Immunophenotyping of putative human B1 B cells in healthy controls and common variable immunodeficiency (CVID) patients.
B1 B cells represent a unique subset of B lymphocytes distinct from conventional B2 B cells, and are important in the production of natural antibodies. A potential human homologue of murine B1 cells was defined recently as a CD20(+) CD27(+) CD43(+) cell. Common variable immunodeficiency (CVID) is a group of heterogeneous conditions linked by symptomatic primary antibody failure. In this preliminary report, we examined the potential clinical utility of introducing CD20(+) CD27(+) CD43(+) B1 cell immunophenotyping as a routine assay in a diagnostic clinical laboratory. Using a whole blood assay, putative B1 B cells in healthy controls and in CVID patients were measured. Peripheral blood from 33 healthy donors and 16 CVID patients were stained with relevant monoclonal antibodies and underwent flow cytometric evaluation. We established a rapid, whole blood flow cytometric assay to investigate putative human B1 B cells. Examination of CD20(+) CD27(+) CD43(+) cells is complicated by CD3(+) CD27(+) CD43(hi) T cell contamination, even when using stringent CD20 gating. These can be excluded by gating on CD27(+) CD43(lo-int) B cells. Although proportions of CD20(+)CD27(+)CD43(lo–int) cells within B cells in CVID patients were decreased by 50% compared to controls (P < 0·01), this was not significant when measured as a percentage of all CD27(+) B cells (P = 0·78) [corrected]. Immunophenotypic overlap of this subset with other innate-like B cells described recently in humans is limited. We have shown that putative B1 B cell immunophenotyping can be performed rapidly and reliably using whole blood. CD20(+) CD27(+) CD43(lo-int) cells may represent a distinct B1 cell subset within CD27(+) B cells. CVID patients were not significantly different from healthy controls when existing CD27(+) B cell deficiencies were taken into account.


A Novel Xanthomonas sp. Causes Bacterial Spot of Rose (Rosa spp.).
A bacterial spot of rose (Rosa spp.) caused by a xanthomonad was observed in Florida and Texas. Ten representative strains collected from the two states between 2004 and 2010 were used to determine the taxonomic position of this rose pathogen. Fatty acid methyl ester analysis was performed and a nearly 2-kb 16S-23S rRNA intergenic spacer along with flanking portions of the 16S and 23S rRNA genes were sequenced for selected strains, showing that they were members of the genus Xanthomonas. Multilocus sequence typing and analysis (MLST/MLSA) and pathogenicity tests were conducted to further characterize the Xanthomonas strains. The MLSA, based on six gene fragments-fusA, gapA, gltA, gyrB, lacF, and lepA-showed that the rose strains fell into Xanthomonas axonopodis subgroup 9.2 and shared the highest similarity values (98.8 to 99.7%) with X. alfalfae subsp. citrumelonis of the subgroup. However, principal coordinate analysis grouped the rose strains into a unique cluster distinct from other members of the subgroup according to virulence phenotypes on 11 plant species belonging to five plant families (Araceae, Euphorbiaceae, Rosaceae, Rutaceae, and Solanaceae). Moreover, the rose strains were aggressive on rose and Indian Hawthorn (Rhaphiolepsis indica). On the basis of the MLSA and virulence phenotypes, the pathovar epithet rosa is proposed.


Hemangiomas of the maxillofacial area: Case Report, Classification and Treatment Planning.
Vascular lesions of the maxillofacial area are even more challenging than in other different body district, because of the high aesthetic and functional relevance of this area for the sense organs presence like eye, brain, tongue, ear and nose. For these reasons, we propose an accurate classification based on hemodynamic and histologic aspects usefulthat is for diagnostic and therapeutic planning. A female, 60 years old patient came to our observation showing a vascular lesion of the lower left lip, and complaining for aesthetical and functional impairment. To confirm the diagnosis of vascular malformation and to detect lesion flow rate or other possible localization, a Tc red blood cell scintigraphy was carried out. Result was a venous low flow lesion; hence, sclerotherapy by a 3% Polidocanol solution (Atossisclerol) followed by surgery was planned. The aim of this work was to propose a diagnostic and therapeutic scheme with an integration of ISSVA and a flow rate classifications for a three-step planning based on 1) the biological findings in an early age at the lesion discover with a pharmacological treatment; 2) Hemodynamic study of the lesions at growing age followed by sclerotherapy or embolization; 3) Imaging study of these lesions for patients candidate to surgery when after step 1 and step 2 diagnostic and therapeutic planning results were incompletely successful.


Second-order spatial analysis of epidermal nerve fibers.
Breakthroughs in imaging of skin tissue reveal new details on the distribution of nerve fibers in the epidermis. Preliminary neurologic studies indicate qualitative differences in the spatial patterns of nerve fibers based on pathophysiologic conditions in the subjects. Of particular interest is the evolution of spatial patterns observed in the progression of diabetic neuropathy. It appears that the spatial distribution of nerve fibers becomes more 'clustered' as neuropathy advances, suggesting the possibility of diagnostic prediction based on patterns observed in skin biopsies. We consider two approaches to establish statistical inference relating to this observation. First, we view the set of locations where the nerves enter the epidermis from the dermis as a realization of a spatial point process. Secondly, we treat the set of fibers as a realization of a planar fiber process. In both cases, we use estimated second-order properties of the observed data patterns to describe the degree and scale of clustering observed in the microscope images of blister biopsies. We illustrate the methods using confocal microscopy blister images taken from the thigh of one normal (disease-free) individual and two images each taken from the thighs of subjects with mild, moderate, and severe diabetes and report measurable differences in the spatial patterns of nerve entry points/fibers associated with disease status.


MAPO-18 Catalysts for the Methanol to Olefins Process: Influence of Catalyst Acidity in a High-Pressure Syngas (CO and H2) Environment.
The transition from integrated petrochemical complexes toward decentralized chemical plants utilizing distributed feedstocks calls for simpler downstream unit operations. Less separation steps are attractive for future scenarios and provide an opportunity to design the next-generation catalysts, which function efficiently with effluent reactant mixtures. The methanol to olefins (MTO) reaction constitutes the second step in the conversion of CO2, CO, and H2 to light olefins. We present a series of isomorphically substituted zeotype catalysts with the AEI topology (MAPO-18s, M = Si, Mg, Co, or Zn) and demonstrate the superior performance of the M(II)-substituted MAPO-18s in the conversion of MTO when tested at 350 °C and 20 bar with reactive feed mixtures consisting of CH3OH/CO/CO2/H2. Co-feeding high pressure H2 with methanol improved the catalyst activity over time, but simultaneously led to the hydrogenation of olefins (olefin/paraffin ratio < 0.5). Co-feeding H2/CO/CO2/N2 mixtures with methanol revealed an important, hitherto undisclosed effect of CO in hindering the hydrogenation of olefins over the Brønsted acid sites (BAS). This effect was confirmed by dedicated ethene hydrogenation studies in the absence and presence of CO co-feed. Assisted by spectroscopic investigations, we ascribe the favorable performance of M(II)APO-18 under co-feed conditions to the importance of the M(II) heteroatom in altering the polarity of the M-O bond, leading to stronger BAS. Comparing SAPO-18 and MgAPO-18 with BAS concentrations ranging between 0.2 and 0.4 mmol/gcat, the strength of the acidic site and not the density was found to be the main activity descriptor. MgAPO-18 yielded the highest activity and stability upon syngas co-feeding with methanol, demonstrating its potential to be a next-generation MTO catalyst.


Temperament and behaviour problems in young children with Down syndrome at 12, 30, and 45 months.
Though cross-sectional studies have yielded important information regarding the trajectory of psychopathology in middle childhood and adolescence in Down syndrome, there has been little exploration of maladaptive behaviour in the earliest years of development. In this study, we explore the emergence of maladaptive behaviour in young children with Down syndrome (n=24) and a mental age-matched comparison group (n=33) of young children with developmental disabilities of mixed etiologies. Behavioural data (Bayley Scales of Infant Development, Infant Temperament Questionnaire, Achenbach's Child Behaviour Checklist) were collected for children in each group at 12 months, 30 months, and 45 months. Findings from this study suggest that the onset of internalising behaviour difficulties in young children with Down syndrome may emerge later in early development than in children with developmental disabilities of mixed/nonspecific etiologies. In addition, temperament at Time I appeared to be a much stronger predictor of maladaptive behaviour outcomes at Time 3 in the mixed comparison group than in the Down syndrome group. Implications of this trajectory of the early development of maladaptive behaviour in Down syndrome for intervention for young children with Down syndrome are discussed.


Potential explanation of limb combination performance differences for two-limb coordination tasks.
Rhythmic two-limb coordinated movements in the sagittal plane are variable and inaccurate when the movements are in the opposite direction as compared with those in the same direction (directional constraint). The magnitude of directional constraint depends on the particular limb combination. It is prominent in ipsilateral hand-foot coordination, but minimal in bimanual hand coordination. The reason for such differences remains unclear. In this study, we investigated the possible mechanisms underlying the production of the difference that depend on limb combination. Subjects performed two-limb rhythmic coordinated movements either in the same or in the opposite direction for three separate limb combinations (bilateral hands, contralateral hand and foot, and ipsilateral hand and foot). For each combination two different tasks were performed. In the first condition, subjects actively moved two limbs (active condition). Second, subjects actively moved one limb in coordination with a passively moved limb (passive condition). In the active condition, the directional constraint was dependent upon the limb combination, as reported in previous studies; the directional constraint was quite prominent in ipsilateral combinations, intermediate in contralateral combinations, and minimal for bilateral combination. However, differences in the directional constraint did not depend on limb combination for any combination in the passive conditions which apparently utilized closed-loop control. In other word, the difference depending on limb combination disappeared when control strategies become uniformly closed-loop. Thus, we speculate that the control strategy utilized depends on limb combination in the active condition. Additionally, different mechanisms other than closed-loop control also would have influence depending on the particular limb combination. This may result in differences in performance depending upon the limb combination.


