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We describe herein a method of recharging used commercial spin columns or assembling homemade spin columns using filter paper as binding material for cost-effective, low throughput nucleic acid purification. The efficiency of filter paper-based spin columns was evaluated for purification of nucleic acids from various sources. Following protocols of commercial kits, we found filter paper to be a useful binding material for purification of nucleic acids, including plant genomic DNA, plant total RNA, PCR products, and DNA from agarose gels. However, filter paper has a weak binding affinity to plasmid DNA in tested miniprep protocols. Protocols for the use of filter paper recharged spin columns or homemade spin columns for low throughput purification of plant genomic DNA and total RNA with unused commercial kit buffers or less expensive homemade buffers are presented.

Introduction \ud Data collection using paper-based questionnaires can be time consuming and return errors affect data accuracy, completeness, and information quality in health surveys. We compared smartphone and paper-based data collection systems in the Burden of Obstructive Lung Disease (BOLD) study in rural Sudan. \ud Methods \ud This exploratory pilot study was designed to run in parallel with the cross-sectional household survey. The Open Data Kit was used to programme questionnaires in Arabic into smartphones. We included 100 study participants (83% women; median age = 41.5 ± 16.4 years) from the BOLD study from 3 rural villages in East-Gezira and Kamleen localities of Gezira state, Sudan. Questionnaire data were collected using smartphone and paper-based technologies simultaneously. We used Kappa statistics and inter-rater class coefficient to test agreement between the two methods. Results Symptoms reported included cough (24%), phlegm (15%), wheezing (17%), and shortness of breath (18%). One in five were or had been cigarette smokers. The two data collection methods varied between perfect to slight agreement across the 204 variables evaluated (Kappa varied between 1.00 and 0.02 and inter-rater coefficient between 1.00 and -0.12). Errors were most commonly seen with paper questionnaires (83% of errors seen) vs smartphones (17% of errors seen) administered questionnaires with questions with complex skip-patterns being a major source of errors in paper questionnaires. Automated checks and validations in smartphone-administered questionnaires avoided skip-pattern related errors. Incomplete and inconsistent records were more likely seen on paper questionnaires. \ud Conclusion \ud Compared to paper-based data collection, smartphone technology worked well for data collection in the study, which was conducted in a challenging rural environment in Sudan. This approach provided timely, quality data with fewer errors and inconsistencies compared to paper-based data collection. We recommend this method for future BOLD studies and other population-based studies in similar settings.

BACKGROUND The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTATM Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo's national surveillance for routine diagnosis were selected for this study. METHODS The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory's standard DNA extraction, and processing the same samples after being dried and stored on FTATM card, and DNA extracted following the manufacturer's instructions. RESULTS The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTATM cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory's standard methodology, results showed high concordance, with Kappa index ranges of 0.9877-1.00 for CSF, and 0.8004-1.00 for serum samples. CONCLUSION The use of FTATM cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost.

Background The use of filter paper as a simple, inexpensive tool for storage and transportation of blood, ‘Dried Blood Spots’ or Guthrie cards, for diagnostic assays is well-established. In contrast, there are a paucity of diagnostic evaluations of dried cerebrospinal fluid (CSF) spots. These have potential applications in low-resource settings, such as Laos, where laboratory facilities for central nervous system (CNS) diagnostics are only available in Vientiane. In Laos, a major cause of CNS infection is Japanese encephalitis virus (JEV). We aimed to develop a dried CSF spot protocol and to evaluate its diagnostic performance using the World Health Organisation recommended anti-JEV IgM antibody capture enzyme-linked immunosorbent assay (JEV MAC-ELISA). Methodology and Principal Findings Sample volumes, spotting techniques and filter paper type were evaluated using a CSF-substitute of anti-JEV IgM positive serum diluted in Phosphate Buffer Solution (PBS) to end-limits of detection by JEV MAC-ELISA. A conventional protocol, involving eluting one paper punch in 200μl PBS, did not detect the end-dilution, nor did multiple punches utilising diverse spotting techniques. However, pre-cut filter paper enabled saturation with five times the volume of CSF-substitute, sufficiently improving sensitivity to detect the end-dilution. The diagnostic accuracy of this optimised protocol was compared with routine, neat CSF in a pilot, retrospective study of JEV MAC-ELISA on consecutive CSF samples, collected 2009–15, from three Lao hospitals. In comparison to neat CSF, 132 CSF samples stored as dried CSF spots for one month at 25–30°C showed 81.6% (65.7–92.3 95%CI) positive agreement, 96.8% (91.0–99.3 95%CI) negative agreement, with a kappa coefficient of 0.81 (0.70–0.92 95%CI). Conclusions/Significance The novel design of pre-cut filter paper saturated with CSF could provide a useful tool for JEV diagnostics in settings with limited laboratory access. It has the potential to improve national JEV surveillance and inform vaccination policies. The saturation of filter paper has potential use in the wider context of pathogen detection, including dried spots for detecting other analytes in CSF, and other body fluids., Author Summary Japanese encephalitis virus infection is widespread in Asia, and primarily affects children in poor, rural areas. The virus spreads to the brain and spinal cord resulting in significant death and disability. Diagnosis requires testing for the immune response ‘antibody’ specific to Japanese encephalitis virus in the cerebrospinal fluid that surrounds the brain and spinal cord. However, in most areas where the infection occurs there are no laboratory facilities. In the absence of laboratory diagnosis, we have poor knowledge of the extent of the problem in these areas. Dried blood spots on filter paper have been used widely for many years as simple, cost-effective tools for transporting samples for testing. However, there have been few attempts at using dried cerebrospinal fluid spots. We developed a technique for using dried spots to store and test cerebrospinal fluid for antibodies to Japanese encephalitis virus. We compared the results of dried spots with routine neat cerebrospinal fluid in 132 patient samples, and demonstrated excellent agreement between the two tests. This novel method for saturating dried cerebrospinal fluid spots has the potential to enhance our knowledge of Japanese encephalitis virus epidemiology, and inform health policies where they are most needed. It could also be transferred for use in diagnosing other infectious diseases, including using other body fluid samples.

