Diffusion tensor image registration using hybrid connectivity and tensor features.
Most existing diffusion tensor imaging (DTI) registration methods estimate structural correspondences based on voxelwise matching of tensors. The rich connectivity information that is given by DTI, however, is often neglected. In this article, we propose to integrate complementary information given by connectivity features and tensor features for improved registration accuracy. To utilize connectivity information, we place multiple anchors representing different brain anatomies in the image space, and define the connectivity features for each voxel as the geodesic distances from all anchors to the voxel under consideration. The geodesic distance, which is computed in relation to the tensor field, encapsulates information of brain connectivity. We also extract tensor features for every voxel to reflect the local statistics of tensors in its neighborhood. We then combine both connectivity features and tensor features for registration of tensor images. From the images, landmarks are selected automatically and their correspondences are determined based on their connectivity and tensor feature vectors. The deformation field that deforms one tensor image to the other is iteratively estimated and optimized according to the landmarks and their associated correspondences. Experimental results show that, by using connectivity features and tensor features simultaneously, registration accuracy is increased substantially compared with the cases using either type of features alone.


Central role of reverting mutations in HLA associations with human immunodeficiency virus set point.
Much uncertainty still exists over what T-cell responses need to be induced by an effective human immunodeficiency virus (HIV) vaccine. Previous studies have hypothesized that the effective CD8(+) T-cell responses are those driving the selection of escape mutations that reduce viral fitness and therefore revert post-transmission. In this study, we adopted a novel approach to define better the role of reverting escape mutations in immune control of HIV infection. This analysis of sequences from 710 study subjects with chronic C-clade HIV type 1 infection demonstrates the importance of mutations that impose a fitness cost in the control of viremia. Consistent with previous studies, the viral set points associated with each HLA-B allele are strongly correlated with the number of Gag-specific polymorphisms associated with the relevant HLA-B allele (r = -0.56, P = 0.0034). The viral set points associated with each HLA-C allele were also strongly correlated with the number of Pol-specific polymorphisms associated with the relevant HLA-C allele (r = -0.67, P = 0.0047). However, critically, both these correlations were dependent solely on the polymorphisms identified as reverting. Therefore, despite the inevitable evolution of viral escape, viremia can be controlled through the selection of mutations that are detrimental to viral fitness. The significance of these results is in highlighting the rationale for an HIV vaccine that can induce these broad responses.


Fluoride Exposure Aggravates the Testicular Damage and Sperm Quality in Diabetic Mice: Protective Role of Ginseng and Banaba.
Fluoride toxicity is known to pose infertility in fluoride-intoxicated animals as well as in people residing in fluoride endemic zones. The present study addresses the degree of impairments caused due to co-exposure of high fluoride toxicity in diabetic mice. Swiss mice, Mus musculus, were subjected to fluoride toxicity by providing fluoride-supplemented drinking water (600 ppm NaF) for a period of 30 days after the confirmation of streptozotocin-induced diabetes(STZ, 50 mg/kgbw). Consequently, aggravated hyperglycemia and tissue fluoride accumulation were witnessed in fluoride-intoxicated diabetic mice; later, these toxicated mice were treated with ginseng extract (GE) and banaba leaf extract, (BLE) at dose of 150 mg/kgbw/day alone and in combination for 15 and 30-day duration to check the efficacy of phytoextracts in reversing the toxicity. The spermatological indices studied, such as sperm density, motility, viability and morphology as well as the testicular biochemical parameters showed enhanced impairment in reproductive status of fluoride-intoxicated diabetic mice. Further, 15-days administration of GE and BLE in combination at a dose of 150 mg/kgbw/day was found to be beneficial in normalizing the alterations observed upon fluoride intoxication to diabetic mice. However, the correlates showed moderate association between blood glucose levels and the spermatological as well as biochemical indices wherein the tissue fluoride levels correlate least.


The effects of age on inflammatory and coagulation-fibrinolysis response in patients hospitalized for pneumonia.
To determine whether inflammatory and hemostasis response in patients hospitalized for pneumonia varies by age and whether these differences explain higher mortality in the elderly. In an observational cohort of subjects with community-acquired pneumonia (CAP) recruited from emergency departments (ED) in 28 hospitals, we divided subjects into 5 age groups (<50, 51-64, 65-74, 75-84, and ≥85). We measured circulating levels of inflammatory (TNF, IL-6, and IL-10), hemostasis (D-dimer, Factor IX, thrombin-antithrombin complex, antithrombin and plasminogen-activator inhibitor-1), and cell-surface markers (TLR-2, TLR-4, and HLA-DR) during the first week of hospitalization and at discharge and compared 90-day mortality. We used logistic regression to compare odds ratios (OR) for 90-day mortality between age groups, adjusting for differences in pre-infection factors alone and then additionally adjusting for immune markers. Of 2,183 subjects, 495, 444, 403, 583, and 258 subjects were <50, 51-64, 65-74, 75-84, and ≥85 years of age, respectively. Large age-related differences were observed in 90-day mortality (0.82% vs. 3.2% vs. 6.4% vs. 12.8% vs. 13.6%, p<0.01). No age-related differences in inflammatory and cell surface markers occurred during the first week. Older subjects had higher pro-coagulant markers on ED presentation and over first week (p ≤ 0.03), but these differences were modest (1.0-1.7-fold differences). Odds of death for older adults changed minimally in models incorporating differences in hemostasis and inflammatory markers (for subjects ≥ 85 compared to those <50, OR = 4.36, when adjusted for pre-infection factors and OR = 3.49 when additionally adjusted for hemostasis markers). At discharge, despite clinical recovery as evidenced by normal vital signs in >85% subjects, older subjects had modestly increased hemostasis markers and IL-6 levels (p<0.01). Modest age-related increases in coagulation response occur during hospitalization for CAP; however these differences do not explain the large differences in mortality. Despite clinical recovery, immune resolution may be delayed in older adults at discharge.


A novel material of cross-linked styrylpyridinium salt intercalated montmorillonite for drug delivery.
A facile synthesis of a styrylpyridinium salt (SbQ)/montmorillonite (MMT) via cationic exchange interactions between styrylpyridinium species (specifically SbQ) and MMT platelets is reported in this work. The SbQ-MMT solutions were irradiated under ultraviolet (UV) light for a specific time to obtain the cross-linked SbQ-MMT materials. Scanning electron microscopy and atomic force microscopy analyses revealed the structures and morphologies of MMT and modified MMT. X-ray diffraction and transmission electron microscope analyses indicated that the basal spacing increased from 1.24 to 1.53 nm compared with the pristine MMT, which proved that SbQ had interacted with MMT. Thermal gravimetric analysis curves showed that the amount of SbQ in the MMT interlayers was 35.71 meq/100 g. Fourier transform infrared spectroscopy also confirmed the intercalation of SbQ species into MMT interlayers, and UV spectroscopy was used to follow up the cross-linking of SbQ-MMT. This novel material has potential applications in drug delivery, and it can also be used as an additive to improve the mechanical properties of polymers.


Fluoxetine interacts with the lipid bilayer of the inner membrane in isolated rat brain mitochondria, inhibiting electron transport and F1F0-ATPase activity.
The effects of fluoxetine on the oxidative phosphorylation of mitochondria isolated from rat brain and on the kinetic properties of submitochondrial particle F1F0-ATPase were evaluated. The state 3 respiration rate supported by pyruvate + malate, succinate, or ascorbate + tetramethyl-p-phenylenediamine (TMPD) was substantially decreased by fluoxetine. The IC50 for pyruvate + malate oxidation was approximately 0.15 mM and the pattern of inhibition was the typical one of the electron-transport inhibitors, in that the drug inhibited both ADP- and carbonyl cyanide m-chlorophenylhydrazone (CCCP)-stimulated respirations and the former inhibition was not released by the uncoupler. Fluoxetine also decreased the activity of submitochondrial particle F1F0-ATPase (IC50 approximately 0.08 mM) even though K0.5 and activity of Triton X-100 solubilized enzyme were not changed substantially. As a consequence of these effects, fluoxetine decreased the rate of ATP synthesis and depressed the phosphorylation potential of mitochondria. Incubation of mitochondria or submitochondrial particles with fluoxetine under the conditions of respiration or F1F0-ATPase assays, respectively, caused a dose-dependent enhancement of 1-anilino-8-naphthalene sulfonate (ANS) fluorescence. These results show that fluoxetine indirectly and nonspecifically affects electron transport and F1F0)-ATPase activity inhibiting oxidative phosphorylation in isolated rat brain mitochondria. They suggest, in addition, that these effects are mediated by the drug interference with the physical state of lipid bilayer of inner mitochondrial membrane.


Enterobacter spp.: pathogens poised to flourish at the turn of the century.
Knowledge of the genus Enterobacter and its role in human disease has expanded exponentially in recent years. The incidence of infection in the hospital and the community has increased. New clinical syndromes have been recognized. Enterobacter spp. have also been implicated as causes of other syndromes that traditionally have been associated almost exclusively with more easily treatable pathogens, such as group A streptococci and staphylococci. Rapid emergence of multiple-drug resistance has been documented in individual patients during therapy and in populations and environments with strong selective pressure from antimicrobial agents, especially the cephalosporins. Therapeutic options for patients infected with multiply resistant strains have become severely limited. Carbapenems or, alternatively, fluoroquinolones are the most predictively active options, although resistance to both classes has been observed on rare occasions. Enterobacter spp. appear well adapted for survival and even proliferation as the turn of the century approaches.


Perspectives of the management of childhood lymphoma: experience at Tygerberg Hospital, Western Cape, South Africa.
Hodgkin's disease (HD) in children corresponds to a large degree to HD in adults. Non-Hodgkin's Lymphoma (NHL) in children, however, differs from NHL in adults with respect to the classification, natural history, management and course. For practical reasons clinicians generally classify and treat NHL in children as either B-cell or T-cell disease. Over the past 22 years, the Paediatric Oncology Unit of the Tygerberg Hospital has treated HD with three different regimens. Use of the CLVPP and MOPP/ABVD regimens resulted in late relapses that adversely affected event free survival (EFS). For the last four years HD has been treated according to the regimen suggested by Schellong with good short term survival rates. Lymphoblastic or T-cell NHL is treated with regimens normally used for acute lymphoblastic leukaemia (e.g. BFM protocols) or modified leukaemia treatments for leukaemia-lymphoma syndromes (e.g. LSA2L2). We lately use a modified BFM regimen with a 70% EFS for all stages. Three consecutive regimens have been used to treat B-cell NHL over the past 22 years. The first was a COMP regimen, followed by the LMB-89 and LMB-96 regimens. Although toxicity has increased with the increased intensity of the treatment regimen, EFS has improved from 25% to 87% for all B-cell NHL. The majority of patients had stage III and IV disease. Although the LMB regimens are toxic, the implementation is manageable provided good laboratory back up and supportive facilities are available.