Determining the role of oxygen vacancies in the photoelectrocatalytic performance of WO3 for water oxidation.
Oxygen vacancies are common to most metal oxides, whether intentionally incorporated or otherwise, and the study of these defects is of increasing interest for solar water splitting. In this work, we examine nanostructured WO3 photoanodes of varying oxygen content to determine how the concentration of bulk oxygen-vacancy states affects the photocatalytic performance for water oxidation. Using transient optical spectroscopy, we follow the charge carrier recombination kinetics in these samples, from picoseconds to seconds, and examine how differing oxygen vacancy concentrations impact upon these kinetics. We find that samples with an intermediate concentration of vacancies (∼2% of oxygen atoms) afford the greatest photoinduced charge carrier densities, and the slowest recombination kinetics across all timescales studied. This increased yield of photogenerated charges correlates with improved photocurrent densities under simulated sunlight, with both greater and lesser oxygen vacancy concentrations resulting in enhanced recombination losses and poorer J-V performances. Our conclusion, that an optimal - neither too high nor too low - concentration of oxygen vacancies is required for optimum photoelectrochemical performance, is discussed in terms of the competing beneficial and detrimental impact these defects have on charge separation and transport, as well as the implications held for other highly doped materials for photoelectrochemical water oxidation.


Hepatitis D: A Review.
Hepatitis D virus (HDV) infection occurs in association with hepatitis B virus (HBV) infection and affects approximately 12 million to 72 million people worldwide. HDV causes more rapid progression to cirrhosis and higher rates of hepatocellular carcinoma than HBV alone or hepatitis C virus. HDV requires HBV to enter hepatocytes and to assemble and secrete new virions. Acute HDV-HBV coinfection is followed by clearance of both viruses in approximately 95% of people, whereas HDV superinfection in an HBV-infected person results in chronic HDV-HBV infection in more than 90% of infected patients. Chronic hepatitis D causes more rapidly progressive liver disease than HBV alone. Approximately 30% to 70% of patients with chronic hepatitis D have cirrhosis at diagnosis and more than 50% die of liver disease within 10 years of diagnosis. However, recent studies suggested that progression is variable and that more than 50% of people may have an indolent course. Only approximately 20% to 50% of people infected by hepatitis D have been diagnosed due to lack of awareness and limited access to reliable diagnostic tests for the HDV antibody and HDV RNA. The HBV vaccine prevents HDV infection by preventing HBV infection, but no vaccines are available to protect those with established HBV infection against HDV. Interferon alfa inhibits HDV replication and reduces the incidence of liver-related events such as liver decompensation, hepatocellular carcinoma, liver transplant, or mortality from 8.5% per year to 3.3% per year. Adverse effects from interferon alfa such as fatigue, depression, and bone marrow suppression are common. HBV nucleos(t)ide analogues, such as entecavir or tenofovir, are ineffective against HDV. Phase 3 randomized clinical trials of bulevirtide, which blocks entry of HDV into hepatocytes, and lonafarnib, which interferes with HDV assembly, showed that compared with placebo or observation, these therapies attained virological and biochemical response in up to 56% of patients after 96 weeks of bulevirtide monotherapy and 19% after 48 weeks of lonafarnib, ritonavir, and pegylated interferon alfa treatment. HDV infection affects approximately 12 million to 72 million people worldwide and is associated with more rapid progression to cirrhosis and liver failure and higher rates of hepatocellular carcinoma than infection with HBV alone. Bulevirtide was recently approved for HDV in Europe, whereas pegylated interferon alfa is the only treatment available in most countries.


A spatial epidemiology case study of mentally unhealthy days (MUDs): air pollution, community resilience, and sunlight perspectives.
The main objective of this spatial epidemiologic research is to gain greater insights into the geographic dimension displayed by the different duration of mentally unhealthy days (MUDs) across U.S. counties. Mentally unhealthy days (MUDs) are studied in entire cross counties for year of 2014. Using Behavioural Risk Factor Surveillance System (BRFSS) data in 2014, we examine main factors of mental health hazard including health behaviour, clinical care, socioeconomic and physical environment, demographic, community resilience, and extreme climatic conditions. In this study, we take complex design factors such as clustering, stratification and sample weight in the BRFSS data into account by using Complex Samples General Linear Model (CSGLM). Then, spatial regression models, spatial lag and error models, are applied to examine spatial dependencies and heteroscedasticity. Results of the geographic analyses indicate that counties with lower air pollution (PM2.5), higher community resilience (social, economic, infrastructure, and institutional resilience), and higher sunlight exposure had significantly lower average number of MUDs reported in the past 30 days. These findings suggest that policy makers should take air pollution, community resilience, and sunlight exposure into account when designing environmental and health policies and allocating resources to more effectively manage mental health problems.


Accuracy of the Kirchhoff-approximation and Kirchhoff-ray-mode fish swimbladder acoustic scattering models.
The acoustic backscatter from pressure release prolate spheroids and a three-dimensional representation of a fish swimbladder (Chilean jack mackerel, Trachurus symmetricus murphyi) was calculated using four target strength models (Kirchhoff-approximation, Kirchhoff-ray-mode, finite element solution of the Helmholtz equation, and prolate-spheroid-modal-series). Smoothly varying errors were found in the Kirchhoff-approximation and Kirchhoff-ray-mode model results when compared to the other models, and provide objective criteria for constraining the use of the KA and KRM models. A generic correction technique is also proposed for the prolate spheroid estimates and tentatively tested on a jack mackerel swimbladder, resulting in improvements to the target strength estimates from the Kirchhoff-approximation and Kirchhoff-ray-mode models.


Masking by vaccines in pediatric drug safety signal detection in the EudraVigilance database.
Postmarketing drug safety surveillance relies upon measures of disproportionate reporting in spontaneous reporting systems. It has been hypothesized that products or events reported frequently may "mask" signals. We analyzed the masking effect of vaccines in pediatrics in the EudraVigilance database by conducting disproportionality analysis in the full database (containing vaccine exposures) and in a restricted set (excluding vaccine exposures). We measured performance of the reporting odds ratio (ROR) in both data sets using a pediatric-specific drug reference set and in the absence of a reference set. We assessed masking effects across age groups and conducted a classification tree (CART) analysis. Removal of vaccines decreased the ROR values both in negative and positive controls. Exceptions were drug-event combinations including outcomes frequent in vaccine reports. When restricted to positive control associations, removal of vaccine-related events resulted in increased ROR values for events commonly reported following vaccination. For events rarely associated with vaccination, ROR values decreased for all age groups, especially infants. Analysis in the absence of a reference set showed decrease in ROR following vaccine removal and CART revealed that change in ROR with vaccine removal depended upon age and proportion of reports including a vaccine. Removal of vaccines for signal detection in a pediatric population has an impact on ROR, dependent upon the reporting frequency of the event of interest in combination with vaccines. We recommend stratification by age and removal of vaccine exposures if the investigated adverse drug reactions include those typically reported in association with vaccines for the age strata.


Laparoscopic Roux-en-Y gastric bypass in adolescents with severe obesity (AMOS): a prospective, 5-year, Swedish nationwide study.
Severe obesity in adolescence is associated with reduced life expectancy and impaired quality of life. Long-term benefits of conservative treatments in adolescents are known to be modest, whereas short-term outcomes of adolescent bariatric surgery are promising. We aimed to compare 5-year outcomes of adolescent surgical patients after Roux-en-Y gastric bypass with those of conservatively treated adolescents and of adults undergoing Roux-en-Y gastric bypass, in the Adolescent Morbid Obesity Surgery (AMOS) study. We did a nationwide, prospective, non-randomised controlled study of adolescents (aged 13-18 years) with severe obesity undergoing Roux-en-Y gastric bypass at three specialised paediatric obesity treatment centres in Sweden. We compared clinical outcomes in adolescent surgical patients with those of matched adolescent controls undergoing conservative treatment and of adult controls undergoing Roux-en-Y gastric bypass. The primary outcome measure was change in BMI over 5 years. We used multilevel mixed-effect regression models to assess longitudinal changes. This trial is registered with ClinicalTrials.gov, number NCT00289705. Between April, 2006, and May, 2009, 100 adolescents were recruited to the study, of whom 81 underwent Roux-en-Y gastric bypass (mean age 16·5 years [SD 1·2], bodyweight 132·8 kg [22·1], and BMI 45·5 kg/m2 [SD 6·1]). 80 matched adolescent controls and 81 matched adult controls were enrolled for comparison of outcomes. The change in bodyweight in adolescent surgical patients over 5 years was -36·8 kg (95% CI -40·9 to -32·8), resulting in a reduction in BMI of -13·1 kg/m2 (95% CI -14·5 to -11·8), although weight loss less than 10% occurred in nine (11%). Mean BMI rose in adolescent controls (3·3 kg/m2, 95% CI 1·1-4·8) over the 5-year study period, whereas the BMI change in adult controls was similar to that in adolescent surgical patients (mean change -12·3 kg/m2, 95% CI -13·7 to -10·9). Comorbidities and cardiovascular risk factors in adolescent surgical patients showed improvement over 5 years and compared favourably with those in adolescent controls. 20 (25%) of 81 adolescent surgical patients underwent additional abdominal surgery for complications of surgery or rapid weight loss and 58 (72%) showed some type of nutritional deficiency; health-care consumption (hospital attendances and admissions) was higher in adolescent surgical patients compared with adolescent controls. 20 (25%) of 81 adolescent controls underwent bariatric surgery during the 5-year follow-up. Adolescents with severe obesity undergoing Roux-en-Y gastric bypass had substantial weight loss over 5 years, alongside improvements in comorbidities and risk factors. However, gastric bypass was associated with additional surgical interventions and nutritional deficiencies. Conventional non-surgical treatment was associated with weight gain and a quarter of patients had bariatric surgery within 5 years. Swedish Research Council; Swedish Governmental Agency for Innovation Systems; National Board of Health and Welfare; Swedish Heart and Lung Foundation; Swedish Childhood Diabetes Foundation; Swedish Order of Freemasons Children's Foundation; Stockholm County Council; Västra Götaland Region; Mrs Mary von Sydow Foundation; Stiftelsen Göteborgs Barnhus; Stiftelsen Allmänna Barnhuset; and the US National Institute of Diabetes, Digestive, and Kidney Diseases (National Institutes of Health).