BackgroundMost research manuscripts are not accepted for publication on first submission. A major part of the resubmission process is reformatting to another journal's specific requirements, a process separate from revising the scientific content. There has been little research to understand the magnitude of the burden imposed by the current resubmission process.MethodsWe analyzed original research article submission requirements from twelve randomly selected journals in each of eight scientific and clinical focus areas from the InCites Journal Citation Reports database. From the 96 journals selected, we randomly identified three recently published manuscripts and sent surveys to those first and/or corresponding authors (288 total) to solicit information on time spent reformatting resubmissions and opinions on the process.FindingsThere was significant variation in manuscript submission requirements for journals within the same scientific focus and only 4% of journals offered a fully format-free initial submission. Of 203 authors responding (71.5% response rate), only 11.8% expressed satisfaction with the resubmission process and 91% desired reforming the current system. Time spent on reformatting delays most publications by at least two weeks and by over three months in about 20% of manuscripts. The effort to comply with submission requirements has significant global economic burden, estimated at over $1.1 billion dollars annually when accounting for a research team's time.InterpretationWe demonstrate that there is significant resource utilization associated with resubmitting manuscripts, heretofore not properly quantified. The vast majority of authors are not satisfied with the current process. Addressing these issues by reconciling reformatting requirements among journals or adopting a universal format-free initial submission policy would help resolve a major subject for the scientific research community and provide more efficient dissemination of findings.

BackgroundThe psychometric properties of an online test are not necessarily identical to its paper and pencil original. The aim of this study is to test whether the factor structure of the Community Assessment of Psychic Experiences (CAPE) is measurement invariant with respect to online vs. paper and pencil assessment.MethodThe factor structure of CAPE items assessed by paper and pencil (N = 796) was compared with the factor structure of CAPE items assessed by the Internet (N = 21,590) using formal tests for Measurement Invariance (MI). The effect size was calculated by estimating the Signed Item Difference in the Sample (SIDS) index and the Signed Test Difference in the Sample (STDS) for a hypothetical subject who scores 2 standard deviations above average on the latent dimensions.ResultsThe more restricted Metric Invariance model showed a significantly worse fit compared to the less restricted Configural Invariance model (χ(2)(23) = 152.75, pConclusionsOur findings did not support measurement invariance with respect to assessment method. Because of the small effect sizes, the measurement differences between the online assessed CAPE and its paper and pencil original can be neglected without major consequences for research purposes. However, a person with a high vulnerability for psychotic symptoms would score 4.80 points lower on the total scale if the CAPE is assessed online compared to paper and pencil assessment. Therefore, for clinical purposes, one should be cautious with online assessment of the CAPE.

The distinction between worker and reproductive castes of social insects is receiving increased attention from a developmental rather than adaptive perspective. In the wasp genus Polistes, colonies are founded by one or more females, and the female offspring that emerge in that colony are either non-reproducing workers or future reproductives of the following generation (gynes). A growing number of studies now indicate that workers emerge with activated reproductive physiology, whereas the future reproductive gynes do not. Low nourishment levels for larvae during the worker-rearing phase of the colony cycle and higher nourishment levels for larvae when gynes are reared are now strongly suspected of playing a major role in this difference. Here, we present the results of a laboratory rearing experiment in which Polistes metricus single foundresses were held in environmental conditions with a higher level of control than in any previously published study, and the amount of protein nourishment made available to feed larvae was the only input variable. Three experimental feeding treatments were tested: restricted, unrestricted, and hand-supplemented. Analysis of multiple response variables shows that wasps reared on restricted protein nourishment, which would be the case for wasps reared in field conditions that subsequently become workers, tend toward trait values that characterize active reproductive physiology. Wasps reared on unrestricted and hand-supplemented protein, which replicates higher feeding levels for larvae in field conditions that subsequently become gynes, tend toward trait values that characterize inactive reproductive physiology. Although the experiment was not designed to test for worker behavior per se, our results further implicate activated reproductive physiology as a developmental response to low larval nourishment as a fundamental aspect of worker behavior in Polistes.

