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Microbial fuel cells (MFCs) can evolve in a viable technology if environmentally sound materials are developed and became available at low cost for these devices. This is especially important not only for the designing of large wastewater treatment systems, but also for the fabrication of low-cost, single-use devices. In this work we synthesized membranes by a simple procedure involving easily-biodegradable and economic materials such as poly (vinyl alcohol) (PVA), chitosan (CS) and the composite PVA:CS. Membranes were chemical and physically characterized and compared to Nafion®. Performance was studied using the membrane as separator in a typical H-Type MFCs showing that PVA:CS membrane outperform Nafion® 4 times (power production) while being 75 times more economic. We found that performance in MFC depends over interactions among several membrane characteristics such as oxygen permeability and ion conductivity. Moreover, we design a paper-based micro-scale MFC, which was used as a toxicity assay using 16 µL samples containing formaldehyde as a model toxicant. The PVA:CS membrane presented here can offer low environmental impact and become a very interesting option for point of need single-use analytical devices, especially in low-income countries where burning is used as disposal method, and toxic fluoride fumes (from Nafion®) can be released to the environment. Fil: Gonzalez Pabon, Maria Jesus. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Figueredo, Federico. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Martínez Casillas, Diana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Corton, Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Humans introduced paper mulberry (Broussonetia papyrifera) from Taiwan into the Pacific over 5000 years ago as a fiber source to make barkcloth textiles that were, and still are, important cultural artifacts throughout the Pacific. We have used B. papyrifera, a species closely associated to humans, as a proxy to understand the human settlement of the Pacific Islands. We report the first genetic analysis of paper mulberry textiles from historical and archaeological contexts (200 to 50 years before present) and compare our results with genetic data obtained from contemporary and herbarium paper mulberry samples. Following stringent ancient DNA protocols, we extracted DNA from 13 barkcloth textiles. We confirmed that the fiber source is paper mulberry in nine of the 13 textiles studied using the nuclear ITS-1 marker and by statistical estimates. We detected high genetic diversity in historical Pacific paper mulberry barkcloth with a set of ten microsatellites, showing new alleles and specific genetic patterns. These genetic signatures allow tracing connections to plants from the Asian homeland, Near and Remote Oceania, establishing links not observed previously (using the same genetic tools) in extant plants or herbaria samples. These results show that historic barkcloth textiles are cultural materials amenable to genetic analysis to reveal human history and that these artifacts may harbor evidence of greater genetic diversity in Pacific B. papyrifera in the past.

Many complex physical systems exhibit a rich variety of discrete behavioural modes. Often, the system complexity limits the applicability of standard modelling tools. Hence, understanding the underlying physics of different behaviours and distinguishing between them is challenging. Although traditional machine learning techniques could predict and classify behaviour well, typically they do not provide any meaningful insight into the underlying physics of the system. In this paper we present a novel method for extracting physically meaningful clusters of discrete behaviour from limited experimental observations. This method obtains a set of physically plausible functions that both facilitate behavioural clustering and aid in system understanding. We demonstrate the approach on the V-shaped falling paper system, a new falling paper type system that exhibits four distinct behavioural modes depending on a few morphological parameters. Using just 49 experimental observations, the method discovered a set of candidate functions that distinguish behaviours with an error of 2.04%, while also aiding insight into the physical phenomena driving each behaviour. © 2019 Howison et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

The Panama Papers comprise one of the most recent influential leaks containing detailed information on intermediary companies (such as law firms), offshore entities and company officers, and serve as a valuable source of insight into the operations of (approximately) 214,000 shell companies incorporated in tax havens around the globe over the past half century. Entities and relations in the papers can be used to construct a network that permits, in principle, a systematic and scientific study at scale using techniques developed in the computational social science and network science communities. In this paper, we propose such a study by attempting to quantify and profile the importance of entities. In particular, our research explores whether intermediaries are significantly more influential than offshore entities, and whether different centrality measures lead to varying, or even incompatible, conclusions. Some findings yield conclusions that resemble Simpson’s paradox. We also explore the role that jurisdictions play in determining entity importance.

In our planned study, we shall empirically study the assessment of cited papers within the framework of the anchoring-and-adjustment heuristic. We are interested in the question whether citation decisions are (mainly) driven by the quality of cited references. The design of our study is oriented towards the study by Teplitskiy, Duede [10]. We shall undertake a survey of corresponding authors with an available email address in the Web of Science database. The authors are asked to assess the quality of papers that they cited in previous papers. Some authors will be assigned to three treatment groups that receive further information alongside the cited paper: citation information, information on the publishing journal (journal impact factor), or a numerical access code to enter the survey. The control group will not receive any further numerical information. In the statistical analyses, we estimate how (strongly) the quality assessments of the cited papers are adjusted by the respondents to the anchor value (citation, journal, or access code). Thus, we are interested in whether possible adjustments in the assessments can not only be produced by quality-related information (citation or journal), but also by numbers that are not related to quality, i.e. the access code. The results of the study may have important implications for quality assessments of papers by researchers and the role of numbers, citations, and journal metrics in assessment processes.

Early industrialization and the development of cheap production processes for paper have led to an exponential accumulation of paper-based documents during the last two centuries. Archives and libraries harbor vast amounts of ancient and modern documents and have to undertake extensive endeavors to protect them from abiotic and biotic deterioration. While services for mechanical preservation such as ex post de-acidification of historic documents are already commercially available, the possibilities for long-term protection of paper-based documents against fungal attack (apart from temperature and humidity control) are very limited. Novel processes for mechanical enhancement of damaged cellulosic documents use Ionic Liquids (IL) as essential process components. With some of these ILs having azole-functionalities similar to well-known fungicides such as Clotrimazole, the possibility of antifungal activities of these ILs was proposed but has not yet been experimentally confirmed.We evaluated the potency of four ILs with potential application in paper restoration for suppression of fungal growth on five relevant paper-infesting molds. The results revealed a general antifungal activity of all ILs, which increased with the size of the non-polar group. Physiological experiments and ultimate elemental analysis allowed to determine the minimal inhibitory concentration of each IL as well as the residual IL concentration in process-treated paper. These results provide valuable guidelines for IL-applications in paper restoration processes with antifungal activity as an added benefit. With azoles remaining in the paper after the process, simultaneous repair and biotic protection in treated documents could be facilitated.