The development of the Starr-Edwards heart valve.
Development of the Starr-Edwards heart valve marked a new era in the treatment of valvular heart disease. Until the development of the Starr-Edwards valve, there were no published reports of patients who had lived longer than 3 months with a prosthetic valve in the mitral position. This valve was the result of a unique partnership between a young surgeon, Dr. Albert Starr, and an experienced engineer, Mr. Lowell Edwards. Working as a team, these 2 men developed and successfully implanted the 1st Starr-Edwards valve within less than 2 years of their 1st meeting. Their key to success was their willingness and ability to make repeated modifications to their design to solve each clinical problem as it arose. Their constant focus on the clinical goal aided the rapid transformation of their design from a leaflet valve to a shielded ball valve, and finally to an unshielded ball valve suitable for implantation in a human being. Along the way, they abandoned the idea of imitating the appearance of native valves, in favor of developing valves that would be clinically successful. Their work has provided help and hope for patients who otherwise would have died from the complications of rheumatic heart disease and other valvular disorders for which valve replacement is the only treatment.


Cholinergic modulation of excitatory synaptic transmission in the CA3 area of the hippocampus.
Cholinergic innervation of the hippocampus has been implicated in memory formation and retrieval. Here we study cholinergic modulation of excitatory transmission in the CA3 area of the rat hippocampus. We used a combination of optical measurements of presynaptic calcium and electrophysiological measurements of synaptic currents to study associational-commissural (A/C) and mossy fiber (MF) synapses in brain slices. Direct synaptic modulation mediated by ACh receptors is only evident at the A/C synapse, where synaptic inhibition primarily reflects presynaptic calcium channel inhibition mediated by muscarinic receptors. MF synapses can, however, be indirectly modulated by muscarinic receptor activation. Muscarine elevates the firing rate of inhibitory cells, which increases GABA release and inhibits MF synapses by activating presynaptic GABA(B) receptors. Muscarine also depolarizes dentate granule cells and elevates their rate of firing. This leads to synaptic enhancement when combined with the use-dependent facilitation of MF synapses. In addition we were unable to evoke an increase in presynaptic calcium levels in MF boutons with local application of nicotinic receptor agonists. This finding does not support a leading hypothesis for MF modulation in which activation of presynaptic nicotinic receptors enhances transmission directly by elevating presynaptic calcium levels. However, indirect synaptic modulation could arise from nicotinic excitation of inhibitory neurons. Thus, to understand cholinergic modulation within the CA3 region, it is necessary to take into account secondary actions on synapses arising from other chemical messengers released by other cell types and to consider effects on firing patterns of presynaptic cells, which in turn influence release via use-dependent synaptic plasticity.


Three-dimensional clinical PET in lung cancer: validation and practical strategies.
The feasibility of 3-dimensional acquisition mode for semiquantitative analysis in thoracic PET studies was compared to the conventional 2-dimensional mode. Several practical considerations were analyzed to propose an optimized scanning protocol for clinical use. Twenty-one patients with focal thoracic abnormalities were evaluated with FDG PET. The acquisition consisted of 3 consecutive static scans for a single bed position: 3-dimensional (10 min), 2-dimensional (15 min), and 3-dimensional (5 min). On the basis of the average and maximum activity values per region of interest, standardized uptake value (SUV) normalized for total body weight (TBW), lean body mass (LBM), body surface area (BSA), and blood glucose level (PGL) were evaluated. The effect of the delay between tracer injection and PET scanning on the SUV, as well as on the relative error of the activity distribution, was studied from 40-134 min after tracer injection. A strong positive correlation was observed among SUVs from 2-dimensional and both 3-dimensional acquisitions. The mean SUV percentage differences between both acquisition modes were about 17%, differences that were not statistically significant when time postinjection was addressed in the analysis of covariance. SUVs provided the greatest variability and differences among studies on experimental periods up to 70 min postinjection. Indeed, the variability of 20% observed on the SUVs from 2 PET scans 13 min apart was reduced to 9% when the acquisitions started at least 70 min after tracer injection. In addition, a two-fold reduction in the relative error of the activity distribution was observed over this period of time. The reproducibility coefficient was increased from 0.87 to 0.95 before and after 70 min postinjection, respectively. No correlation was found between different normalization procedures of SUV and LBM, BSA, TBW, or height, whereas a weak correlation was found between SUV and PGL. 18F-FDG 3-dimensional PET is a realistic alternative to the gold standard 2-dimensional for clinical nonkinetic studies. A short, 5-min 3-dimensional acquisition at 70 min postinjection is proposed as the best protocol for the clinical evaluation of thoracic pathologies.


[Heritability and environment in psychiatric disorders].
In recent years quantitative genetic research has addressed all the major psychiatric disorders. In order to interpret the results of this type of research one needs to be aware of its potential and its limitations. To discuss the basic concepts and the main results of quantitative genetic research and to consider how this can help us to better understand the aetiology of psychiatric disorders. Using Medline (1990-February 2006) we reviewed the literature on the subject of quantitative genetic and psychiatric disorders. In addition we studied the standard books on the subject. A fairly large number of psychiatric disorders, namely about 30 to 85%, can be inherited. In addition, the non-shared environment has a considerable influence on the phenotype. The influence of the shared environment seems to have only a limited influence or it is totally absent. The results of quantitative genetic research are specific to a particular time or environment and therefore may not be applicable to other populations. There may be a correlation or interaction between genetic factors and the environment while the phenotype is being formed. However, because of the analytical methods used, this is only partly visible in the results. Quantitative genetic research has made an impressive contribution to our knowledge about the heritability of psychiatric disorders. By definition quantitative genetic research always provides information about environmental influences.


Neuron-glia interactions in glutamatergic neurotransmission: roles of oxidative and glycolytic adenosine triphosphate as energy source.
Glutamatergic neurotransmission accounts for a considerable part of energy consumption related to signaling in the brain. Chemical energy is provided by adenosine triphosphate (ATP) formed in glycolysis and tricarboxylic acid (TCA) cycle combined with oxidative phosphorylation. It is not clear whether ATP generated in these pathways is equivalent in relation to fueling of the energy-requiring processes, i.e., vesicle filling, transport, and enzymatic processing in the glutamatergic tripartite synapse (the astrocyte and pre- and postsynapse). The role of astrocytic glycogenolysis in maintaining theses processes also has not been fully elucidated. Cultured astrocytes and neurons were utilized to monitor these processes related to glutamatergic neurotransmission. Inhibitors of glycolysis and TCA cycle in combination with pathway-selective substrates were used to study glutamate uptake and release monitored with D-aspartate. Western blotting of glyceraldehyde-3-P dehydrogenase (GAPDH) and phosphoglycerate kinase (PGK) was performed to determine whether these enzymes are associated with the cell membrane. We show that ATP formed in glycolysis is superior to that generated by oxidative phosphorylation in providing energy for glutamate uptake both in astrocytes and in neurons. The neuronal vesicular glutamate release was less dependent on glycolytic ATP. Dependence of glutamate uptake on glycolytic ATP may be at least partially explained by a close association in the membrane of GAPDH and PGK and the glutamate transporters. It may be suggested that these enzymes form a complex with the transporters and the Na(+) /K(+) -ATPase, the latter providing the sodium gradient required for the transport process.


A novel ATP-dependent inward rectifier potassium channel expressed predominantly in glial cells.
We have isolated a novel inward rectifier K+ channel predominantly expressed in glial cells of the central nervous system. Its amino acid sequence exhibited 53% identity with ROMK1 and approximately 40% identity with other inward rectifier K+ channels. Xenopus oocytes injected with cRNA derived from this clone expressed a K+ current, which showed classical inward rectifier K+ channel characteristics. Intracellular Mg.ATP was required to sustain channel activity in excised membrane patches, which is consistent with a Walker type-A ATP-binding domain on this clone. We designate this new clone as KAB-2 (the second type of inward rectifying K+ channel with an ATP-binding domain). In situ hybridization showed KAB-2 mRNA to be expressed predominantly in glial cells of the cerebellum and forebrain. This is the first description of the cloning of a glial cell inward rectifier potassium channel, which may be responsible for K+ buffering action of glial cells in the brain.


[Artificial oxygen carriers as an alternative to red blood cell transfusion].
The expected cost-explosion in transfusion medicine (increasing imbalance between donors and recipients, treatment of transfusion-associated complications) increases the socio-economic significance of the development of safe and effective synthetic oxygen carriers as an alternative to the transfusion of allogeneic red blood cells. Currently two types of artificial oxygen carriers have been tested for safety and efficacy in cases of severe anemia otherwise requiring transfusion. Solutions based on human or bovine hemoglobin (HBOC) possess vasoconstrictor properties in addition to their oxygen transport capacity. The impact of vasoconstriction on tissue perfusion and organ function is however not yet fully understood. Nevertheless, in 2001 the bovine HBOC Hemopure was approved in South Africa for treatment of acutely anemic surgical patients. The purely synthetic perfluorocarbon (PFC) emulsions increase the physically dissolved portion of arterial oxygen content. Due to their particulate nature (emulsion droplets) PFCs may only be infused in low doses to avoid overload and malfunction of phagocytic cells of the reticulo-endothelial system. As part of a multimodal blood conservation program (including normovolemic hemodilution and hyperoxia) the low-dose administration of Oxygent effectively increases intraoperative anemia tolerance. Although reduction of perioperative allogeneic blood transfusion has already been demonstrated for HBOC and PFC, the global clinical establishment of artificial oxygen carriers is not to be expected in the near future.


An inverse approach for elucidating dendritic function.
We outline an inverse approach for investigating dendritic function-structure relationships by optimizing dendritic trees for a priori chosen computational functions. The inverse approach can be applied in two different ways. First, we can use it as a "hypothesis generator" in which we optimize dendrites for a function of general interest. The optimization yields an artificial dendrite that is subsequently compared to real neurons. This comparison potentially allows us to propose hypotheses about the function of real neurons. In this way, we investigated dendrites that optimally perform input-order detection. Second, we can use it as a "function confirmation" by optimizing dendrites for functions hypothesized to be performed by classes of neurons. If the optimized, artificial, dendrites resemble the dendrites of real neurons the artificial dendrites corroborate the hypothesized function of the real neuron. Moreover, properties of the artificial dendrites can lead to predictions about yet unmeasured properties. In this way, we investigated wide-field motion integration performed by the VS cells of the fly visual system. In outlining the inverse approach and two applications, we also elaborate on the nature of dendritic function. We furthermore discuss the role of optimality in assigning functions to dendrites and point out interesting future directions.