India has the highest burden of Tuberculosis (TB) and multidrug-resistant TB (MDR-TB) worldwide. Innovative technology is the need of the hour to identify these cases that remain either undiagnosed or inadequately diagnosed due to the unavailability of appropriate tools at primary healthcare settings. We developed and evaluated 3 kits, namely ‘TB Detect’ (containing BioFM-Filter device), ‘TB Concentration and Transport’ (containing Trans-Filter device) and ‘TB DNA Extraction’ kits. These kits enable bio-safe equipment-free concentration of sputum on filters and improved fluorescence microscopy at primary healthcare centres, ambient temperature transport of dried inactivated sputum filters to central laboratories and molecular detection of drug resistance by PCR and DNA sequencing (Mol-DST). In a 2-site evaluation (n = 1190 sputum specimens) on presumptive TB patients, BioFM-Filter smear exhibited a significant increase in positivity of 7% and 4% over ZN smear and LED-FM smear (p

Poor research reporting is a major contributing factor to low study reproducibility, financial and animal waste. The ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines were developed to improve reporting quality and many journals support these guidelines. The influence of this support is unknown. We hypothesized that papers published in journals supporting the ARRIVE guidelines would show improved reporting compared with those in non-supporting journals. In a retrospective, observational cohort study, papers from 5 ARRIVE supporting (SUPP) and 2 non-supporting (nonSUPP) journals, published before (2009) and 5 years after (2015) the ARRIVE guidelines, were selected. Adherence to the ARRIVE checklist of 20 items was independently evaluated by two reviewers and items assessed as fully, partially or not reported. Mean percentages of items reported were compared between journal types and years with an unequal variance t-test. Individual items and sub-items were compared with a chi-square test. From an initial cohort of 956, 236 papers were included: 120 from 2009 (SUPP; n = 52, nonSUPP; n = 68), 116 from 2015 (SUPP; n = 61, nonSUPP; n = 55). The percentage of fully reported items was similar between journal types in 2009 (SUPP: 55.3 ± 11.5% [SD]; nonSUPP: 51.8 ± 9.0%; p = 0.07, 95% CI of mean difference -0.3-7.3%) and 2015 (SUPP: 60.5 ± 11.2%; nonSUPP; 60.2 ± 10.0%; p = 0.89, 95%CI -3.6-4.2%). The small increase in fully reported items between years was similar for both journal types (p = 0.09, 95% CI -0.5-4.3%). No paper fully reported 100% of items on the ARRIVE checklist and measures associated with bias were poorly reported. These results suggest that journal support for the ARRIVE guidelines has not resulted in a meaningful improvement in reporting quality, contributing to ongoing waste in animal research.

Background There is widespread evidence that statistical methods play an important role in original research articles, especially in medical research. The evaluation of statistical methods and reporting in journals suffers from a lack of standardized methods for assessing the use of statistics. The objective of this study was to develop and evaluate an instrument to assess the statistical intensity in research articles in a standardized way. Methods A checklist-type measure scale was developed by selecting and refining items from previous reports about the statistical contents of medical journal articles and from published guidelines for statistical reporting. A total of 840 original medical research articles that were published between 2007–2015 in 16 journals were evaluated to test the scoring instrument. The total sum of all items was used to assess the intensity between sub-fields and journals. Inter-rater agreement was examined using a random sample of 40 articles. Four raters read and evaluated the selected articles using the developed instrument. Results The scale consisted of 66 items. The total summary score adequately discriminated between research articles according to their study design characteristics. The new instrument could also discriminate between journals according to their statistical intensity. The inter-observer agreement measured by the ICC was 0.88 between all four raters. Individual item analysis showed very high agreement between the rater pairs, the percentage agreement ranged from 91.7% to 95.2%. Conclusions A reliable and applicable instrument for evaluating the statistical intensity in research papers was developed. It is a helpful tool for comparing the statistical intensity between sub-fields and journals. The novel instrument may be applied in manuscript peer review to identify papers in need of additional statistical review.

SettingSouth Africa has the third highest tuberculosis (TB) burden in the world. Intensified case finding, recommended by WHO, is one way to control TB.ObjectiveTo evaluate the effectiveness and acceptability of a paper slip method for TB contact tracing.MethodTB patients were offered paper slips to give to their contacts, inviting them for TB screening. The number of contacts screened and the proportion diagnosed with TB was calculated. Contacts that returned to the clinic after receiving the slips were interviewed. A focus group discussion (FGD) with TB patients was held to determine their acceptability.ResultsFrom 718 paper slips issued, a 26% TB contact tracing rate was found, with a 12% case detection rate. The majority (68%) of contacts were screened within 2 weeks of receiving the slip. Age and gender were not significantly associated with time to screening. 16% of the contacts screened did not reside with the TB patients. 98% of the contacts said the method was acceptable. FGD findings show that this method is acceptable and may prevent stigma associated with TB/HIV.ConclusionThis simple, inexpensive method yields high contact tracing and case detection rates and potentially would yield additional benefits outside households.

