Underestimating the frequency, strength and cost of antipredator responses with data from GPS collars: an example with wolves and elk.
Field studies that rely on fixes from GPS-collared predators to identify encounters with prey will often underestimate the frequency and strength of antipredator responses. These underestimation biases have several mechanistic causes. (1) Step bias: The distance between successive GPS fixes can be large, and encounters that occur during these intervals go undetected. This bias will generally be strongest for cursorial hunters that can rapidly cover large distances (e.g., wolves and African wild dogs) and when the interval between GPS fixes is long relative to the duration of a hunt. Step bias is amplified as the path travelled between successive GPS fixes deviates from a straight line. (2) Scatter bias: Only a small fraction of the predators in a population typically carry GPS collars, and prey encounters with uncollared predators go undetected unless a collared group-mate is present. This bias will generally be stronger for fission-fusion hunters (e.g., spotted hyenas, wolves, and lions) than for highly cohesive hunters (e.g., African wild dogs), particularly when their group sizes are large. Step bias and scatter bias both cause underestimation of the frequency of antipredator responses. (3) Strength bias: Observations of prey in the absence of GPS fix from a collared predator will generally include a mixture of cases in which predators were truly absent and cases in which predators were present but not detected, which causes underestimation of the strength of antipredator responses. We quantified these biases with data from wolves and African wild dogs and found that fixes from GPS collars at 3-h intervals underestimated the frequency and strength of antipredator responses by a factor >10. We reexamined the results of a recent study of the nonconsumptive effects of wolves on elk in light of these results and confirmed that predation risk has strong effects on elk dynamics by reducing the pregnancy rate.


A pooled analysis of post-diagnosis lifestyle factors in association with late estrogen-receptor-positive breast cancer prognosis.
Lifestyle factors have been well studied in relation to breast cancer prognosis overall; however, associations of lifestyle and late outcomes (>5 years after diagnosis) have been much less studied, and no studies have focused on estrogen receptor-positive (ER+) breast cancer survivors, who may have high risk of late recurrence and mortality. We utilized a large prospective pooling study to evaluate the associations of lifestyle factors with late recurrence and all-cause mortality among 6,295 5-year ER+ Stage I-III breast cancer survivors. Pooled and harmonized data were available on clinical factors and lifestyle factors (pre- to post-diagnosis weight change, body mass index (BMI) (kg/m(2)), recreational physical activity, alcohol intake and smoking history), measured on average 2.1 years after diagnosis. Updated information for weight only was available. Study heterogeneity was evaluated by the Q-statistic. Multivariable Cox regression models were stratified by study. Adjusting for clinical factors and potential confounders, ≥ 10% weight gain and obesity (BMI, 30-34.99 and ≥ 35) were associated with increased risk of late recurrence (hazard ratios (95% confidence intervals): 1.24 (1.00-1.53), 1.40 (1.05-1.86) and 1.41 (1.02-1.93), respectively). Daily alcohol intake was associated with late recurrence, 1.28 (1.01-1.62). Physical activity was inversely associated with late all-cause mortality (0.81 (0.71-0.93) and 0.71 (0.61-0.82) for 4.9 to <17.4 and ≥ 17.4 metabolic equivalent-hr/week). A U-shaped association was observed for late all-cause mortality and BMI using updated weight (1.42 (1.15-1.74) and 1.40 (1.09-1.81), <21.5 and ≥ 35, respectively). Smoking was associated with increased risk of late outcomes. In this large prospective pooling project, modifiable lifestyle factors were associated with late outcomes among long-term ER+ breast cancer survivors.


The response of Isidorella newcombi to copper exposure: Using an integrated biological framework to interpret transcriptomic responses from RNA-seq analysis.
This study describes the transcriptomic response of the Australian endemic freshwater gastropod Isidorella newcombi exposed to 80±1μg/L of copper for 3days. Analysis of copper tissue concentration, lysosomal membrane destabilisation and RNA-seq were conducted. Copper tissue concentrations confirmed that copper was bioaccumulated by the snails. Increased lysosomal membrane destabilisation in the copper-exposed snails indicated that the snails were stressed as a result of the exposure. Both copper tissue concentrations and lysosomal destabilisation were significantly greater in snails exposed to copper. In order to interpret the RNA-seq data from an ecotoxicological perspective an integrated biological response model was developed that grouped transcriptomic responses into those associated with copper transport and storage, survival mechanisms and cell death. A conceptual model of expected transcriptomic changes resulting from the copper exposure was developed as a basis to assess transcriptomic responses. Transcriptomic changes were evident at all the three levels of the integrated biological response model. Despite lacking statistical significance, increased expression of the gene encoding copper transporting ATPase provided an indication of increased internal transport of copper. Increased expression of genes associated with endocytosis are associated with increased transport of copper to the lysosome for storage in a detoxified form. Survival mechanisms included metabolic depression and processes associated with cellular repair and recycling. There was transcriptomic evidence of increased cell death by apoptosis in the copper-exposed organisms. Increased apoptosis is supported by the increase in lysosomal membrane destabilisation in the copper-exposed snails. Transcriptomic changes relating to apoptosis, phagocytosis, protein degradation and the lysosome were evident and these processes can be linked to the degradation of post-apoptotic debris. The study identified contaminant specific transcriptomic markers as well as markers of general stress. From an ecotoxicological perspective, the use of a framework to group transcriptomic responses into those associated with copper transport, survival and cell death assisted with the complex process of interpretation of RNA-seq data. The broad adoption of such a framework in ecotoxicology studies would assist in comparison between studies and the identification of reliable transcriptomic markers of contaminant exposure and response.


Zinc-induced neuronal death in cortical neurons.
Although Zn2+ is normally stored and released in the brain, excessive exposure to extracellular Zn2+ can be neurotoxic. The purpose of the present study was to determine the type of neuronal cell death, necrosis versus apoptosis, induced by Zn2+ exposure. Addition of 10-50 microM ZnCl2 to the bathing medium of murine neuronal and glial cell cultures induced, over the next 24 hrs., Zn2+-concentration-dependent neuronal death; some glial death also occurred with Zn2+ concentrations above 30 microM. The neuronal death induced by 20 microM Zn2+ was characterized by coarse chromatin condensation, the formation of apoptotic bodies, and internucleosomal DNA fragmentation. It was attenuated in cortical cell cultures prepared from mice null for the bax gene, and by the caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-CH2F (ZVAD, 100 microM), but not by the NMDA receptor antagonist, D-2-amino-5-phosphonovalerate (D-APV, 200 microM ). In contrast, the neuronal death induced by 50 microM Zn2+ was characterized by plasma membrane disruption and random DNA fragmentation; this death was attenuated by D-APV, but exhibited little sensitivity to ZVAD or deletion of bax. These results suggest that Zn2+ can induce cell death with characteristics of either apoptosis or necrosis, depending on the intensity of the Zn2+ exposure.


Content validity and readability of patient-reported questionnaire instruments of hearing disability.
Objective: This study evaluates the content validity (i.e. domains assessed) and readability levels of patient-reported questionnaire instruments using internationally recognised procedures and tools. Design: A review of the literature to identify candidate instruments and a synthesis of information including mapping extracted items onto the World Health Organisation's - International Classification of Functioning, Disability, and Health (WHO-ICF) and estimating readability. Study sample: 14 patient-reported questionnaire instruments. Results: In general, item content focussed on body function and on activity limitations and participation restrictions, with less emphasis on environmental and personal factors and with different emphases across instruments. Many items did not clearly map onto any of the WHO-ICF categories (i.e. not coded items ranged from 3.7 to 39.1% across the 14 questionnaires). All 14 instruments exceeded the sixth-grade reading level when calculated according to the FORCAST formula which is appropriate for assessing a non-narrative text. Conclusions: Clinical assessment of hearing disability is only as comprehensive as the items covered by the chosen measurement instrument. Our findings confirmed the diversity of domains covered by hearing disability instruments and gaps in assessment. Some concern is raised about whether the item content is appropriate for those respondents with poor literacy.


D-Dimer Levels before HIV Seroconversion Remain Elevated Even after Viral Suppression and Are Associated with an Increased Risk of Non-AIDS Events.
The mechanism underlying the excess risk of non-AIDS diseases among HIV infected people is unclear. HIV associated inflammation/hypercoagulability likely plays a role. While antiretroviral therapy (ART) may return this process to pre-HIV levels, this has not been directly demonstrated. We analyzed data/specimens on 249 HIV+ participants from the US Military HIV Natural History Study, a prospective, multicenter observational cohort of >5600 active duty military personnel and beneficiaries living with HIV. We used stored blood specimens to measure D-dimer and Interleukin-6 (IL-6) at three time points: pre-HIV seroconversion, ≥6 months post-HIV seroconversion but prior to ART initiation, and ≥6 months post-ART with documented HIV viral suppression on two successive evaluations. We evaluated the changes in biomarker levels between time points, and the association between these biomarker changes and future non-AIDS events. During a median follow-up of 3.7 years, there were 28 incident non-AIDS diseases. At ART initiation, the median CD4 count was 361cells/mm3; median duration of documented HIV infection 392 days; median time on ART was 354 days. Adjusted mean percent increase in D-dimer levels from pre-seroconversion to post-ART was 75.1% (95% confidence interval 24.6-148.0, p = 0.002). This increase in D-dimer was associated with a significant 22% increase risk of future non-AIDS events (p = 0.03). Changes in IL-6 levels across time points were small and not associated with future non-AIDS events. In conclusion, ART initiation and HIV viral suppression does not eliminate HIV associated elevation in D-dimer levels. This residual pathology is associated with an increased risk of future non-AIDS diseases.