Evidence for significant overlap between common risk variants for Crohn's disease and ankylosing spondylitis.
A multicenter genome-wide association scan for Crohn's Disease (CD) has recently reported 40 CD susceptibility loci, including 29 novel ones (19 significant and 10 putative). To gain insight into the genetic overlap between CD and ankylosing spondylitis (AS), these markers were tested for association in AS patients. Two previously established associations, namely with the MHC and IL23R loci, were confirmed. In addition, rs2872507, which maps to a locus associated with asthma and influences the expression of the ORMDL3 gene in lymphoblastoid cells, showed a significant association with AS (p = 0.03). In gut biopsies of AS and CD patients, ORMDL3 expression was not significantly different from controls and no correlation was found with the rs2872507 genotype (Spearman's rho: -0.067). The distribution of p-values for the remaining 36 SNPs was significantly skewed towards low p-values unless the top 5 ranked SNPs (ORMDL3, NKX2-3, PTPN2, ICOSLG and MST1) were excluded from the analysis. Association analysis using risk variants for CD led to the identification of a new risk variant associated with AS (ORMDL3), underscoring a role for ER stress in AS. In addition, two known and five potentially relevant associations were detected, contributing to common susceptibility of CD and AS.


Dielectrowetting manipulation for digital microfluidics: creating, transporting, splitting, and merging of droplets.
Generating, splitting, transporting, and merging droplets are fundamental and critical unit operations for digital (droplet-based) microfluidics. State-of-the-art digital microfluidics performs such operations commonly using electrowetting-on-dielectric (EWOD) in the typical configuration of two parallel channel plates. This paper presents such operations using dielectrowetting (derived from liquid dielectrophoresis), not EWOD, with an array of interdigitated electrodes. The major and unique feature is that the present droplet manipulations are effective for conductive (water with/without surfactant) and non-conductive (propylene carbonate) fluids. An equally important aspect is that the manipulations are performed in an open space without the covering top plate. This behavior is attributed to the intrinsic nature of dielectrowetting to generate stronger wetting forces than EWOD (with the ability to achieve complete wetting with contact angle = 0° to form a thin film). Using dielectrowetting, micro-droplets of various volumes are created from a large droplet and transported. Splitting a single droplet as well as multiple droplets and merging them are also achieved, even when the droplets are smaller than the electrode pads. The above splitting, transport, and merging operations are effective for propylene carbonate as well as DI water with/without surfactant, though the creating operation is proven only for propylene carbonate at this moment. All the above manipulations are successfully carried out on a single plate, which not only simplifies the structure and operation procedure, but could also eliminate the restriction to the volume of fluid handled.


The Uppsala Electron-Density Server.
The Uppsala Electron Density Server (EDS; http://eds.bmc.uu.se/) is a web-based facility that provides access to electron-density maps and statistics concerning the fit of crystal structures and their maps. Maps are available for approximately 87% of the crystallographic Protein Data Bank (PDB) entries for which structure factors have been deposited and for which straightforward map calculations succeed in reproducing the published R value to within five percentage points. Here, an account is provided of the methods that are used to generate the information contained in the server. Some of the problems that are encountered in the map-generation process as well as some spin-offs of the project are also discussed.


Accounting for probe-level noise in principal component analysis of microarray data.
Principal Component Analysis (PCA) is one of the most popular dimensionality reduction techniques for the analysis of high-dimensional datasets. However, in its standard form, it does not take into account any error measures associated with the data points beyond a standard spherical noise. This indiscriminate nature provides one of its main weaknesses when applied to biological data with inherently large variability, such as expression levels measured with microarrays. Methods now exist for extracting credibility intervals from the probe-level analysis of cDNA and oligonucleotide microarray experiments. These credibility intervals are gene and experiment specific, and can be propagated through an appropriate probabilistic downstream analysis. We propose a new model-based approach to PCA that takes into account the variances associated with each gene in each experiment. We develop an efficient EM-algorithm to estimate the parameters of our new model. The model provides significantly better results than standard PCA, while remaining computationally reasonable. We show how the model can be used to 'denoise' a microarray dataset leading to improved expression profiles and tighter clustering across profiles. The probabilistic nature of the model means that the correct number of principal components is automatically obtained.


Society of Gynecologic Oncology statement on risk assessment for inherited gynecologic cancer predispositions.
Women with germline mutations in the cancer susceptibility genes, BRCA1 or BRCA2, associated with Hereditary Breast & Ovarian Cancer syndrome, have up to an 85% lifetime risk of breast cancer and up to a 46% lifetime risk of ovarian, tubal, and peritoneal cancers. Similarly, women with mutations in the DNA mismatch repair genes, MLH1, MSH2, MSH6, or PMS2, associated with the Lynch/Hereditary Non-Polyposis Colorectal Cancer (HNPCC) syndrome, have up to a 40-60% lifetime risk of both endometrial and colorectal cancers as well as a 9-12% lifetime risk of ovarian cancer. Mutations in other genes including TP53, PTEN, and STK11 are responsible for hereditary syndromes associated with gynecologic, breast, and other cancers. Evaluation of the likelihood of a patient having one of these gynecologic cancer predisposition syndromes enables physicians to provide individualized assessments of cancer risk, as well as the opportunity to provide tailored screening and prevention strategies such as surveillance, chemoprevention, and prophylactic surgery that may reduce the morbidity and mortality associated with these syndromes. Evaluation for the presence of a hereditary cancer syndrome is a process that includes assessment of clinical and tumor characteristics, education and counseling conducted by a provider with expertise in cancer genetics, and may include genetic testing after appropriate consent is obtained. This commentary provides guidance on identification of patients who may benefit from assessment for the presence of a hereditary breast and/or gynecologic cancer syndrome.


Juvenile mice show greater flexibility in multiple choice reversal learning than adults.
We hypothesized that decision-making strategies in juvenile animals, rather than being immature, are optimized to navigate the uncertainty and instability likely to be encountered in the environment at the time of the animal's transition to independence. We tested juvenile and young adult mice on discrimination and reversal of a 4-choice and 2-choice odor-based foraging task. Juvenile mice (P26–27) learned a 4-choice discrimination and reversal faster than adults (P60–70), making fewer perseverative and distraction errors. Juvenile mice had shorter choice latencies and more focused search strategies. In both ages, performance of the task was significantly impaired by a lesion of the dorsomedial frontal cortex. Our data show that the frontal cortex can support highly flexible behavior in juvenile mice at a time coincident with weaning and first independence. The unexpected developmental decline in flexibility of behavior one month later suggests that frontal cortex based executive function may not inevitably become more flexible with age, but rather may be developmentally tuned to optimize exploratory and exploitative behavior for each life stage.


In-vivo31P-MRS of skeletal muscle and liver: A way for non-invasive assessment of their metabolism.
In addition to direct assessment of high energy phosphorus containing metabolite content within tissues, phosphorus magnetic resonance spectroscopy (31P-MRS) provides options to measure phospholipid metabolites and cellular pH, as well as the kinetics of chemical reactions of energy metabolism in vivo. Even though the great potential of 31P-MR was recognized over 30 years ago, modern MR systems, as well as new, dedicated hardware and measurement techniques provide further opportunities for research of human biochemistry. This paper presents a methodological overview of the 31P-MR techniques that can be used for basic, physiological, or clinical research of human skeletal muscle and liver in vivo. Practical issues of 31P-MRS experiments and examples of potential applications are also provided. As signal localization is essential for liver 31P-MRS and is important for dynamic muscle examinations as well, typical localization strategies for 31P-MR are also described.


Precision of sensing cell length via concentration gradients.
Unicellular organisms are typically found to have a characteristic cell size. To achieve a homeostatic distribution of cell sizes over many generations requires that cell length is actively sensed and regulated. However, the mechanisms by which cell size is controlled remain poorly understood. Recent experiments in fission yeast have shown that cell length is controlled in part by polar gradients of the protein Pom1 together with localized measurement of concentration at midcell. Dilution as the cell grows leads to a reduction in the midcell protein concentration, which lifts a block on mitosis. Here we analyze the precision of this mechanism for length sensing in the presence of inevitable intrinsic noise in the processes leading to formation and measurement of this gradient. We find that the use of concentration gradients allows for more robust length sensing than a comparable spatially uniform system, and allows for reliable length determination even if the average protein concentration throughout the cell remains constant as the cell grows. Optimal values for the gradient decay length and receptor dissociation constant emerge from maximizing sensitivity while minimizing the impact of density fluctuations.


Lesser magnitudes of lower extremity variability during terminal swing characterizes walking patterns in children with autism.
Anecdotally, children with Autism Spectrum Disorder have highly variable lower extremity walking patterns, yet, this has not been sufficiently quantified. As such, the purpose of this study was to examine walking pattern variability by way of lower extremity coordination and spatio-temporal characteristics in children with autism compared with individuals with typical development during over-ground walking. Bilateral continuous relative phase variability was computed for the thigh-leg, leg-foot, and thigh-foot segment couples for 11 children with autism and 9 children with typical development at each gait sub-phase. Furthermore, left and right stride lengths and stride width were computed and compared. The Model Statistic was utilized to test for statistical differences in variability between each child with autism to an aggregate group with typical development. Effect sizes were computed to determine the meaningfulness between responses for children with autism and typical development. Coefficient of variation and effect sizes were computed for stride lengths and stride width. Analysis revealed that children with autism exhibited differences in variability in each gait sub-phase. Notably, all but two children with autism exhibited lesser variability in all segment couples during terminal swing. Differences in stride lengths were relatively minimal, however, greater coefficient of variation magnitudes in stride width were observed in children with autism. This finding reveals that children with autism may have limited or a preferred movement strategy when preparing the foot for ground contact. The findings from this study suggest variability may be an identifiable characteristic during movement in children with autism.


