Microscale mesoarrays created by dip-pen nanolithography for screening of protein-protein interactions

Using microarrays to probe protein-protein interactions is becoming increasingly attractive due to their compatibility with highly sensitive detection techniques, selectivity of interaction, robustness and capacity for examining multiple proteins simultaneously. The major drawback to using this approach is the relatively large volumes and high concentrations necessary. Reducing the protein array spot size should allow for smaller volumes and lower concentrations to be used as well as opening the way for combination with more sensitive detection technologies. Dip-Pen Nanolithography (DPN) is a recently developed technique for structure creation on the nano to microscale with the capacity to create biological architectures. Here we describe the creation of miniaturised microarrays, 'mesoarrays', using DPN with protein spots 400× smaller by area compared to conventional microarrays. The mesoarrays were then used to probe the ERK2-KSR binding event of the Ras/Raf/MEK/ERK signalling pathway at a physical scale below that previously reported. Whilst the overall assay efficiency was determined to be low, the mesoarrays could detect KSR binding to ERK2 repeatedly and with low non-specific binding. This study serves as a first step towards an approach that can be used for analysis of proteins at a concentration level comparable to that found in the cellular environment.

Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors

Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost-effective manufacturing processes. Microcarriers enable the culture of anchorage-dependent cells in stirred-tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow-derived MSC (hBM-MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM-MSC1 underwent more cumulative population doublings over three passages in comparison to hBM-MSC2 and hBM-MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM-MSC expansion. HBM-MSCs were successfully harvested and characterised, demonstrating hBM-MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier-based allogeneic cell therapy manufacture. Large-scale production of human bone-marrow derived mesenchymal stem cells (hBM-MSCs) requires expansion on microcarriers in agitated systems. This study demonstrates the importance of microcarrier selection and presents a systematic methodology for selection of an optimal microcarrier. The study also highlights the impact of an agitated culture environment in comparison to a static system, resulting in a significantly higher hBM-MSC yield under agitated conditions.

Developing and Validating a Perinatal Depression Screening Tool in Bungoma County, Kenya

Background: Depression-screening tools exist and are widely used in Western settings. There have been few studies done to explore whether or not existing tools are valid and effective to use in sub-Saharan Africa. Our study aimed to develop and validate a perinatal depression-screening tool in rural Kenya.

Methods: We utilized conducted free listing and card sorting exercises with a purposive sample of 12 women and 38 CHVs living in a rural community to explore the manifestations of perinatal depression in that setting. We used the information obtained to produce a locally relevant depression-screening tool that comprised of existing Western psychiatric concepts and locally derived items. Subsequently, we administered the novel depression-screening tool and two existing screening tools (the Edinburgh Postnatal Depression Scale and the Patient Health Questionnaire-9) to 193 women and compared the results of the screening tool with that of a gold standard structured clinical interview to determine validity.

Results: The free listing and card sorting exercise produced a set of 60 screening items. Of the items in this set, we identified the 10 items that most accurately classified cases and non-cases. This 10-item scale had a sensitivity of 100.0 and specificity of 81.2. This compared to 90.0, 31.5 and 90.0, 49.7 for the EPDS and the PHQ-9, respectively. Overall, we found a prevalence of depression of 5.2 percent.

Conclusions: The new scale does very well in terms of diagnostic validity, having the highest scores in this domain compared to the EPDS, EPDS-R and PHQ-9. The adapted scale does very well with regards to convergent validity-illustrating clear distinction between mean scores across the different categories. It does well with regards to discriminant validity, internal consistency reliability, and test-retest reliability- not securing top scores in those domains but still yielding satisfactory results.