Respiratory muscle activity and pulmonary function during acutely induced airways obstruction.
Patients with airway obstruction may spontaneously breathe with a positive expiratory pressure by pursing their lips during expiration, especially in case of dyspnea. Dyspnea seems to be related to increased activity of the respiratory muscles. A potential explanation for the use of pursed lips breathing (PLB) is that this type of breathing leads to a reduction of respiratory muscle activity. The purpose of this study was to assess the effect of breathing with a positive expiratory pressure of 5 cm H2O, simulating pursed lips breathing (SPLB), on respiratory muscle activity and pulmonary function during induced airway obstruction. In twelve asthmatic patients, tonic and phasic electromyographic (EMG) activity of the following muscles was obtained: scalene muscle, parasternal muscle, and abdominal muscles. Pulmonary function and EMG measurements were performed before and after propranolol induced airway obstruction. Simulated pursed lips breathing resulted in a significant increase of functional residual capacity and tidal volume both at baseline and during airway obstruction. Phasic respiratory muscle activity during PEP breathing increased especially at baseline. We conclude that beneficial effects of breathing with a positive expiratory pressure of 5 cm H2O, which is similar to pursed lips breathing, cannot be explained by changes in respiratory muscle activity or pulmonary function.


Interexaminer agreement on palpatory diagnosis and patient self-assessment of disability: a pilot study.
To evaluate interexaminer agreement on palpatory diagnostic findings, we compared interexaminer results, patient disability self-evaluations, and assessment of patients' progress by referring physicians. Three examiners (two clinicians and a third-year osteopathic medical student fellow) monitored patients' progress using negotiated examination procedures. The patients were examined independently by each examiner at each of eight weekly visits. Patients completed a disability assessment form at each visit, and the referring physicians recorded changes in their patients' condition. The patients' disability self-rating and examiner test results did not show clear correlation. There was a 62.2% agreement between the two physician examiners when the general descriptors "improvement," "no change," or "aggravation" were used. The student examiner's agreement with the clinician examiners was 60.2% and 51.8%. Interexaminer agreement of findings from osteopathic testing procedures appears to depend on general clinical experience and specific experience with the testing procedures.


LCMV-specific, class II-restricted cytotoxic T cells in beta 2-microglobulin-deficient mice.
Intracranial infection of normal mice with lymphocytic choriomeningitis virus (LCMV) causes meningitis and death mediated by CD8+ major histocompatibility complex (MHC) class I-restricted cytotoxic T lymphocytes (CTLs). beta 2-Microglobulin-deficient mice (beta 2M-/-) do not express functional MHC class I proteins and do not produce significant numbers of CD8+ T cells. When beta 2M-/- mice were infected with LCMV, many died from LCMV disease and produced a specific response to LCMV mediated by CD4+ CTLs that were class II-restricted. In these mice, CD4+ CTLs may compensate for the lack of CD8+ CTLs.


Niemann-Pick type C2 protein regulates liver cancer progression via modulating ERK1/2 pathway: Clinicopathological correlations and therapeutical implications.
Primary hepatocellular carcinoma (HCC) is the fifth most common malignancy worldwide and the third leading cause of cancer-related death. It is important to identify new targets for early diagnosis and treatment of HCC. Niemann-Pick type C2 (NPC2) plays an important role in the regulation of intracellular cholesterol homeostasis via direct binding with free cholesterol. However, little is known about the significance of NPC2 in HCC tumorigenesis. In this study, we showed that NPC2 is abundantly expressed in normal liver, but is downregulated in human HCC tissues. The patients with NPC2 downregulation expressed much higher α-fetoprotein, multiple tumor type, vascular invasion, later pathological stage and shorter survival rate. Knockdown NPC2 in liver cancer cell lines promote cell proliferation, migration and xenograft tumorigenesis. In contrast, NPC2 overexpression inhibits HuH7 promoted tumor growth. Furthermore, administration of hepatotropic adeno-associated virus 8 (AAV8) delivered NPC2 decreased the inflammatory infiltration, the expression of two early HCC markers-glypican 3 and survivin and suppressed the spontaneous HCC development in mice. To identify the NPC2-dependent mechanism, we emphasized on the status of MAPK/ERK signaling. MEK1/2 inhibitor treatment demonstrated that the expression of NPC2 affected the activation of ERK1/2 but not MEK1/2. In addition, cholesterol trafficking inhibitor treatment did not alter the cell proliferation and the activation of MEK/ERK. In conclusion, our study demonstrates that NPC2 may play an important role in negatively regulate cell proliferation and ERK1/2 activation that were independent of cholesterol accumulation. AAV-NPC2 may thus represent a new treatment strategy for liver cancer.


RB regulates the stability and the apoptotic function of p53 via MDM2.
The binding of RB to MDM2 is shown to be essential for RB to overcome both the antiapoptotic function of MDM2 and the MDM2-dependent degradation of p53. The RB-MDM2 interaction does not prevent MDM2 from inhibiting p53-dependent transcription, but the RB-MDM2 complex still binds to p53. Since RB specifically rescues the apoptotic function but not the transcriptional activity of p53 from negative regulation by MDM2, transactivation by wild-type p53 is not required for the apoptotic function of p53. However, an RB-MDM2-p53 trimeric complex is active in p53-mediated transrepression. These data link directly the function of two tumor suppressor proteins and demonstrate a novel role of RB in regulating the apoptotic function of p53.


Functional and biological diversity of foliar spectra in tree canopies throughout the Andes to Amazon region.
Spectral properties of foliage express fundamental chemical interactions of canopies with solar radiation. However, the degree to which leaf spectra track chemical traits across environmental gradients in tropical forests is unknown. We analyzed leaf reflectance and transmittance spectra in 2567 tropical canopy trees comprising 1449 species in 17 forests along a 3400-m elevation and soil fertility gradient from the Amazonian lowlands to the Andean treeline. We developed quantitative links between 21 leaf traits and 400-2500-nm spectra, and developed classifications of tree taxa based on spectral traits. Our results reveal enormous inter-specific variation in spectral and chemical traits among canopy trees of the western Amazon. Chemical traits mediating primary production were tightly linked to elevational changes in foliar spectral signatures. By contrast, defense compounds and rock-derived nutrients tracked foliar spectral variation with changing soil fertility in the lowlands. Despite the effects of abiotic filtering on mean foliar spectral properties of tree communities, the spectra were dominated by phylogeny within any given community, and spectroscopy accurately classified 85-93% of Amazonian tree species. Our findings quantify how tropical tree canopies interact with sunlight, and indicate how to measure the functional and biological diversity of forests with spectroscopy.


Effects of vitamin D and quercetin, alone and in combination, on cardiorespiratory fitness and muscle function in physically active male adults.
Vitamin D and the antioxidant quercetin, are promising agents for improving physical performance because of their possible beneficial effects on muscular strength and cardiorespiratory fitness. The purpose of this study was to determine the effects of increased intakes of vitamin D, quercetin, and their combination on antioxidant status, the steroid hormone regulators of muscle function, and measures of physical performance in apparently healthy male adults engaged in moderate-to-vigorous-intensity exercise training. A total of 40 adult male participants were randomized to either 4,000 IU vitamin D/d, 1,000 mg/d quercetin, vitamin D plus quercetin, or placebo for 8 weeks. Measures of cardiorespiratory fitness and muscle function, blood markers for antioxidant and vitamin D status, and hormones 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and testosterone were measured pre- and postsupplementation. At enrollment, 88.6% of participants were vitamin D sufficient (serum 25-hydroxyvitamin D >50 nmol/L) and had normal serum testosterone levels. Supplementation with vitamin D significantly increased serum 25(OH)D concentration (by 87.3% in the vitamin D group, P<0.001) and was associated with an increasing trend of testosterone concentration. There were no changes in concentration of 1,25(OH)2D3 and markers of antioxidant status associated with vitamin D or quercetin supplementation. No improvements in physical performance measures associated with vitamin D and quercetin supplementation were found. The findings obtained demonstrate that long-term vitamin D and quercetin supplementation, alone or in combination, does not improve physical performance in male adults with adequate vitamin D, testosterone, and antioxidant status.


SU-E-T-575: To Analyze the Clinical Impact of Esophageal Sparing on Treatment Plans for Patients with Grade 3 Esophagitis.
To analyze the clinical impact of esophageal sparing on treatment plans for patients with grade 3 esophagitis. The treatment plans of 8 patients (project total: 20 patients) who were treated with IMRT and exhibited stage 3 esophagitis were re-planned to give a simulated clinical plan with dose distribution that mirrored our current clinical practice (74Gy to the target, and 5mm margins), and a plan that emphasized esophageal sparing. Doses to the esophagus, heart, cord, lung and PTV were compared. Comparing the esophageal sparing plan to the simulated clinical plan, the mean reduction in esophageal volume receiving 50, 55, 60, 65, and 70Gy were 2.0, 3.2, 5.0, 7.2, and 10.9 cm3 , respectively. The mean reduction in the continuous length of esophagus receiving 50, 55, 60, 65, and 70Gy were 12, 24, 38, 40, and 47mm, respectively. The associated reduction in dose to 90% and 95% of the PTV was 2.2 and 3.8Gy, respectively. Of the 8 patients examined, 2 showed a significant decrease in PTV coverage (4.6Gy, 12.3Gy for 90% of PTV), 4 showed decreases under 1.1Gy, but 2 showed an increase of 1.4Gy and 0.5Gy for 90% PTV. Cord dose was maintained below 50Gy, and there was a slight increase in mean heart dose and mean lung dose of 2.4Gy, and 2.7Gy, respectively. Data will also be presented comparing these plans with the actual treated plans (for which the patients had grade 3 esophagitis) and plans that emphasize PTV coverage. Treatment planning to emphasize esophageal sparing can reduce the volume and continuous length of the esophagus which receives high doses. There is some associated modest reduction in PTV coverage. In summary, in many cases esophageal sparing can be accomplished for lung cancer cases while maintaining adequate PTV coverage, although there is variability between patients.


Relationships between bacterial community composition, functional trait composition and functioning are context dependent--but what is the context?
Bacterial communities are immensely diverse and drive many fundamental ecosystem processes. However, the role of bacterial community composition (BCC) for functioning is still unclear. Here we evaluate the relative importance of BCC (from 454-sequencing), functional traits (from Biolog Ecoplates) and environmental conditions for per cell biomass production (BPC; 3H-leucine incorporation) in six data sets of natural freshwater bacterial communities. BCC explained significant variation of BPC in all six data sets and most variation in four. BCC measures based on 16S rRNA (active bacteria) did not consistently explain more variation in BPC than measures based on the 16S rRNA-gene (total community), and adding phylogenetic information did not, in general, increase the explanatory power of BCC. In contrast to our hypothesis, the importance of BCC for BPC was not related to the anticipated dispersal rates in and out of communities. Functional traits, most notably the ability to use cyclic and aromatic compounds, as well as local environmental conditions, i.e. stoichiometric relationships of nutrients, explained some variation in all six data sets. In general there were weak associations between variation in BCC and variation in the functional traits contributing to productivity. This indicates that additional traits may be important for productivity as well. By comparing several data sets obtained in a similar way we conclude that no single measure of BCC was obviously better than another in explaining BPC. We identified some key functional traits for productivity, but although there was a coupling between BCC, functional traits and productivity, the strength of the coupling seems context dependent. However, the exact context is still unresolved.