Control of lupus nephritis by changes of gut microbiota.
Systemic lupus erythematosus, characterized by persistent inflammation, is a complex autoimmune disorder with no known cure. Immunosuppressants used in treatment put patients at a higher risk of infections. New knowledge of disease modulators, such as symbiotic bacteria, can enable fine-tuning of parts of the immune system, rather than suppressing it altogether. Dysbiosis of gut microbiota promotes autoimmune disorders that damage extraintestinal organs. Here we report a role of gut microbiota in the pathogenesis of renal dysfunction in lupus. Using a classical model of lupus nephritis, MRL/lpr, we found a marked depletion of Lactobacillales in the gut microbiota. Increasing Lactobacillales in the gut improved renal function of these mice and prolonged their survival. We used a mixture of 5 Lactobacillus strains (Lactobacillus oris, Lactobacillus rhamnosus, Lactobacillus reuteri, Lactobacillus johnsonii, and Lactobacillus gasseri), but L. reuteri and an uncultured Lactobacillus sp. accounted for most of the observed effects. Further studies revealed that MRL/lpr mice possessed a "leaky" gut, which was reversed by increased Lactobacillus colonization. Lactobacillus treatment contributed to an anti-inflammatory environment by decreasing IL-6 and increasing IL-10 production in the gut. In the circulation, Lactobacillus treatment increased IL-10 and decreased IgG2a that is considered to be a major immune deposit in the kidney of MRL/lpr mice. Inside the kidney, Lactobacillus treatment also skewed the Treg-Th17 balance towards a Treg phenotype. These beneficial effects were present in female and castrated male mice, but not in intact males, suggesting that the gut microbiota controls lupus nephritis in a sex hormone-dependent manner. This work demonstrates essential mechanisms on how changes of the gut microbiota regulate lupus-associated immune responses in mice. Future studies are warranted to determine if these results can be replicated in human subjects.


Nucleation and Early Stage Growth of Li Electrodeposits.
The morphologies that metal electrodeposits form during the earliest stages of electrodeposition are known to play a critical role in the recharge of electrochemical cells that use metals as anodes. Here we report results from a combined theoretical and experimental study of the early stage nucleation and growth of electrodeposited lithium at liquid-solid interfaces. The spatial characteristics of lithium electrodeposits are studied via scanning electron microscopy (SEM) in tandem with image analysis. Comparisons of Li nucleation and growth in multiple electrolytes provide a comprehensive picture of the initial nucleation and growth dynamics. We report that ion diffusion in the bulk electrolyte and through the solid electrolyte interphase (SEI) formed spontaneously on the metal play equally important roles in regulating Li nucleation and growth. We show further that the underlying physics dictating bulk and surface diffusion are similar across a range of electrolyte chemistries and measurement conditions, and that fluorinated electrolytes produce a distinct flattening of Li electrodeposits at low rates. These observations are rationalized using X-ray photoelectron spectroscopy (XPS), electrochemical impedance spectroscopy (EIS), and contact angle goniometry to probe the interfacial chemistry and dynamics. Our results show that high interfacial energy and high surface ion diffusivity are necessary for uniform Li plating.


Impact of co-adsorbed oxygen on crotonaldehyde adsorption over gold nanoclusters: a computational study.
Crotonaldehyde (2-butenal) adsorption over gold sub-nanometer particles, and the influence of co-adsorbed oxygen, has been systematically investigated by computational methods. Using density functional theory, the adsorption energetics of crotonaldehyde on bare and oxidised gold clusters (Au13, d = 0.8 nm) were determined as a function of oxygen coverage and coordination geometry. At low oxygen coverage, sites are available for which crotonaldehyde adsorption is enhanced relative to bare Au clusters by 10 kJ mol(-1). At higher oxygen coverage, crotonaldehyde is forced to adsorb in close proximity to oxygen weakening adsorption by up to 60 kJ mol(-1) relative to bare Au. Bonding geometries, density of states plots and Bader analysis, are used to elucidate crotonaldehyde bonding to gold nanoparticles in terms of partial electron transfer from Au to crotonaldehyde, and note that donation to gold from crotonaldehyde also becomes significant following metal oxidation. At high oxygen coverage we find that all molecular adsorption sites have a neighbouring, destabilising, oxygen adatom so that despite enhanced donation, crotonaldehyde adsorption is always weakened by steric interactions. For a larger cluster (Au38, d = 1.1 nm) crotonaldehyde adsorption is destabilized in this way even at a low oxygen coverage. These findings provide a quantitative framework to underpin the experimentally observed influence of oxygen on the selective oxidation of crotyl alcohol to crotonaldehyde over gold and gold-palladium alloys.


Nanoparticle-mediated measurement of target-drug binding in cancer cells.
Responses to molecularly targeted therapies can be highly variable and depend on mutations, fluctuations in target protein levels in individual cells, and drug delivery. The ability to rapidly quantitate drug response in cells harvested from patients in a point-of-care setting would have far reaching implications. Capitalizing on recent developments with miniaturized NMR technologies, we have developed a magnetic nanoparticle-based approach to directly measure both target expression and drug binding in scant human cells. The method involves covalent conjugation of the small-molecule drug to a magnetic nanoparticle that is then used as a read-out for target expression and drug-binding affinity. Using poly(ADP-ribose) polymerase (PARP) inhibition as a model system, we developed an approach to distinguish differential expression of PARP in scant cells with excellent correlation to gold standards, the ability to mimic drug pharmacodynamics ex vivo through competitive target-drug binding, and the potential to perform such measurements in clinical samples.


Predicting Daily Activities From Egocentric Images Using Deep Learning.
We present a method to analyze images taken from a passive egocentric wearable camera along with the contextual information, such as time and day of week, to learn and predict everyday activities of an individual. We collected a dataset of 40,103 egocentric images over a 6 month period with 19 activity classes and demonstrate the benefit of state-of-the-art deep learning techniques for learning and predicting daily activities. Classification is conducted using a Convolutional Neural Network (CNN) with a classification method we introduce called a late fusion ensemble. This late fusion ensemble incorporates relevant contextual information and increases our classification accuracy. Our technique achieves an overall accuracy of 83.07% in predicting a person's activity across the 19 activity classes. We also demonstrate some promising results from two additional users by fine-tuning the classifier with one day of training data.


Dramatic acceleration of the pd-catalyzed [4+2] benzannulation reaction of enynes and diynes in the presence of lewis acids and bases: expanded scope and new mechanistic insights.
Dramatic acceleration of the palladium catalyzed [4+2] benzannulation and sequential [2+2+2] trimerization reactions in the presence of a Lewis acid/phosphine combination or in the presence of Bronsted bases has been explored. These novel sets of conditions allowed for remarkable enhancement of reaction rates and broadening of the substrate scope and for a significant improvement of reaction yields, particularly for problematic pentasubstituted benzenes. It was found that the real nature of Lewis acid acceleration does not only lie in the isomerization of conjugated enynes but also in the direct acceleration of the [4+2] benzannulation reaction. These experimental findings, combined with the deuterium-labeling studies and DFT calculations, led to a mechanistic rationale, which (a) reasonably accounts for the observed acceleration of the reaction by Lewis acid and bases; (b) provides a viable alternative route for the ring-closing step in the mechanism of benzannulation; (c) clarifies the mechanism of hydrogen migration; and (d) for the first time provides a rationale for the origins of the remarkable stereoselectivity of the hydrogen migration during the benzannulation reaction.


Fit for high altitude: are hypoxic challenge tests useful?
Altitude travel results in acute variations of barometric pressure, which induce different degrees of hypoxia, changing the gas contents in body tissues and cavities. Non ventilated air containing cavities may induce barotraumas of the lung (pneumothorax), sinuses and middle ear, with pain, vertigo and hearing loss. Commercial air planes keep their cabin pressure at an equivalent altitude of about 2,500 m. This leads to an increased respiratory drive which may also result in symptoms of emotional hyperventilation. In patients with preexisting respiratory pathology due to lung, cardiovascular, pleural, thoracic neuromuscular or obesity-related diseases (i.e. obstructive sleep apnea) an additional hypoxic stress may induce respiratory pump and/or heart failure. Clinical pre-altitude assessment must be disease-specific and it includes spirometry, pulsoximetry, ECG, pulmonary and systemic hypertension assessment. In patients with abnormal values we need, in addition, measurements of hemoglobin, pH, base excess, PaO2, and PaCO2 to evaluate whether O2- and CO2-transport is sufficient.Instead of the hypoxia altitude simulation test (HAST), which is not without danger for patients with respiratory insufficiency, we prefer primarily a hyperoxic challenge. The supplementation of normobaric O2 gives us information on the acute reversibility of the arterial hypoxemia and the reduction of ventilation and pulmonary hypertension, as well as about the efficiency of the additional O2-flow needed during altitude exposure. For difficult judgements the performance of the test in a hypobaric chamber with and without supplemental O2-breathing remains the gold standard. The increasing numbers of drugs to treat acute pulmonary hypertension due to altitude exposure (acetazolamide, dexamethasone, nifedipine, sildenafil) or to other etiologies (anticoagulants, prostanoids, phosphodiesterase-5-inhibitors, endothelin receptor antagonists) including mechanical aids to reduce periodical or insufficient ventilation during altitude exposure (added dead space, continuous or bilevel positive airway pressure, non-invasive ventilation) call for further randomized controlled trials of combined applications.


Design of the Advance Directives Cohort: a study of end-of-life decision-making focusing on Advance Directives.
ADs are documents in which one can state one's preferences concerning end-of-life care, aimed at making someone's wishes known in situations where he/she is not able to do so in another manner. There is still a lot unclear about ADs. We designed a study aimed at investigating the whole process from the formulating of an AD to its actual use at the end of life. The study has mixed methods: it's longitudinal, consisting of a quantitative cohort-study which provides a framework for predominantly qualitative sub-studies. The members of the cohort are persons owning an AD, recruited through two Dutch associations who provide the most common standard ADs in the Netherlands, the NVVE (Right to Die-NL), of which 5561 members participate, and the NPV (Dutch Patient Organisation), of which 1263 members participate. Both groups were compared to a sample of the Dutch general public. NVVE-respondents are more often single, higher educated and non-religious, while amongst NPV-respondents there are more Protestants compared to the Dutch public. They are sent a questionnaire every 1,5 year with a follow-up of at least 7,5 years. The response rate after the second round was 88% respectively 90% for the NVVE and NPV. Participants were asked if we were allowed to approach close-ones after their possible death in the future. In this way we can get insight in the actual use of ADs at the end of life, also by comparing our data to that from the Longitudinal Aging Study Amsterdam, whose respondents generally do not have an AD. The cohort is representative for people with an AD as is required to study the main research questions. The longitudinal nature of the study as well as the use of qualitative methods makes it has a broad scope, focusing on the whole course of decision-making involving ADs. It is possible to compare the end of life between patients with and without an AD with the use of data from another cohort.