The reliability and factorial validity of the Adolescent Alcohol Involvement Scale in a large sample in the United Kingdom

The assessment of adolescent drinking behavior is a complex task, complicated by variability in drinking patterns, the transitory and developmental nature of the behavior and the reliance (for large scale studies) on self-report questionnaires. The Adolescent Alcohol Involvement Scale (Mayer & Filstead, 1979) is a 14-item screening tool designed to help to identify alcohol misusers or more problematic drinkers. The present study utilized a large sample (n = 4066) adolescents from Northern Ireland. Results of Confirmatory Factor Analyses and reliability estimates revealed that the 14-items share sufficient common variance that scores can be considered to be reliable and that the 14 items can be scored to provide a composite alcohol use score.

STOPPFrail (Screening Tool of Older Persons Prescriptions in Frail Adults with Limited Life Expectancy): Consensus Validation

Background: To validate STOPPFrail, a list of explicit criteria for potentially inappropriate medications (PIMs) in frailer older adults with limited life expectancy. A Delphi consensus survey of an expert panel (n = 17) comprising specialists in geriatric medicine, clinical pharmacology, palliative care, psychiatry of old age, clinical pharmacy and general practice.
Methods: STOPPFrail criteria was initially created by the authors based on clinical
experience and appraisal of the available literature. Criteria were organised according to physiological system. Each criterion was accompanied by an explanation. Panellists ranked their agreement with each criterion on a 5-point Likert scale and invited to provide written feedback. Criteria with a median Likert response of 4/5 (agree/strongly agree) and a 25th centile of ≥4 were included in the final criteria.
Results: Three Delphi rounds were required. All panellists completed all rounds. Thirty criteria were proposed for inclusion; 26 were accepted. No new criteria were added. The first two criteria suggest deprescribing medications with no indication or where compliance is poor. The remaining 24 criteria include lipid-lowering therapies, alpha-blockers for hypertension, anti-platelets, neuroleptics, proton pump inhibitors, H-2 receptor antagonists, anti-spasmodics, theophylline, leukotriene antagonists, calcium supplements, bone anti-resorptive therapy, selective oestrogen receptor modulators, non-steroidal antiinflammatories, corticosteroids, 5-alpha reductase inhibitors, alpha-1 selective blockers, muscarinic antagonists, oral diabetic agents, ACE-inhibitors, angiotensin receptor blockers, systemic oestrogens, multivitamins, nutritional supplements and prophylactic antibiotics. Anticoagulants and anti-depressants were excluded. Despite incorporation of panellists’ suggestions, memantine and acetyl-cholinesterase inhibitors remained inconclusive.
Conclusion: STOPPFrail comprises 26 criteria, which have been judged by broad consensus, to be potentially inappropriate in frailer older patients with limited life expectancy. STOPPFrail may assist in deprescribing medications in these patients.

Modeling long-term variability and change of soil moisture and groundwater level - from catchment to global scale

The water stored in and flowing through the subsurface is fundamental for sustaining human activities and needs, feeding water and its constituents to surface water bodies and supporting the functioning of their ecosystems. Quantifying the changes that affect the subsurface water is crucial for our understanding of its dynamics and changes driven by climate change and other changes in the landscape, such as in land-use and water-use. It is inherently difficult to directly measure soil moisture and groundwater levels over large spatial scales and long times. Models are therefore needed to capture the soil moisture and groundwater level dynamics over such large spatiotemporal scales. This thesis develops a modeling framework that allows for long-term catchment-scale screening of soil moisture and groundwater level changes. The novelty in this development resides in an explicit link drawn between catchment-scale hydroclimatic and soil hydraulics conditions, using observed runoff data as an approximation of soil water flux and accounting for the effects of snow storage-melting dynamics on that flux. Both past and future relative changes can be assessed by use of this modeling framework, with future change projections based on common climate model outputs. By direct model-observation comparison, the thesis shows that the developed modeling framework can reproduce the temporal variability of large-scale changes in soil water storage, as obtained from the GRACE satellite product, for most of 25 large study catchments around the world. Also compared with locally measured soil water content and groundwater level in 10 U.S. catchments, the modeling approach can reasonably well reproduce relative seasonal fluctuations around long-term average values. The developed modeling framework is further used to project soil moisture changes due to expected future climate change for 81 catchments around the world. The future soil moisture changes depend on the considered radiative forcing scenario (RCP) but are overall large for the occurrence frequency of dry and wet events and the inter-annual variability of seasonal soil moisture. These changes tend to be higher for the dry events and the dry season, respectively, than for the corresponding wet quantities, indicating increased drought risk for some parts of the world.