A study on the student pilot's mental workload due to personality types of both instructor and student.
The main purpose of this paper is to provide the ideal flight crew combination for instructor and student pilots in order to enhance the flight training effects based on Myers Briggs Type Indicator (MBTI) theory. In addition to personality, various levels of flight difficulty are considered in order to investigate their potential interaction effect in terms of student pilots' mental workload measured in heart rate, altitude deviation, NASA-TLX (Task Load Index) and subjective degree of personality harmony. Based on an experiment performed in a real flight situation, we found significant effects of personality combinations in terms of all four outcome measures. Both group C types of instructors and students who are concrete, realistic and have mechanical skills turn out to be the ideal flight crew combination. A structural equation model, fitted to analyse causality among the four response variables, implied that as the heart rate increased, the altitude deviation increased. In addition, as the altitude deviation and personality harmony increased, NASA-TLX increased. The results of this study are expected to provide a theoretical basis for manning the flight crew combinations and thereby enhancing the efficiency of flight training.


Photo-induced H2 production by [NiFe]-hydrogenase from T. roseopersicina covalently linked to a Ru(II) photosensitizer.
The potential of hydrogen as a clean renewable fuel source and the finite reserves of platinum metal to be utilized in hydrogen production catalysts have provided the motivation for the development of non-noble metal-based solutions for catalytic hydrogen production. There are a number of microorganisms that possess highly efficient hydrogen production catalysts termed hydrogenases that generate hydrogen under certain metabolic conditions. Although hydrogenases occur in photosynthetic microorganisms, the oxygen sensitivity of these enzymes represents a significant barrier in directly coupling hydrogen production to oxygenic photosynthesis. To overcome this barrier, there has been considerable interest in identifying or engineering oxygen tolerant hydrogenases or generating mimetic systems that do not rely on oxygen producing photocatalysts. In this work, we demonstrate photo-induced hydrogen production from a stable [NiFe]-hydrogenase coupled to a [Ru(2,2'-bipyridine)(2)(5-amino-1,10-phenanthroline)](2+) photocatalyst. When the Ru(II) complex is covalently attached to the hydrogenase, photocatalytic hydrogen production occurs more efficiently in the presence of a redox mediator than if the Ru(II) complex is simply present in solution. Furthermore, sustained hydrogen production occurs even in the presence of oxygen by presumably creating a local anoxic environment through the reduction of oxygen similar to what is proposed for oxygen tolerant hydrogenases. These results provide a strong proof of concept for engineering photocatalytic hydrogen production in the presence of oxygen using biohybrid mimetic systems.


Use of Network Analysis and Spread Models to Target Control Actions for Bovine Tuberculosis in a State from Brazil.
Livestock movements create complex dynamic interactions among premises that can be represented, interpreted, and used for epidemiological purposes. These movements are a very important part of the production chain but may also contribute to the spread of infectious diseases through the transfer of infected animals over large distances. Social network analysis (SNA) can be used to characterize cattle trade patterns and to identify highly connected premises that may act as hubs in the movement network, which could be subjected to targeted control measures in order to reduce the transmission of communicable diseases such as bovine tuberculosis (TB). Here, we analyzed data on cattle movement and slaughterhouse surveillance for detection of TB-like lesions (TLL) over the 2016-2018 period in the state of Rio Grande do Sul (RS) in Brazil with the following aims: (i) to characterize cattle trade describing the static full, yearly, and monthly snapshots of the network contact trade, (ii) to identify clusters in the space and contact networks of premises from which animals with TLL originated, and (iii) to evaluate the potential of targeted control actions to decrease TB spread in the cattle population of RS using a stochastic metapopulation disease transmission model that simulated within-farm and between-farm disease spread. We found heterogeneous densities of premises and animals in the study area. The analysis of the contact network revealed a highly connected (~94%) trade network, with strong temporal trends, especially for May and November. The TLL cases were significantly clustered in space and in the contact network, suggesting the potential for both local (e.g., fence-to-fence) and movement-mediated TB transmission. According to the disease spread model, removing the top 7% connected farms based on degree and betweenness could reduce the total number of infected farms over three years by >50%. In conclusion, the characterization of the cattle network suggests that highly connected farms may play a role in TB dissemination, although being close to infected farms was also identified as a risk factor for having animals with TLL. Surveillance and control actions based on degree and betweenness could be useful to break the transmission cycle between premises in RS.


A review of data abstraction.
It is well-known that Artificial Intelligence (AI), and in particular Machine Learning (ML), is not effective without good data preparation, as also pointed out by the recent wave of data-centric AI. Data preparation is the process of gathering, transforming and cleaning raw data prior to processing and analysis. Since nowadays data often reside in distributed and heterogeneous data sources, the first activity of data preparation requires collecting data from suitable data sources and data services, often distributed and heterogeneous. It is thus essential that providers describe their data services in a way to make them compliant with the FAIR guiding principles, i.e., make them automatically Findable, Accessible, Interoperable, and Reusable (FAIR). The notion of data abstraction has been introduced exactly to meet this need. Abstraction is a kind of reverse engineering task that automatically provides a semantic characterization of a data service made available by a provider. The goal of this paper is to review the results obtained so far in data abstraction, by presenting the formal framework for its definition, reporting about the decidability and complexity of the main theoretical problems concerning abstraction, and discuss open issues and interesting directions for future research.


Rapid treatment of PTSD: why psychological exposure with acupoint tapping may be effective.
Combining brief psychological exposure with the manual stimulation of acupuncture points (acupoints) in the treatment of posttraumatic stress disorder (PTSD) and other emotional conditions is an intervention strategy that integrates established clinical principles with methods derived from healing traditions of Eastern cultures. Two randomized controlled trials and six outcome studies using standardized pre- and posttreatment measures with military veterans, disaster survivors, and other traumatized individuals corroborate anecdotal reports and systematic clinical observation in suggesting that (a) tapping on selected acupoints, (b) during imaginal exposure, (c) quickly and permanently reduces maladaptive fear responses to traumatic memories and related cues. The approach has been controversial. This is in part because the mechanisms by which stimulating acupoints can contribute to the treatment of serious or longstanding psychological disorders have not been established. Speculating on such mechanisms, the current paper suggests that adding acupoint stimulation to brief psychological exposure is unusually effective in its speed and power because deactivating signals are sent directly to the amygdala, resulting in the rapid attenuation of threat responses to innocuous stimuli. This formulation and the preliminary evidence supporting it could, if confirmed, lead to more powerful exposure protocols for treating PTSD.


Effects of a 20 year rain event: a quantitative microbial risk assessment of a case of contaminated bathing water in Copenhagen, Denmark.
Quantitative microbial risk assessments (QMRAs) often lack data on water quality leading to great uncertainty in the QMRA because of the many assumptions. The quantity of waste water contamination was estimated and included in a QMRA on an extreme rain event leading to combined sewer overflow (CSO) to bathing water where an ironman competition later took place. Two dynamic models, (1) a drainage model and (2) a 3D hydrodynamic model, estimated the dilution of waste water from source to recipient. The drainage model estimated that 2.6% of waste water was left in the system before CSO and the hydrodynamic model estimated that 4.8% of the recipient bathing water came from the CSO, so on average there was 0.13% of waste water in the bathing water during the ironman competition. The total estimated incidence rate from a conservative estimate of the pathogenic load of five reference pathogens was 42%, comparable to 55% in an epidemiological study of the case. The combination of applying dynamic models and exposure data led to an improved QMRA that included an estimate of the dilution factor. This approach has not been described previously.


[Infant mortality in a midwestern municipality of the State of São Paulo, Brazil, 1990 to 1992].
Infant mortality was studied in an urban area of Southeastern Brazil in the period from 1990 to 1992 using data from death certificates collected at the registry office, by the application of methods for obtaining a collective diagnosis which will assist in the identification and choice of strategies for the control of local problems. The original data were corrected using documental research into health services and household interviews. Data of the Live Birth Information System (SINASC) was used to study variables such as maternal age and birthweight. The quality of original death certificates was initially analyzed using the amount of information, sensitivity, specificity and Kappa value. The global sensitivity for the underlying cause was 78.84% and Kappa 71.32% for the total of causes. One hundred and eighty-nine deaths occurred, 66.15% of them in the neonatal period, (41.28% during the first day of life) and 33.85% in late childhood. The birthweight of 58.28% of deaths was less than 2.500 g. The underlying causes of death were studied the by possibility, of their avoidance (a method developed by Erica Taucher), by a "reduced" group of causes (utilized in International Collaborative Effort (ICE)), multiple causes statistics and geographical distribution. It was observed that in the deaths occurring up to 27 days, 22.23% could have been avoided by adequate care during labour, and 20.64% could have been avoided by early diagnosis and early treatment, 13.75% by good pregnancy care and only 7.94% were unavoidable. Of the deaths occurring in late childhood, 12.17% were classified as of avoidable causes and 4.23% were considered as unavoidable. Using ICE groups, 58.74% died of immaturity or asphyxia, 19.58 of infection and 12.17% from congenital abnormalities. The results suggest that priority be given to obstetrical care at delivery and during labour and to the pediatric care of low birth weight, among others. The analysis using multiple causes statistics shows that 76.05% of the deaths have underlying causes related to neonatal disorders and confirms the relationship with the weight deficiencies of the newborn. The maternal complications were also related to weight deficiencies. Great differences were identified in infant mortality rates in urban zones not only restricted to the value of the rates but also to the diseases responsible for the occurrence of deaths. We therefore conclude that there is an advantage to be gained in using the four methods, which are complementary, for studying or planning actions with a view to prevent infant mortality.