Pyocyanin is a toxin produced by Pseudomonas aeruginosa. Here we describe a novel paper-based electrochemical sensor for pyocyanin detection, manufactured with a simple and inexpensive approach based on electrode printing on paper. The resulting sensors constitute an effective electrochemical method to quantify pyocyanin in bacterial cultures without the conventional time consuming pretreatment of the samples. The electrochemical properties of the paper-based sensors were evaluated by ferri/ferrocyanide as a redox mediator, and showed reliable sensing performance. The paper-based sensors readily allow for the determination of pyocyanin in bacterial cultures with high reproducibility, achieving a limit of detection of 95 nM and a sensitivity of 4.30 μA/μM in standard culture media. Compared to the similar commercial ceramic based sensors, it is a 2.3-fold enhanced performance. The simple in-house fabrication of sensors for pyocyanin quantification allows researchers to understand in vitro adaptation of P. aeruginosa infections via rapid screenings of bacterial cultures that otherwise are expensive and time-consuming.

Objective To survey the knowledge, attitude, and practices of Vietnamese pharmacists regarding the COVID-19 pandemic. Method This cross-sectional, paper-based study was conducted from June to August 2020. A validated questionnaire (Cronbach’s alpha = 0.84) was used to interview 1,023 pharmacists in nine provinces of Vietnam. Analysis of covariance was employed to identify factors associated with the knowledge of pharmacists. The best model was chosen by using the Bayesian Model Averaging method in R software version 4.0.4. Results The mean knowledge score was 12.02 ± 1.64 (range: 6–15), which indicated that 93.4% of pharmacists had good knowledge of COVID-19. There was no difference in the average score between males and females (p > 0.05). The multivariate linear regression model revealed that the knowledge was significantly associated with pharmacists’ age, education level, and residence (p < 0.001). About attitude and practices, pharmacists daily sought and updated information on the COVID-19 pandemic through mass media and the internet (social network and online newspapers). Nearly 48% of them conceded that they communicated with customers when at least one person did not wear a face mask at the time of the COVID-19 outbreak. At medicine outlets, many measures were applied to protect pharmacists and customers, such as equipping pharmacists with face masks and hand sanitizers (95.0%), using glass shields (83.0%), and maintaining at least one-meter distance between two people (85.2%). Conclusion The pharmacists’ knowledge of COVID-19 transmission, symptoms, and prevention was good. Many useful measures against the spread of this perilous virus were applied in medicine outlets. However, pharmacists should restrict forgetting to wear face masks in communication with medicine purchasers. The government and health agencies should have practical remedies to reduce the significant differences in the COVID-19 knowledge of pharmacists among provinces and education-level groups.

We propose to use Twitter data as social-spatial sensors. This study deals with the question whether research papers on certain diseases are perceived by people in regions (worldwide) that are especially concerned by these diseases. Since (some) Twitter data contain location information, it is possible to spatially map the activity of Twitter users referring to certain papers (e.g., dealing with tuberculosis). The resulting maps reveal whether heavy activity on Twitter is correlated with large numbers of people having certain diseases. In this study, we focus on tuberculosis, human immunodeficiency virus (HIV), and malaria, since the World Health Organization ranks these diseases as the top three causes of death worldwide by a single infectious agent. The results of the social-spatial Twitter maps (and additionally performed regression models) reveal the usefulness of the proposed sensor approach. One receives an impression of how research papers on the diseases have been perceived by people in regions that are especially concerned by these diseases. Our study demonstrates a promising approach for using Twitter data for research evaluation purposes beyond simple counting of tweets.

Background A paper-based, multiplexed, microfluidic assay has been developed to visually measure alanine aminotransferase (ALT) in a fingerstick sample, generating rapid, semi-quantitative results. Prior studies indicated a need for improved accuracy; the device was subsequently optimized using an FDA-approved automated platform (Abaxis Piccolo Xpress) as a comparator. Here, we evaluated the performance of the optimized paper test for measurement of ALT in fingerstick blood and serum, as compared to Abaxis and Roche/Hitachi platforms. To evaluate feasibility of remote results interpretation, we also compared reading cell phone camera images of completed tests to reading the device in real time. Methods 96 ambulatory patients with varied baseline ALT concentration underwent fingerstick testing using the paper device; cell phone images of completed devices were taken and texted to a blinded off-site reader. Venipuncture serum was obtained from 93/96 participants for routine clinical testing (Roche/Hitachi); subsequently, 88/93 serum samples were captured and applied to paper and Abaxis platforms. Paper test and reference standard results were compared by Bland-Altman analysis. Findings For serum, there was excellent agreement between paper test and Abaxis results, with negligible bias (+4.5 U/L). Abaxis results were systematically 8.6% lower than Roche/Hitachi results. ALT values in fingerstick samples tested on paper were systematically lower than values in paired serum tested on paper (bias -23.6 U/L) or Abaxis (bias -18.4 U/L); a correction factor was developed for the paper device to match fingerstick blood to serum. Visual reads of cell phone images closely matched reads made in real time (bias +5.5 U/L). Conclusions The paper ALT test is highly accurate for serum testing, matching the reference method against which it was optimized better than the reference methods matched each other. A systematic difference exists between ALT values in fingerstick and paired serum samples, and can be addressed by application of a correction factor to fingerstick values. Remote reading of this device is feasible.