Glasgow Psychosis Screening tool for use in adults with Intellectual Disabilities (GPS-ID): development and psychometric properties

Background: Prevalence of psychosis is known to be higher in adults with intellectual disabilities (ID) than in the general adult population. However, there have been no attempts to develop a psychosis screening tool specifically for the adult ID population. The present study describes the development and preliminary evaluation of a new measure, the Glasgow Psychosis Screening tool for use in Adults with Intellectual Disabilities (GPS-ID). Method: An item pool was generated following: 1) focus groups with adults with ID and psychosis, and their carers and/or workers; 2) expert input from clinicians. A draft scale was compiled and refined following expert feedback. The new scale, along with the Psychotic Symptom Rating Scales was administered to 20 adults with ID (10 with and 10 without psychosis) and their relative or carers. Results: The GPS-ID total score, self-report subscale and informant rating-subscale differentiated psychosis and non-psychosis groups. The tool had good internal consistency (Cronbach’s α=0.91), and a cut-off score ≥4 yielded high sensitivity (90%) and specificity (100%). The method of tool development supports face and content validity. Criterion validity was not supported. Conclusions: Preliminary investigation of the tool’s psychometric properties is positive, although further investigation is required. The tool is accessible to adults with mild to moderate ID and can be completed in 15-30 minutes. The GPS-ID is not a diagnostic tool, therefore any adult exceeding the cut-off score of ≥4 should receive further assessment.

Screening of esophageal varices by esophageal capsule endoscopy: results of a French multicenter prospective study

International audience

Study of depressive symptoms according to Zung's self-rating scale on men deprived of freedom in a city of Colombia

Objective: To assess depressive symptoms in men deprived from freedom in a prison in a Colombian intermediate city. Material and Method: A cross sectional study was performed on a sample of three hundred and three patients in the Medium Security penitentiary and Prison Facility of the city of Manizales between April and May 2014. The information was collected through the Zung self-rating depression scale (SDS), subsequently there were established the positive results for depression screening according to the final score of the scale. Results: 303 men deprived from freedom were evaluated, mean age of 32.96 years +/- 10.8 years, 43.5% were living in cohabitation, 38% were single and 10.2% married; 33.7% had a primary education, 58% had secondary or incomplete secondary education, 5.6% reported higher studies; 38.6% (95% CI: 35.8; 41.4) reported symptoms of depression, predominating in ages between 18 to 44 years, no statistically significant differences p> 0.05 between the variables analyzed were found. Conclusions: The results of this study give rise to clinical evaluation, by specialized staff in the area of psychiatry and his intervention, given the characteristics of self-reported depression for this population.

ESR/ERS white paper on lung cancer screening

Lung cancer is the most frequently fatal cancer, with poor survival once the disease is advanced. Annual low-dose computed tomography has shown a survival benefit in screening individuals at high risk for lung cancer. Based on the available evidence, the European Society of Radiology and the European Respiratory Society recommend lung cancer screening in comprehensive, quality-assured, longitudinal programmes within a clinical trial or in routine clinical practice at certified multidisciplinary medical centres. Minimum requirements include: standardised operating procedures for low-dose image acquisition, computer-assisted nodule evaluation, and positive screening results and their management; inclusion/exclusion criteria; expectation management; and smoking cessation programmes. Further refinements are recommended to increase quality, outcome and cost-effectiveness of lung cancer screening: inclusion of risk models, reduction of effective radiation dose, computer-assisted volumetric measurements and assessment of comorbidities (chronic obstructive pulmonary disease and vascular calcification). All these requirements should be adjusted to the regional infrastructure and healthcare system, in order to exactly define eligibility using a risk model, nodule management and a quality assurance plan. The establishment of a central registry, including a biobank and an image bank, and preferably on a European level, is strongly encouraged. Key points: • Lung cancer screening using low dose computed tomography reduces mortality. • Leading US medical societies recommend large scale screening for high-risk individuals. • There are no lung cancer screening recommendations or reimbursed screening programmes in Europe as of yet. • The European Society of Radiology and the European Respiratory Society recommend lung cancer screening within a clinical trial or in routine clinical practice at certified multidisciplinary medical centres. • High risk, eligible individuals should be enrolled in comprehensive, quality-controlled longitudinal programmes.