Mitigation of ammonia, nitrous oxide and methane emissions from manure management chains: a meta-analysis and integrated assessment.
Livestock manure contributes considerably to global emissions of ammonia (NH3 ) and greenhouse gases (GHG), especially methane (CH4 ) and nitrous oxide (N2 O). Various measures have been developed to mitigate these emissions, but most of these focus on one specific gas and/or emission source. Here, we present a meta-analysis and integrated assessment of the effects of mitigation measures on NH3 , CH4 and (direct and indirect) N2 O emissions from the whole manure management chain. We analysed the effects of mitigation technologies on NH3 , CH4 and N2 O emissions from individual sources statistically using results of 126 published studies. Whole-chain effects on NH3 and GHG emissions were assessed through scenario analysis. Significant NH3 reduction efficiencies were observed for (i) housing via lowering the dietary crude protein (CP) content (24-65%, compared to the reference situation), for (ii) external slurry storages via acidification (83%) and covers of straw (78%) or artificial films (98%), for (iii) solid manure storages via compaction and covering (61%, compared to composting), and for (iv) manure application through band spreading (55%, compared to surface application), incorporation (70%) and injection (80%). Acidification decreased CH4 emissions from stored slurry by 87%. Significant increases in N2 O emissions were found for straw-covered slurry storages (by two orders of magnitude) and manure injection (by 26-199%). These side-effects of straw covers and slurry injection on N2 O emission were relatively small when considering the total GHG emissions from the manure chain. Lowering the CP content of feed and acidifying slurry are strategies that consistently reduce NH3 and GHG emissions in the whole chain. Other strategies may reduce emissions of a specific gas or emissions source, by which there is a risk of unwanted trade-offs in the manure management chain. Proper farm-scale combinations of mitigation measures are important to minimize impacts of livestock production on global emissions of NH3 and GHG.


The influence of cold treatment on properties of temperature-sensitive poly(N-isopropylacrylamide) hydrogels.
Poly(N-isopropylacrylamide) (PNIPAAm) hydrogel exhibits a response to external temperature variation and shrinks in volume abruptly as the temperature is increased above its lower critical solution temperature. It has great potential applications in biomedical fields. A rapid response rate is essential, especially when this material is designed as an on-off switch for targeted drug delivery. However, due to the appearance of a thick, dense skin layer on the hydrogel surface during the shrinking process, the deswelling rate of conventional PNIPAAm gels is low. In this article, a novel method is proposed to modify the surface morphology of PNIPAAm gel, in which the swollen gels are frozen at low temperature (-20 degrees C). The scanning electron micrographs revealed that a fishnet-like skin layer appeared on the surfaces of the cold-treated gels. Dramatically rapid deswelling was achieved with the cold-treated gels since the fishnet-like structure with numerous small pores prevented the formation of a dense, thick skin layer during the deswelling process, which commonly occurs in normal PNIPAAm hydrogels. Prolonging the cold treatment from 1 day to 10 days resulted in a slightly higher deswelling rate. Rearrangement of the hydrogel matrix structure during the freezing process might contribute to the formation of the fishnet-like skin layer. The water uptake of the hydrogels increased nearly in proportion to the square root of time, indicating that the reswelling rate of hydrogels was controlled predominantly by water diffusion into the network. However, there were no significant differences in the equilibrated swelling ratio and reswelling kinetics at room temperature (22 degrees C) between normal gels and cold-treated gels, which implied that cold treatment did not change bulk porosity and gel tortuosity much.


Guinea pigs sublethally infected with aerosolized Legionella pneumophila develop humoral and cell-mediated immune responses and are protected against lethal aerosol challenge. A model for studying host defense against lung infections caused by intracellular pathogens.
We have employed the guinea pig model of L. pneumophila infection, which mimics Legionnaires' disease in humans both clinically and pathologically, to study humoral and cell-mediated immune responses to L. pneumophila and to examine protective immunity after aerosol exposure, the natural route of infection. Guinea pigs exposed to sublethal concentrations of L. pneumophila by aerosol developed strong humoral immune responses. By the indirect fluorescent antibody assay, exposed guinea pigs had a median serum antibody titer (expressed as the reciprocal of the highest positive dilution) of 32, whereas control guinea pigs had a median titer of less than 1. Sublethally infected (immunized) guinea pigs also developed strong cell-mediated immune responses. In response to L. pneumophila antigens, splenic lymphocytes from immunized but not control animals proliferated strongly in vitro, as measured by their capacity to incorporate [3H]thymidine. Moreover, immunized but not control guinea pigs developed strong cutaneous delayed-type hypersensitivity to intradermally injected L. pneumophila antigens. Sublethally infected (immunized) guinea pigs exhibited strong protective immunity to L. pneumophila. In two independent experiments, all 22 immunized guinea pigs survived aerosol challenge with one or three times the lethal dose of L. pneumophila whereas none of 16 sham-immunized control guinea pigs survived (p less than 0.0001 in each experiment). Immunized guinea pigs were not protected significantly from challenge with 10 times the lethal dose. Immunized but not control animals cleared the bacteria from their lungs. This study demonstrates that guinea pigs sublethally infected with L. pneumophila by the aerosol route develop strong humoral immune responses to this pathogen, develop strong cell-mediated immune responses and cutaneous delayed-type hypersensitivity to L. pneumophila antigens, are protected against subsequent lethal aerosol challenge, and are able to clear the bacteria from their lungs. The guinea pig model of L. pneumophila pulmonary infection is as an excellent one for studying general principles of host defense against pulmonary infections caused by intracellular pathogens.


Effects of summer internship and follow-up distance mentoring programs on middle and high school student perceptions and interest in health careers.
Minorities are underrepresented in health professions and efforts to recruit minority students into health careers are considered a way to reduce health disparities. There is little research about the effectiveness of these programs, other than satisfaction. This study aimed to measure program effects on student understanding of and interest in health careers. Students took a career interest inventory, completed a scale measuring their self-reported understanding and interest in health careers, and wrote essays about health careers before and after completing a 1 week on campus internship on health careers and after a 9 month follow up distance mentoring program where they continued to interact with university faculty by videoconference about career options. Changes in inventory, scale, and essay scores were analyzed for changes over time using Wilcoxon and Mann-Whitney tests. Inventory scores were unchanged over time, but scale and essay scores trended upward significantly post internship and mentoring. Health career education and mentoring programs can positively affect student knowledge of health careers and their attitudes about them. The study's methods extend measures of program impact beyond satisfaction.


Content analysis of age verification, purchase and delivery methods of internet e-cigarette vendors, 2013 and 2014.
Identify the population of internet e-cigarette vendors (IEVs) and conduct content analyses of their age verification, purchase and delivery methods in 2013 and 2014. We used multiple sources to identify IEV websites, primarily complex search algorithms scanning more than 180 million websites. In 2013, we manually screened 32 446 websites, identifying 980 IEVs, selecting the 281 most popular for content analysis. This methodology yielded 31 239 websites for screening in 2014, identifying 3096 IEVs, with 283 selected for content analysis. The proportion of vendors that sold online-only, with no retail store, dropped significantly from 2013 (74.7%) to 2014 (64.3%) (p<0.01), with a corresponding significant decrease in US-based vendors (71.9% in 2013 and 65% in 2014). Most vendors did little to prevent youth access in either year, with 67.6% in 2013 and 63.2% in 2014 employing no age verification or relying exclusively on strategies that cannot effectively verify age. Effective age verification strategies such as online age verification services (7.1% in 2013 and 8.5% in 2014), driving licences (1.8% in 2013 and 7.4% in 2014, p<0.01) or age verification at delivery (6.4% in 2013 and 8.1% in 2104) were rarely advertised on IEV websites. Nearly all vendors advertised accepting credit cards, and about ¾ shipping via United States Postal Service, similar to the internet cigarette industry prior to federal bans. The number of IEVs grew sharply from 2013 to 2014, with poor age verification practices. New and expanded regulations for online e-cigarette sales are needed, including strict age and identity verification requirements.


Genetic and epigenetic alterations of the cell cycle regulators and tumor suppressor genes in pediatric osteosarcomas.
To analyze the genetic and epigenetic alterations affecting the RB1, TP53, p16INK4, and p21WAF1 tumor suppressor genes, loss of heterozygosity (LOH) at 3q and 18q, and the clinical variables of a series of Spanish children with osteosarcoma. These genetic changes were tested for an association with prognosis. Peripheral blood samples and clinical data were available from 76 patients with osteosarcoma. Paired tissue was available from 41 of them. The mutation and methylation status of p16INK4, p21WAF1, TP53, and RB1 was screened as well as LOH at 3q and 18q. Loss of heterozygosity affecting RB1 (37.2%), TP53 (42.3%), and 18q (30.8%) and TP53 mutation (39%) were frequently encountered. TP53 mutation was associated with diagnosis at a later age. RB1 alteration was associated with reduced survival and event-free survival. The clinical variables associated with poor prognosis were the presence of metastasis at diagnosis (P = 0.035) or during treatment (P = 0.016) and the chondroblastic histologic subtype (P = 0.007); the response to induction chemotherapy (<90% necrosis) also tended to be related to poor prognosis (P = 0.08). RB1, TP53, and possibly other tumor suppressor genes located at 18q and other localizations are involved in pediatric osteosarcoma carcinogenesis, together with other genetic alterations not fully understood to date. Based on these results, the presence of an altered RB1 gene should be regarded as a poor prognostic factor for pediatric osteosarcoma.