Raman spectroscopy has proven to be an indispensable technique for the identification of various types of analytes due to the fingerprint vibration spectrum obtained. Paper microfluidics has also emerged as a low cost, easy to fabricate and portable approach for point of care testing. However, due to inherent background fluorescence, combining Raman spectroscopy with paper microfluidics is to date an unmet challenge in the absence of using surface enhanced mechanisms. We describe the first use of wavelength modulated Raman spectroscopy (WMRS) for analysis on a paper microfluidics platform. This study demonstrates the ability to suppress the background fluorescence of the paper using WMRS and the subsequent implementation of this technique for pharmaceutical analysis. The results of this study demonstrate that it is possible to discriminate between both paracetamol and ibuprofen, whilst, also being able to detect the presence of each analyte quantitatively at nanomolar concentrations. Publisher PDF

Type 1 diabetes (T1D) is increasing in incidence and predictable with measurement of serum islet autoantibodies (iAb) years prior to clinical disease onset. Identifying iAb positive individuals reduces diabetic ketoacidosis and identifies individuals for T1D prevention trials. However, large scale screening for iAb remains challenging as assays have varying sensitivities and specificities, insulin autoantibodies remain difficult to measure and venipuncture is generally required to obtain serum. We developed an approach to reliably measure all four major iAb, including insulin autoantibodies, from dried blood spots (DBS) on filter-paper. By spiking iAb positive serum into iAb negative whole blood in a dose titration, we optimized the conditions for autoantibody elution from filter paper as measured by fluid phase radioimmunoassays. After assessing stability of measuring iAb from DBS over time, we then screened iAb from DBS and the corresponding serum in new-onset T1D (n = 52), and controls (n = 72) which included first-degree relatives of T1D patients. iAb measured from eluted DBS in new-onset T1D strongly correlated with serum measurements (R2 = 0.96 for mIAA, GADA = 0.94, IA-2A = 0.85, ZnT8A = 0.82, p

AimsTo evaluate the relative efficiencies of five Internet-based digital and three paper-based scientific surveys and to estimate the costs for different-sized cohorts.MethodsInvitations to participate in a survey were distributed via e-mail to employees of two university hospitals (E1 and E2) and to members of a medical association (E3), as a link placed in a special text on the municipal homepage regularly read by the administrative employees of two cities (H1 and H2), and paper-based to workers at an automobile enterprise (P1) and college (P2) and senior (P3) students. The main parameters analyzed included the numbers of invited and actual participants, and the time and cost to complete the survey. Statistical analysis was descriptive, except for the Kruskal-Wallis-H-test, which was used to compare the three recruitment methods. Cost efficiencies were compared and extrapolated to different-sized cohorts.ResultsThe ratios of completely answered questionnaires to distributed questionnaires were between 81.5% (E1) and 97.4% (P2). Between 6.4% (P1) and 57.0% (P2) of the invited participants completely answered the questionnaires. The costs per completely answered questionnaire were $0.57-$1.41 (E1-3), $1.70 and $0.80 for H1 and H2, respectively, and $3.36-$4.21 (P1-3). Based on our results, electronic surveys with 10, 20, 30, or 42 questions would be estimated to be most cost (and time) efficient if more than 101.6-225.9 (128.2-391.7), 139.8-229.2 (93.8-193.6), 165.8-230.6 (68.7-115.7), or 188.2-231.5 (44.4-72.7) participants were required, respectively.ConclusionsThe study efficiency depended on the technical modalities of the survey methods and engagement of the participants. Depending on our study design, our results suggest that in similar projects that will certainly have more than two to three hundred required participants, the most efficient way of conducting a questionnaire-based survey is likely via the Internet with a digital questionnaire, specifically via a centralized e-mail.

There is demand from science funders, industry, and the public that science should become more risk-taking, more out-of-the-box, and more interdisciplinary. Is it possible to tell how interdisciplinary and out-of-the-box scientific papers are, or which papers are mainstream? Here we use the bibliographic coupling network, derived from all physics papers that were published in the Physical Review journals in the past century, to try to identify them as mainstream, out-of-the-box, or interdisciplinary. We show that the network clusters into scientific fields. The position of individual papers with respect to these clusters allows us to estimate their degree of mainstreamness or interdisciplinary. We show that over the past decades the fraction of mainstream papers increases, the fraction of out-of-the-box decreases, and the fraction of interdisciplinary papers remains constant. Studying the rewards of papers, we find that in terms of absolute citations, both, mainstream and interdisciplinary papers are rewarded. In the long run, mainstream papers perform less than interdisciplinary ones in terms of citation rates. We conclude that to avoid a trend towards mainstreamness a new incentive scheme is necessary., 12 pages, 9 figures

Background Adiponectin is an adipocyte-derived hormone that acts as a marker of insulin sensitivity. Bloodspot sampling by fingerstick onto filter paper may increase the feasibility of large-scale studies of the determinants of insulin sensitivity. We first describe the validation of an enzyme-linked immunoassay (ELISA) for quantifying adiponectin from dried blood spots and then demonstrate its application in a large trial (PROBIT). Methods We quantified adiponectin from 3-mm diameter discs (≈3 µL of blood) punched from dried blood spots obtained from: i) whole blood standards (validation); and ii) PROBIT trial samples (application) in which paediatricians collected blood spots from 13,879 children aged 11.5 years from 31 sites across Belarus. We examined the distribution of bloodspot adiponectin by demographic and anthropometric factors, fasting insulin and glucose. Results In the validation study, mean intra-assay coefficients of variation (n = 162) were 15%, 13% and 10% for ‘low’ (6.78 µg/ml), ‘medium’ (18.18 µg/ml) and 'high’ (33.13 µg/ml) internal quality control (IQC) samples, respectively; the respective inter-assay values (n = 40) were 23%, 21% and 14%. The correlation coefficient between 50 paired whole bloodspot versus plasma samples, collected simultaneously, was 0.87 (95% CI: 0.78 to 0.93). Recovery of known quantities of adiponectin (between 4.5 to 36 µg/ml) was 100.3–133%. Bloodspot adiponectin was stable for at least 30 months at −80°C. In PROBIT, we successfully quantified fasting adiponectin from dried blood spots in 13,329 of 13,879 (96%) children. Mean adiponectin (standard deviation) concentrations were 17.34 µg/ml (7.54) in boys and 18.41 µg/ml (7.92) in girls and were inversely associated with body mass index, fat mass, triceps and subscapular skin-fold thickness, waist circumference, height and fasting glucose. Conclusions Bloodspot ELISA is suitable for measuring adiponectin in very small volumes of blood collected on filter paper and can be applied to large-scale studies.