Portuguese Version of the Suicidal Behaviors Questionnaire-Revised: Validation Data and the Establishment of a Cut-Score for Screening Purposes

The aim of the present study is to provide validation data regarding the Portuguese version of the Suicidal Behaviors Questionnaire Revised in nonclinical individuals. Two studies were undertaken with two different nonclinical samples in order to demonstrate reliability, concurrent, predictive, and construct validity, and in order to establish an appropriate cut-score for nonclinical individuals. A sample of 810 community adults participated in Study 1. Results from this study provided information regarding scale internal consistency, exploratory and confirmatory factor analysis, and concurrent validity. Receiver operating characteristic curve analysis established a cut-off score to be used for screening purposes with nonclinical individuals. A sample of 440 young adults participated in Study 2, which demonstrated scale score internal consistency and 5-month predictive validity. Further, 5-month test-retest reliability was also evaluated and the correlations of SBQ-R scale scores with two other measures that assess constructs related to suicidality, depression and psychache, were also performed. In addition, confirmatory factor analysis was undertaken to demonstrate the robustness of the result obtained in Study 1. Overall, findings supported the psychometric appropriateness of the Portuguese Suicidal Behaviors Questionnaire-Revise

Large scale underground energy storage for renewables integration: general criteria for reservoir identification and viable technologies.

The increasing integration of renewable energies in the electricity grid contributes considerably to achieve the European Union goals on energy and Greenhouse Gases (GHG) emissions reduction. However, it also brings problems to grid management. Large scale energy storage can provide the means for a better integration of the renewable energy sources, for balancing supply and demand, to increase energy security, to enhance a better management of the grid and also to converge towards a low carbon economy. Geological formations have the potential to store large volumes of fluids with minimal impact to environment and society. One of the ways to ensure a large scale energy storage is to use the storage capacity in geological reservoir. In fact, there are several viable technologies for underground energy storage, as well as several types of underground reservoirs that can be considered. The geological energy storage technologies considered in this research were: Underground Gas Storage (UGS), Hydrogen Storage (HS), Compressed Air Energy Storage (CAES), Underground Pumped Hydro Storage (UPHS) and Thermal Energy Storage (TES). For these different types of underground energy storage technologies there are several types of geological reservoirs that can be suitable, namely: depleted hydrocarbon reservoirs, aquifers, salt formations and caverns, engineered rock caverns and abandoned mines. Specific site screening criteria are applicable to each of these reservoir types and technologies, which determines the viability of the reservoir itself, and of the technology for any particular site. This paper presents a review of the criteria applied in the scope of the Portuguese contribution to the EU funded project ESTMAP – Energy Storage Mapping and Planning.

Newborn Hearing Screening And Early Diagnostic In The Nicu.