Altered erythrocyte cation permeability in familial pseudohyperkalaemia.
1. Erythrocyte cation transport pathways have been investigated in a family with pseudohyperkalaemia. 2. Ouabain- and bumetanide-resistant Na+ and K+ effluxes in three pseudohyperkalaemic patients were not different from those of control subjects when assessed at 37 degrees C. 3. When the temperature was decreased to 20 degrees C and 9 degrees C, K+ passive permeability markedly increased and Na+ permeability remained unchanged in these patients. In contrast, in control subjects a reduction in temperature caused a marked reduction in Na+ and K+ passive permeability. 4. These findings could account for the marked increase in plasma K+ concentration observed at subphysiological temperatures. 5. The Na+-K+ co-transport pathway was reduced in all members of the family, but the Na+-K+ pump was reduced in only two of them. These alterations were independent from the pseudohyperkalaemic state.


Prevalence of human papillomaviruses in the mouths of New Zealand women.
Human papillomavirus (HPV) in the oral cavity has been retrospectively associated with an increased risk of developing HPV-positive head and neck squamous cell carcinoma (HNSCC). The aim of this study was to determine the prevalence of oral HPV infection in a local population of New Zealand women aged 18 to 25 years, including determination of HPV genotypes, and to assess potential risk factors for oral HPV infection using participant questionnaire responses. Oral brushings and questionnaire responses were collected from 234 women recruited from sexual health and student health centres. Questions covered age, ethnicity, sexual partners, alcohol consumption and smoking. PGMY primers were used for HPV detection by PCR, and results confirmed by sequencing and the cobas® 4800 HPV system. The prevalence of HPV infection was 3.2% of 216 women (95% CI: 1.6%-6.5%). Samples from two women (0.9%, 95% CI: 0.3%-3.3%) contained oncogenic HPV, and another five (2.3%, 95% CI: 1.0%-5.3%) were positive for HPV 13. No significant associations were found between putative risk factors and the presence of oral HPV infection. The prevalence of HPV in the oral cavity of New Zealand woman was comparable to results of other studies, but showed an unusual distribution of HPV types. The comparatively high detection rate of HPV 13 suggests that further work into clinical significance of oral HPV 13 infection is warranted.


An efficient asymmetric synthesis of (-)-lupinine.
The asymmetric synthesis of (-)-lupinine was achieved in 8 steps, 15% overall yield and >99 : 1 dr from commercially available starting materials. The strategy used for the construction of the quinolizidine scaffold involved reaction of an enantiopure tertiary dibenzylamine via two sequential ring-closures which both occurred with concomitant N-debenzylation.


Transcription factors responsive to cAMP.
In eukaryotes, transcriptional regulation upon stimulation of the adenylate cyclase signaling pathway is mediated by a family of cAMP-responsive nuclear factors. This family consists of a large number of members that may act as activators or repressors. These factors contain the basic domain/leucine zipper motifs and bind as dimers to cAMP-response elements (CRE). The function of CRE-binding proteins is modulated by phosphorylation by several kinases. The ICER (inducible cAMP early repressor) protein is the only inducible member of this family. The induction of this powerful repressor is likely to be important for the transient nature of cAMP-induced gene expression. CRE-binding proteins have been found to play an important role in the physiology of the pituitary gland, in regulating spermatogenesis, in the response to circadian rhythms, and in the molecular basis of memory.


Natural Products from Actinobacteria Associated with Fungus-Growing Termites.
The chemical analysis of insect-associated Actinobacteria has attracted the interest of natural product chemists in the past years as bacterial-produced metabolites are sought to be crucial for sustaining and protecting the insect host. The objective of our study was to evaluate the phylogeny and bioprospecting of Actinobacteria associated with fungus-growing termites. We characterized 97 Actinobacteria from the gut, exoskeleton, and fungus garden (comb) of the fungus-growing termite Macrotermes natalensis and used two different bioassays to assess their general antimicrobial activity. We selected two strains for chemical analysis and investigated the culture broth of the axenic strains and fungus-actinobacterium co-cultures. From these studies, we identified the previously-reported PKS-derived barceloneic acid A and the PKS-derived rubterolones. Analysis of culture broth yielded a new dichlorinated diketopiperazine derivative and two new tetracyclic lanthipeptides, named rubrominins A and B. The discussed natural products highlight that insect-associated Actinobacteria are highly prolific natural product producers yielding important chemical scaffolds urgently needed for future drug development programs.


Transient brain activity disentangles fMRI resting-state dynamics in terms of spatially and temporally overlapping networks.
Dynamics of resting-state functional magnetic resonance imaging (fMRI) provide a new window onto the organizational principles of brain function. Using state-of-the-art signal processing techniques, we extract innovation-driven co-activation patterns (iCAPs) from resting-state fMRI. The iCAPs' maps are spatially overlapping and their sustained-activity signals temporally overlapping. Decomposing resting-state fMRI using iCAPs reveals the rich spatiotemporal structure of functional components that dynamically assemble known resting-state networks. The temporal overlap between iCAPs is substantial; typically, three to four iCAPs occur simultaneously in combinations that are consistent with their behaviour profiles. In contrast to conventional connectivity analysis, which suggests a negative correlation between fluctuations in the default-mode network (DMN) and task-positive networks, we instead find evidence for two DMN-related iCAPs consisting the posterior cingulate cortex that differentially interact with the attention network. These findings demonstrate how the fMRI resting state can be functionally decomposed into spatially and temporally overlapping building blocks using iCAPs.


Proteinases in chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) is a major health problem worldwide, and we have little specific therapy to offer these patients. One potential strategy to limit loss of lung function in COPD would be to inhibit matrix-degrading proteinases. Several serine proteinases and matrix metalloproteinases are expressed in association with COPD in humans. Application of gene-targeted macrophage elastase and neutrophil elastase to a mouse model of cigarette-smoke-induced emphysema has uncovered roles for these proteinases in airspace enlargement, and has identified many interactions between these proteolytic systems.


Energetics of carbohydrate binding by a 14 kDa S-type mammalian lectin.
The thermodynamics of the binding of derivatives of galactose and lactose to a 14 kDa beta-galactoside-binding lectin (L-14) from sheep spleen has been studied in 10 nM phosphate/150 mM NaCl/10 mM beta-mercaptoethanol buffer, pH 7.4, and in the temperature range 285-300 K using titration calorimetry. The single-site binding constants of various sugars for the lectin were in the following order: N-acetyl-lactosamine thiodigalactoside > 4-methylumbelliferyl lactoside > lactose > 4-methylumbelliferyl alpha-D-galactoside > methyl-alpha-galactose > methyl-beta-galactose. Reactions were essentially enthalpically driven with the binding enthalpies ranging from -53.8 kJ/mol for thiodigalactoside at 301 K to -2.2 kJ/mol for galactose at 300 K, indicating that hydrogen-bonding and van der Waals interactions provide the major stabilization for these reactions. However, the binding of 4-methylumbelliferyl-alpha-D-galactose displays relatively favourable entropic contributions, indicating the existence of a non-polar site adjacent to the galactose-binding subsite. From the increments in the enthalpies for the binding of lactose, N-acetyl-lactosamine and thiodigalactoside relative to methyl-beta-galactose, the contribution of glucose binding in the subsite adjacent to that for galactose shows that glucose makes a major contribution to the stability of L-14 disaccharide complexes. Observation of enthalpy-entropy compensation for the recognition of saccharides such as lactose by L-14 and the absence of it for monosaccharides such as galactose, together with the lack of appreciable changes in the heat capacity (delta Cp), indicate that reorganization of water plays an important role in these reactions.


Intermittent ambiguous stimuli: implicit memory causes periodic perceptual alternations.
When viewing a stimulus that has multiple plausible real-world interpretations, perception alternates between these interpretations every few seconds. Alternations can be halted by intermittently removing the stimulus from view. The same interpretation dominates over many successive presentations, and perception stabilizes. Here we study perception during long sessions of such intermittent presentation. We demonstrate that, rather than causing truly stable perception, intermittent presentation gives rise to a perceptual alternation cycle with its own characteristics and dependencies, different from those during continuous presentation. Alternations during intermittent viewing typically occur once every few minutes--much less frequently than the seconds-scale alternations during continuous viewing. Strikingly, alternations during intermittent viewing occur at fairly regular intervals, making for a surprisingly periodic alternation cycle. The duration of this cycle becomes longer as the blank duration between presentations is increased, reaching dozens of minutes in some cases. We interpret our findings in terms of a mathematical model that describes a neural network with competition between alternative interpretations. Network sensitivities depend on prior dominance, thus providing a memory for past perception. Slow changes in sensitivity produce both perceptual stabilization and the regular but infrequent alternations, meaning that the same memory traces are responsible for both. This model provides a good description of psychophysical findings, and offers several indications regarding their neural basis.


Meeting the Iron Needs of Low and Very Low Birth Weight Infants.
Low birth weight (LBW), defined as a birth weight of <2,500 g, affects 16% of all newborns and is a risk factor for impaired neurodevelopment as well as adverse cardiovascular and metabolic outcomes, including hypertension. LBW infants include both term, small for gestational age infants and preterm infants. Most LBW infants have only marginally LBW (2,000-2,500 g). Recent advances in neonatal care have significantly improved the survival of very LBW (VLBW) infants (<1,500 g). LBW infants are at high risk of iron deficiency due to low iron stores at birth and higher iron requirements due to rapid growth. Using a factorial approach, iron requirements of LBW infants have been estimated to be 1-2 mg/kg/day, which is much higher than the requirements of term, normal birth weight infants, who need almost no dietary iron during the first 6 months of life. In VLBW infants, blood losses and blood transfusions related to neonatal intensive care, as well as erythropoietin treatment, will greatly influence iron status and iron requirements. The timing of umbilical cord clamping at birth is of great importance for the amount of blood transfused from the placenta to the newborn and thereby total body iron. Delayed cord clamping of LBW infants is associated with less need for blood transfusion, less intraventricular hemorrhage, and less necrotizing enterocolitis. Randomized controlled trials have shown that an iron intake of 1-3 mg/kg/day (1-2 mg for marginally LBW and 2-3 mg for VLBW) is needed to effectively prevent iron deficiency. There is some recent evidence that these levels of iron intake will prevent some of the negative health consequences associated with LBW, especially behavioral problems and other neurodevelopmental outcomes and possibly even hypertension. However, it is also important to avoid excessive iron intakes which have been associated with adverse effects in LBW infants.