E-readers are fast rivaling print as a dominant method for reading. Because they offer accessibility options that are impossible in print, they are potentially beneficial for those with impairments, such as dyslexia. Yet, little is known about how the use of these devices influences reading in those who struggle. Here, we observe reading comprehension and speed in 103 high school students with dyslexia. Reading on paper was compared with reading on a small handheld e-reader device, formatted to display few words per line. We found that use of the device significantly improved speed and comprehension, when compared with traditional presentations on paper for specific subsets of these individuals: Those who struggled most with phoneme decoding or efficient sight word reading read more rapidly using the device, and those with limited VA Spans gained in comprehension. Prior eye tracking studies demonstrated that short lines facilitate reading in dyslexia, suggesting that it is the use of short lines (and not the device per se) that leads to the observed benefits. We propose that these findings may be understood as a consequence of visual attention deficits, in some with dyslexia, that make it difficult to allocate attention to uncrowded text near fixation, as the gaze advances during reading. Short lines ameliorate this by guiding attention to the uncrowded span.

BackgroundThe objectives of the study were to develop web-based Spanish and Catalan versions of the KIDSCREEN, and to compare scores and psychometric properties with the paper version.MethodsInternet and paper Spanish and Catalan versions of the KIDSCREEN-52 were included in a cross-sectional study in school-age children. Web-based and paper Spanish or Catalan versions of the KIDSCREEN-52 were administered to students aged 8 to 18 years from primary and secondary schools in Palafolls (Barcelona, Spain, n = 923). All students completed both web-based and paper versions during school time with an interval of at least 2 hours between administrations. The order of administration was randomized. The KIDSCREEN-52, the Strengths and Difficulties Questionnaire (SDQ), and sociodemographic variables were collected. Missing values, floor and ceiling effects, and internal consistency were compared between both versions, as well as mean score differences, level of agreement, and known groups and construct validity.ResultsParticipation rate was 77% (n = 715). Web-based and paper versions showed low percentage of missing values and similar high ceiling effect (range 0 to 44%). Mean score differences showed an effect size (ES) lower than 0.2 in all dimensions. Internal consistency ranged from 0.7 to 0.88, and degree of agreement was excellent (Intraclass correlation coefficient [ICC] range 0.75 to 0.87). Expected differences were seen by sex, age, socioeconomic status and mental health status.ConclusionsThe web-based KIDSCREEN-52 showed similar scale score and reliability and validity than the paper version. It will incorporate the child population in the assessment of quality of life providing a more attractive format.

Moderate-to-severe psoriasis is associated with significant comorbidity, an impaired quality of life, and increased medical costs, including those associated with treatments. Systematic reviews (SRs) and meta-analyses (MAs) of randomized clinical trials are considered two of the best approaches to the summarization of high-quality evidence. However, methodological bias can reduce the validity of conclusions from these types of studies and subsequently impair the quality of decision making. As co-authorship is among the most well-documented forms of research collaboration, the present study aimed to explore whether authors' collaboration methods might influence the methodological quality of SRs and MAs of psoriasis. Methodological quality was assessed by two raters who extracted information from full articles. After calculating total and per-item Assessment of Multiple Systematic Reviews (AMSTAR) scores, reviews were classified as low (0-4), medium (5-8), or high (9-11) quality. Article metadata and journal-related bibliometric indices were also obtained. A total of 741 authors from 520 different institutions and 32 countries published 220 reviews that were classified as high (17.2%), moderate (55%), or low (27.7%) methodological quality. The high methodological quality subnetwork was larger but had a lower connection density than the low and moderate methodological quality subnetworks; specifically, the former contained relatively fewer nodes (authors and reviews), reviews by authors, and collaborators per author. Furthermore, the high methodological quality subnetwork was highly compartmentalized, with several modules representing few poorly interconnected communities. In conclusion, structural differences in author-paper affiliation network may influence the methodological quality of SRs and MAs on psoriasis. As the author-paper affiliation network structure affects study quality in this research field, authors who maintain an appropriate balance between scientific quality and productivity are more likely to develop higher quality reviews.

Paper mulberry (Broussonetia papyrifera (L.) L'Hér. ex Vent) is a dioecious tree native to East Asia and mainland Southeast-Asia, introduced prehistorically to Polynesia as a source of bark fiber by Austronesian-speaking voyagers. In Oceania, trees are coppiced and harvested for production of bark-cloth, so flowering is generally unknown. A survey of botanical records of paper mulberry revealed a distributional disjunction: the tree is apparently absent in Borneo and the Philippines. A subsequent study of chloroplast haplotypes linked paper mulberry of Remote Oceania directly to a population in southern Taiwan, distinct from known populations in mainland Southeast-Asia.We describe the optimization and use of a DNA marker designed to identify sex in paper mulberry. We used this marker to determine the sex distribution in selected localities across Asia, Near and Remote Oceania. We also characterized all samples using the ribosomal internal transcribed spacer sequence (ITS) in order to relate results to a previous survey of ITS diversity.In Near and Remote Oceania, contemporary paper mulberry plants are all female with the exception of Hawaii, where plants of both sexes are found. In its natural range in Asia, male and female plants are found, as expected. Male plants in Hawaii display an East Asian ITS genotype, consistent with modern introduction, while females in Remote Oceania share a distinctive variant.Most paper mulberry plants now present in the Pacific appear to be descended from female clones introduced prehistorically. In Hawaii, the presence of male and female plants is thought to reflect a dual origin, one a prehistoric female introduction and the other a modern male introduction by Japanese/Chinese immigrants. If only female clones were dispersed from a source-region in Taiwan, this may explain the absence of botanical records and breeding populations in the Philippines and Borneo, and Remote Oceania.