Retinal oxygen distribution and the role of neuroglobin.
The retina is the tissue layer at the back of the eye that is responsible for light detection. Whilst equipped with a rich supply of oxygen, it has one of the highest oxygen demands of any tissue in the body and, as such, supply and demand are finely balanced. It has been suggested that the protein neuroglobin (Ngb), which is found in high concentrations within the retina, may help to maintain an adequate supply of oxygen via the processes of transport and storage. We construct mathematical models, formulated as systems of reaction-diffusion equations in one-dimension, to test this hypothesis. Numerical simulations show that Ngb may play an important role in oxygen transport, but not in storage. Our models predict that the retina is most susceptible to hypoxia in the regions of the photoreceptor inner segment and inner plexiform layers, where Ngb has the potential to prevent hypoxia and increase oxygen uptake by 30-40 %. Analysis of a simplified model confirms the utility of Ngb in transport and shows that its oxygen affinity ([Formula: see text] value) is near optimal for this process. Lastly, asymptotic analysis enables us to identify conditions under which the piecewise linear and quadratic approximations to the retinal oxygen profile, used in the literature, are valid.


How many treatment sessions and patients are needed to create a stable score of adherence and competence in the treatment of cocaine dependence?
The study utilized a generalizability theory analysis of adherence and competence ratings to evaluate the number of sessions and patients needed to yield dependable scores at the patient and therapist levels. Independent judges' ratings of supportive expressive therapy (n = 94), cognitive therapy (n = 103), and individual drug counseling (n = 98) were obtained on tapes of sessions from the NIDA Collaborative Cocaine Treatment Study. Generalizability coefficients revealed that, for all three treatments, ratings made on approximately five to 10 sessions per patient are needed to achieve sufficient dependability at the patient level. At the therapist level, four to 14 patients need to be evaluated (depending on the modality), to yield dependable scores. Many studies today use fewer numbers.


Vaccines for preventing influenza in healthy adults.
Three different types of influenza vaccines are currently produced worldwide. None is traditionally targeted to healthy adults. Despite the publication of a large number of clinical trials, there is still substantial uncertainty about the clinical effectiveness of influenza vaccines and this has negative impact on the vaccines acceptance and uptake. To assess the effects of vaccines on influenza in healthy adults. To assess the effectiveness of vaccines in preventing cases of influenza in healthy adults. To estimate the frequency of adverse effects associated with influenza vaccination in healthy adults. We searched the Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library, Issue 1, 2004) which contains the Cochrane Acute Respiratory Infections Group trials register; MEDLINE (January 1966 to December 2003); and EMBASE (1990 to December 2003). We wrote to vaccine manufacturers and first or corresponding authors of studies in the review. Any randomised or quasi-randomised studies comparing influenza vaccines in humans with placebo, control vaccines or no intervention, or comparing types, doses or schedules of influenza vaccine. Live, attenuated or killed vaccines or fractions thereof administered by any route, irrespective of antigenic configuration were considered. Only studies assessing protection from exposure to naturally occurring influenza in healthy individuals aged 14 to 60 (irrespective of influenza immune status) were considered. Two reviewers independently assessed trial quality and extracted data. Twenty five reports of studies involving 59,566 people were included. The recommended live aerosol vaccines reduced the number of cases of serologically confirmed influenza by 48% (95% confidence interval (CI) 24% to 64%), whilst recommended inactivated parenteral vaccines had a vaccine efficacy of 70% (95% CI 56% to 80%). The yearly recommended vaccines had low effectiveness against clinical influenza cases: 15%(95% CI 8% to 21%) and 25% (95% CI 13% to 35%) respectively. Overall the percentage of participants experiencing clinical influenza decreased by 6%. Use of the vaccine significantly reduced time off work but only by 0.16 days for each influenza episode (95% CI 0.04 to 0.29 days); Analysis of vaccines matching the circulating strain gave higher estimates of efficacy, whilst inclusion of all other vaccines reduced the efficacy. Influenza vaccines are effective in reducing serologically confirmed cases of influenza. However, they are not as effective in reducing cases of clinical influenza and number of working days lost. Universal immunisation of healthy adults is not supported by the results of this review.


Overexpression of manganese superoxide dismutase mRNA may correlate with aggressiveness in gastric and colorectal adenocarcinomas.
The expression or activity of manganese superoxide dismutase (Mn-SOD) is reduced in a variety of malignant tumors and Mn-SOD may act as a new type of tumor suppressor gene. On the other hand, increased expression of Mn-SOD can diminish the cytotoxic effects of several anticancer modalities, including tumor necrosis factor alpha, ionizing radiation, certain chemotherapeutic agents and hyperthermia. Although Mn-SOD expression and its role in various cancers are intensely studied, little is known about its function in gastrointestinal carcinomas. To examine the expression level and significance of Mn-SOD in gastrointestinal carcinomas, Mn-SOD mRNA expression was examined in 53 gastric carcinoma and 38 colorectal carcinoma by reverse transcription-polymerase chain reaction and was compared with those in the corresponding normal mucosal tissues. The tumor/normal (T/N) ratio was calculated and the data were clinicopathologically analyzed. The average T/N ratios of Mn-SOD mRNA expression in gastric and colorectal carcinomas were 2.19 and 3. 72, respectively. Clinicopathologic analyses revealed positive correlation between the Mn-SOD expression level and venous invasion in both gastric and colorectal carcinomas (p<0.05 and p<0.05, respectively). Furthermore, the colorectal carcinoma with lymph node metastasis showed significantly higher Mn-SOD expression than those without it (p<0.05). Our results suggest that Mn-SOD mRNA overexpression can occur in gastric and colorectal carcinomas and may be related to increased aggressiveness.


SmartTots Update Regarding Anesthetic Neurotoxicity in the Developing Brain.
SmartTots (http://smarttots.org/) represents a public-private partnership between the International Anesthesia Research Society and the US Food and Drug Administration. Over the past 7 years, SmartTots has worked in collaboration with various stakeholders to determine whether anesthetic drugs have detrimental effects on the developing brain. SmartTots has funded clinical and preclinical studies, organized meetings, served as a repository of peer-reviewed information, and facilitated the development of consensus-based statements. Here, we report advances in the field of anesthetic neurotoxicity and provide an update on SmartTots' activities. Clinical studies have provided some reassurance that a brief exposure to anesthetic drugs does not cause overt, persistent cognitive deficits. New recommendations aim to increase the reproducibility and "clinical relevance" of data from studies of laboratory animals. Overall, the field has advanced substantially; however, it remains paramount to definitively resolve whether anesthetic drugs are neurotoxic to the immature brain. The results of SmartTots efforts will either ally unwarranted fears or substantially change pediatric anesthetic practice and prompt studies to identify neuroprotective strategies.


Meanings and purposes of caring for a family member: an autoethnography.
Engagement in two interlinked areas of occupation, familial social participation and caring for my father at the end of his life, led to a journey of self-discovery. A qualitative research methodology, autoethnography, is used to develop a narrative that examines engagement in these two occupations before, during, and after my father's illness and death. I discuss meanings and purposes of familial social participation and caregiving, suggesting that transforming fear of death to awareness of death is a central purpose of caregiving. Implications for therapists and caregivers include considerations about the value of occupation, discussion of a continuum of caregiving, examination of boundaries when caring for a parent, thoughts about the roles of altruism, love, anger, and "bad faith" in caregiving, and analysis of sons as caregivers for fathers. Further research on meanings and purposes of caregiving is proposed.


Persistent primary hyperparathyroidism: an uncommon location for an ectopic gland--case report and review.
Primary hyperparathyroidism (PHPT) is a common endocrine disorder that mainly affects middle-aged women. Patients are usually asymptomatic. The disease might be ascribable to hyperplasia, carcinoma, and single or multiple adenomas. PHPT may be sporadic or familial, the latter comprising multiple endocrine neoplasia type 1 or 2A, familial benign hypocalciuria hypercalcemia, and hyperparathyroidism-jaw tumor syndrome. The most common causes for persistent PHPT are multiglandular disease, and missed abnormal ectopic or orthotopic parathyroid glands. Imaging localization studies should precede a new surgical intervention. Ectopic parathyroid glands are rarely located at the aortopulmonary window. For diagnosis confirmation, (99m)Tc-sestamibi SPECT/CT seems to be an advantageous test. Another possibility is to perform (99m)Tc-sestamibi followed by thoracic CT or MRI. Parathyroidectomy may be performed by means of median sternotomy, thoracotomy, or video-assisted thoracoscopy. We describe a case of persistent primary hyperparathyroidism due to the presence of an ectopic parathyroid gland found at the aortopulmonary window. As the investigation necessary to clarify the etiology of recurrent nephrolithiasis proceeded, the diagnosis of PHPT was determined. The patient underwent subtotal parathyroidectomy; nevertheless, PHPT persisted. Genetic syndromes that could account for this condition were excluded. Imaging studies available at that time were not able to locate abnormal glands; moreover, the patient refused to undergo surgical exploration. Later, the patient underwent (99m)Tc-sestamibi SPECT/CT, which revealed a parathyroid gland at the aortopulmonary window.


CagA-ASPP2 complex mediates loss of cell polarity and favors H. pylori colonization of human gastric organoids.
The main risk factor for stomach cancer, the third most common cause of cancer death worldwide, is infection with Helicobacter pylori bacterial strains that inject cytotoxin-associated gene A (CagA). As the first described bacterial oncoprotein, CagA causes gastric epithelial cell transformation by promoting an epithelial-to-mesenchymal transition (EMT)-like phenotype that disrupts junctions and enhances motility and invasiveness of the infected cells. However, the mechanism by which CagA disrupts gastric epithelial cell polarity to achieve its oncogenicity is not fully understood. Here we found that the apoptosis-stimulating protein of p53 2 (ASPP2), a host tumor suppressor and an important CagA target, contributes to the survival of cagA-positive H. pylori in the lumen of infected gastric organoids. Mechanistically, the CagA-ASPP2 interaction is a key event that promotes remodeling of the partitioning-defective (PAR) polarity complex and leads to loss of cell polarity of infected cells. Blockade of cagA-positive H. pylori ASPP2 signaling by inhibitors of the EGFR (epidermal growth factor receptor) signaling pathway-identified by a high-content imaging screen-or by a CagA-binding ASPP2 peptide, prevents the loss of cell polarity and decreases the survival of H. pylori in infected organoids. These findings suggest that maintaining the host cell-polarity barrier would reduce the detrimental consequences of infection by pathogenic bacteria, such as H. pylori, that exploit the epithelial mucosal surface to colonize the host environment.