When young children do not receive adequate amounts of the micronutrient iodine in their diet, their growth and cognitive development can be impaired. Nearly every country in the world has programs in place to track iodine intake and provide supplemental iodine if needed, usually in the form of fortified salt. The iodine nutrition status of a population can be tracked by monitoring iodine levels in urine samples to see if the median value falls in the range of 100-300 micrograms of iodine per liter of urine (μg I/L), which indicates adequate or more than adequate iodine nutrition. Many low and middle-income countries (LMIC) do not have a laboratory capable of carrying out this challenging assay, so samples must be sent out for assay in external labs, which is expensive and time-consuming. In most LMIC, population iodine surveys are carried out every 5-10 years, which limits the utility of the data for program monitoring and evaluation. To solve this problem, we developed a field-friendly paper test card that uses the Sandell-Kolthoff reaction to measure urinary iodine levels. A blind internal validation study showed that 93% of samples (n = 60) of iodide in an artificial urine matrix were categorized correctly by visual analysis as deficient, adequate, or excessive for levels set forth by the World Health Organization. Quantitative measurements based on computer image analysis had an error of 40 ± 20 μg I/L (n = 35 for samples in the calibration range) and these results categorized 88% of the samples (n = 60) correctly. We employed lifecycle analysis principles to address the known toxicity of arsenic, which is an obligatory reagent in the Sandell-Kolthoff reaction. Disposal of the cards in a landfill (their most likely destination after use) could let arsenic leach into groundwater; toxicity characteristic leaching procedure (TCLP) tests showed that the level of arsenic leached from the cards was 28.78 ppm, which is above the United States Environmental Protection Agency's limit of 5 parts per million for solid waste. We integrated a remediation module into the card. This module contains oxone, to oxidize As(III) to As(V) oxyacids, and the iron oxide goethite. TCLP testing showed that the leachable amount of arsenic was reduced by at least 97.6%-from 28.8 ppm to lower than 0.7 ± 0.7 ppm (n = 20). This upstream intervention rendered the test card suitable for landfilling while retaining its functionality to perform a critical public health evaluation.

Authorship on peer-reviewed journal articles and abstracts has become the main currency and reward unit in academia. Such a reward has become a crucial component for students and postdocs who are often under-compensated and thus value authorship as their primary reward mechanism. While numerous scientific and publishing organizations have attempted to write guidelines for defining who gets to be an author and what rank they should be listed in, there remains much ambiguity when it comes to how the various criteria are weighed by research faculty. Here we sought to quantify the relative importance of each of the 11 criteria we defined as being significant contributions for scientific authorship. We sent out an anonymous survey to approximately 564 faculty members at ten different research institutions across the United States. The faculty were from the biomedical engineering, biology, and bioengineering departments. The response rate was approximately 18% with a final sample size of 102 faculty members. We found that there was a consensus on some criteria as being crucial, such as time spent conducting experiments, but there was a lack of consensus regarding the role of obtaining funding. This study provides one of the first quantitative assessments of how faculty members in the biomedical sciences evaluated these 11 authorship criteria. We believe researchers will find this insightful and will narrow down the disparity between what they assume as being important and what faculty value. This understanding will also bring us closer to establishing a more standardized system for determining authorship and rank in the biosciences.

Background Loop mediated isothermal amplification (LAMP) provides an opportunity for improved, field-friendly detection of malaria infections in endemic areas. However data on the diagnostic accuracy of LAMP for active case detection, particularly low-density parasitaemias, are lacking. We therefore evaluated the performance of a new LAMP kit compared with PCR using DNA from filter paper blood spots. Methods and Findings Samples from 865 fever patients and 465 asymptomatic individuals collected in Zanzibar were analysed for Pan (all species) and Pf (P. falciparum) DNA with the Loopamp MALARIA Pan/Pf kit. Samples were amplified at 65°C for 40 minutes in a real-time turbidimeter and results were compared with nested PCR. Samples with discordant results between LAMP and nested PCR were analysed with real-time PCR. The real-time PCR corrected nested PCR result was defined as gold standard. Among the 117 (13.5%) PCR detected P. falciparum infections from fever patients (mean parasite density 7491/µL, range 6–782,400) 115, 115 and 111 were positive by Pan-LAMP, Pf-LAMP and nested PCR, respectively. The sensitivities were 98.3% (95%CI 94–99.8) for both Pan and Pf-LAMP. Among the 54 (11.6%) PCR positive samples from asymptomatic individuals (mean parasite density 10/µL, range 0–4972) Pf-LAMP had a sensitivity of 92.7% (95%CI 80.1–98.5) for detection of the 41 P. falciparum infections. Pan-LAMP had sensitivities of 97% (95%CI 84.2–99.9) and 76.9% (95%CI 46.2–95) for detection of P. falciparum and P. malariae, respectively. The specificities for both Pan and Pf-LAMP were 100% (95%CI 99.1–100) in both study groups. Conclusion Both components of the Loopamp MALARIA Pan/Pf detection kit revealed high diagnostic accuracy for parasite detection among fever patients and importantly also among asymptomatic individuals of low parasite densities from minute blood volumes preserved on filter paper. These data support LAMPs potential role for improved detection of low-density malaria infections in pre-elimination settings.

The Maternal Health Task Force (MHTF) and PLOS Medicine issue the call for papers for Year 3 of the MHTF-PLOS Collection: Integrating Health Care to Meet the Needs of the Mother–Infant Pair. Please see later in the article for the Editors' Summary

We describe a method that uses crowdsourced postcode coordinates and Google maps to estimate average distance and travel time for inhabitants of a municipality to a casualty clinic in Norway. The new method was compared with methods based on population centroids, median distance and town hall location, and we used it to examine how distance affects the utilisation of out-of-hours primary care services. At short distances our method showed good correlation with mean travel time and distance. The utilisation of out-of-hours services correlated with postcode based distances similar to previous research. The results show that our method is a reliable and useful tool for estimating average travel distances and travel times. publishedVersion

The PLOS Medicine editors issue a call for papers on addressing global disparities in non-communicable diseases–CVD, cancer, COPD, asthma, diabetes, mental health disorders, and substance abuse–for 2013.