Urinary N-acetyl-beta-d-glucosaminidase (NAG) with neutrophil gelatinase-associated lipocalin (NGAL) improves the diagnostic value for proximal tubule damage in diabetic kidney disease.
Screening for diabetic kidney disease (DKD) remains a challenge; however, there has been an ongoing research to investigate the diagnostic value of different biomarkers to identify DKD. The aim of this study was to assess the diagnostic value of both N-acetyl-beta-d-glucosaminidase (NAG) and neutrophil gelatinase-associated lipocalin (NGAL) in the progression of DKD. This cross-sectional case-control study included 92 type 2 diabetic patients with or without DKD. Urinary NAG and NGAL were measured to evaluate their diagnostic values as biochemical markers related to DKD. Both urinary NAG and NGAL levels were significantly higher among patients with DKD. In multiple linear regression analysis, NAG showed a positive significant association with NGAL in the three different adjusted models, while no significant correlation with fasting blood glucose, glycated hemoglobin, serum creatinine, estimated glomerular filtration rate, and albumin creatinine ratio were observed. The area under the curve for NGAL was 0.659 (p = 0.01) and 0.564 (p = 0.297) for NAG in DKD patients. This study demonstrates the association between urinary NAG and NGAL as a tubular damage marker for DKD although longitudinal studies are needed to evaluate its diagnostic value.


Aqueous photochemical degradation of mefenamic acid and triclosan: role of wastewater effluent matrices.
In this study, photochemical degradation of two emerging pharmaceutical chemicals, mefenamic acid (MF) and triclosan (TCS), was investigated to clarify the role of treated wastewater effluent matrices on their environmental photolysis. Target compounds were individually exposed to simulated sunlight in different media: ultrapure buffered water and synthetic field water with treated municipal wastewater effluent. The results in ultrapure buffered water showed that the direct photolysis processes in aquatic environments are not relevant to the elimination of MF. However, in samples containing treated wastewater effluent, photochemical degradation of MF was clearly enhanced. Our results indicate that MF undergoes indirect photolysis by reactive intermediates produced in an effluent matrix. Further quenching experiments suggested that photochemically produced hydroxyl radicals and excited triplet state dissolved organic matter drive the degradation of MF. In contrast to MF, TCS photochemical degradation proceeds through rapid direct photolysis. TCS was quickly degraded in ultrapure buffered water but it is considerably hampered in samples containing wastewater effluent. The declined degradation of TCS in the synthetic field water was discussed in terms of underlying optical filter effects by coexisting chromophoric substances. Results emphasize the importance of taking local water chemistry into consideration when predicting natural attenuation of pharmaceutical chemicals in receiving areas.


Complex amplitude reconstruction for dynamic beam quality M2 factor measurement with self-referencing interferometer wavefront sensor.
A real-time complex amplitude reconstruction method for determining the dynamic beam quality M2 factor based on a Mach-Zehnder self-referencing interferometer wavefront sensor is developed. By using the proposed complex amplitude reconstruction method, full characterization of the laser beam, including amplitude (intensity profile) and phase information, can be reconstructed from a single interference pattern with the Fourier fringe pattern analysis method in a one-shot measurement. With the reconstructed complex amplitude, the beam fields at any position z along its propagation direction can be obtained by first utilizing the diffraction integral theory. Then the beam quality M2 factor of the dynamic beam is calculated according to the specified method of the Standard ISO11146. The feasibility of the proposed method is demonstrated with the theoretical analysis and experiment, including the static and dynamic beam process. The experimental method is simple, fast, and operates without movable parts and is allowed in order to investigate the laser beam in inaccessible conditions using existing methods.


Building capacity in palliative care for personal support workers in long-term care through experiential learning.
Providing palliative care in long-term care (LTC) homes is an area of growing importance. As a result, attention is being given to exploring effective palliative care learning strategies for personal support workers (PSWs) who provide the most hands-on care to LTC residents. The purpose of this intervention was to explore hospice visits as an experiential learning strategy to increase the capacity of PSWs in palliative care, specifically related to their new learning, and how they anticipated this experience changed their practices in LTC. This study utilised a qualitative descriptive design. Eleven PSWs from four Ontario LTC homes were sent to their local hospice to shadow staff for one to two days. After the visit, PSWs completed a questionnaire with open-ended questions based on critical reflection. Data were analysed using thematic content analysis. PSWs commented on the extent of resident-focused care at the hospice and how palliative care interventions were tailored to meet the needs of residents. PSWs were surprised with the lack of routine at the hospice but felt that hospice staff prioritised their time effectively in order to meet family and client care needs. Some PSWs were pleased to see how well integrated the PSW role is on the community hospice team without any hierarchical relationships. Finally, PSWs felt that other LTC staff would benefit from palliative care education and becoming more comfortable with talking about death and dying with other staff, residents and family members. This study highlighted the benefits of PSWs attending a hospice as an experiential learning strategy. Future work is needed to evaluate this strategy using more rigorous designs as a way to build capacity within PSWs to provide optimal palliative care for LTC residents and their family members. PSWs need to be recognised as important members within the interdisciplinary team. PSWs who shadow staff at hospices view this experience as a positive strategy to meet their learning needs related to palliative care.


Mode selectivity and mirror misalignment effects in unstable laser resonators.
It is known that single transverse mode operation of an unstable laser resonator can be achieved using large output coupling and large Fresnel number operation. However, it has not been recognized that there is an upper limit on usable Fresnel number, determined by such effects as mirror alignment stability. We have analyzed a variable-reflectivity unstable resonator using mode solutions obtained by Zucker. The differential loss between the lowest order modes leads to conditions on the Fresnel number and the output coupling if single mode operation is to be realized. The Zucker solution is extended by a perturbation analysis to examine mode distortion caused by mirror misalignment. The on-axis intensity in the far field of the laser output is shown to be a sensitive function of mirror misalignment, and the mirror alignment tolerance permitted varies inversely as the Fresnel number. In practice, this places an upper limit on the usable Fresnel number. The results of this theory are shown to be consistent with laser experiments using unstable resonators.


In vitro activity of posaconazole against clinical isolates of dermatophytes.
A broth macrodilution method following the recommendations established by the National Committee for Clinical Laboratory Standards was used to compare the in vitro activity of posaconazole (PCZ) with that of itraconazole (ITC) against 30 clinical isolates of dermatophytes belonging to six different species. In terms of MICs, PCZ showed an activity equal to that of ITC. MICs of PCZ at which 50% (MIC(50)) and 90% (MIC(90)) of the isolates were inhibited were 0.5 and > 4.0 microg/ml, respectively. The MIC(50) and MIC(90) of ITC were 1.0 and > 4.0 microg/ml, respectively. However, PCZ showed a more potent fungicidal activity than that of ITC against isolates belonging to the genus Microsporum (P = 0.03). PCZ merits further investigation as a potentially useful agent for treatment of dermatophytosis.


Common variants in NLRP2 and NLRP3 genes are strong prognostic factors for the outcome of HLA-identical sibling allogeneic stem cell transplantation.
The inflammasomes are macromolecular cytosolic complexes involved in the production of interleukin-1beta (IL-1beta) and IL-18 in response to several pathogen-derived stimuli. Such interleukins have been implicated in the origin of severe allogeneic stem cell transplant (allo-SCT) complications. We analyzed the relationship between the interindividual variability in inflammasome protein-encoding genes in donors and patients and clinical outcome after allo-SCT. Fourteen common genetic variants in 5 genes of the inflammasome, namely, NLRP1, NLRP2, NLRP3, CARD8, and CASP5, were genotyped in 133 human leukocyte antigen-identical sibling pairs undergoing allo-SCT. In the multivariate analysis, donor variants in NLRP2 and NLRP3 were the most important prognostic factors for the clinical outcome after allo-SCT. Thus, donor TT genotype at rs10925027 in NLRP3 was associated with disease relapse (odds ratio (OR) = 6.3, P = 1 x 10(-7)), and donor GG genotype at rs1043684 in NLRP2 was associated with nonrelapse mortality (OR = 4.4, P = 6 x 10(-4)) and overall survival (OR = 3.1, P = .001). In addition, patient AA genotype at rs5862 in NLRP1 was associated with nonrelapse mortality (OR = 2.8, P = .005) and overall survival (OR = 2.0, P = .009). These results suggest that inflammasome genetic variants are important prognostic factors for the outcome of allo-SCT. If validated in larger studies, including unrelated allo-SCT, NLRPs genotype would become an important factor in donor selection.


Epidemiologic, clinical, and laboratory features of Coxsackie B1-B5 infections in the United States, 1970-79.
In the period 1970 through 1979, the Coxsackie B1, B2, B3, B4, and B5 viruses constituted 24 percent of more than 18,000 enteroviruses isolated and reported through national surveillance. Young children, especially males, were most frequently affected: 48 percent of the national surveillance population were less than 5 years of age, including 30 percent who were less than 1 year old. Among the most frequently reported clinical syndromes associated with B infection were meningitis (in 56 percent of patients with B1-B5 infections), encephalitis (in 15 percent), and respiratory tract disease (in 14 percent). Carditis, a well-known B syndrome, was reported with only 2 percent of B1-B5 infections. Like most enteroviral agents, Group B viruses were isolated primarily during the summer: 87 percent of all these isolations were made during the 5 months from June through October. Although B2, B3, and B4 viruses were isolated at relatively uniform levels each year, B1 and B5 viral illnesses occurred nationwide as explosive epidemics only in certain years. A separate population of B-infected patients, identified by the Nassau County Medical Center (NCMC) Virus Laboratory, East Meadow, N.Y., during the same 10-year period, was studied to compare epidemiologic characteristics and to evaluate in greater detail clinical and laboratory features of B infections. Because of more active solicitation of specimens for testing, ascertainment in the NCMC system was more complete. The most frequently reported clinical findings at NCMC included fever (97 percent of cases), which was biphasic in 27 percent; pharyngitis (85 percent); vomiting (56 percent); headache (49 percent); other respiratory signs and symptoms (44 percent); diarrhea (40 percent); abdominal pain (33 percent); rash (31 percent); and otitis (28 percent). Rash was more frequently associated with younger than with older age groups (P < .01) for all B agents. Overall, throat (T) and rectal (R) swabs had the highest B-positivity rates among known infected patients(83 percent for T and 78 percent for R). Only for T was the positivity rate correlated with the interval between onset of illness and obtaining the specimen (P < .05). B agents grew most quickly from T specimens, but most reliably from R specimens. On the basis of these data,the authors recommend that both T and R specimens be obtained from every patient for whom prompt and reliable laboratory diagnosis of B infection is sought.To the authors' knowledge, these results from 10 years of national surveillance represent the largest surveillance summary of Coxsackie B viruses to date in the literature. Comparison of these results with those reported over the same 10 years by NCMC reflects differences that arise mostly because of differences in ascertainment systems.