The SM protein Car/Vps33A regulates SNARE-mediated trafficking to lysosomes and lysosome-related organelles.
The SM proteins Vps33A and Vps33B are believed to act in membrane fusions in endosomal pathways, but their specific roles are controversial. In Drosophila, Vps33A is the product of the carnation (car) gene. We generated a null allele of car to test its requirement for trafficking to different organelles. Complete loss of car function is lethal during larval development. Eye-specific loss of Car causes late, light-independent degeneration of photoreceptor cells. Earlier in these cells, two distinct phenotypes were detected. In young adults, autophagosomes amassed indicating that their fusion with lysosomes requires Car. In eye discs, endocytosed receptors and ligands accumulate in Rab7-positive prelysosomal compartments. The requirement of Car for late endosome-to-lysosome fusion in imaginal discs is specific as early endosomes are unaffected. Furthermore, lysosomal delivery is not restored by expression of dVps33B. This specificity reflects the distinct pattern of binding to different Syntaxins in vitro: dVps33B predominantly binds the early endosomal Avl and Car to dSyntaxin16. Consistent with a role in Car-mediated fusion, dSyntaxin16 is not restricted to Golgi membranes but also present on lysosomes.


A new diagnostic algorithm for Burkitt and diffuse large B-cell lymphomas based on the expression of CSE1L and STAT3 and on MYC rearrangement predicts outcome.
Aggressive mature B-cell non-Hodgkin's lymphomas (BCL) sharing features of Burkitt's lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) (intermediate BL/DLBCL) but deviating with respect to one or more characteristics are increasingly recognized. The limited knowledge about these biologically heterogeneous lymphomas hampers their assignment to a known entity, raising incertitude about optimal treatment approaches. We therefore searched for discriminative, prognostic, and predictive factors for their better characterization. We analyzed 242 cytogenetically defined aggressive mature BCL for differential protein expression. Marker selection was based on recent gene-expression profile studies. Predictive models for diagnosis were established and validated by a different set of lymphomas. CSE1L- and inhibitor of DNA binding-3 (ID3)-overexpression was associated with the diagnosis of BL and signal transduction and transcription-3 (STAT3) with DLBCL (P<0.001 for all markers). All three markers were associated with patient outcome in DLBCL. A new algorithm discriminating BL from DLBCL emerged, including the expression of CSE1L, STAT3, and MYC translocation. This 'new classifier' enabled the identification of patients with intermediate BL/DLBCL who benefited from intensive chemotherapy regimens. The proposed algorithm, which is based on markers with reliable staining properties for routine diagnostics, represents a novel valid tool in separating BL from DLBCL. Most interestingly, it allows segregating intermediate BL/DLBCL into groups with different treatment requirements.


Aortofemoral bypass in young patients with premature atherosclerosis: is superficial femoral vein superior to Dacron?
Previous studies have documented poor patency rates in "young" patients (age 55 years or younger) with premature atherosclerosis undergoing aortofemoral bypass (AFB) to treat aortoiliac occlusive disease. Given the high reported graft patency rates with superficial femoral vein (SFV) grafts performed because of aortic graft infection, we evaluated the role of SFV grafts for AFB as primary therapy for premature atherosclerosis in a case-control study. Over 10 years 31 patients aged 55 year or younger underwent AFB with use of SFV (V-AFB). Case controls consisted of all patients 55 years of age or younger who underwent AFB with use of Dacron (D-AFB) during the same period (n = 80). In all cases this was the initial therapy (no repeat operations). The two groups were well matched for age, sex, weight, preoperative ankle-brachial index, and the comorbid conditions of smoking, coronary artery disease, chronic obstructive pulmonary disease, hyperlipidemia, hypertension, and renal insufficiency. There were more patients with diabetes in the V-AFB group (34% vs 16%; P =.05). Patients in the V-AFB group had more advanced disease, and the surgical indication was more frequently critical ischemia compared with the D-AFB group (90% vs 46%; P <.001). There was only one perioperative death in each group. There were no differences in cardiac, pulmonary, or gastrointestinal complications. However, fasciotomy occurred more frequently with V-AFB (44% vs 1%; P <.001). Surgery time was longer with V-AFB (7.3 vs 4.5 hours; P <.001). Despite these short-term drawbacks, V-AFB proved superior at long-term follow-up. The 5-year primary patency rate was significantly higher with V-AFB than with D-AFB (100% vs 56%; P =.013). There was also a trend for higher limb salvage at 5 years (90% vs 62%). Four graft infections occurred with D-AFB, and none with V-AFB (P =.32). AFB performed with SFV grafts is a far more durable operation than standard D-AFB in young patients with aortoiliac occlusive disease. However, V-AFB is far more likely to require lower extremity fasciotomy, and takes almost twice as long to perform.


Focal segmental glomerulosclerosis. A common entity in nephrotic black adults.
We studied 100 renal biopsy specimens from adults with the primary nephrotic syndrome in an inner city hospital serving mostly black patients and found that 47 had focal segmental glomerulosclerosis. Most of the men presented in the third decade of life, a peak distribution not seen in women. Half of the patients were hypertensive at presentation. Two thirds of the patients had not used intravenous drugs. The addicts were younger than nonaddicts (mean +/- SD age, 27 +/- 4 years vs 35 +/- 13 years), had greater proteinuria (10 +/- 5 g/d vs 6.3 +/- 5 g/d), and exhibited more glomerulosclerosis and tubulointerstitial fibrosis on biopsy. Of the 18 patients (8 addicts) remaining under our care, 4 addicts and 4 nonaddicts became uremic within 3 years. We conclude that even in the absence of drug addiction, focal segmental glomerulosclerosis is a common cause of primary glomerular disease in black adults, in whom it may represent a nonspecific glomerular reaction to injury. The prognosis in the nonaddict may not be different from that in the addict, but more patients need to be studied.


Calcium-activated chloride conductance in frog olfactory cilia.
We have measured the effects of cytoplasmic Ca2+ on the conductance of single cilia excised from frog olfactory receptor neurons. When free cytoplasmic Ca2+ is buffered at 0.1 microM, ciliary conductance is low. As Ca2+ is increased, ciliary conductance increases. Maximal conductance averages sevenfold higher than that measured in the absence of Ca2+. We estimate that the K1/2 for Ca2+ activation is 5 microM; the dose-response curve indicates some positive cooperativity of Ca2+ binding. Activation by Ca2+ is rapid and fully reversible. Most of the Ca(2+)-activated current is carried by Cl- and persists in the absence of Na+ and K+. The Cl- channel inhibitor 3',5-dichlorodiphenylamine-2-carboxylate (300 microM) reduces the Ca(2+)-activated current by 90%. Odorants induce a Ca2+ influx in some olfactory receptor neurons, but the consequences of this influx for neuronal function are not well understood. Our findings allow us to predict that a Ca2+ influx would increase the permeability of the olfactory cilia to Cl-. How this would affect the neuronal potential is uncertain, since the equilibrium potential for Cl- in olfactory receptor neurons is unknown.


Proteomic profiling reveals autoimmune targets in sarcoidosis.
There is a need to further characterize the antibody repertoire in relation to sarcoidosis and potentially related autoantigens. We investigated bronchoalveolar lavage (BAL) and serum samples from patients with sarcoidosis and healthy and diseased control subjects to discover sarcoidosis-associated autoantigens. Antigen microarrays built on 3,072 protein fragments were used to screen for IgG reactivity in 73 BAL samples from subjects with sarcoidosis, subjects with asthma, and healthy subjects. A set of 131 targets were selected for subsequent verification on suspension bead arrays using 272 additional BAL samples and 141 paired sera. Reactivity to four antigens was furthermore analyzed in 22 unprocessed BAL samples from patients with fibrosis and 269 plasma samples from patients diagnosed with myositis. Reactivity toward zinc finger protein 688 and mitochondrial ribosomal protein L43 were discovered with higher frequencies in patients with sarcoidosis, for mitochondrial ribosomal protein L43 especially in patients with non-Löfgren syndrome. Increased reactivity toward nuclear receptor coactivator 2 was also observed in patients with non-Löfgren syndrome as compared with patients with Löfgren syndrome. The antigen representing adenosine diphosphate-ribosylation factor GTPase activating protein 1 revealed high reactivity frequency in all sample groups but with significantly higher level of IgG reactivities in patients with sarcoidosis. Autoantigen reactivity was present in most BAL and serum samples analyzed, and the results revealed high interindividual heterogeneity, with most of the reactivities observed in single individuals only. Four proteins are here proposed as sarcoidosis-associated autoimmune targets and of interest for further validation in independent cohorts.


Children's interpretive understanding, moral judgments, and emotion attributions: relations to social behaviour.
The study investigated interpretive understanding, moral judgments, and emotion attributions in relation to social behaviour in a sample of 59 5-year-old, 123 7-year-old, and 130 9-year-old children. Interpretive understanding was assessed by two tasks measuring children's understanding of ambiguous situations. Moral judgments and emotion attributions were measured using two moral rule transgressions. Social behaviour was assessed using teachers' ratings of aggressive and prosocial behaviour. Aggressive behaviour was positively related to interpretive understanding and negatively related to moral reasoning. Prosocial behaviour was positively associated with attribution of fear. Moral judgments and emotion attributions were related, depending on age. Interpretive understanding was unrelated to moral judgments and emotion attributions. The findings are discussed in regard to the role of interpretive understanding and moral and affective knowledge in understanding children's social behaviour.