Excretion and tissue distribution of selenium following treatment of male F344 rats with benzylselenocyanate or sodium selenite.
Benzylselenocyanate (BSC), a synthetic organoselenium compound less toxic than sodium selenite (Na2SeO3), has been demonstrated to inhibit the development of neoplasia in several experimental animal models. We examined the excretion and tissue distribution of total Se after acute administration of BSC compared to Na2SeO3. Male F344 rats were treated po with approximately one-tenth of the LD50 values, our estimate of highest non-toxic dose. The doses administered were 9.85 mg/kg in the case of BSC and 4.35 mg/kg in the case of Na2SeO3. The rats were sacrificed at 1, 6, 24, 72, or 120 hr to obtain biological samples. The levels of total Se were determined by graphite furnace atomic absorption spectrophotometry following microwave digestion. In serum, the highest Se level was observed at 6 hr after administration of either BSC or Na2SeO3: 1.34 +/- 0.07 (mean +/- SE), or 2.09 +/- 0.11 micrograms/ml of serum, respectively. In urine and feces, the cumulative percentages of doses excreted within 3 days of BSC or Na2SeO3 treatment were, respectively, as follows: 11.36 +/- 0.82% and 18.33 +/- 0.77% in urine; and 6.67 +/- 0.66% and 31.14 +/- 4.66% in feces. Among the tissues of BSC-treated rats, the kidneys were found to have the highest Se levels throughout the experimental period (as much as 29 micrograms/g of tissue at 72 hr), followed by liver, small intestine, large intestine, lung, pancreas, heart, and spleen. The results indicate that Se from BSC-treated animals is excreted very slowly and is retained in the organs for a much longer period compared to rats treated with Na2SeO3. Whether the slow excretion and prolonged retention of BSC and/or its metabolites play a role in its chemopreventive action is currently under investigation.


Treadmill walking during vocabulary encoding improves verbal long-term memory.
Moderate physical activity improves various cognitive functions, particularly when it is applied simultaneously to the cognitive task. In two psychoneuroendocrinological within-subject experiments, we investigated whether very low-intensity motor activity, i.e. walking, during foreign-language vocabulary encoding improves subsequent recall compared to encoding during physical rest. Furthermore, we examined the kinetics of brain-derived neurotrophic factor (BDNF) in serum and salivary cortisol. Previous research has associated both substances with memory performance.In both experiments, subjects performed better when they were motorically active during encoding compared to being sedentary. BDNF in serum was unrelated to memory performance. In contrast we found a positive correlation between salivary cortisol concentration and the number of correctly recalled items. In summary, even very light physical activity during encoding is beneficial for subsequent recall.


Apoptosis in lymphoid and parenchymal cells during sepsis: findings in normal and T- and B-cell-deficient mice.
To determine if apoptosis (programmed cell death) occurs systemically in lymphoid and parenchymal cells during sepsis. To examine the potential role of T and B cells in the apoptotic process using knockout mice deficient in mature T and B lymphocytes. Prospective, randomized, controlled trial. Animal laboratory in a university medical setting. Cecal ligation and puncture (CLP) (n = 34) or sham surgery (n = 13) was performed in female ND4 mice and, 15 to 22 hrs postoperatively, thymus, lung, heart, spleen, ileum, colon, liver, kidney, brain, and muscle were obtained and examined for apoptosis. A second group of mice (Rag-1) which are totally deficient in mature T and B cells also underwent CLP (n = 14) or sham surgery (n = 14) and had examination of tissues for apoptosis. Four methods with varying sensitivities and specificities were used to detect apoptosis, including: a) DNA agarose gel electrophoresis; b) terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL); c) electron microscopy; and d) light microscopy. In CLP mice, multiple methods demonstrated apoptosis in lymphocytes in thymus, spleen, ileum, colon, lung, and skeletal muscle. In addition to lymphocytes, parenchymal cells in ileum, colon, lung, and to a lesser extent, in skeletal muscle and kidney were apoptotic in CLP mice. There was no evidence of apoptosis by any method of detection in liver, brain, or heart. Results in Rag-1 mice which are deficient in T and B cells demonstrated extensive apoptosis in thymus, spleen, and ileum with less degrees of apoptosis in colon and lung. Both lymphoid cells and parenchymal cells were apoptotic. Rag-1 mice which underwent CLP did not die prematurely and there were no apparent observable differences in the physical response (tachypnea, piloerection, lethargy, etc), or intra-abdominal bowel inflammation/adhesions compared with CLP mice with normal T and B cells. Apoptosis is an important mechanism of cell death in lymphocytes and parenchymal cells in sepsis and occurs systemically in many organs. Apoptosis may be an important cause of immunologic suppression in sepsis by inducing widespread lymphocyte depletion. Alternately, apoptosis may be beneficial to host survival by down-regulating the inflammatory response which accompanies sepsis. The degree to which parenchymal cell apoptosis is contributing to multiple organ failure cannot be determined from the present study. Findings in Rag-1 mice demonstrate that mature T and B cells and their secretory products are not necessary for apoptosis to occur during sepsis and that apoptotic cell death is not restricted to T or B cells. Apoptosis may be a key regulator of the balance between the pro- and anti-inflammatory process.


Genetic linkage between von Hippel-Lindau disease and three microsatellite polymorphisms refines the localisation of the VHL locus.
Von Hippel-Lindau (VHL) disease is a dominantly inherited familial cancer syndrome characterised by the development of retinal and central nervous system haemangioblastomas, renal cell carcinoma and phaeochromocytoma. The gene for VHL disease has been mapped to chromosome 3p25-p26 and presymptomatic diagnosis using linked DNA markers is available. We have previously mapped the VHL disease gene to a 4 cM interval between D3S1250 and D3S18. To increase access to presymptomatic diagnosis and to accelerate progress towards isolating the VHL disease gene we attempted to identify microsatellite DNA markers linked to the disease gene by genetic linkage analysis in 29 families. We found significant linkage between the VHL disease gene and dinucleotide (CA) repeat polymorphisms at D3S1038 (Zmax = 22.24 at theta = 0.01, CI 0.0001-0.06), D3S1110 (Zmax = 11.32 at theta = 0.07, CI 0.03-0.14) and D3S651 (Zmax = 7.73 at theta = 0.04, CI 0.008-0.13). We localised D3S1038 between D3S1250 and D3S601, and mapped D3S1110 and D3S651 centromeric to D3S1250. Multipoint linkage analysis mapped the VHL disease locus between D3S1038 and D3S18 with the maximum likelihood at D3S601. There was no evidence of locus heterogeneity. This study has (i) identified three microsatellite DNA markers in chromosome 3p25 linked to the VHL disease gene and (ii) narrowed the target region for the isolation of the VHL disease gene by positional cloning techniques. These findings will improve the management of families with VHL disease by improving the accuracy and availability of presymptomatic diagnosis using linked DNA markers, and will accelerate progress towards isolating the VHL disease gene.


Limb myorhythmia in association with hypertrophy of the inferior olive: report of two cases.
We report on 2 patients who uncommonly developed isolated limb myorhythmia in association with inferior olive hypertrophy (IOH) after an acute stroke in the brain stem. A slow tremor presented in the proximal upper limbs predominantly when at rest. It was aggravated by outstretched arms and by active hand movements. The surface electromyogram (EMG) recorded simultaneous activities over the agonist and antagonist muscles with a rate of 3.5 Hz and 2.5 Hz in 2 patients respectively. In the first patient, bilateral limb myorhythmia presented 12 months after the brain stem stroke, and both inferior olives were hypertrophic. In the second patient, unilateral limb myorhythmia developed in the left hand 7 months after right pontine hemorrhage, and only the right inferior olive was hypertrophic. These findings indicate that limb myorhythmia commencing after brain stem insult is anatomically and temporally related to hypertrophy of the contralateral inferior olive. Based on our 2 patients and previously reported cases, we propose that a possible causal relationship exists between limb myorhythmia and contralateral IOH, although its pathophysiological mechanisms remain to be established. We suggest that, similar to palatal myoclonus, isolated limb myorhythmia is within the clinical spectrum of IOH.


Use of fibrin glue in fixation of amniotic membranes in sterile corneal ulceration.
To describe a method of fibrin glue fixation of single or multilayered amniotic membranes in sterile corneal ulceration. A modified microsurgical procedure. The corneal ulcer is cleaned and the components of fibrin glue are applied separately. Amniotic membranes are rotated through the fibrinogen component and subsequently into the site of the ulceration where thrombin is already applied. An overlay membrane is sutured additionally on top. The results for one patient are illustrated as an example. The application technique prevents development of foam and leads to a thin fibrin film, which minimizes any irregularities of the fixed membranes and creates extra time to adjust the membranes' position. This method presents as an effective procedure for the treatment of sterile corneal ulceration and creates valuable time if any elective corneal graft procedure is required.


A molecular palaeobiological exploration of arthropod terrestrialization.
Understanding animal terrestrialization, the process through which animals colonized the land, is crucial to clarify extant biodiversity and biological adaptation. Arthropoda (insects, spiders, centipedes and their allies) represent the largest majority of terrestrial biodiversity. Here we implemented a molecular palaeobiological approach, merging molecular and fossil evidence, to elucidate the deepest history of the terrestrial arthropods. We focused on the three independent, Palaeozoic arthropod terrestrialization events (those of Myriapoda, Hexapoda and Arachnida) and showed that a marine route to the colonization of land is the most likely scenario. Molecular clock analyses confirmed an origin for the three terrestrial lineages bracketed between the Cambrian and the Silurian. While molecular divergence times for Arachnida are consistent with the fossil record, Myriapoda are inferred to have colonized land earlier, substantially predating trace or body fossil evidence. An estimated origin of myriapods by the Early Cambrian precedes the appearance of embryophytes and perhaps even terrestrial fungi, raising the possibility that terrestrialization had independent origins in crown-group myriapod lineages, consistent with morphological arguments for convergence in tracheal systems.This article is part of the themed issue 'Dating species divergences using rocks and clocks'.