Relationship between subtypes of irritable bowel syndrome and severity of symptoms associated with panic disorder.
To investigate the relationship between subtypes of irritable bowel syndrome (IBS) and severity of symptoms associated with panic disorder (PD). The study comprised 178 consecutive new PD outpatients. Sixty-four patients met the Rome-II criteria for IBS (IBS[+]; 29 diarrhea-predominant IBS (IBSD), 14 constipation-predominant IBS (IBSC), 21 other types of IBS). IBSD patients with agoraphobia avoided a greater number of scenes owing to fear of panic attack than did PD patients without IBS (IBS[-]) and with agoraphobia. IBS[+] patients with avoidant behavior due to fear of IBS symptoms had significantly higher Beck Depression Inventory (BDI) scores and avoided a larger number of scenes owing to fear of panic attack than IBS[+] patients with agoraphobia and without avoidant behavior due to fear of IBS symptoms or IBS[-] patients with agoraphobia. The results suggest that the presence of IBSD or avoidant behavior because of fear of IBS symptoms may be associated with a more severe form of agoraphobia, and the latter may also be associated with depression.


Impact of single phase CT angiography collateral status on functional outcome over time: results from the MR CLEAN Registry.
Collateral status modified the effect of endovascular treatment (EVT) for stroke in several randomized trials. We assessed the association between collaterals and functional outcome in EVT treated patients and investigated if this association is time dependent. We included consecutive patients from the Multicenter Randomized Clinical Trial of Endovascular Treatment for Acute Ischemic Stroke in The Netherlands (MR CLEAN) Registry (March 2014-June 2016) with an anterior circulation large vessel occlusion undergoing EVT. Functional outcome was measured on the modified Rankin Scale (mRS) at 90 days. We investigated the association between collaterals and mRS in the MR CLEAN Registry with ordinal logistic regression and if this association was time dependent with an interaction term. Additionally, we determined modification of EVT effect by collaterals compared with MR CLEAN controls, and also investigated if this was time dependent with multiplicative interaction terms. 1412 patients were analyzed. Functional independence (mRS score of 0-2) was achieved in 13% of patients with grade 0 collaterals, in 27% with grade 1, in 46% with grade 2, and in 53% with grade 3. Collaterals were significantly associated with mRS (adjusted common OR 1.5 (95% CI 1.4 to 1.7)) and significantly modified EVT benefit (P=0.04). None of the effects were time dependent. Better collaterals corresponded to lower mortality (P<0.001), but not to lower rates of symptomatic intracranial hemorrhage (P=0.14). In routine clinical practice, better collateral status is associated with better functional outcome and greater treatment benefit in EVT treated acute ischemic stroke patients, independent of time to treatment. Within the 6 hour time window, a substantial proportion of patients with absent and poor collaterals can still achieve functional independence.


The effects of different pre-game motivational interventions on athlete free hormonal state and subsequent performance in professional rugby union matches.
We examined the effect of different pre-match motivational interventions on athlete free testosterone (T) and cortisol (C) concentrations and subsequent match performance in professional rugby union. Male participants (n=12) playing at a senior or academy level in rugby union were recruited and each completed three interventions (15 min each) before a competitive game; 1) watching a video clip of successful skill execution by the player with positive coach feedback [VPCF1]; 2) watching a video clip of successful skill execution by an opposing player with cautionary coach feedback [VCCF], 3) the player left alone to self-motivate [SM1]. The first and last interventions were retested [VPCF2 and SM2]. Salivary free T and C measures were taken pre-intervention and pre-game. Within each game, players were rated by coaching staff on a key performance indicator (KPI) from identified skills and an overall performance indicator (OPI), where 1 = best performance to 5 = worst performance. The VPCF1 and VPCF2 interventions both promoted significant T responses (11.8% to 12.5%) before each game and more so than SM1, SM2 and VCCF. The VCCF approach produced the largest C response (17.6%) and this differed from all other treatments. The VPCF interventions were also associated with better game KPI (1.5 to 1.8) and OPI ratings (1.7 to 1.8) than SM1, SM2 and/or VCCF. Across all treatments, greater individual T responses and lower C responses were associated with better KPI and OPI outcomes. In conclusion, the pre-game presentation of motivational strategies to athletes involving specific video footage and coach feedback produced different outcomes on two indicators of match performance, which were also associated with changes in free hormonal state.


Different extraction methodologies and their influence on the bioactivity of the wild edible mushroom Laetiporus sulphureus (Bull.) Murrill.
Laetiporus sulphureus is an edible wood-rooting basidiomycete. The nutritional and medicinal properties of this mushroom have long been known by traditional practitioners. The aim of this study was to determine the proximate composition, total phenol antioxidant capacity and antimicrobial activities of different extracts of L. sulphureus. Different extraction methodologies, including high energy techniques, were employed and their effect was examined on the activity of the extracts. Optimum extraction methodologies (classical and ultrasound-assisted) provided one fraction containing neutral and polar lipids and the other fraction containing fungal carotenoids and pigments. Fatty acid analysis indicated a predominant level of polyunsaturated fatty acids followed by saturated and mono-unsaturated fatty acids. Both the aqueous methanolic and water extracts contained higher TPC and showed better antioxidant capacity than the ethanolic extract. Irrespective of the type of extraction applied, L. sulphureus showed good antimicrobial activity against all the tested bacteria and fungi, being in some cases stronger than the used antibiotics and mycotics. Therefore, this edible mushroom could be considered as a positive candidate to be utilised by the food industry, not only for obtaining bioactive compounds to be used as natural antioxidants/antimicrobial agents, but possibly also for its nutritional value and health benefits.


Spectra of "real-world" graphs: beyond the semicircle law.
Many natural and social systems develop complex networks that are usually modeled as random graphs. The eigenvalue spectrum of these graphs provides information about their structural properties. While the semicircle law is known to describe the spectral densities of uncorrelated random graphs, much less is known about the spectra of real-world graphs, describing such complex systems as the Internet, metabolic pathways, networks of power stations, scientific collaborations, or movie actors, which are inherently correlated and usually very sparse. An important limitation in addressing the spectra of these systems is that the numerical determination of the spectra for systems with more than a few thousand nodes is prohibitively time and memory consuming. Making use of recent advances in algorithms for spectral characterization, here we develop methods to determine the eigenvalues of networks comparable in size to real systems, obtaining several surprising results on the spectra of adjacency matrices corresponding to models of real-world graphs. We find that when the number of links grows as the number of nodes, the spectral density of uncorrelated random matrices does not converge to the semicircle law. Furthermore, the spectra of real-world graphs have specific features, depending on the details of the corresponding models. In particular, scale-free graphs develop a trianglelike spectral density with a power-law tail, while small-world graphs have a complex spectral density consisting of several sharp peaks. These and further results indicate that the spectra of correlated graphs represent a practical tool for graph classification and can provide useful insight into the relevant structural properties of real networks.


Disruptive de novo mutations of DYRK1A lead to a syndromic form of autism and ID.
Dual-specificity tyrosine-(Y)-phosphorylation-regulated kinase 1 A (DYRK1A) maps to the Down syndrome critical region; copy number increase of this gene is thought to have a major role in the neurocognitive deficits associated with Trisomy 21. Truncation of DYRK1A in patients with developmental delay (DD) and autism spectrum disorder (ASD) suggests a different pathology associated with loss-of-function mutations. To understand the phenotypic spectrum associated with DYRK1A mutations, we resequenced the gene in 7162 ASD/DD patients (2446 previously reported) and 2169 unaffected siblings and performed a detailed phenotypic assessment on nine patients. Comparison of our data and published cases with 8696 controls identified a significant enrichment of DYRK1A truncating mutations (P=0.00851) and an excess of de novo mutations (P=2.53 × 10(-10)) among ASD/intellectual disability (ID) patients. Phenotypic comparison of all novel (n=5) and recontacted (n=3) cases with previous case reports, including larger CNV and translocation events (n=7), identified a syndromal disorder among the 15 patients. It was characterized by ID, ASD, microcephaly, intrauterine growth retardation, febrile seizures in infancy, impaired speech, stereotypic behavior, hypertonia and a specific facial gestalt. We conclude that mutations in DYRK1A define a syndromic form of ASD and ID with neurodevelopmental defects consistent with murine and Drosophila knockout models.


Highly sensitive fluorescence detection technology currently available for HTS.
Homogeneous fluorescence methods are providing an important tool for HTS technologies. A wide range of different techniques have been established on the market, with read-outs ranging from total fluorescence intensity to statistical analysis of fluorescence fluctuations for biochemical assays or fluorescence imaging techniques for cellular systems. Each method has its own advantages and limitations, which have to be accounted for when designing a specific assay. Here, recently developed fluorescence techniques and some of their applications, with a particular focus on sensitivity, are summarized and their principles are presented.