Regulation of Kv4 channel expression in failing rat heart by the thioredoxin system.
Redox imbalance elicited by oxidative stress contributes to pathogenic remodeling of ion channels that underlies arrhythmogenesis and contractile dysfunction in the failing heart. This study examined whether the expression of K(+) channels in the remodeled ventricle is controlled by the thioredoxin system, a principal oxidoreductase network regulating redox-sensitive proteins. Ventricular dysfunction was induced in rats by coronary artery ligation, and experiments were conducted 6-8 wk postinfarction. Biochemical assays of tissue extracts from infarcted hearts showed that thioredoxin reductase activity was decreased by 32% from sham-operated controls (P < 0.05), whereas thioredoxin activity was 51% higher postinfarction (P < 0.05). These differences in activities paralleled changes in protein abundance as determined by Western blot analysis. However, whereas real-time PCR showed thioredoxin reductase mRNA levels to be significantly decreased postinfarction, thioredoxin mRNA was not altered. In voltage-clamp studies of myocytes from infarcted hearts, the characteristic downregulation of transient-outward K(+) current density was reversed by exogenous pyruvate (5 mmol/l), and this effect was blocked by the specific inhibitors of the thioredoxin system: auranofin or 13-cis-retinoic acid. Real-time PCR and Western blot analyses of myocyte suspensions from infarcted hearts showed that pyruvate increased mRNA and protein abundance of Kv4.2 and Kv4.3 channel alpha-subunits as well as the accessory protein KChIP2 when compared with time-matched, untreated cells (P < 0.05). The pyruvate-induced increase in Kv4.x expression was blocked by auranofin, but the upregulation of KChIP2 expression was not affected. These data suggest that the expression of Kv4.x channels is redox-regulated by the thioredoxin system, which may be a novel therapeutic target to reverse or limit electrical remodeling of the failing heart.


Multiple primary melanomas among 16,570 patients with melanoma diagnosed at Kaiser Permanente Northern California, 1996 to 2011.
Published rates of cutaneous multiple primary melanoma (MPM) vary widely. We examined incidence of and risk factors associated with MPMs among Kaiser Permanente Northern California members. We estimated MPM incidence among 16,570 patients with melanoma from 1996 through 2011. We compared characteristics between patients with MPMs and single primary melanomas and estimated crude and adjusted hazard ratios of MPMs using Cox models. In all, 15,448 patients had a single melanoma and 1122 had MPMs. Patients with MPMs were older and more often male, non-Hispanic white, and partnered. Subsequent primary melanomas were diagnosed after a mean of 3.83 (SD 3.61, median 2.82) years and were more likely in situ and thinner than initial tumors. The risk of a subsequent melanoma decreased from 2% in the first year after diagnosis to a stable approximately 1% rate through 15 years of follow-up. We lacked data on some known melanoma risk factors and had small numbers of non-white patients and certain tumor subtypes. The risk of MPMs, although highest in the first year after diagnosis, remains stable thereafter. Those at highest risk of MPMs are older, male, white, and partnered. Clinicians should be aware of the rate of MPMs and recognize high-risk subgroups.


Replicating phages in the epidermal mucosa of the eel (Anguilla anguilla).
In this work, we used the eel (Anguilla anguilla) as an animal model to test the hypothesis of Barr et al. (2013a,b) about the putative role of the epidermal mucosa as a phage enrichment layer. To this end, we analyzed the microbial content of the skin mucus of wild and farmed eels by using a metagenomic approach. We found a great abundance of replicating phage genomes (concatemers) in all the samples. They were assembled in four complete genomes of three Myovirus and one Podovirus. We also found evidences that ΦKZ and Podovirus phages could be part of the resident microbiota associated to the eel mucosal surface and persist on them over the time. Moreover, the viral abundance estimated by epiflorescent counts and by metagenomic recruitment from eel mucosa was higher than that of the surrounding water. Taken together, our results support the hypothesis that claims a possible role of phages in the animal mucus as agents controlling bacterial populations, including pathogenic species, providing a kind of innate immunity.


Neonatal hypothalamic androgenization in the female rat induces changes in peripheral insulin sensitivity and adiposity function at adulthood.
It is recognized that there exists a link between hyperandrogenicity and insulin resistance. By using the neonatally androgenized female rat we explored whether this treatment modifies peripheral insulin sensitivity and visceral fat function at adulthood. On day 5 of age, female Sprague-Dawley pups were injected, sub cutaneous, with either 50 ml of sterile corn oil alone (CT) or containing 1.25 mg of testosterone propionate (TP) and further used for experimentation on day 100 of age. CT and TP rats were killed by decapitation in non-fasting condition and blood samples were kept frozen for measurement of different metabolites. Immediately after sacrifice, freshly dissected visceral fat pads were used for isolation of adipocytes, these cells were then incubated with medium alone or containing different concentrations of insulin in order to determine leptin secreted into the medium. Additionally, in vivo metabolic responses to intravenous high glucose load were performed in, 24 hour-fasting, CT and TP rats. We found that neonatal androgenization induced adult animals displaying higher visceral adiposity mass, body weight and leptinemia than CT rats. No group differences were found in basal circulating levels of several hormones and metabolic parameters. The results of the high glucose load 90-min test indicated that TP and CT rats developed similar glycemia but this accounted because of an early significantly higher peak values of circulating insulin in TP than in CT rats, regardless of similar enhancement in circulating glucocorticoid concentrations in both groups. While high glucose load significantly increased, over the baseline, circulating leptin concentrations as early as 30 min post-glucose in CT rats, in TP animals, it significantly enhanced leptinemia only by the end of the test. Finally, results of in vitro incubations of isolated visceral adipocytes indicated that cells from androgenized rats spontaneously released more leptin than control cells, although they were less responsive than CT cells to insulin-induced leptin output. Our study strongly supports the hypothesis that development of insulin resistance seems to be dependent on early hyperandrogenicity.


Isoform-selective inactivation of human arylamine N-acetyltransferases by reactive metabolites of carcinogenic arylamines.
Human arylamine N-acetyltransferases (NATs) are expressed as two polymorphic isoforms, NAT1 and NAT2, that have toxicologically significant functions in the detoxification of xenobiotic arylamines by N-acetylation and in the bioactivation of N-arylhydroxylamines by O-acetylation. NAT1 also catalyzes the N-acetylation of 4-aminobenzoylglutamic acid, a product of folic acid degradation, and is associated with endogenous functions in embryonic development. On the basis of earlier studies with hamster NAT1, hamster NAT2, and human NAT1, we proposed that human NAT2 would be more susceptible than NAT1 to inactivation by N-arylhydroxamic acid metabolites of arylamines. Kinetic analyses of the inactivation of recombinant NAT1 and NAT2 by the N-arylhydroxamic acid, N-hydroxy-2-acetylaminofluorene (N-OH-AAF), as well as the inactivation of NAT2 by N-hydroxy-4-acetylaminobiphenyl (N-OH-4-AABP), resulted in second-order inactivation rate constants (k(inact)/K(I)) that were several fold greater for NAT2 than for NAT1. Mass spectrometric analysis showed that inactivation of NAT2 in the presence of the N-arylhydroxamic acids was due to formation of a sulfinamide adduct with Cys68. Treatment of HeLa cells with N-OH-4-AABP and N-OH-AAF revealed that the compounds were less potent inactivators of intracellular NAT activity than the corresponding nitrosoarenes, but unexpectedly, the hydroxamic acids caused a significantly greater loss of NAT1 activity than of NAT2 activity. Nitrosoarenes are the electrophilic products responsible for NAT inactivation upon interaction of the enzymes with N-arylhydroxamic acids, as well as being metabolic products of arylamine oxidation. Treatment of recombinant NAT2 with the nitrosoarenes, 4-nitrosobiphenyl (4-NO-BP) and 2-nitrosofluorene (2-NO-F), caused rapid and irreversible inactivation of the enzyme by sulfinamide adduct formation with Cys68, but the k(inact)/K(I) values for inactivation of recombinant NAT2 and NAT1 did not indicate significant selectivity for either isoform. Also, the IC(50) values for inactivation of HeLa cell cytosolic NAT1 and NAT2 by 4-NO-BP were similar, as were the IC(50) values obtained with 2-NO-F. Treatment of HeLa cells with low concentrations (1-10 microM) of either 4-NO-BP or 2-NO-F resulted in preferential and more rapid loss of NAT1 activity than NAT2 activity. Because of its wide distribution in human tissues and its early expression in developing tissues, the apparent high sensitivity of intracellular NAT1 to inactivation by reactive metabolites of environmental arylamines may have important toxicological consequences.


Structural basis for high-affinity HER2 receptor binding by an engineered protein.
The human epidermal growth factor receptor 2 (HER2) is specifically overexpressed in tumors of several cancers, including an aggressive form of breast cancer. It is therefore a target for both cancer diagnostics and therapy. The 58 amino acid residue Zher2 affibody molecule was previously engineered as a high-affinity binder of HER2. Here we determined the structure of Zher2 in solution and the crystal structure of Zher2 in complex with the HER2 extracellular domain. Zher2 binds to a conformational epitope on HER2 that is distant from those recognized by the therapeutic antibodies trastuzumab and pertuzumab. Its small size and lack of interference may provide Zher2 with advantages for diagnostic use or even for delivery of therapeutic agents to HER2-expressing tumors when trastuzumab or pertuzumab are already employed. Biophysical characterization shows that Zher2 is thermodynamically stable in the folded state yet undergoing conformational interconversion on a submillisecond time scale. The data suggest that it is the HER2-binding conformation that is formed transiently prior to binding. Still, binding is very strong with a dissociation constant K(D) = 22 pM, and perfect conformational homogeneity is therefore not necessarily required in engineered binding proteins. A comparison of the original Z domain scaffold to free and bound Zher2 structures reveals how high-affinity binding has evolved during selection and affinity maturation and suggests how a compromise between binding surface optimization and stability and dynamics of the unbound state has been reached.


Black pepper essential oil to enhance intravenous catheter insertion in patients with poor vein visibility: a controlled study.
To evaluate the effect of topically applied black pepper essential oil on easing intravenous catheter insertion (IVC) in patients with no palpable or visible veins compared to a control group (standard nursing practice). Randomized, controlled study. One hundred twenty hospitalized patients, who were referred to a hospital vascular team because of difficulty in accessing veins for IVC insertion. Topical application of 20% essential oil of black pepper in aloe vera gel or standard nursing care (hot packs with or without vigorous tactile stimulation). Pre- and post-test vein visibility and/or palpability and number of attempts at IVC insertion. A higher percentage of patients achieved optimal scoring (vein score=2) or improved scoring (vein score of 1 or 2) to black pepper intervention than standard nursing care. The black pepper group also reduced the number of patients whose veins were still not visible or palpable after the intervention to nearly half that of the control group (p<0.05). The number of IVC attempts following black pepper was also half that of the control group. Topical application of black pepper is a viable and effective way to enhance vein visibility and palpability prior to intravenous insertion in patients with limited vein accessibility; it also improves ease of IVC insertion.