The paper mulberry is one of the multifunctional tree species in agroforestry systems and is also commonly utilized in traditional medicine in China and other Asian countries. However, little is known about its molecular genetics, which hinders research on and exploitation of this valuable resource. To discern the correlation between gene expression and the essential properties of the paper mulberry, we performed a transcriptomics analysis, assembling a total of 37,725 unigenes from 54,638,676 reads generated by RNA-seq. Among these, 22,692 unigenes showed greater than 60% similarity with genes from other species. The lengths of 13,566 annotated unigenes were longer than 1,000 bp. Functional clustering analysis with COG (Cluster of Orthologous Groups) revealed that 17,184 unigenes are primarily involved in transcription, translation, signal transduction, carbohydrate metabolism, secondary metabolism, and energy metabolism. GO (Gene Ontology) annotation suggests enrichment of genes encoding antioxidant activity, transporter activity, biosynthesis, metabolism and stress response, with a total of 30,659 unigenes falling in these categories. KEGG (Kyoto Encyclopedia of Genes and Genomes) metabolic pathway analysis showed that 7,199 unigenes are associated with 119 metabolic pathways. In addition to the basic metabolism, these genes are enriched for plant pathogen interaction, flavonoid metabolism and other secondary metabolic processes. Furthermore, differences in the transcriptomes of leaf, stem and root tissues were analyzed and 7,233 specifically expressed unigenes were identified. This global expression analysis provided novel insights about the molecular mechanisms of the biosynthesis of flavonoid, lignin and cellulose, as well as on the response to biotic and abiotic stresses including the remediation of contaminated soil by the paper mulberry.

BackgroundAlthough researchers have worked in collaboration since the origins of modern science and the publication of the first scientific journals in the eighteenth century, this phenomenon has acquired exceptional importance in the last several decades. Since the mid-twentieth century, new knowledge has been generated from within an ever-growing network of investigators, working cooperatively in research groups across countries and institutions. Cooperation is a crucial determinant of academic success.ObjectiveThe aim of the present paper is to analyze the evolution of scientific collaboration at the micro level, with regard to the scientific production generated on psoriasis research.MethodsA bibliographic search in the Medline database containing the MeSH terms "psoriasis" or "psoriatic arthritis" was carried out. The search results were limited to articles, reviews and letters. After identifying the co-authorships of documents on psoriasis indexed in the Medline database (1942-2013), various bibliometric indicators were obtained, including the average number of authors per document and degree of multi-authorship over time. In addition, we performed a network analysis to study the evolution of certain features of the co-authorship network as a whole: average degree, size of the largest component, clustering coefficient, density and average distance. We also analyzed the evolution of the giant component to characterize the changing research patterns in the field, and we calculated social network indicators for the nodes, namely betweenness and closeness.ResultsThe main active research clusters in the area were identified, along with their authors of reference. Our analysis of 28,670 documents sheds light on different aspects related to the evolution of scientific collaboration in the field, including the progressive increase in the mean number of co-authors (which stood at 5.17 in the 2004-2013 decade), and the rise in multi-authored papers signed by many different authors (in the same decade, 25.77% of the documents had between 6 and 9 co-authors, and 10.28% had 10 or more). With regard to the network indicators, the average degree gradually increased up to 10.97 in the study period. The percentage of authors pertaining to the largest component also rose to 73.02% of the authors. The clustering coefficient, on the other hand, remained stable throughout the entire 70-year period, with values hovering around 0.9. Finally, the average distance peaked in the decades 1974-1983 (8.29) and 1984-2003 (8.12) then fell over the next two decades, down to 5.25 in 2004-2013. The construction of the co-authorship network (threshold of collaboration ≥ 10 co-authored works) revealed a giant component of 161 researchers, containing 6 highly cohesive sub-components.ConclusionsOur study reveals the existence of a growing research community in which collaboration is increasingly important. We can highlight an essential feature associated with scientific collaboration: multi-authored papers, with growing numbers of collaborators contributing to them, are becoming more and more common, therefore the formation of research groups of increasing depth (specialization) and breadth (multidisciplinarity) is now a cornerstone of research success.

Background In the context of a randomized controlled trial (RCT) on HIV testing in the emergency department (ED) setting, we evaluated preferences for survey modality and data quality arising from each modality. Methods Enrolled participants were offered the choice of answering a survey via audio computer assisted self-interview (ACASI) or pen and paper self-administered questionnaire (SAQ). We evaluated factors influencing choice of survey modality. We defined unusable data for a particular survey domain as answering fewer than 75% of the questions in the domain. We then compared ACASI and SAQ with respect to unusable data for domains that address sensitive topics. Results Of 758 enrolled ED patients, 218 (29%) chose ACASI, 343 chose SAQ (45%) and 197 (26%) opted not to complete either. Results of the log-binomial regression indicated that older (RR = 1.08 per decade) and less educated participants (RR = 1.25) were more likely to choose SAQ over ACASI. ACASI yielded substantially less unusable data than SAQ. Conclusions In the ED setting there may be a tradeoff between increased participation with SAQ versus better data quality with ACASI. Future studies of novel approaches to maximize the use of ACASI in the ED setting are needed.

Scholars traditionally receive career credit for a paper based on where in the author list they appear, but position in an author list often carries little information about what the contribution of each researcher was. “Contributorship” refers to a movement to formally document the nature of each researcher’s contribution to a project. We discuss the emerging CRediT standard for documenting contributions and describe a web-based app and R package called tenzing that is designed to facilitate its use. tenzing can make it easier for researchers on a project to plan and record their planned contributions and to document those contributions in a journal article.