Cleaton-Jones, chair of the ethics committee that approved the trial of circumcision for preventing HIV, shares with us the discussions that the committee had ahead of granting approval.

The highly contagious nature of SARS-CoV-2 has led to several studies on the transmission of the virus. A little studied potential fomite of great concern in the community is currency, which has been shown to harbor microbial pathogens in several studies. Since the onset of the COVID-19 pandemic, many businesses in the United States have limited the use of banknotes in favor of credit cards. However, SARS-CoV-2 has shown greater stability on plastic in several studies. Herein, the stability of SARS-CoV-2 at room temperature on banknotes, money cards and coins was investigated. In vitro studies with live virus suggested SARS-CoV-2 was highly unstable on banknotes, showing an initial rapid reduction in viable virus and no viral detection by 24 hours. In contrast, SARS-CoV-2 displayed increased stability on money cards with live virus detected after 48 hours. Environmental swabbing of currency and money cards on and near the campus of Brigham Young University supported these results, with no detection of SARS-CoV-2 RNA on banknotes, and a low level on money cards. However, no viable virus was detected on either. These preliminary results suggest that the use of money cards over banknotes in order to slow the spread of this virus may be ill-advised. These findings should be investigated further through larger environmental studies involving more locations.

This study, based in San Bernardino County, Southern California, collected and examined tap water samples within the area to explore the feasibility of adopting non-industrial equipment and methods to reduce water hardness and total dissolved solids(TDS). We investigated how water quality could be improved by utilizing water boiling, activated carbon and sodium bicarbonate additives, as well as electrolysis methods. The results show that heating is effective at lower temperatures rather than long boils, as none of the boiling tests were lower than the original value. Activated carbon is unable to lower TDS, because it is unable to bind to any impurities present in the water. This resulted in an overall TDS increase of 3.5%. However, adding small amounts of sodium bicarbonate(NaHCO3) will further eliminate water hardness by reacting with magnesium ions and improve taste, while increasing the pH. When added to room temperature tap water, there is a continuous increase in TDS of 24.8% at the 30 mg/L mark. The new findings presented in this study showed that electrolysis was the most successful method in eliminating TDS, showing an inverse proportion where an increasing electrical current and duration of electrical lowers more amounts of solids. This method created a maximum decrease in TDS by a maximum of 22.7%, with 3 tests resulting in 15.3–16.6% decreases. Furthermore, when water is heated to a temperature around 50°C (122°F), a decrease in TDS of around 16% was also shown. The reduction of these solids will help lower water hardness and improve the taste of tap water. These results will help direct residents to drink more tap water rather than bottled water with similar taste and health benefits for a cheaper price as well as a reduction on plastic usage.

Synthetic fibers are increasingly seen to dominate microplastic pollution profiles in aquatic environments, with evidence pointing to textiles as a potentially important source. However, the loss of microfibers from textiles during laundry is poorly understood. We evaluated microfiber release from a variety of synthetic and natural consumer apparel textile samples (n = 37), with different material types, constructions, and treatments during five consecutive domestic laundry cycles. Microfiber loss ranged from 9.6 mg to 1,240 mg kg-1 of textile per wash, or an estimated 8,809 to > 6,877,000 microfibers. Mechanically-treated polyester samples, dominated by fleeces and jerseys, released six times more microfibers (161 ± 173 mg kg-1 per wash) than did nylon samples with woven construction and filamentous yarns (27 ± 14 mg kg-1 per wash). Fiber shedding was positively correlated with fabric thickness for nylon and polyester. Interestingly, cotton and wool textiles also shed large amounts of microfibers (165 ± 44 mg kg-1 per wash). The similarity between the average width of textile fibers here (12.4 ± 4.5 μm) and those found in ocean samples provides support for the notion that home laundry is an important source of microfiber pollution. Evaluation of two marketed laundry lint traps provided insight into intervention options for the home, with retention of up to 90% for polyester fibers and 46% for nylon fibers. Our observation of a > 850-fold difference in the number of microfibers lost between low and high shedding textiles illustrates the strong potential for intervention, including more sustainable clothing design.

Phytoseiids are predatory mites that prey on other mites and small arthropods, and several species are used in commercial agriculture for biological control of pests. To optimize phytoseiid mites' use in biocontrol, an efficient method for oral delivery of test compounds is required to assess their sensitivities to pesticides, RNAi for gene functional analysis and artificial diets. Here we developed four methods for oral delivery of a solution of xenobiotics to different life stages of the commercially available generalist predatory mite Neoseiulus californicus: (i) soaking mites in the solution, or allowing them to feed on (ii) spider mites soaked in the solution, (iii) a solution droplet, or (iv) solution-saturated filter paper. As measured by ingestion of a tracer dye, the droplet-based feeding system was most efficient; the dye was observed in the alimentary canal of >90% test mites of all life stages, with no mortality. The droplet-based feeding system was also effective for the commercially available specialist predatory mite Phytoseiulus persimilis, with >80% delivery efficiency. This study paves the way for development of methods for high-throughput RNAi and for toxicological or nutritional assays in phytoseiid mites.