The aim was to describe the outcome of neonatal hearing screening (NHS) and audiological diagnosis in neonates in the NICU. The sample was divided into Group I: neonates who underwent NHS in one step and Group II: neonates who underwent a test and retest NHS. NHS procedure was automated auditory brainstem response. NHS was performed in 82.1% of surviving neonates. For GI, referral rate was 18.6% and false-positive was 62.2% (normal hearing in the diagnostic stage). In GII, with retest, referral rate dropped to 4.1% and false-positive to 12.5%. Sensorineural hearing loss was found in 13.2% of infants and conductive in 26.4% of cases. There was one case of auditory neuropathy spectrum (1.9%). Dropout rate in whole process was 21.7% for GI and 24.03% for GII. We concluded that it was not possible to perform universal NHS in the studied sample or, in many cases, to apply it within the first month of life. Retest reduced failure and false-positive rate and did not increase evasion, indicating that it is a recommendable step in NHS programs in the NICU. The incidence of hearing loss was 2.9%, considering sensorineural hearing loss (0.91%), conductive (1.83%) and auditory neuropathy spectrum (0.19%).

[why Does The Prevalence Of Cytopathological Results Of Cervical Cancer Screening Can Vary Significantly Between Two Regions Of Brazil?].

To analyze the prevalence of cervical cytopathological results for the screening of cervical cancer with regard to women's age and time since the last examination in Maceió and Rio de Janeiro, Brazil, among those assisted by the Brazilian Unified Health System. Cervical cytopathological results available in the Information System of Cervical Cancer Screening for the year 2011 were analyzed, corresponding to 206,550 for Rio de Janeiro and 45,243 for Maceió. In Rio de Janeiro, examination at one and two year intervals predominated, while in Maceió examination at one and three year intervals had a higher predominance. Women who underwent cervical smear screening in Maceió were older than those in Rio de Janeiro. The prevalence of invasive squamous cell carcinoma was similar for the two cities, but all the other results presented a higher prevalence in Rio de Janeiro: ASCUS (PR=5.32; 95%CI 4.66-6.07); ASCH (PR=4.27; 95%CI 3.15-5.78); atypical glandular cells (PR=10.02; 95%CI 5.66-17.76); low-grade squamous intraepithelial lesions (PR=6.10; 95%CI 5.27-7.07); high-grade squamous intraepithelial lesions (PR=8.90; 95%CI 6.50-12.18) and adenocarcinoma (PR=3.00; 95%CI 1.21-7.44). The rate of unsatisfactory cervical samples was two times higher in Maceió and that of rejected samples for analysis was five times higher in Maceió when compared to Rio de Janeiro. The prevalence rates of altered cervical cytopathological results was significantly higher in Rio de Janeiro than in Maceió. There is no objective information that may justify this difference. One hypothesis is that there may be a difference in the diagnostic performance of the cervical cancer screening, which could be related to the quality of the Pap smear. Thus, these findings suggest that it would be necessary to perform this evaluation at national level, with emphasis on the performance of cervical cancer screening in order to improve the effectiveness of cervical cancer control.

Microbial Lipid Production: Screening With Yeasts Grown On Brazilian Molasses.

Rhodotorula glutinis CCT 2182, Rhodosporidium toruloides CCT 0783, Rhodotorula minuta CCT 1751 and Lipomyces starkeyi DSM 70296 were evaluated for the conversion of sugars from Brazilian molasses into single-cell oil (SCO) feedstock for biodiesel. Pulsed fed-batch fermentations were performed in 1.65 l working volume bioreactors. The maximum specific growth rate (µmax), lipid productivity (Pr) and cellular lipid content were, respectively, 0.23 h(-1), 0.41 g l(-1) h(-1), and 41% for Rsp. toruloides; 0.20 h(-1), 0.27 g l(-1) h(-1), and 36% for Rta. glutinis; 0.115 h(-1), 0.135 g l(-1) h(-1), and 27 % for Rta. minuta; and 0.11 h(-1), 0.13 g l(-1) h(-1), and 32% for L. starkeyi. Based on their microbial lipid productivity, content, and profile, Rsp. toruloides and Rta. glutinis are promising candidates for biodiesel production from Brazilian molasses. All the oils from the yeasts were similar to the composition of plant oils (rapeseed and soybean) and could be used as raw material for biofuels, as well as in food and nutraceutical products.