Chronic nicotine exposure in preadolescence enhances later spontaneous recovery of fear memory.
Preadolescent mice have been shown to be differentially susceptible to the effects of both acute and chronic nicotine exposure on contextual fear learning relative to adults. For this study, we tested the effects of chronic nicotine exposure in preadolescence on adulthood extinction and spontaneous recovery of fear memory in a model in which contextual fear acquisition occurred prior to nicotine exposure. Preadolescent (postnatal day 23) and adult (postnatal day 53) male C57BL/6J mice underwent contextual fear conditioning and were then exposed to chronic nicotine at 12.6 mg/kg/day for 12 days via osmotic minipump. Eighteen days following the removal of nicotine, both groups of mice underwent fear extinction, followed by a spontaneous recovery session a week later. History of chronic nicotine did not affect later extinction of fear memory in adult-trained mice, whereas adolescent-trained mice exhibited a global impairment in retention of fear memory that precluded detection of effects of early nicotine on later fear extinction. However, it was found that adult spontaneous recovery of fear memory was impaired in mice exposed to nicotine as adults and enhanced in mice exposed to nicotine as preadolescents. These results may indicate greater vulnerability to recurrence of traumatic memory as well as compromised inhibitory control in young smokers. (PsycINFO Database Record


Age-related changes in physical and perceptual markers of recovery following high-intensity interval cycle exercise.
The purpose of this study was to compare physical performance, perceptual and haematological markers of recovery in well-trained masters and young cyclists across 48 h following a bout of repeated high-intensity interval exercise. Nine masters (mean ± SD; age = 55.6 ± 5.0 years) and eight young (age = 25.9 ± 3.0 years) cyclists performed a high-intensity interval exercise session consisting of 6 × 30 s intervals at 175% peak power output with 4.5 min rest between efforts. Maximal voluntary contraction (MVC), 10 s sprint (10SST), 30-min time trial (30TT) performance, creatine kinase concentration (CK) and perceptual measures of motivation, total recovery, fatigue and muscle soreness were collected at baseline and at standardised time points across the 48 h recovery period. No significant group-time interactions were observed for performance of MVC, 10SST, 30TT and CK (P > 0.05). A significant reduction in 10SST peak power was found in both masters (P = 0.002) and young (P = 0.003) cyclists at 1 h post exercise, however, both groups physically recovered at similar rates. Neither group showed significant (P > 0.05) or practically meaningful increases in CK (%∆ < 10%). A significant age-related difference was found for perceptual fatigue (P = 0.01) and analysis of effect size (ES) showed that perceptual recovery was delayed with masters cyclists reporting lower motivation (ES ±90%CI = 0.69 ± 0.77, moderate), greater fatigue (ES = 0.75 ± 0.93, moderate) and muscle soreness (ES = 0.61 ± 0.70, moderate) after 48 h of recovery. The delay in perceived recovery may have negative effects on long-term participation to systematic training.


E-type asteroid (2867) Steins as imaged by OSIRIS on board Rosetta.
The European Space Agency's Rosetta mission encountered the main-belt asteroid (2867) Steins while on its way to rendezvous with comet 67P/Churyumov-Gerasimenko. Images taken with the OSIRIS (optical, spectroscopic, and infrared remote( )imaging system) cameras on board Rosetta show that Steins is an oblate body with an effective spherical diameter of 5.3 kilometers. Its surface does not show color variations. The morphology of Steins is dominated by linear faults and a large 2.1-kilometer-diameter crater near its south pole. Crater counts reveal a distinct lack of small craters. Steins is not solid rock but a rubble pile and has a conical appearance that is probably the result of reshaping due to Yarkovsky-O'Keefe-Radzievskii-Paddack (YORP) spin-up. The OSIRIS images constitute direct evidence for the YORP effect on a main-belt asteroid.


Measles virus receptors: SLAM and CD46.
The success of vaccination against measles in developed countries has significantly reduced the incidence of measles-related morbidity and mortality. However, measles is still the leading cause of mortality in children from underdeveloped countries due to low vaccination coverage, high transmissibility of the measles virus as well as primary and secondary vaccine failure. As with any viral disease, the identification of the host molecule to which the measles virus binds and gains entry into the host cell is a major step in understanding the molecular pathology of the disease. Two cell surface receptors, CD46 and signaling lymphocyte-activation molecule (SLAM), have been identified as measles virus receptors. CD46 is ubiquitously expressed on all nucleated cells and acts as a receptor for the Edmonston strain and all vaccine strains derived from it. SLAM is selectively expressed on some T and B cells and is utilised by the Edmonston strain and wild-type strains that cannot use CD46 for cell entry. Understanding the structural and functional variations in measles virus receptors with regard to host response can facilitate the development of new vaccines as well as provide new insights into measles virus tropism and pathogenesis and, importantly, into possible mechanisms for vaccine non-response. Our review focuses on the structure of measles virus receptors, measles virus receptor function, isoforms and polymorphic forms.


Photonic jet driven non-linear optics: example of two-photon fluorescence enhancement by dielectric microspheres.
The two-photon excited fluorescence from a dye solution is enhanced when a small amount of micro-meter sized silica beads are added. This observation is made in the simple scattering regime (inter-sphere distance four times larger than their radius) and is shown to depend on the concentration of the silica spheres. For a solution of rhodamine B, the enhancement can reach more than 30 %. As complementary experiments show that the fluorescence efficiency is unchanged, we argue that the non-linear absorption is enhanced due to focussing of the incident beam in the near-field of the spheres, a situation previously referred to as photonic (nano-)jets [3]. Our calculations indeed show that for the parameters of the spheres studied near-field focussing leads to an intensity concentration close to the sphere surface. We suggest that these photonic jets could be used to enhance other non-linear optical effects.


Anisakis simplex: from obscure infectious worm to inducer of immune hypersensitivity.
Infection of humans with the nematode worm parasite Anisakis simplex was first described in the 1960s in association with the consumption of raw or undercooked fish. During the 1990s it was realized that even the ingestion of dead worms in food fish can cause severe hypersensitivity reactions, that these may be more prevalent than infection itself, and that this outcome could be associated with food preparations previously considered safe. Not only may allergic symptoms arise from infection by the parasites ("gastroallergic anisakiasis"), but true anaphylactic reactions can also occur following exposure to allergens from dead worms by food-borne, airborne, or skin contact routes. This review discusses A. simplex pathogenesis in humans, covering immune hypersensitivity reactions both in the context of a living infection and in terms of exposure to its allergens by other routes. Over the last 20 years, several studies have concentrated on A. simplex antigen characterization and innate as well as adaptive immune response to this parasite. Molecular characterization of Anisakis allergens and isolation of their encoding cDNAs is now an active field of research that should provide improved diagnostic tools in addition to tools with which to enhance our understanding of pathogenesis and controversial aspects of A. simplex allergy. We also discuss the potential relevance of parasite products such as allergens, proteinases, and proteinase inhibitors and the activation of basophils, eosinophils, and mast cells in the induction of A. simplex-related immune hypersensitivity states induced by exposure to the parasite, dead or alive.


Alterations in the leaf lipidome of Brassica carinata under high-temperature stress.
Brassica carinata (A) Braun has recently gained increased attention across the world as a sustainable biofuel crop. B. carinata is grown as a summer crop in many regions where high temperature is a significant stress during the growing season. However, little research has been conducted to understand the mechanisms through which this crop responds to high temperatures. Understanding traits that improve the high-temperature adaption of this crop is essential for developing heat-tolerant varieties. This study investigated lipid remodeling in B. carinata in response to high-temperature stress. A commercial cultivar, Avanza 641, was grown under sunlit-controlled environmental conditions in Soil-Plant-Atmosphere-Research (SPAR) chambers under optimal temperature (OT; 23/15°C) conditions. At eight days after sowing, plants were exposed to one of the three temperature treatments [OT, high-temperature treatment-1 (HT-1; 33/25°C), and high-temperature treatment-2 (HT-2; 38/30°C)]. The temperature treatment period lasted until the final harvest at 84 days after sowing. Leaf samples were collected at 74 days after sowing to profile lipids using electrospray-ionization triple quadrupole mass spectrometry. Temperature treatment significantly affected the growth and development of Avanza 641. Both high-temperature treatments caused alterations in the leaf lipidome. The alterations were primarily manifested in terms of decreases in unsaturation levels of membrane lipids, which was a cumulative effect of lipid remodeling. The decline in unsaturation index was driven by (a) decreases in lipids that contain the highly unsaturated linolenic (18:3) acid and (b) increases in lipids containing less unsaturated fatty acids such as oleic (18:1) and linoleic (18:2) acids and/or saturated fatty acids such as palmitic (16:0) acid. A third mechanism that likely contributed to lowering unsaturation levels, particularly for chloroplast membrane lipids, is a shift toward lipids made by the eukaryotic pathway and the channeling of eukaryotic pathway-derived glycerolipids that are composed of less unsaturated fatty acids into chloroplasts. The lipid alterations appear to be acclimation mechanisms to maintain optimal membrane fluidity under high-temperature conditions. The lipid-related mechanisms contributing to heat stress response as identified in this study could be utilized to develop biomarkers for heat tolerance and ultimately heat-tolerant varieties.


Analyzing patterns of drug use in clinical notes for patient safety.
Doctors prescribe drugs for indications that are not FDA approved. Research indicates that 21% of prescriptions filled are for off-label indications. Of those, more than 73% lack supporting scientific evidence. Traditional drug safety alerts may not cover usages that are not FDA approved. Therefore, analyzing patterns of off-label drug usage in the clinical setting is an important step toward reducing the incidence of adverse events and for improving patient safety. We applied term extraction tools on the clinical notes of a million patients to compile a database of statistically significant patterns of drug use. We validated some of the usage patterns learned from the data against sources of known on-label and off-label use. Given our ability to quantify adverse event risks using the clinical notes, this will enable us to address patient safety because we can now rank-order off-label drug use and prioritize the search for their adverse event profiles.


High-resolution mapping of open chromatin in the rice genome.
Gene expression is controlled by the complex interaction of transcription factors binding to promoters and other regulatory DNA elements. One common characteristic of the genomic regions associated with regulatory proteins is a pronounced sensitivity to DNase I digestion. We generated genome-wide high-resolution maps of DNase I hypersensitive (DH) sites from both seedling and callus tissues of rice (Oryza sativa). Approximately 25% of the DH sites from both tissues were found in putative promoters, indicating that the vast majority of the gene regulatory elements in rice are not located in promoter regions. We found 58% more DH sites in the callus than in the seedling. For DH sites detected in both the seedling and callus, 31% displayed significantly different levels of DNase I sensitivity within the two tissues. Genes that are differentially expressed in the seedling and callus were frequently associated with DH sites in both tissues. The DNA sequences contained within the DH sites were hypomethylated, consistent with what is known about active gene regulatory elements. Interestingly, tissue-specific DH sites located in the promoters showed a higher level of DNA methylation than the average DNA methylation level of all the DH sites located in the promoters. A distinct elevation of H3K27me3 was associated with intergenic DH sites. These results suggest that epigenetic modifications play a role in the dynamic changes of the numbers and DNase I sensitivity of DH sites during development.


The health of men: structured inequalities and opportunities.
I have summarized in this article data on the magnitude of health challenges faced by men in the United States. Across a broad range of indicators, men report poorer health than women. Although men in all socioeconomic groups are doing poorly in terms of health, some especially high-risk groups include men of low socioeconomic status (SES) of all racial/ethnic backgrounds, low-SES minority men, and middle-class Black men. Multiple factors contribute to the elevated health risks of men. These include economic marginality, adverse working conditions, and gendered coping responses to stress, each of which can lead to high levels of substance use, other health-damaging behaviors, and an aversion to health-protective behaviors. The forces that adversely affect men's health are interrelated, unfold over the life course, and are amenable to change.