Ultrasonographic findings in patients with peristomal bulging.
The aim of this study was to obtain a classification of peristomal bulging based on findings at ultrasonography in patients with a sigmoid colostomy. The patient material comprised 30 men and 33 women. The median age was 69 years (28-90) and the median time between stoma creation and investigation was 68 months (3-426). Any bulging was measured, and the abdominal opening for the stoma bowel was evaluated at the clinical examination. At the ultrasonographic investigation, the patients were first investigated in the supine position to measure the transverse and vertical diameter of the abdominal opening and the thickness of the abdominal muscles. Three types of ultrasonographic findings were identified. In ultra-I, the stoma bowel was completely fixed or showed telescoping-like movement through the abdominal opening. In ultra-II, fatty tissue was prolapsed together with the stoma bowel forming a bend in the subcutaneous tissue. In ultra-III, another bowel segment or fatty tissue passed beside the stoma bowel through the abdominal opening into the abdominal wall. A normal finding without any bulging at the clinical examination was associated with a smaller area and a smaller diameter of the abdominal opening than the area and diameter in patients with a visible peristomal bulging. There was no difference in the thickness of the muscle layer of the abdominal wall between patients with and without bulging. Ultrasonography can make a dynamic diagnosis of parastomal hernia. In patients with visible peristomal bulging, the area of the abdominal opening is increased but there is no decrease in the thickness of the muscles of the abdominal wall.


Independent prognostic impact of lymphatic vessel density and presence of low-grade lymphangiogenesis in cutaneous melanoma.
The aim of this study was to determine lymphatic vessel density (LVD) in a series of nodular melanoma and correlate the findings with the expression of several angiogenic factors, including vascular endothelial growth factor-C, basic fibroblast growth factor (bFGF), patient survival, and clinico-pathologic data. Patients with nodular melanoma and complete follow-up information were included. Lymphatic vessels were immunostained with the LYVE-1 and Podoplanin antibodies, and LVD was evaluated in both intra- and peri-tumoral (LVDpt) areas. Median LVD was 6.3 and 12.5 vessels/mm(2) in intra- and peri-tumoral areas, and coexpression of LYVE-1 and Ki-67/MIB-1 in lymphatic endothelial cells within the tumor was demonstrated, indicating active but low-grade lymphangiogenesis. Increased LVDpt was significantly associated with localization on the extremities (P = 0.005), decreased tumor thickness (P = 0.036), absence of vascular invasion (P = 0.004), brisk lymphocytic infiltration (P = 0.018), low proliferative rate by Ki-67 (P = 0.011), increased bFGF expression in tumor cells (P = 0.01) as well as in endothelial cells (P = 0.008), and decreased tumor cell expression of Ephrin-A1 (P = 0.009). Decreased LVD in intra-tumoral areas and LVDpt both predicted improved survival rates in multivariate analyses (for LVDpt, Hazard ratio: 2.1, P = 0.009). We found that decreased LVD was present in thicker and more proliferative tumors (Ki-67) and that increased LVD was significantly associated with improved patient survival in multivariate analysis. In addition, our data suggest the presence of low-grade intra-tumoral lymphangiogenesis in melanoma and a stimulating role of bFGF in lymphangiogenesis.


Testosterone replacement therapy improves insulin resistance, glycaemic control, visceral adiposity and hypercholesterolaemia in hypogonadal men with type 2 diabetes.
Low levels of testosterone in men have been shown to be associated with type 2 diabetes, visceral adiposity, dyslipidaemia and metabolic syndrome. We investigated the effect of testosterone treatment on insulin resistance and glycaemic control in hypogonadal men with type 2 diabetes. This was a double-blind placebo-controlled crossover study in 24 hypogonadal men (10 treated with insulin) over the age of 30 years with type 2 diabetes. Patients were treated with i.m. testosterone 200 mg every 2 weeks or placebo for 3 months in random order, followed by a washout period of 1 month before the alternate treatment phase. The primary outcomes were changes in fasting insulin sensitivity (as measured by homeostatic model index (HOMA) in those not on insulin), fasting blood glucose and glycated haemoglobin. The secondary outcomes were changes in body composition, fasting lipids and blood pressure. Statistical analysis was performed on the delta values, with the treatment effect of placebo compared against the treatment effect of testosterone. Testosterone therapy reduced the HOMA index (-1.73 +/- 0.67, P = 0.02, n = 14), indicating an improved fasting insulin sensitivity. Glycated haemoglobin was also reduced (-0.37 +/- 0.17%, P = 0.03), as was the fasting blood glucose (-1.58 +/- 0.68 mmol/l, P = 0.03). Testosterone treatment resulted in a reduction in visceral adiposity as assessed by waist circumference (-1.63 +/- 0.71 cm, P = 0.03) and waist/hip ratio (-0.03 +/- 0.01, P = 0.01). Total cholesterol decreased with testosterone therapy (-0.4 +/- 0.17 mmol/l, P = 0.03) but no effect on blood pressure was observed. Testosterone replacement therapy reduces insulin resistance and improves glycaemic control in hypogonadal men with type 2 diabetes. Improvements in glycaemic control, insulin resistance, cholesterol and visceral adiposity together represent an overall reduction in cardiovascular risk.


Factors associated with medical students speaking-up about medical errors: A cross-sectional study.
Training medical students to speak up when they witness a potential error is an important competency for patient safety, but details regarding the barriers that prevent medical students from effectively communicating are lacking. Therefore, this study aimed at exploring the factors affecting medical students' willingness to speak up for patient safety when a medical error was observed. This is a cross-sectional study at a medical university in Taiwan, and 151 medical students in clinical clerkship completed a survey including demographic characteristics, conflict of interests/social relationship, personal capability, and personality and characteristics of senior staff domains. Data were analyzed using t-test. Three of five items in the conflict of interests/social relationship domain showed statistically significant importance, including 'I am afraid of being punished' (Mean difference, MD = 0.37; p < 0.01), 'I do not want to break unspoken rules' (MD = 0.55; p < 0.01), and 'I do not want to have bad team relationship' (MD = 0.58; p < 0.01). Two items (perception of knowledge/understanding and communication skills) in the personal capability domain were significantly important to speaking up. Six of 10 items in personality and characteristics of senior staff domain were rated significantly important in deciding to speak up. The top three factors of them were senior personnel with 'Grumpy' personality (MD = 1.20; p < 0.01), 'hierarchy gap' (MD = 1.12; p < 0.01), and senior personnel with 'Stubborn' personality (MD = 1.06; p < 0.01). Our findings demonstrated medical students' perspectives on barriers to speaking up in the event of medical error. Some factors related to characteristics of senior staff could compromise medical students' ability to speak up in the event of medical error. These results might be important for medical educators in designing personalized educational activities related to medical students' ability to speak up for patient safety.


Large-scale genomic analysis of antimicrobial resistance in the zoonotic pathogen Streptococcus suis.
Antimicrobial resistance (AMR) is among the gravest threats to human health and food security worldwide. The use of antimicrobials in livestock production can lead to emergence of AMR, which can have direct effects on humans through spread of zoonotic disease. Pigs pose a particular risk as they are a source of zoonotic diseases and receive more antimicrobials than most other livestock. Here we use a large-scale genomic approach to characterise AMR in Streptococcus suis, a commensal found in most pigs, but which can also cause serious disease in both pigs and humans. We obtained replicated measures of Minimum Inhibitory Concentration (MIC) for 16 antibiotics, across a panel of 678 isolates, from the major pig-producing regions of the world. For several drugs, there was no natural separation into 'resistant' and 'susceptible', highlighting the need to treat MIC as a quantitative trait. We found differences in MICs between countries, consistent with their patterns of antimicrobial usage. AMR levels were high even for drugs not used to treat S. suis, with many multidrug-resistant isolates. Similar levels of resistance were found in pigs and humans from regions associated with zoonotic transmission. We next used whole genome sequences for each isolate to identify 43 candidate resistance determinants, 22 of which were novel in S. suis. The presence of these determinants explained most of the variation in MIC. But there were also interesting complications, including epistatic interactions, where known resistance alleles had no effect in some genetic backgrounds. Beta-lactam resistance involved many core genome variants of small effect, appearing in a characteristic order. We present a large dataset allowing the analysis of the multiple contributing factors to AMR in S. suis. The high levels of AMR in S. suis that we observe are reflected by antibiotic usage patterns but our results confirm the potential for genomic data to aid in the fight against AMR.


Three-dimensional imaging in aberration-corrected electron microscopes.
This article focuses on the development of a transparent and uniform understanding of possibilities for three-dimensional (3D) imaging in scanning transmission and confocal electron microscopes (STEMs and SCEMs), with an emphasis on the annular dark-field STEM (ADF-STEM), bright-field SCEM (BF-SCEM), and ADF-SCEM configurations. The incoherent imaging approximation and a 3D linear imaging model for ADF-STEM are reviewed. A 3D phase contrast model for coherent-SCEM as well as a pictorial way to find boundaries of information transfer in reciprocal space are reviewed and applied to both BF- and ADF-SCEM to study their 3D point spread functions and contrast transfer functions (CTFs). ADF-STEM is capable of detecting the depths of dopant atoms in amorphous materials but can fail for crystalline materials when channeling substantially modifies the electron propagation. For the imaging of extended (i.e., nonpointlike) features, ADF-STEM and BF-SCEM exhibit strong elongation artifacts due to the missing cone of information. ADF-SCEM shows an improvement over ADF-STEM/BF-SCEM due to its differential phase contrast eliminating slowly varying backgrounds, an effect that partially suppresses the elongation artifacts. However, the 3D CTF still has a cone of missing information that will result in some residual feature elongation as has been observed in A. Hashimoto et al., J Appl Phys 160(8), 086101 (2009).


Induction of specific human cytotoxic T cells using dendritic cells transduced with an adenovector encoding rat epidermal growth factor receptor 2.
This study demonstrates the ability to generate antigen-specific cytotoxic T cells (CTLs) against HER2 using a xenoantigenic immune stimulation strategy. Dendritic cells (DCs) were transduced with an adenovirus vector incorporating full-length cDNA for rat (xenoantigen) epidermal growth factor receptor 2 (Adv-HER2). Stimulation of autologous T cells with Adv-HER2 infected DCs led to enhanced HER2-specific reactivity as assessed by quantitative real-time polymerase chain reaction (qRT-PCR) for T cell IFN-γ mRNA. In ELISPOT and intracellular cytokine staining (ICS) assays, CD8+ CTLs induced by Adv-HER2 transduced DCs released IFN-γ following stimulation with irradiated autologous DCs infected with Adv-HER2 or loaded with a human prostate cancer cell line (LNCaP) lysate. DCs pulsed with HER2 peptides were less stimulatory than Adv-HER2 transduced DCs. HER2 DC induced CTL lysed HER2+ HLA-A2+ tumor cells (MCF-7); significantly reduced lysis occurred in HER2+ HLA-A2- tumor cells (SKOV-3), and the NK cell sensitive cell line K-562.