In newborn screening, samples suspected for congenital adrenal hyperplasia (CAH), a potentially lethal inborn error of steroid biosynthesis, need to be confirmed using liquid chromatography–tandem mass spectrometry. Daily quality controls (QCs) for the 2nd-tier CAH assay are not commercially available and are therefore generally prepared within the laboratory. For the first time, we aimed to compare five different QC preparation approaches used in routine diagnostics for CAH on the concentrations of cortisol, 21-deoxycortisol, 11-deoxycortisol, 4-androstenedione and 17-hydroxyprogesterone in dried blood spots. The techniques from Prep1 to Prep5 were tested at two analyte concentrations by spiking aliquots of a steroid-depleted blood, derived from washed erythrocyte suspension and steroid-depleted serum. The preparation processes differed in the sequence of the preparation steps and whether freeze-thaw cycles were used to facilitate blood homogeneity. The five types of dried blood spot QCs were assayed and quantitated in duplicate on five different days using a single calibration row per day. Inter-assay variations less than 15% and concentrations within ±15% of the nominal values were considered acceptable. Results obtained by means of the four dried blood spot QC preparation techniques (Prep1, Prep2, Prep4 and Prep5) were statistically similar and remained within the ±15% ranges in terms of both reproducibility and nominal values. However, concentration results for Prep3 (spiking prior to three freeze-thaw cycles) were significantly lower than the nominal values in this setting, with differences exceeding the ±15% range in many cases despite acceptable inter-assay variations. These findings have implications for the in-house preparation of QC samples in laboratory developed tests for CAH, including 2nd-tier assays in newborn screening.

Among the challenges of sustainable management of meat production, the key issue is to improve the energy efficiency of production processes, which will consequently affect the reduction of greenhouse gas emissions. Such effects are achieved by combining various chilling systems with electrical stimulation that determines the quality of meat at the slaughter stage. The novelties of the research undertaken included determining the impact of various variants of meat production (chilling method: slow, fast, accelerated + HVES/NES) on changes in the basic (industrial) quality indicators (pH and temperature) of beef produced from Polish Holstein-Friesian breed cattle, and then indicating the optimal variant for energy-efficient (sustainable) beef production. The HVES and the fast chilling method yielded positive economic (meat weight loss), technological (high quality, hot-boning), energetic (lower electricity consumption), and organizational effects (reduced chilling and storage surfaces and expenditures for staff wages) compared to the slow and accelerated methods. Reaching the desired final temperature with an increased amount of chilled meat enables obtaining a few-fold decrease in the specific energy consumption and a higher energy efficiency of the process. This allows recommending the above actions to be undertaken by entrepreneurs in the pursuit of sustainable meat production.

The use of personal protective equipment (PPE) has been considered the most effective way to avoid the contamination of healthcare workers by different microorganisms, including SARS-CoV-2. A spray disinfection technology (chamber) was developed, and its efficacy in instant decontamination of previously contaminated surfaces was evaluated in two exposure times. Seven test microorganisms were prepared and inoculated on the surface of seven types of PPE (respirator mask, face shield, shoe, glove, cap, safety glasses and lab coat). The tests were performed on previously contaminated PPE using a manikin with a motion device for exposure to the chamber with biocidal agent (sodium hypochlorite) for 10 and 30s. In 96.93% of the experimental conditions analyzed, the percentage reduction was >99% (the number of viable cells found on the surface ranged from 4.3x106 to E. faecalis collected from the glove showed the lowest percentages reduction, with 86.000 and 86.500% for exposure times of 10 and 30 s, respectively. The log10 reduction values varied between 0.85 log10 (E. faecalis at 30 s in glove surface) and 9.69 log10 (E. coli at 10 and 30 s in lab coat surface). In general, E. coli, S. aureus, C. freundii, P. mirabilis, C. albicans and C. parapsilosis showed susceptibility to the biocidal agent under the tested conditions, with >99% reduction after 10 and 30s, while E. faecalis and P. aeruginosa showed a lower susceptibility. The 30s exposure time was more effective for the inactivation of the tested microorganisms. The results show that the spray disinfection technology has the potential for instant decontamination of PPE, which can contribute to an additional barrier for infection control of healthcare workers in the hospital environment.

Faecal Sludge Management (FSM) has become a prominent environmental concern in the today’s world. Dewatering of sludge and the treatment of wastewater (WW) are the prime spiny issue because of the deleterious essence of faecal sludge (FS) and WW in the environment. The main focus of this study was on FSM by ‘Deebag’ and ‘Jute Bag’ through dewatering and filtering. Deebag is a dewatering as well as filtering media which is made with geotextile and polypropylene. Contrariwise, three types of jute bags were made of jute fiber for using as the same purposes of Deebag. A polyacrylamide polymer was used in this study and both filtering and dewatering were done in two ways—with and without the presence of polymer. Biochemical Oxygen Demand at 5 days (BOD5), Chemical Oxygen Demand (COD), pH, Electrical Conductivity (EC), Chloride (Cl-), Phosphate (PO43-), Nitrate (NO3-), Total Suspended Solid (TSS) and Total Dissolved Solid (TDS) parameters of raw and filtering samples were analyzed to assess the performance of WW treatment by Deebag and jute bags. Only using polymer was observed as one kind of treatment of WW. Deebag has been found to show the maximum dewatering capacity as well as treatment efficiency comparing with the jute bags. However, among three types of jute bags, double jute layered bag has shown the best performance. Maximum dewatering for Deebag and jute bags were found 88% and 83% respectively while using the polymer.