Novel chemistry for vector control is urgently needed to counter insecticide resistance in mosquitoes. Here a new meta-diamide insecticide, broflanilide (TENEBENALTM), was evaluated in East African experimental huts in Moshi, northern Tanzania. Two consecutive experimental hut trials with broflanilide 50WP were conducted; the first evaluating the efficacy of three concentrations, 50 mg/m2, 100 mg/m2, and 200 mg/m2 using a prototype formulation, and the second trial evaluating an improved formulation. The IRS treatments were applied on both mud and concrete surfaces and efficacy was monitored over time. The mortality, blood-feeding inhibition and exiting behaviour of free-flying wild mosquitoes was compared between treatment arms. Additionally, cone assays with pyrethroid-susceptible and resistant mosquito strains were conducted in the huts to determine residual efficacy. The first trial showed a dosage-mortality response of the prototype formulation and 3–8 months of residual activity, with longer activity on concrete than mud. The second trial with an improved formulation showed prolonged residual efficacy of the 100 mg/m2 concentration to 5–6 months on mud, and mosquito mortality on the concrete surface ranged between 94–100% for the full duration of the trial. In both trials, results with free-flying, wild Anopheles arabiensis echoed the mortality trend shown in cone assays, with the highest dose inducing the highest mortality and the improved formulation showing increased mortality rates. No blood-feeding inhibition or insecticide-induced exiting effects were observed with broflanilide. Broflanilide 50WP was effective against both susceptible and pyrethroid-resistant mosquito strains, demonstrating an absence of cross resistance between broflanilide and pyrethroids. The improved formulation, which has now been branded VECTRONTM T500, resulted in a prolonged residual efficacy. These results indicate the potential of this insecticide as an addition to the arsenal of IRS products needed to maintain both control of malaria and resistance management of malaria-transmitting mosquitoes.

As a means of obtaining reproducible and accurate numbers of larvae for laboratory rearing, we tested a large-particle flow-cytometer type device called the ‘Automated Particle Counter’ (APC). The APC is a gravity-fed, self-contained unit that detects changes in light intensity caused by larvae passing the detector in a water stream and controls dispensing by stopping the flow when the desired number has been reached. We determined the accuracy (number dispensed compared to the target value) and precision (distribution of number dispensed) of dispensing at a variety of counting sensitivity thresholds and larva throughput rates (larvae per second) using < 1-day old Anopheles gambiae and Aedes aegypti larvae. All measures were made using an APC algorithm called the ‘Smoothed Z-Score’ which allows the user to define how many standard deviations (Z scores) from the baseline light intensity a particle’s absorbance must exceed to register a count. We dispensed a target number of 100 An. gambiae larvae using Z scores from 2.5–8 and observed no difference among them in the numbers dispensed for scores from 2.5–6, however, scores of 7 and 8 under-counted (over-dispensed) larvae. Using a Z score ≤ 6, we determined the effect of throughput rate on the accuracy of the device to dispense An. gambiae larvae. For rates ≤ 98 larvae per second, the accuracy of dispensing a target of 100 larvae was - 2.29% ± 0.72 (95% CI of the mean) with a mode of 99 (49 of 348 samples). When using a Z score of 3.5 and rates ≤ 100 larvae per second, the accuracy of dispensing a target of 100 Ae. aegypti was - 2.43% ± 1.26 (95% CI of the mean) with a mode of 100 (6 of 42 samples). No effect on survival was observed on the number of An. gambiae first stage larvae that reached adulthood as a function of dispensing.

Previous studies have demonstrated that skin vibration is an important factor affecting the roughness perception of fine textures. For coarse textures, the determining physical factor is much less clear and there are indications that this might be participant-dependent. In this paper, we focused on roughness perception of both coarse and fine textures of different materials (glass particle surfaces and sandpapers). We investigated the relationship between subjective roughness ratings and three physical parameters (skin vibration, friction coefficient, and particle size) within a group of 30 participants. Results of the glass particle surfaces showed both spatial information (particle size) and temporal information (skin vibration) had a high correlation with subjective roughness ratings. The former correlation was slightly but significantly higher than the latter. The results also indicated different weights of temporal information and spatial information for roughness ratings among participants. Roughness ratings of a different material (sandpaper versus glass particles) could be either larger, similar or smaller, indicating differences among individuals. The best way to describe our results is that in their perceptual evaluation of roughness, different individuals weight temporal information, spatial information, and other mechanical properties differently.

The application of photocatalyst in pavements has received comprehensive attention in recent years due to its ability to decontaminate nitrogen oxides (NOx). However, it's remarkable that NOx also accumulated extensively in parking lots. The purpose of this study is to develop a movable photocatalytic plate (remarked photocatalytic KT plates) coupled with high activity to purify NOx. Firstly, the type of photocatalytic KT plates was determined according to NO removal experiment in laboratory. Then the plates were employed in the parking lots for removing NOx. One sample T-test, normality test and paired sample T-test methods for NOx concentration variation were conducted to determine the appropriate comparative means of dates under both dark and illuminated conditions. The difference of NOX concentration between dark and illuminated conditions was obtained to evaluate the photocatalytic removal efficiency. The results indicated that NO removal efficiency in laboratory and parking lots were 51.31% and 9.2%, respectively.