The MRI Sepsis Score: An Innovative Tool for the Evaluation of Septic Peritonitis in Mice Using 7-Tesla Small Animal MRI.
Magnetic resonance imaging (MRI) techniques are rarely used in the context of abdominal sepsis and in sepsis research. This study investigates the impact of MRI for monitoring septic peritonitis in an animal model (colon ascendens stent-induced peritonitis, CASP). The CASP model closely mimics that of human disease and is highly standardized. The most frequently employed readout parameter in mouse CASP studies is prolonged or decreased rate of survival. Monitoring the progression of peritonitis via MRI could provide a helpful tool in the evaluation of severity. The use of alternative readout systems could very well reduce the number of research animals. Perspectively, clinical improvement after certain treatment could be classified. This study describes for the first time MRI findings following the induction of septic peritonitis in mice using the CASP model. Two sublethal groups of mice with septic peritonitis were investigated. Each had received one of two differing stent diameters in order to control the leakage of feces into the abdominal cavity. Each mouse served as its own control. Imaging and analyses were performed blinded. Gut diameters, stomach volume, abdominal organ wall diameters, and volume of the adrenal glands were measured. Serum corticosterone levels were detected using ELISA. Serum IL-6, TNF-α, IL-1β, and IL-10 levels were screened by cytometric bead array. Statistical analysis was performed using the Mann-Whitney U test for nonparametric probes and the Kruskal-Wallis and t tests. Using a 7-tesla MRI scanner 24 and 48 h after induction of septic peritonitis, interenteric fluid, organ swelling of spleen and adrenal glands, as well as dilatation of the stomach were compared to nonseptic conditions. Swelling of adrenal glands resulted in an increased serum corticosterone level. In addition, the wall of the intestine bowel was thickened. Based upon these findings, an MRI score (MRI sepsis score, MSS) for abdominal sepsis in mice was established. Reduced stent sizes led to reduced severity of the abdominal sepsis, which could be reproduced in the MSS, which is described here for the first time. Intraabdominal variations during septic peritonitis are detectable by MRI techniques. MRI methods should become a more important tool for the evaluation of abdominal peritonitis. MSS could provide an interesting tool for the evaluation of therapeutic strategies.


Phase stabilization of Al:HfO2 grown on In(x)Ga(1-x)As substrates (x = 0, 0.15, 0.53) via trimethylaluminum-based atomic layer deposition.
Al:HfO2 is grown on III-V compound substrates by atomic layer deposition after an in situ trimethylaluminum-based preconditioning treatment of the III-V surface. After post-deposition rapid thermal annealing at 700 °C, the cubic/tetragonal crystalline phase is stabilized and the chemical composition of the stack is preserved. The observed structural evolution of Al:HfO2 on preconditioned III-V substrates shows that the in-diffusion of semiconductor species from the substrate through the oxide is inhibited. Al-induced stabilization of the Al:HfO2 crystal polymorphs up to 700 °C can be used as a permittivity booster methodology with possibly important implications in the stack scaling issues of high-mobility III-V based logic applications.


Essential role of Smad3 in infarct healing and in the pathogenesis of cardiac remodeling.
Postinfarction cardiac repair is regulated through timely activation and repression of inflammatory pathways, followed by transition to fibrous tissue deposition and formation of a scar. The transforming growth factor-beta/Smad3 pathway is activated in healing infarcts and may regulate cellular events critical for the inflammatory and the fibrotic responses. We examined the effects of Smad3 gene disruption on infarct healing and the pathogenesis of cardiac remodeling. In the absence of injury, Smad3-null hearts had comparable function to and similar morphology as wild-type hearts. Smad3-null animals had suppressed peak chemokine expression and decreased neutrophil recruitment in the infarcted myocardium but showed timely repression of inflammatory gene synthesis and resolution of the inflammatory infiltrate. Although myofibroblast density was higher in Smad3-null infarcts, interstitial deposition of collagen and tenascin-C in the remodeling myocardium was markedly reduced. Compared with wild-type animals, Smad3-/- mice exhibited decreased dilative remodeling and attenuated diastolic dysfunction; however, infarct size was comparable between groups. Transforming growth factor-beta-mediated induction of procollagen type III and tenascin-C in isolated cardiac fibroblasts was dependent on Smad3, which suggests that decreased fibrotic remodeling in infarcted Smad3-null hearts may be due to abrogation of the profibrotic transforming growth factor-beta responses. Smad3 loss does not alter the time course of resolution of inflammation in healing infarcts, but it prevents interstitial fibrosis in the noninfarcted myocardium and attenuates cardiac remodeling. Thus, the Smad3 cascade may be a promising therapeutic target for the treatment of myocardial infarction.


Productivity of Clinical Trials Conducted in the Gulf Cooperative Council Region.
This study aimed to assess the productivity of clinical trials in the Gulf Cooperation Council (GCC) region, including Oman, Saudi Arabia, United Arab Emirates, Qatar, Bahrain and Kuwait and to address the existing research gap. Specifically, this paper aimed to estimate the number of clinical trials conducted and estimate the proportion of clinical trials conducted in the GCC countries published in peer-reviewed journals. The clinical trials registry of the US National Library of Medicine was searched for clinical trials conducted from January 2000 to October 2019. The productivity was assessed by the publication status of the trials in the registry and through the search in MEDLINE indexed journals. A total of 682 trials were found from the GCC region, with an overall trend of 4.1 trials each year. However, the clinical trial productivity from the region contributes to only 0.37% of the trials globally. When comparing the raw data, Saudi Arabia showed the highest proportion, contributing 66.6% of the clinical trials from the region (P <0.001), while Oman contributed 3.5%. After normalisation to population, Qatar was found to be the highest contributor, with 42.78 trials per million. A total of 238 trials were conducted before 2016, of which 46.6% were published. Saudi Arabia is the leading country in clinical trial productivity in the GCC region. Countries in the GCC should utilise the triple helix model to form partnerships with the industry and improve their contributions to science.


Amantadine treatment is an independent predictor of improved survival in Parkinson's disease.
Amantadine has been used for more than 20 years in the symptomatic treatment of Parkinson's disease (PD). Several recent discoveries suggest that amantadine could also have a neuroprotective effect in PD. We studied survival in all parkinsonism (including PD and other parkinsonian syndromes) patients attending a single clinic, employing standard survival curves and a Cox regression model, to identify independent predictive variables for survival (while taking into account factors potentially associated with both outcome and treatment selection). Amantadine-treated patients (n = 250) were similar to the patients not treated with amantadine (n = 586) in terms of age, gender, type of parkinsonism, Hoehn and Yahr stage and dementia status at initial neurological visit. Amantadine use was an independent predictor of improved survival (p < 0.01). Improved survival was also associated with a higher 10-year expected survival (based on age, gender, and birth year), absence of dementia, type of parkinsonism = PD, and low Hoehn and Yahr stage (I or II) at initial neurologic visit (all p < 0.01); these additional factors occurred in statistically similar proportions in the groups that were and were not treated with amantadine. The association of improved survival with amantadine use may stem from symptomatic benefit or may reflect a "neuroprotective" effect, mediated through N-methyl-D-aspartate (NMDA) receptor antagonism, dopamine uptake blockade activity, or other mechanisms. Our preliminary findings suggest that a prospective, controlled, randomized trial of amantadine's effects on PD progression is warranted.


Serotonin syndrome in the intensive care unit: clinical presentations and precipitating medications.
Serotonin syndrome (SS) is becoming a more frequent diagnosis in the intensive care unit (ICU). We sought to determine the clinical presentation, drug exposures, and outcomes of SS in critically ill patients. A retrospective study of 33 consecutive ICU patients with SS between March 2007 and March 2012 in ICUs in a large teaching hospital. SS was defined using the Hunter Serotonin Toxicity Criteria. Seventeen patients (52%) were admitted for mental status changes, including seven patients (21%) with drug overdose and four cases (12%) in which SS was considered the primary admission diagnosis. In 13 patients (39%) the features of SS developed only after a mean of 6.8 ± 9 days of hospitalization. Most received multiple serotonergic drugs upon diagnosis (median three drugs, range 1-5). Antidepressants were the serotonergic medications most often used before admission, and opioids (principally fentanyl) and antiemetics were the most frequently prescribed new serotonin-enhancing medications. Altered mental status was present in all patients and myoclonus, rigidity, and hyperreflexia were the most prevalent examination signs. All but one patient had documented recovery. The mean time to neurological improvement was 56 ± 5 h, but ranged from 8 to 288 h. There were no cases of renal failure related to rhabdomyolysis, or death or persistent disability caused by SS. SS in the ICU occurs most often because of exposure to multiple serotonergic agents. Continuation of antidepressants plus the addition of opioids and antiemetics during hospitalization are most commonly responsible for this complication.


How epigenomics contributes to the understanding of gene regulation in Toxoplasma gondii.
How apicomplexan parasites regulate their gene expression is poorly understood. The complex life cycle of these parasites implies tight control of gene expression to orchestrate the appropriate expression pattern at the right moment. Recently, several studies have demonstrated the role of epigenetic mechanisms for control of coordinated expression of genes. In this review, we discuss the contribution of epigenomics to the understanding of gene regulation in Toxoplasma gondii. Studying the distribution of modified histones on the genome links chromatin modifications to gene expression or gene repression. In particular, coincident trimethylated lysine 4 on histone H3 (H3K4me3), acetylated lysine 9 on histone H3 (H3K9ac), and acetylated histone H4 (H4ac) mark promoters of actively transcribed genes. However, the presence of these modified histones at some non-expressed genes and other histone modifications at only a subset of active promoters implies the presence of other layers of regulation of chromatin structure in T. gondii. Epigenomics analysis provides a powerful tool to characterize the activation state of genomic loci of T. gondii and possibly of other Apicomplexa including Plasmodium or Cryptosporidium. Further, integration of epigenetic data with expression data and other genome-wide datasets facilitates refinement of genome annotation based upon experimental data.