Molecular genetics of human cancer predisposition and progression.
The development of human cancer is generally thought to entail a series of events that cause a progressively more malignant phenotype. Such a hypothesis predicts that tumor cells of the ultimate stage will carry each of the events, cells of the penultimate stage will carry each of the events less the last one and so on. A dissection of the pathway from a normal cell to a fully malignant tumor may thus be viewed as the unraveling of a nested set of aberrations. In experiments designed to elucidate these events we have compared genotypic combinations at genomic loci defined by restriction endonuclease recognition site variation in normal and tumor tissues from patients with various forms and stages of cancer. The first step, inherited predisposition, is best described for retinoblastoma in which a recessive mutation of a locus residing in the 13q14 region of the genome is unmasked by aberrant, but specific, mitotic chromosomal segregation. Similar mechanisms involving the distal short arm of chromosome 17 are apparent in astrocytic tumors and the events are shared by cells in each malignancy state. DNA sequencing indicates that these events accomplish the homozygosis of mutant alleles of the p53 gene. Copy number amplification of the epidermal growth factor receptor gene occurs in intermediate and late-stage tumors whereas loss of heterozygosity for loci on chromosome 10 is restricted to the ultimate stage, glioblastoma multiforme. These results suggest a genetic approach to defining degrees of tumor progression and the locations of genes involved in the pathway as a prelude to their molecular isolation and characterization.


Colorimetry as Quality Control Tool for Individual Inkjet-Printed Pediatric Formulations.
Printing technologies were recently introduced to the pharmaceutical field for manufacturing of drug delivery systems. Printing allows on demand manufacturing of flexible pharmaceutical doses in a personalized manner, which is critical for a successful and safe treatment of patient populations with specific needs, such as children and the elderly, and patients facing multimorbidity. Printing of pharmaceuticals as technique generates new demands on the quality control procedures. For example, rapid quality control is needed as the printing can be done on demand and at the point of care. This study evaluated the potential use of a handheld colorimetry device for quality control of printed doses of vitamin Bs on edible rice and sugar substrates. The structural features of the substrates with and without ink were also compared. A multicomponent ink formulation with vitamin B1, B2, B3, and B6 was developed. Doses (4 cm2) were prepared by applying 1-10 layers of yellow ink onto the white substrates using thermal inkjet technology. The colorimetric method was seen to be viable in detecting doses up to the 5th and 6th printed layers until color saturation of the yellow color parameter (b*) was observed on the substrates. Liquid chromatography mass spectrometry was used as a reference method for the colorimetry measurements plotted against the number of printed layers. It was concluded that colorimetry could be used as a quality control tool for detection of different doses. However, optimization of the color addition needs to be done to avoid color saturation within the planned dose interval.


Protein profiling in hepatocellular carcinoma by label-free quantitative proteomics in two west African populations.
Hepatocellular Carcinoma is the third most common cause of cancer related death worldwide, often diagnosed by measuring serum AFP; a poor performance stand-alone biomarker. With the aim of improving on this, our study focuses on plasma proteins identified by Mass Spectrometry in order to investigate and validate differences seen in the respective proteomes of controls and subjects with LC and HCC. Mass Spectrometry analysis using liquid chromatography electro spray ionization quadrupole time-of-flight was conducted on 339 subjects using a pooled expression profiling approach. ELISA assays were performed on four significantly differentially expressed proteins to validate their expression profiles in subjects from the Gambia and a pilot group from Nigeria. Results from this were collated for statistical multiplexing using logistic regression analysis. Twenty-six proteins were identified as differentially expressed between the three subject groups. Direct measurements of four; hemopexin, alpha-1-antitrypsin, apolipoprotein A1 and complement component 3 confirmed their change in abundance in LC and HCC versus control patients. These trends were independently replicated in the pilot validation subjects from Nigeria. The statistical multiplexing of these proteins demonstrated performance comparable to or greater than ALT in identifying liver cirrhosis or carcinogenesis. This exercise also proposed preliminary cut offs with achievable sensitivity, specificity and AUC statistics greater than reported AFP averages. The validated changes of expression in these proteins have the potential for development into high-performance tests usable in the diagnosis and or monitoring of HCC and LC patients. The identification of sustained expression trends strengthens the suggestion of these four proteins as worthy candidates for further investigation in the context of liver disease. The statistical combinations also provide a novel inroad of analyses able to propose definitive cut-offs and combinations for evaluation of performance.


Guanylate binding proteins enable rapid activation of canonical and noncanonical inflammasomes in Chlamydia-infected macrophages.
Interferon (IFN)-inducible guanylate binding proteins (GBPs) mediate cell-autonomous host resistance to bacterial pathogens and promote inflammasome activation. The prevailing model postulates that these two GBP-controlled activities are directly linked through GBP-dependent vacuolar lysis. It was proposed that the rupture of pathogen-containing vacuoles (PVs) by GBPs destroyed the microbial refuge and simultaneously contaminated the host cell cytosol with microbial activators of inflammasomes. Here, we demonstrate that GBP-mediated host resistance and GBP-mediated inflammatory responses can be uncoupled. We show that PVs formed by the rodent pathogen Chlamydia muridarum, so-called inclusions, remain free of GBPs and that C. muridarum is impervious to GBP-mediated restrictions on bacterial growth. Although GBPs neither bind to C. muridarum inclusions nor restrict C. muridarum growth, we find that GBPs promote inflammasome activation in C. muridarum-infected macrophages. We demonstrate that C. muridarum infections induce GBP-dependent pyroptosis through both caspase-11-dependent noncanonical and caspase-1-dependent canonical inflammasomes. Among canonical inflammasomes, we find that C. muridarum and the human pathogen Chlamydia trachomatis activate not only NLRP3 but also AIM2. Our data show that GBPs support fast-kinetics processing and secretion of interleukin-1β (IL-1β) and IL-18 by the NLRP3 inflammasome but are dispensable for the secretion of the same cytokines at later times postinfection. Because IFN-γ fails to induce IL-1β transcription, GBP-dependent fast-kinetics inflammasome activation can drive the preferential processing of constitutively expressed IL-18 in IFN-γ-primed macrophages in the absence of prior Toll-like receptor stimulation. Together, our results reveal that GBPs control the kinetics of inflammasome activation and thereby shape macrophage responses to Chlamydia infections.


Asymmetric adhesion of rod-shaped bacteria controls microcolony morphogenesis.
Surface colonization underpins microbial ecology on terrestrial environments. Although factors that mediate bacteria-substrate adhesion have been extensively studied, their spatiotemporal dynamics during the establishment of microcolonies remains largely unexplored. Here, we use laser ablation and force microscopy to monitor single-cell adhesion during the course of microcolony formation. We find that adhesion forces of the rod-shaped bacteria Escherichia coli and Pseudomonas aeruginosa are polar. This asymmetry induces mechanical tension, and drives daughter cell rearrangements, which eventually determine the shape of the microcolonies. Informed by experimental data, we develop a quantitative model of microcolony morphogenesis that enables the prediction of bacterial adhesion strength from simple time-lapse measurements. Our results demonstrate how patterns of surface colonization derive from the spatial distribution of adhesive factors on the cell envelope.


Competing discrete interfacial effects are critical for amyloidogenesis.
Amyloid accumulation is associated with pathological conditions, including type II diabetes and Alzheimer's disease. Lipids influence amyloidogenesis and are themselves targets for amyloid-mediated cell membrane disruption. Amyloid precursors are surface-active, accumulating at hydrophobic-hydrophilic interfaces (e.g., air-water), where their biophysical and kinetic behaviors differ from those in the bulk solution with significant and underappreciated consequences. Biophysical modeling predicted the probability and rate of beta-sheet amyloid dimer formation to be higher and faster at the air-water interface (AWI) than in the bulk (by 14 and approximately 1500 times, respectively). Time-course staining experiments with a typical amyloid dye verified our predictions by demonstrating that without AWI, islet amyloid polypeptide (IAPP) fibrilization was abolished or slowed, depending on the conditions. Our controls included undisturbed IAPP reactions, and we ascertained that the AWI removal process (technical or material) did not itself affect the reaction. Furthermore, we showed that the role of membranes in amyloidogenesis has been previously underestimated; in an in vivo-like situation (with no AWI), anionic liposomes (containing dioleoylphosphatidylglycerol) enhanced IAPP fibrilogenesis far more than described previously in conventional assay conditions (in the presence of an AWI). These findings have implications for the protein misfolding field and in assay design to target toxic protein aggregation.


Clinical Outcomes and Prognostic Factors of Up-Front Autologous Stem Cell Transplantation in Patients with Extranodal Natural Killer/T Cell Lymphoma.
Limited data exist on up-front autologous stem cell transplantation (ASCT) in extranodal natural killer/T cell lymphoma (ENKTL). Sixty-two patients (43 men and 19 women) with newly diagnosed ENKTL who underwent up-front ASCT after primary therapy were identified. Poor-risk characteristics included advanced stage (50%), high-intermediate to high-risk International Prognostic Index (25.8%), and group 3 to 4 of NK/T Cell Lymphoma Prognostic Index (NKPI, 67.7%). Pretransplant responses included complete remission in 61.3% and partial remission in 38.7% of patients, and final post-transplantation response included complete remission in 78.3%. Early progression occurred in 12.9%. At a median follow-up of 43.3 months (range, 3.7 to 114.6), 3-year progression-free survival (PFS) was 52.4% and 3-year overall survival (OS) was 60.0%. Patients with limited disease had significantly better 3-year PFS (64.5% versus 40.1%, P = .017) and OS (67.6% versus 52.3%, P = .048) than those with advanced disease. Multivariate analysis showed NKPI and pretransplant response were independent prognostic factors influencing survival, particularly NKPI in limited disease and pretransplant response in advanced disease. Radiotherapy was an independent factor for reduced progression and survival in patients with limited disease, but anthracycline-based chemotherapy was a poor prognostic factor for progression in patients with advanced disease. Up-front ASCT is an active treatment in ENKTL patients responding to primary therapy.