Determination of the mutation spectrum of the EXT1/EXT2 genes in British Caucasian patients with multiple osteochondromas, and exclusion of six candidate genes in EXT negative cases.
We describe here the spectrum and distribution of mutations in the EXT1 and EXT2 genes in the largest reported British Caucasian multiple osteochondromas (MO) population. Furthermore, we report for the first time the screening of the EXT1 and EXT2 promoters, 5'UTRs, and 3'UTRs, and exclude six potential MO candidate genes in individuals without a detectable mutation within the coding region of EXT1 and EXT2. The coding exons of EXT1 and EXT2 were screened in 72 unrelated probands affected with MO. Forty-six different mutations were identified in 56 probands, of which 29 were novel. Mutation in the EXT1 and EXT2 genes each accounted for 50% of the mutations identified. Of the 72 probands, 42 were of British Caucasian descent, which when added to the 41 British Caucasian families previously reported from our total cohort, gave a total of 83 families. This cohort's proportional frequency for EXT1/EXT2 mutation was 53%/47%. We also validated the technique of high-resolution melting analysis in a blind study using 27 unique EXT1 or EXT2 mutations. This technique was found to be sensitive with a detection rate of 100% regarding heterozygote detection for EXT mutation scanning. Furthermore, this technique has a very high throughput and is very cost-effective.


CD28/B7 interactions deliver a unique signal to naive T cells that regulates cell survival but not early proliferation.
CD28/B7 ligation provides costimulatory signals important for the development of T cell responses. In the present study, we examined whether CD28/B7 interactions have a specialized role in the regulation of cell cycle progression and sustained T cell proliferative responses in naive T cell populations using TCR transgenic mice. CD28-mediated signaling was shown to be uniquely capable of regulating cell survival compared with TCR-mediated signaling. Increasing the strength of the TCR-mediated signal 1 increased early proliferative responses, but had no effect on sustained cell survival. In contrast, CD28 ligation, signal 2, was not required for early proliferative responses, but dramatically influenced long term T cell survival. The increased cell survival after CD28 ligation was not due to increased IL-2 production, but was linked to up-regulation of Bcl-xL. The Bcl-xL protein could not be induced following increased TCR cross-linking in the absence of CD28 signaling. In addition, survival of T cells from Bcl-xL transgenic mice was not inhibited by blocking CD28 ligation, suggesting that CD28-induced T cell survival is regulated by Bcl-xL expression. Together, these results suggest that the unique role of CD28 signaling is not to costimulate the initial activation of naive T cells, but is, in fact, to sustain the late proliferative response and enhance long term cell survival.


Perceptual enhancement: persistent effects of an experience.
Presenting a word enhances its later perceptual identification. This article focuses on the relation between this effect on perception and recognition memory. Prior experiments have revealed that perceptual enhancement is independent of recognition memory and have led to the two types of task being identified with separate memory systems. In contrast, the present experiments reveal parallel effects on the two types of task. Perceptual enhancement persists over days but, like recognition memory, is influenced by manipulations of retrieval conditions. I conclude that both perceptual and memory tasks rely on the retrieval of memory for whole prior processing episodes but can differ in terms of the number and nature of retrieval cues that they provide. I describe perception and memory within a common framework.


LINNAEUS: a species name identification system for biomedical literature.
The task of recognizing and identifying species names in biomedical literature has recently been regarded as critical for a number of applications in text and data mining, including gene name recognition, species-specific document retrieval, and semantic enrichment of biomedical articles. In this paper we describe an open-source species name recognition and normalization software system, LINNAEUS, and evaluate its performance relative to several automatically generated biomedical corpora, as well as a novel corpus of full-text documents manually annotated for species mentions. LINNAEUS uses a dictionary-based approach (implemented as an efficient deterministic finite-state automaton) to identify species names and a set of heuristics to resolve ambiguous mentions. When compared against our manually annotated corpus, LINNAEUS performs with 94% recall and 97% precision at the mention level, and 98% recall and 90% precision at the document level. Our system successfully solves the problem of disambiguating uncertain species mentions, with 97% of all mentions in PubMed Central full-text documents resolved to unambiguous NCBI taxonomy identifiers. LINNAEUS is an open source, stand-alone software system capable of recognizing and normalizing species name mentions with speed and accuracy, and can therefore be integrated into a range of bioinformatics and text-mining applications. The software and manually annotated corpus can be downloaded freely at http://linnaeus.sourceforge.net/.


GTP signaling links metabolism, DNA repair, and responses to genotoxic stress.
How cell metabolism regulates DNA repair is incompletely understood. Here, we define a GTP-mediated signaling cascade that links metabolism to DNA repair and has significant therapeutic implications. GTP, but not other nucleotides, regulates the activity of Rac1, a G protein, that promotes the dephosphorylation of serine 323 on Abl-interactor 1 (Abi-1) by protein phosphatase 5 (PP5). Dephosphorylated Abi-1, a protein previously not known to activate DNA repair, promotes non-homologous end joining. In patients and mouse models of glioblastoma, Rac1 and dephosphorylated Abi-1 mediate DNA repair and resistance to standard of care genotoxic treatments. The GTP-Rac1-PP5-Abi-1 signaling axis is not limited to brain cancer, as GTP supplementation promotes DNA repair and Abi-1-S323 dephosphorylation in non-malignant cells and protects mouse tissues from genotoxic insult. This unexpected ability of GTP to regulate DNA repair independently of deoxynucleotide pools has important implications for normal physiology and cancer treatment.


Evolution of the EKA family of powdery mildew avirulence-effector genes from the ORF 1 of a LINE retrotransposon.
The Avrk1 and Avra10 avirulence (AVR) genes encode effectors that increase the pathogenicity of the fungus Blumeria graminis f.sp. hordei (Bgh), the powdery mildew pathogen, in susceptible barley plants. In resistant barley, MLK1 and MLA10 resistance proteins recognize the presence of AVRK1 and AVRA10, eliciting the hypersensitive response typical of gene for gene interactions. Avrk1 and Avra10 have more than 1350 homologues in Bgh genome, forming the EKA (Effectors homologous to Avr k 1 and Avr a 10) gene family. We tested the hypothesis that the EKA family originated from degenerate copies of Class I LINE retrotransposons by analysing the EKA family in the genome of Bgh isolate DH14 with bioinformatic tools specially developed for the analysis of Transposable Elements (TE) in genomes. The Class I LINE retrotransposon copies homologous to Avrk1 and Avra10 represent 6.5 % of the Bgh annotated genome and, among them, we identified 293 AVR/effector candidate genes. We also experimentally identified peptides that indicated the translation of several predicted proteins from EKA family members, which had higher relative abundance in haustoria than in hyphae. Our analyses indicate that Avrk1 and Avra10 have evolved from part of the ORF1 gene of Class I LINE retrotransposons. The co-option of Avra10 and Avrk1 as effectors from truncated copies of retrotransposons explains the huge number of homologues in Bgh genome that could act as dynamic reservoirs from which new effector genes may evolve. These data provide further evidence for recruitment of retrotransposons in the evolution of new biological functions.


Protective role of kallistatin in renal fibrosis via modulation of Wnt/β-catenin signaling.
Kallistatin is a multiple functional serine protease inhibitor that protects against vascular injury, organ damage and tumor progression. Kallistatin treatment reduces inflammation and fibrosis in the progression of chronic kidney disease (CKD), but the molecular mechanisms underlying this protective process and whether kallistatin plays an endogenous role are incompletely understood. In the present study, we observed that renal kallistatin levels were significantly lower in patients with CKD. It was also positively correlated with estimated glomerular filtration rate (eGFR) and negatively correlated with serum creatinine level. Unilateral ureteral obstruction (UUO) in animals also led to down-regulation of kallistatin protein in the kidney, and depletion of endogenous kallistatin by antibody injection resulted in aggravated renal fibrosis, which was accompanied by enhanced Wnt/β-catenin activation. Conversely, overexpression of kallistatin attenuated renal inflammation, interstitial fibroblast activation and tubular injury in UUO mice. The protective effect of kallistatin was due to the suppression of TGF-β and β-catenin signaling pathways and subsequent inhibition of epithelial-to-mesenchymal transition (EMT) in cultured tubular cells. In addition, kallistatin could inhibit TGF-β-mediated fibroblast activation via modulation of Wnt4/β-catenin signaling pathway. Therefore, endogenous kallistatin protects against renal fibrosis by modulating Wnt/β-catenin-mediated EMT and fibroblast activation. Down-regulation of kallistatin in the progression of renal fibrosis underlies its potential as a valuable clinical biomarker and therapeutic target in CKD.


Complete Genome Sequences for Three Chromosomes of the Burkholderia stabilis Type Strain (ATCC BAA-67).
We report here the complete annotated genome sequence of the Burkholderia stabilis type strain ATCC BAA-67. There were three circular chromosomes with a combined size of 8,527,947 bp and G+C composition of 66.4%. These characteristics closely resemble the genomes of other sequenced members of the Burkholderia cepacia complex.


Shame and guilt in the postnatal period: a systematic review.
Objective: The aim of this review was to explore the unique contribution of shame (negative evaluation of the self) and guilt (negative evaluation of behaviour) to postnatal psychological symptoms. Background: Although shame and guilt are related to psychological symptoms, the separate effect of each in postnatal psychological symptoms are not yet known. Methods: Seven electronic databases were systematically reviewed for articles on: (1) quantitative measures of shame, guilt, and psychological symptoms (2) in the postnatal period for infants under two years of age (3) published in English. Results: Of the 1,615 articles retrieved using PRISMA guidelines, five met criteria and were analysed independently by two reviewers using the STROBE criteria. In mothers, shame was significantly related to stress and postnatal depression. Shame significantly predicted postnatal depression. Guilt was significantly related to postnatal depression; however, the relationship was substantially reduced when included with shame. In fathers, shame, but not guilt, was significantly related to stress, anxiety, and depression. Conclusion: Shame and guilt are trans-diagnostic phenomena, negatively impacting on postnatal psychological health, and potentially the parent-child relationship. More research is needed to develop awareness of the unique effects of shame and guilt to optimise perinatal intervention.