Since the innovation of our new half-piece elastometric respirator, this type of filtering facepiece respirator (FFR) has been used widely in Thailand. Decontamination methods including ultraviolet C (UVC) germicidal irradiation and 70% alcohol have been implemented to decontaminate these respirators. We then examined the inactivation potential of different decontamination processes on porcine epidemic diarrhea virus (PEDV) and numerous bacterial strains, most of which were skin-derived. To enable rigorous integrity of the masks after repeated decontamination processes, fit tests by the Bitrex test, tensile strength and elongation at break were also evaluated. Our results showed that UVC irradiation at a dose of 3 J/cm2 can eradicate bacteria after 60 min and viruses after 10 min. No fungi were found on the mask surface before decontamination. The good fit test results, tensile strength and elongation at break were still maintained after multiple cycles of decontamination. No evidence of physical degradation was found by gross visual inspection. Alcohol (70%) is also an easy and effective way to eradicate microorganisms on respirators. As the current pandemic is expected to continue for months to years, the need to supply adequate reserves of personnel protective equipment (PPE) and develop effective PPE reprocessing methods is crucial. Our studies demonstrated that the novel silicone mask can be safely reprocessed and decontaminated for many cycles by UVC irradiation, which will help ameliorate the shortage of important protective devices in the COVID-19 pandemic era.

Bacterial expansin-like proteins have synergistically increased cellulose hydrolysis by cellulolytic enzymes during the initial stages of biofuel production, but they have not been tested on livestock feeds. The objectives of this study were to: isolate and express an expansin-like protein (BsEXLX1), to verify its disruptive activity (expansion) on cotton fibers by immunodetection (Experiment 1), and to determine the effect of dose, pH and temperature for BsEXLX1 and cellulase to synergistically hydrolyze filter paper (FP) and carboxymethyl cellulose (CMC) under laboratory (Experiment 2) and simulated ruminal (Experiment 3) conditions. In addition, we determined the ability of BsEXLX1 to synergistically increase hydrolysis of corn and bermudagrass silages by an exogenous fibrolytic enzyme (EFE) (Experiment 4) and how different doses of BsEXLX1 and EFE affect the gas production (GP), in vitro digestibility and fermentation of a diet for dairy cows (Experiment 5). In Experiment 1, immunofluorescence-based examination of cotton microfiber treated without or with recombinant expansin-like protein expressed from Bacillus subtilis (BsEXLX1) increased the surface area by > 100% compared to the untreated control. In Experiment 2, adding BsEXLX1 (100 μg/g FP) to cellulase (0.0148 FPU) increased release of reducing sugars compared to cellulase alone by more than 40% (P < 0.01) at optimal pH (4.0) and temperature (50°C) after 24 h. In Experiment 3 and 4, adding BsEXLX1 to cellulase or EFE, synergistically increased release of reducing sugars from FP, corn and bermudagrass silages under simulated ruminal conditions (pH 6.0, 39°C). In Experiment 5, increasing the concentration of BsEXLX1 linearly increased (P < 0.01) GP from fermentation of a diet for dairy cows by up to 17.8%. Synergistic effects between BsEXLX1 and EFE increased in vitro NDF digestibility of the diet by 23.3% compared to the control. In vitro digestibility of hemicellulose and butyrate concentration were linearly increased by BsEXLX1 compared to the control. This study demonstrated that BsEXLX1 can improve the efficacy of cellulase and EFE at hydrolyzing pure substrates and dairy cow feeds, respectively.

Fundamental ecological principles of ecosystem-level respiration are extensively applied in greenhouse gas and elemental cycle studies. A laboratory system termed CEMS (Carbon Dioxide Evolution Measurement System), developed to explore microbial biofilm growth and metabolic responses, was evaluated as an early-warning system for microbial disturbances in industrial settings: in (a) potable water system contamination, and (b) bioreactor inhibition. Respiration was detected as CO2 production, rather than O2 consumption, including aerobic and anaerobic metabolism. Design, thresholds, and benefits of the remote CO2 monitoring technology were described. Headspace CO2 correlated with contamination levels, as well as chemical (R2 > 0.83–0.96) and microbiological water quality indicators (R2 > 0.78–0.88). Detection thresholds were limiting factors in monitoring drinking water to national and international standards (0 CFU/100 mL fecal coliforms) in both open- (>1500 CFU/mL) and closed-loop CO2 measuring regimes (>100 CFU/100 mL). However, closed-loop detection thresholds allow for the detection of significant contamination events, and monitoring less stringent systems such as irrigation water (2 fluctuations from chemical CO2 dynamics, to optimize this real-time, sustainable, low-waste technology, facilitating timeous responses to biological disturbances in bioreactors.

Due to the inefficient reproduction of microorganisms in oxygen-deprived environments of the reservoir, the applications of microbial enhanced oil recovery (MEOR) are restricted. To overcome this problem, a new type of air-assisted MEOR process was investigated. Three compounding oil degradation strains were screened using biochemical experiments. Their performances in bacterial suspensions with different amounts of dissolved oxygen were evaluated. Water flooding, microbial flooding and air-assisted microbial flooding core flow experiments were carried out. Carbon distribution curve of biodegraded oil with different oxygen concentration was determined by chromatographic analysis. The long-chain alkanes are degraded by microorganisms. A simulation model was established to take into account the change in oxygen concentration in the reservoir. The results showed that the optimal dissolved oxygen concentration for microbial growth was 4.5~5.5mg/L. The main oxygen consumption in the reservoir happened in the stationary and declining phases of the microbial growth systems. In order to reduce the oxygen concentration to a safe level, the minimum radius of oxygen consumption was found to be about 145m. These results demonstrate that the air-assisted MEOR process can overcome the shortcomings of traditional microbial flooding techniques. The findings of this study can help for better understanding of microbial enhanced oil recovery and improving the efficiency of microbial oil displacement.

