher fitness levels were associated with attenuated differential methylation and transcription during aging. Furthermore, both epigenetic and transcriptomic profiles shifted toward a younger state after exercise training interventions, while the transcriptome shifted toward an older state after forced muscle disuse. We demonstrate that exercise training targets many of the age-related transcripts and DNA methylation loci to maintain younger methylome and transcriptome profiles, specifically in genes related to muscle structure, metabolism, and mitochondrial function. Our comprehensive analysis will inform future studies aiming to identify the best combination of therapeutics and exercise regimes to optimize longevity.


Sensitivity of the periodontal pathogen Aggregatibacter actinomycetemcomitans at mildly acidic pH.
Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), a capnophilic facultative anaerobe, is associated with localized aggressive periodontitis and endocarditis. When grown in broth, the cells begin to die rapidly after overnight growth. The cells also often lose viability on plates within a few days. The aim of this study is to identify the cause of the rapid loss of cell viability. Cell viabilities, as measured by colony forming units, were determined for cells obtained from isolated colonies and from the dense part of a streak on plates. The effect of pH on cell viability was determined by growing cells in broth at various initial glucose concentrations and with or without added bicarbonate. A. actinomycetemcomitans cells were highly sensitive to even a mildly acidic pH of ≈6. Because the bacteria grew at a glucose concentration that is commonly used in many laboratories, there was a dramatic decrease in cell viability as the pH went <6, which happened long before the culture reached saturation. This was easily avoided by using a lower initial glucose concentration, and under these conditions, the addition of bicarbonate to the growth medium was not necessary. Cells resuspended in buffer without nutrients lost viability much faster at pH 6 than at a higher pH. On plates, the cell viability was much higher in isolated colonies than in the dense area of the streak. A. actinomycetemcomitans cells rapidly lost viability at even a mildly acidic pH. The problem was easily rectified by growing cells at a low glucose concentration.


Flame front tracking by laser induced fluorescence spectroscopy and advanced image analysis.
This paper presents advanced image analysis methods for extracting information from high speed Planar Laser Induced Fluorescence (PLIF) data obtained from turbulent flames. The application of non-linear anisotropic diffusion filtering and of Active Contour Models (Snakes) is described to isolate flame boundaries. In a subsequent step, the detected flame boundaries are tracked in time using a frequency domain contour interpolation scheme. The implementations of the methods are described and possible applications of the techniques are discussed.


Emotional experience in parents of children with Zellweger spectrum disorders: A qualitative study.
Zellweger spectrum disorders (ZSDs) are rare, debilitating genetic diseases of peroxisome biogenesis that require constant management and lifelong care. Nevertheless, the experience of family caregivers for children diagnosed with ZSD is not well understood. In this study, we sought to characterize the emotional experience of ZSD family caregivers. Three 90-min focus groups were conducted with thirty-seven parents (25 mothers and 12 fathers) of children with ZSD during a family advocacy conference. Focus groups were arranged by age of proband (Group 1: 0-4 years, Group 2: 5-10 years, Group 3: >11 years). Audio recordings of focus groups were transcribed and analyzed using software for coding purposes. Analyzed content was validated using peer debriefing, member checking, and method triangulation. Focus group results showed that nearly a third of ZSD caregivers described their overall emotional experience as a "rollercoaster." Additionally, three interconnected themes were identified: 1) range of emotions, 2) stressors, and 3) coping. Feeling overwhelmed and devastated were the most frequently described emotional responses. Corresponding stressors to these emotions included the burden of caregiver tasks associated with ZSD, and negative interactions with healthcare professionals. The most common coping strategies were acceptance of limitations of the diseases, redefining "normal" in the parenting experience, and advocating on behalf of the child and the patient community. This study underscores the profound emotional impact on parents who are caregivers for children with ZSDs, highlighting the utility of patient community feedback and qualitative approaches to fully characterize the overall family experience. Simple, targeted approaches focusing on improved communication between healthcare professionals and families, as well as offering resources for emotional support may greatly improve the lives of families living with ZSD and other rare pediatric diseases.