Mechanisms of immune tolerance to allergens in children.
Because the prevalence of allergic diseases has significantly increased in recent years, understanding the causes and mechanisms of these disorders is of high importance, and intense investigations are ongoing. Current knowledge pinpoints immune tolerance mechanisms as indispensable for healthy immune response to allergens in daily life. It is evident that development and maintenance of allergens-pecific T cell tolerance is of vital importance for a healthy immune response to allergens. Such tolerance can be gained spontaneously by dose-dependent exposures to allergens in nature or by allergen-specific immunotherapy. Allergen-specific immunotherapy induces regulatory T cells with the capacity to secrete interleukin-10 and transforming growth factor-β, limits activation of effector cells of allergic inflammation (such as mast cells and basophils), and switches antibody isotype from IgE to the noninflammatory type IgG4. Although allergen-specific immunotherapy is the only method of tolerance induction in allergic individuals, several factors, such as long duration of treatment, compliance problems, and life-threatening side effects, have limited widespread applicability of this immunomodulatory treatment. To overcome these limitations, current research focuses on the introduction of allergens in more efficient and safer ways. Defining the endotypes and phenotypes of allergic diseases might provide the ability to select ideal patients, and novel biomarkers might ensure new custom-tailored therapy modalities.


Analysis of Cyberincivility in Posts by Health Professions Students: Descriptive Twitter Data Mining Study.
Health professions students use social media to communicate with other students and health professionals, discuss career plans or coursework, and share the results of research projects or new information. These platforms allow students to share thoughts and perceptions that are not disclosed in formal education settings. Twitter provides an excellent window through which health professions educators can observe students' sociocultural and learning needs. However, despite its merits, cyberincivility on Twitter among health professions students has been reported. Cyber means using electronic technologies, and incivility is a general term for bad manners. As such, cyberincivility refers to any act of disrespectful, insensitive, or disruptive behavior in an electronic environment. This study aims to describe the characteristics and instances of cyberincivility posted on Twitter by self-identified health professions students. A further objective of the study is to analyze the prevalence of tweets perceived as inappropriate or potentially objectionable while describing patterns and differences in the instances of cyberincivility posted by those users. We used a cross-sectional descriptive Twitter data mining method to collect quantitative and qualitative data from August 2019 to February 2020. The sample was taken from users who self-identified as health professions students (eg, medicine, nursing, dental, pharmacy, physician assistant, and physical therapy) in their user description. Data management and analysis were performed with a combination of SAS 9.4 for descriptive and inferential statistics, including logistic regression, and NVivo 12 for descriptive patterns of textual data. We analyzed 20 of the most recent tweets for each account (N=12,820). A total of 639 user accounts were analyzed for quantitative analysis, including 280 (43.8%) medicine students and 329 (51.5%) nursing students in 22 countries: the United States (287/639, 44.9%), the United Kingdom (197/639, 30.8%), unknown countries (104/639, 16.3%), and 19 other countries (51/639, 8.0%). Of the 639 accounts, 193 (30.2%) were coded as having instances of cyberincivility. Of these, 61.7% (119/193), 32.6% (63/193), and 5.7% (11/193) belonged to students in nursing, medicine, and other disciplines, respectively. Among 502 instances of cyberincivility identified from 641 qualitative analysis samples, the largest categories were profanity and product promotion. Several aggressive or biased comments toward other users, politicians, or certain groups of people were also found. Cyberincivility is a multifaceted phenomenon that must be considered in its complexity if health professions students are to embrace a culture of mutual respect and collaboration. Students' perceptions and reports of their Twitter experiences offer insights into behavior on the web and the evolving role of cyberspace, and potentially problematic posts provide opportunities for teaching digital professionalism. Our study indicates that there is a continued need to provide students with guidance and training regarding the importance of maintaining a professional persona on the web.


EST sequencing and phylogenetic analysis of the model grass Brachypodium distachyon.
Brachypodium distachyon (Brachypodium) is a temperate grass with the physical and genomic attributes necessary for a model system (small size, rapid generation time, self-fertile, small genome size, diploidy in some accessions). To increase the utility of Brachypodium as a model grass, we sequenced 20,440 expressed sequence tags (ESTs) from five cDNA libraries made from leaves, stems plus leaf sheaths, roots, callus and developing seed heads. The ESTs had an average trimmed length of 650 bp. Blast nucleotide alignments against SwissProt and GenBank non-redundant databases were performed and a total of 99.9% of the ESTs were found to have some similarity to existing protein or nucleotide sequences. Tentative functional classification of 77% of the sequences was possible by association with gene ontology or clusters of orthologous group's index descriptors. To demonstrate the utility of this EST collection for studying cell wall composition, we identified homologs for the genes involved in the biosynthesis of lignin subunits. A subset of the ESTs was used for phylogenetic analysis that reinforced the close relationship of Brachypodium to wheat and barley.


Challenges for nurse managers in China.
To critically review the challenges facing nurse leaders in China during healthcare reform. China is now undergoing a major reform aimed at enhancing the accessibility and quality of its healthcare at a level that is affordable to the people. Nurses have a key role to play in this reform. Key documents produced by the government of China were critically reviewed using the strengths, weaknesses, opportunities and threats (SWOT) framework. A number of factors, including the insufficient number of nurses and the medical orientation of the health system, have hindered the development of nursing. However, healthcare reform has created new opportunities for nurses and nursing. This paper reviews the strengths, weaknesses, opportunities and threats faced by nurse managers in China. It identifies barriers but also possibilities for nurse leaders to advance nursing and make nurses visible in this era of transforming healthcare. Many of the issues identified in this review, such as the enhancement of quality and accessibility of care, are important to nurse leaders around the world. However, this article reveals the particular challenges faced by nurse leaders in China, with its unique social and historical background.


Diagnosis and management of migraine in ten steps.
Migraine is a disabling primary headache disorder that directly affects more than one billion people worldwide. Despite its widespread prevalence, migraine remains under-diagnosed and under-treated. To support clinical decision-making, we convened a European panel of experts to develop a ten-step approach to the diagnosis and management of migraine. Each step was established by expert consensus and supported by a review of current literature, and the Consensus Statement is endorsed by the European Headache Federation and the European Academy of Neurology. In this Consensus Statement, we introduce typical clinical features, diagnostic criteria and differential diagnoses of migraine. We then emphasize the value of patient centricity and patient education to ensure treatment adherence and satisfaction with care provision. Further, we outline best practices for acute and preventive treatment of migraine in various patient populations, including adults, children and adolescents, pregnant and breastfeeding women, and older people. In addition, we provide recommendations for evaluating treatment response and managing treatment failure. Lastly, we discuss the management of complications and comorbidities as well as the importance of planning long-term follow-up.


Fabricating versatile cell supports from nano- and micro-sized graphene oxide flakes.
Micro-sized structures made from graphene oxide (GO) attract high interest for their extensive use in tissue engineering. The fabrication and cytotoxicity of 3D graphene-based scaffolds so far have not been extensively discussed with relation to the flake sizes used. In this work we considered GO flakes of two different lognormal size distributions (GO: 4.9 ± 3.8 μm and GO 1 h: 151.6 ± 1.9 nm) as model flakes for fabrication of 3D graphene-based cell culture supports: paper (i.e. 3D layered film structure) and reduced graphene oxide (rGO) microfiber using hydrothermal methods. We then used two model cell lines of neuronal origin (SH-SY5Y and HEK-293) to study subsequent scaffolds surface-cells interactions. In particular, the adhesion of HEK cells to the formed structures was much higher than for SH-SY5Y cells, as evidenced by various atomic force, electron and optical microscopy techniques. Formed rGO microfibers had more desired nano-topography (surface roughness) for cell adhesion and growth than simple GO paper, making it ideal scaffold for neural tissue engineering. This work provides insights into the fundamental rules for fabrication of graphene oxide-based cell supports and their subsequently differing interactions with malignant and non-malignant cells.


The New England Air Quality Forecasting Pilot Program: development of an evaluation protocol and performance benchmark.
The National Oceanic and Atmospheric Administration recently sponsored the New England Forecasting Pilot Program to serve as a "test bed" for chemical forecasting by providing all of the elements of a National Air Quality Forecasting System, including the development and implementation of an evaluation protocol. This Pilot Program enlisted three regional-scale air quality models, serving as prototypes, to forecast ozone (O3) concentrations across the northeastern United States during the summer of 2002. A suite of statistical metrics was identified as part of the protocol that facilitated evaluation of both discrete forecasts (observed versus modeled concentrations) and categorical forecasts (observed versus modeled exceedances/nonexceedances) for both the maximum 1-hr (125 ppb) and 8-hr (85 ppb) forecasts produced by each of the models. Implementation of the evaluation protocol took place during a 25-day period (August 5-29), utilizing hourly O3 concentration data obtained from over 450 monitors from the U.S. Environment Protection Agency's Air Quality System network.


Delayed response to animate implied motion in human motion processing areas.
Viewing static photographs of objects in motion evokes higher fMRI activation in the human medial temporal complex (MT+) than looking at similar photographs without this implied motion. As MT+ is traditionally thought to be involved in motion perception (and not in form perception), this finding suggests feedback from object-recognition areas onto MT+. To investigate this hypothesis, we recorded extracranial potentials evoked by the sight of photographs of biological agents with and without implied motion. The difference in potential between responses to pictures with and without implied motion was maximal between 260 and 400 msec after stimulus onset. Source analysis of this difference revealed one bilateral, symmetrical dipole pair in the occipital lobe. This area also showed a response to real motion, but approximately 100 msec earlier than the implied motion response. The longer latency of the implied motion response in comparison to the real motion response is consistent with a feedback projection onto MT+ following object recognition in higher-level temporal areas.