Intelligent biorefinery design that addresses both the composition of the biomass feedstock as well as fermentation microorganisms could benefit from dedicated tools for computational simulation and computer-assisted optimization. Here we present the BioLego Vn2.0 framework, based on Microsoft Azure Cloud, which supports large-scale simulations of biomass serial fermentation processes by two different organisms. BioLego enables the simultaneous analysis of multiple fermentation scenarios and the comparison of fermentation potential of multiple feedstock compositions. Thanks to the effective use of cloud computing it further allows resource intensive analysis and exploration of media and organism modifications. We use BioLego to obtain biological and validation results, including (1) exploratory search for the optimal utilization of corn biomasses-corn cobs, corn fiber and corn stover-in fermentation biorefineries; (2) analysis of the possible effects of changes in the composition of K. alvarezi biomass on the ethanol production yield in an anaerobic two-step process (S. cerevisiae followed by E. coli); (3) analysis of the impact, on the estimated ethanol production yield, of knocking out single organism reactions either in one or in both organisms in an anaerobic two-step fermentation process of Ulva sp. into ethanol (S. cerevisiae followed by E. coli); and (4) comparison of several experimentally measured ethanol fermentation rates with the predictions of BioLego.

BACKGROUND: Different setups and protocols have been developed for investigating insecticide effects on Anopheles (An.) mosquitoes, vectors of malaria. However, chemical uptake resulting from their tarsal contact with insecticide-treated material has seldom been investigated. To address the challenges encountered in the interpretation of bioassay data, a high throughput method for chemical analysis on malaria vectors was developed and validated for five selected insecticides including alpha-cypermethrin (aCYP), deltamethrin (DM), etofenprox (EPX), permethrin (PM), pirimiphos-methyl (PPM). METHODS: The method includes a single chemical extraction step via an ultrasound probe on mosquito samples and analysis via liquid chromatography coupled to high-resolution tandem mass spectrometry (UHPLC-MS/MS). The protocol was established for two malaria vector species, Anopheles gambiae senso stricto (s.s.) and An. stephensi, both males and females. Recovery rates ranged from 70 to 100% without any influence of sex or species. The method was efficiently applied to female An. gambiae s.s. of the KISUMU1 reference strain, after susceptibility tests using the World Health Organization´s standard protocol. RESULTS: Susceptibility tests revealed 13.4-18.4 minutes knockdown times for 50% mosquitoes during exposure to EPX and pyrethroids. The mortality rates 24 hours post-exposure to insecticides were mostly 99-100%, except in two PM and three PPM assays suggesting possible or confirmed resistance to these insecticides. The mean insecticide uptake in dead mosquitoes ranged from 23 pg (aCYP) to 1812 pg (EPX) per specimen. However, the mean uptake in survivors to PM and PPM was reduced by at least 25%, suggesting that acute doses were not achieved in these specimens during bioassays. CONCLUSIONS: The developed and validated UHPLC-MS/MS method could be used to address some limitations of bioassays or to assess the penetration of insecticides in mosquito matrix with reference to cuticle thickness and other insecticide resistance mechanisms.

Quantifying and comparing the scientific output of researchers has become critical for governments, funding agencies and universities. Comparison by reputation and direct assessment of contributions to the field is no longer possible, as the number of scientists increases and traditional definitions about scientific fields become blurred. The h-index is often used for comparing scientists, but has several well-documented shortcomings. In this paper, we introduce a new index for measuring and comparing the publication records of scientists: the pagerank-index (symbolised as π). The index uses a version of pagerank algorithm and the citation networks of papers in its computation, and is fundamentally different from the existing variants of h-index because it considers not only the number of citations but also the actual impact of each citation. We adapt two approaches to demonstrate the utility of the new index. Firstly, we use a simulation model of a community of authors, whereby we create various ‘groups’ of authors which are different from each other in inherent publication habits, to show that the pagerank-index is fairer than the existing indices in three distinct scenarios: (i) when authors try to ‘massage’ their index by publishing papers in low-quality outlets primarily to self-cite other papers (ii) when authors collaborate in large groups in order to obtain more authorships (iii) when authors spend most of their time in producing genuine but low quality publications that would massage their index. Secondly, we undertake two real world case studies: (i) the evolving author community of quantum game theory, as defined by Google Scholar (ii) a snapshot of the high energy physics (HEP) theory research community in arXiv. In both case studies, we find that the list of top authors vary very significantly when h-index and pagerank-index are used for comparison. We show that in both cases, authors who have collaborated in large groups and/or published less impactful papers tend to be comparatively favoured by the h-index, whereas the pagerank-index highlights authors who have made a relatively small number of definitive contributions, or written papers which served to highlight the link between diverse disciplines, or typically worked in smaller groups. Thus, we argue that the pagerank-index is an inherently fairer and more nuanced metric to quantify the publication records of scientists compared to existing measures.