Optimization of a pain model: effects of body temperature and anesthesia on bladder nociception in mice.
Interstitial cystitis/bladder pain syndrome (IC/BPS) is a debilitating urological condition that is resistant to treatment and poorly understood. To determine novel molecular treatment targets and to elucidate the contribution of the nervous system to IC/BPS, many rodent bladder pain models have been developed. In this study we evaluated the effects of anesthesia induction and temperature variation in a mouse model of bladder pain known as urinary bladder distension (UBD). In this model compressed air is used to distend the bladder to distinct pressures while electrodes record the reflexive visceromotor response (VMR) from the overlying abdominal muscle. Two isoflurane induction models are commonly used before UBD: a short method lasting approximately 30 minutes and a long method lasting approximately 90 minutes. Animals were anesthetized with one of the methods then put through three sets of graded bladder distensions. Distensions performed following the short anesthesia protocol were significantly different from one another despite identical testing parameters; this same effect was not observed when the long anesthesia protocol was used. In order to determine the effect of temperature on VMRs, animals were put through three graded distension sets at 37.5 (normal mouse body temperature), 35.5, and 33.5°C. Distensions performed at 33.5 and 35.5°C were significantly lower than those performed at 37.5°C. Additionally, Western blot analysis revealed significantly smaller increases in spinal levels of phosphorylated extracellular-signal regulated kinase 2 (pERK2) following bladder distension in animals whose body temperature was maintained at 33.5°C as opposed to 37.5°C. These results highlight the significance of the dynamic effects of anesthesia on pain-like changes and the importance of close monitoring of temperature while performing UBD. For successful interpretation of VMRs and translation to human disease, body temperature should be maintained at 37.5°C and isoflurane induction should gradually decrease over the course of 90 minutes.


Substance use disorders and co-morbidities among Asian Americans and Native Hawaiians/Pacific Islanders.
Asian Americans (AAs) and Native Hawaiians/Pacific Islanders (NHs/PIs) are the fastest growing segments of the US population. However, their population sizes are small, and thus AAs and NHs/PIs are often aggregated into a single racial/ethnic group or omitted from research and health statistics. The groups' substance use disorders (SUDs) and treatment needs have been under-recognized. We examined recent epidemiological data on the extent of alcohol and drug use disorders and the use of treatment services by AAs and NHs/PIs. NHs/PIs on average were less educated and had lower levels of household income than AAs. Considered as a single group, AAs and NHs/PIs showed a low prevalence of substance use and disorders. Analyses of survey data that compared AAs and NHs/PIs revealed higher prevalences of substance use (alcohol, drugs), depression and delinquency among NHs than among AAs. Among treatment-seeking patients in mental healthcare settings, NHs/PIs had higher prevalences of DSM-IV diagnoses than AAs (alcohol/drug, mood, adjustment, childhood-onset disruptive or impulse-control disorders), although co-morbidity was common in both groups. AAs and NHs/PIs with an SUD were unlikely to use treatment, especially treatment for alcohol problems, and treatment use tended to be related to involvement with the criminal justice system. Although available data are limited by small sample sizes of AAs and NHs/PIs, they demonstrate the need to separate AAs and NHs/PIs in health statistics and increase research into substance use and treatment needs for these fast-growing but understudied population groups.


Weak responses to auditory feedback perturbation during articulation in persons who stutter: evidence for abnormal auditory-motor transformation.
Previous empirical observations have led researchers to propose that auditory feedback (the auditory perception of self-produced sounds when speaking) functions abnormally in the speech motor systems of persons who stutter (PWS). Researchers have theorized that an important neural basis of stuttering is the aberrant integration of auditory information into incipient speech motor commands. Because of the circumstantial support for these hypotheses and the differences and contradictions between them, there is a need for carefully designed experiments that directly examine auditory-motor integration during speech production in PWS. In the current study, we used real-time manipulation of auditory feedback to directly investigate whether the speech motor system of PWS utilizes auditory feedback abnormally during articulation and to characterize potential deficits of this auditory-motor integration. Twenty-one PWS and 18 fluent control participants were recruited. Using a short-latency formant-perturbation system, we examined participants' compensatory responses to unanticipated perturbation of auditory feedback of the first formant frequency during the production of the monophthong [ε]. The PWS showed compensatory responses that were qualitatively similar to the controls' and had close-to-normal latencies (∼150 ms), but the magnitudes of their responses were substantially and significantly smaller than those of the control participants (by 47% on average, p<0.05). Measurements of auditory acuity indicate that the weaker-than-normal compensatory responses in PWS were not attributable to a deficit in low-level auditory processing. These findings are consistent with the hypothesis that stuttering is associated with functional defects in the inverse models responsible for the transformation from the domain of auditory targets and auditory error information into the domain of speech motor commands.


TLR4 mutation reduces microglial activation, increases Aβ deposits and exacerbates cognitive deficits in a mouse model of Alzheimer's disease.
Amyloid plaques, a pathological hallmark of Alzheimer's disease (AD), are accompanied by activated microglia. The role of activated microglia in the pathogenesis of AD remains controversial: either clearing Aβ deposits by phagocytosis or releasing proinflammatory cytokines and cytotoxic substances. Microglia can be activated via toll-like receptors (TLRs), a class of pattern-recognition receptors in the innate immune system. We previously demonstrated that an AD mouse model homozygous for a loss-of-function mutation of TLR4 had increases in Aβ deposits and buffer-soluble Aβ in the brain as compared with a TLR4 wild-type AD mouse model at 14-16 months of age. However, it is unknown if TLR4 signaling is involved in initiation of Aβ deposition as well as activation and recruitment of microglia at the early stage of AD. Here, we investigated the role of TLR4 signaling and microglial activation in early stages using 5-month-old AD mouse models when Aβ deposits start. Microglial activation and amyloid deposition in the brain were determined by immunohistochemistry in the AD models. Levels of cerebral soluble Aβ were determined by ELISA. mRNA levels of cytokines and chemokines in the brain and Aβ-stimulated monocytes were quantified by real-time PCR. Cognitive functions were assessed by the Morris water maze. While no difference was found in cerebral Aβ load between AD mouse models at 5 months with and without TLR4 mutation, microglial activation in a TLR4 mutant AD model (TLR4M Tg) was less than that in a TLR4 wild-type AD model (TLR4W Tg). At 9 months, TLR4M Tg mice had increased Aβ deposition and soluble Aβ42 in the brain, which were associated with decrements in cognitive functions and expression levels of IL-1β, CCL3, and CCL4 in the hippocampus compared to TLR4W Tg mice. TLR4 mutation diminished Aβ-induced IL-1β, CCL3, and CCL4 expression in monocytes. This is the first demonstration of TLR4-dependent activation of microglia at the early stage of β-amyloidosis. Our results indicate that TLR4 is not involved in the initiation of Aβ deposition and that, as Aβ deposits start, microglia are activated via TLR4 signaling to reduce Aβ deposits and preserve cognitive functions from Aβ-mediated neurotoxicity.


Hypericin-bearing magnetic iron oxide nanoparticles for selective drug delivery in photodynamic therapy.
Combining the concept of magnetic drug targeting and photodynamic therapy is a promising approach for the treatment of cancer. A high selectivity as well as significant fewer side effects can be achieved by this method, since the therapeutic treatment only takes place in the area where accumulation of the particles by an external electromagnet and radiation by a laser system overlap. In this article, a novel hypericin-bearing drug delivery system has been developed by synthesis of superparamagnetic iron oxide nanoparticles (SPIONs) with a hypericin-linked functionalized dextran coating. For that, sterically stabilized dextran-coated SPIONs were produced by coprecipitation and crosslinking with epichlorohydrin to enhance stability. Carboxymethylation of the dextran shell provided a functionalized platform for linking hypericin via glutaraldehyde. Particle sizes obtained by dynamic light scattering were in a range of 55-85 nm, whereas investigation of single magnetite or maghemite particle diameter was performed by transmission electron microscopy and X-ray diffraction and resulted in approximately 4.5-5.0 nm. Surface chemistry of those particles was evaluated by Fourier transform infrared spectroscopy and ζ potential measurements, indicating successful functionalization and dispersal stabilization due to a mixture of steric and electrostatic repulsion. Flow cytometry revealed no toxicity of pure nanoparticles as well as hypericin without exposure to light on Jurkat T-cells, whereas the combination of hypericin, alone or loaded on particles, with light-induced cell death in a concentration and exposure time-dependent manner due to the generation of reactive oxygen species. In conclusion, the combination of SPIONs' targeting abilities with hypericin's phototoxic properties represents a promising approach for merging magnetic drug targeting with photodynamic therapy for the treatment of cancer.


Factors protective against retinopathy in insulin-dependent diabetics free of retinopathy for 30 years.
To investigate which factors might protect against the development of retinopathy 40 insulin-dependent diabetics who had remained free from retinopathy despite diabetes of long duration (mean+/-1 SD 30+/-10 years) were compared with 40 patients who had background and 47 who had proliferative retinopathy (mean durations of disease 16+/-5 and 19+/-5 years respectively). The three groups had had similar mean ages at onset of diabetes. The mean of all postprandial blood glucose measurements at hospital clinics from diagnosis of diabetes to detection of retinopathy, or to the most recent negative eye examination, was 9.9+/-2.1 mmol/l (178+/-38 mg/100 ml) in the group with no retinopathy, 11.8+/-2.1 mmol/l (213+/-38 mg/100 ml) in those with background retinopathy, and 12.4+/-2.1 mmol/l (223+/-38 mg/100 ml) in those with proliferative retinopathy (p <0.0001). This difference was not reflected in present concentrations of haemoglobin A(1C), probably because glycaemic control had been improved after the development of retinopathy. In the groups with background and proliferative retinopathy there were significant negative correlations between mean blood glucose concentrations and the number of years that had elapsed from diagnosis of diabetes to detection of retinopathy, suggesting that the development of both grades of retinopathy depends on the degree and duration of glycaemic exposure.The patients with no retinopathy had attended clinic more frequently (p <0.025), more of them had required emergency hospital treatment for hypoglycaemia (p <0.0025), and they tended to have had a lower prevalence of hyperglycaemic coma than the other groups. Although mean percentage ideal body weight and diastolic blood pressure were lower in the patients with no retinopathy at the time of study, mean body weight, blood pressure, and the prevalence of smoking in the years before the development of retinopathy had been similar in all groups, suggesting that these did not influence the development of retinopathy.