Despite widespread support from policy makers, funding agencies, and scientific journals, academic researchers rarely make their research data available to others. At the same time, data sharing in research is attributed a vast potential for scientific progress. It allows the reproducibility of study results and the reuse of old data for new research questions. Based on a systematic review of 98 scholarly papers and an empirical survey among 603 secondary data users, we develop a conceptual framework that explains the process of data sharing from the primary researcher’s point of view. We show that this process can be divided into six descriptive categories: Data donor, research organization, research community, norms, data infrastructure, and data recipients. Drawing from our findings, we discuss theoretical implications regarding knowledge creation and dissemination as well as research policy measures to foster academic collaboration. We conclude that research data cannot be regarded a knowledge commons, but research policies that better incentivize data sharing are needed to improve the quality of research results and foster scientific progress. ratswd, ratswd working paper, Data Sharing, Academia, Systematic Review, Research Policy, Knowledge Commons, Crowd Science, Commons-based Peer Production

Deconstruction of the cellulose in plant cell walls is critical for carbon flow through ecosystems and for the production of sustainable cellulosic biofuels. Our understanding of cellulose deconstruction is largely limited to the study of microbes in isolation, but in nature, this process is driven by microbes within complex communities. In Neotropical forests, microbes in leaf-cutter ant refuse dumps are important for carbon turnover. These dumps consist of decaying plant material and a diverse bacterial community, as shown here by electron microscopy. To study the portion of the community capable of cellulose degradation, we performed enrichments on cellulose using material from five Atta colombica refuse dumps. The ability of enriched communities to degrade cellulose varied significantly across refuse dumps. 16S rRNA gene amplicon sequencing of enriched samples identified that the community structure correlated with refuse dump and with degradation ability. Overall, samples were dominated by Bacteroidetes, Gammaproteobacteria, and Betaproteobacteria. Half of abundant operational taxonomic units (OTUs) across samples were classified within genera containing known cellulose degraders, including Acidovorax, the most abundant OTU detected across samples, which was positively correlated with cellulolytic ability. A representative Acidovorax strain was isolated, but did not grow on cellulose alone. Phenotypic and compositional analyses of enrichment cultures, such as those presented here, help link community composition with cellulolytic ability and provide insight into the complexity of community-based cellulose degradation. Biological and Environmental Research/[DE-FC02-07ER64494]/BER/Estados Unidos National Science Foundation/[DGE-1256259]/NSF/Estados Unidos National Science Foundation/[DEB-0747002]/NSF/Estados Unidos National Science Foundation/[MCB-0702025]/NSF/Estados Unidos National Institutes of Health/[T32 GM07215]/NIH/Estados Unidos Universidad de Costa Rica/[]/UCR/Costa Rica Ministerio de Ciencia, Tecnología y Telecomunicaciones/[]/MICITT/Costa Rica University of Wisconsin-Madison's Hilldale Undergraduate Faculty Research Fellowship/[]//Estados Unidos UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)

In this paper, a digital 3D model which allows for visualisation in three dimensions and interactive manipulation is explored as a tool to help us understand the structural morphology and elucidate the functions of morphological structures of fragile microorganisms which defy live studies. We developed a deformable generic 3D model of haptoral anchor of dactylogyridean monogeneans that can subsequently be deformed into different desired anchor shapes by using direct manipulation deformation technique. We used point primitives to construct the rectangular building blocks to develop our deformable 3D model. Point primitives are manually marked on a 2D illustration of an anchor on a Cartesian graph paper and a set of Cartesian coordinates for each point primitive is manually extracted from the graph paper. A Python script is then written in Blender to construct 3D rectangular building blocks based on the Cartesian coordinates. The rectangular building blocks are stacked on top or by the side of each other following their respective Cartesian coordinates of point primitive. More point primitives are added at the sites in the 3D model where more structural variations are likely to occur, in order to generate complex anchor structures. We used Catmull-Clark subdivision surface modifier to smoothen the surface and edge of the generic 3D model to obtain a smoother and more natural 3D shape and antialiasing option to reduce the jagged edges of the 3D model. This deformable generic 3D model can be deformed into different desired 3D anchor shapes through direct manipulation deformation technique by aligning the vertices (pilot points) of the newly developed deformable generic 3D model onto the 2D illustrations of the desired shapes and moving the vertices until the desire 3D shapes are formed. In this generic 3D model all the vertices present are deployed for displacement during deformation.

Background Accurate and efficient detection of sub-microscopic malaria infections is crucial for enabling rapid treatment and interruption of transmission. Commercially available malaria LAMP kits have excellent diagnostic performance, though throughput is limited by the need to prepare samples individually. Here, we evaluate the clinical performance of a newly developed high throughput (HTP) sample processing system for use in conjunction with the Eiken malaria LAMP kit. Methods The HTP system utilised dried blood spots (DBS) and liquid whole blood (WB), with parallel sample processing of 94 samples per run. The system was evaluated using 699 samples of known infection status pre-determined by gold standard nested PCR. Results The sensitivity and specificity of WB-HTP-LAMP was 98.6% (95% CI, 95.7–100), and 99.7% (95% CI, 99.2–100); sensitivity of DBS-HTP-LAMP was 97.1% (95% CI, 93.1–100), and specificity 100% against PCR. At parasite densities greater or equal to 2 parasites/μL, WB and DBS HTP-LAMP showed 100% sensitivity and specificity against PCR. At densities less than 2 p/μL, WB-HTP-LAMP sensitivity was 88.9% (95% CI, 77.1–100) and specificity was 99.7% (95% CI, 99.2–100); sensitivity and specificity of DBS-HTP-LAMP was 77.8% (95% CI, 54.3–99.5) and 100% respectively. Conclusions The HTP-LAMP system is a highly sensitive diagnostic test, with the potential to allow large scale population screening in malaria elimination campaigns.