Starch based gluten-free bread (formulations containing mixture of corn and potato starch with hydrocolloids) are deficient in nutrients and do not contain health promoting compounds. Therefore they could be supplemented with raw materials rich in such components, especially antioxidants. Among them pseudo-cereals, seeds, fruits and vegetables are often applied to this purpose. Potato pulp produced by processing red fleshed (Magenta Love) and purple fleshed (Violetta) varieties could become a new innovative substrate for gluten-free bread enrichment, because of high levels of endogenous polyphenols, namely flavonoids, flavonols, phenolic acids and especially anthocyanins with high antioxidant potential, as well as dietary fiber. Study material consisted of gluten-free bread enriched in the pulp. Dietary fiber, acrylamide content and antioxidant and antiradical potential of the bread were determined. Sensory evaluation included crumb elasticity, porosity and other characteristics, taste and smell. Among all analyzed gluten-free breads, the sample containing 7.5% share of freeze-dried red potato pulp Magenta Love was characterized by high content of phenolic compounds and dietary fiber, pronounced antioxidant activity, low levels of potentially dangerous acrylamide and good physical and sensory characteristics. Therefore such an addition (7.5% Magenta Love) could be recommended for industrial production of gluten-free bread.

Waste biomass from the palm oil industry is currently burned as a means of disposal and solutions are required to reduce the environmental impact. Whilst some waste biomass can be recycled to provide green energy such as biogas, this investigation aimed to optimise experimental conditions for recycling palm waste into substrate for insects, farmed as a sustainable high-protein animal feed. NMR spectroscopy and LC-HRMS were used to analyse the composition of palm empty fruit bunches (EFB) under experimental conditions optimised to produce nutritious substrate rather than biogas. Statistical pattern recognition techniques were used to investigate differences in composition for various combinations of pre-processing and anaerobic digestion (AD) methods. Pre-processing methods included steaming, pressure cooking, composting, microwaving, and breaking down the EFB using ionic liquids. AD conditions which were modified in combination with pre-processing methods were ratios of EFB:digestate and pH. Results show that the selection of pre-processing method affects the breakdown of the palm waste and subsequently the substrate composition and biogas production. Although large-scale insect feeding trials will be required to determine nutritional content, we found that conditions can be optimised to recycle palm waste for the production of substrate for insect rearing. Pre-processing EFB using ionic liquid before AD at pH6 with a 2:1 digestate:EFB ratio were found to be the best combination of experimental conditions.

In recent years, spread of invasive alien plant species (IAPS) has been a major concern in Nepal. One such IAPS is Ageratum houstonianum, an Asteraceae, that is a prolific seed producer and difficult-to-control in farmland and various ecological regions causing crop yield and biodiversity losses. However, very little information is available on the germination biology and ecology of this species. Therefore, experiments were conducted to assess the effect of water stress, pH level, and light requirement on seed germination, and the effect of seed burial depth on seedling emergence. Water stress was simulated by polyethylene glycol solutions ranging from 0-5.56 MPa and pH solutions ranging from 4 to 9 were prepared using hydrochloric acid and sodium hydroxide. Germination tests were conducted in petri dishes lined with filter paper and placed in a controlled environment chamber set at 20° C. Light requirement comparisons were made by having petri dishes wrapped with aluminum foil or left unwrapped. Seedling emergence was evaluated by placing seeds at depths ranging from 0 to 20 mm in the soil. Results indicated that this species was moderately drought-tolerant because germination ceased beyond 0.51 MPa. Greater germination occurred at neutral to acidic than at alkaline pH levels. The seeds were positively photoblastic because no germination occurred under dark condition. No seedlings emerged from seeds placed more than 2 mm deep in the soil, indicating that this is a primarily surface germinating species. These findings will help predict future invasions and in development of management strategies for this IAPS.

The safety of highly aggregated tourist crowds is a challenging and important issue. This paper not only provided a comprehensive analysis of the accidents of highly aggregated tourist crowds but also determined the occurrence mechanism and coping paths. Based on the analysis of multiple cases, we found that the variable status of highly aggregated tourist crowds was the result of the interaction of three main elements: multisource pressure, state mutations and management responses. A series of factors interact and result in accidents, and the lack of a management response or a low-quality management response is the root cause of such accidents. A high-quality management response is a basic safety precaution for highly aggregated tourist crowds. Therefore, forming a virtuous circle of multisource pressure, state mutations and management responses is an effective path for coping with accidents.

The response to the COVID-19 epidemic is generating severe shortages of personal protective equipment around the world. In particular, the supply of N95 respirator masks has become severely depleted, with supplies having to be rationed and health care workers having to use masks for prolonged periods in many countries. We sought to test the ability of 7 different decontamination methods: autoclave treatment, ethylene oxide gassing (ETO), low temperature hydrogen peroxide gas plasma (LT-HPGP) treatment, vaporous hydrogen peroxide (VHP) exposure, peracetic acid dry fogging (PAF), ultraviolet C irradiation (UVCI) and moist heat (MH) treatment to decontaminate a variety of different N95 masks following experimental contamination with SARS-CoV-2 or vesicular stomatitis virus as a surrogate. In addition, we sought to determine whether masks would tolerate repeated cycles of decontamination while maintaining structural and functional integrity. All methods except for UVCI were effective in total elimination of viable virus from treated masks. We found that all respirator masks tolerated at least one cycle of all treatment modalities without structural or functional deterioration as assessed by fit testing; filtration efficiency testing results were mostly similar except that a single cycle of LT-HPGP was associated with failures in 3 of 6 masks assessed. VHP, PAF, UVCI, and MH were associated with preserved mask integrity to a minimum of 10 cycles by both fit and filtration testing. A similar result was shown with ethylene oxide gassing to the maximum 3 cycles tested. Pleated, layered non-woven fabric N95 masks retained integrity in fit testing for at least 10 cycles of autoclaving but the molded N95 masks failed after 1 cycle; filtration testing however was intact to 5 cycles for all masks. The successful application of autoclaving for layered, pleated masks may be of particular use to institutions globally due to the virtually universal accessibility of autoclaves in health care settings. Given the ability to modify widely available heating cabinets on hospital wards in well-resourced settings, the application of moist heat may allow local processing of N95 masks.

Honey bees (genusApis)are well known for the impressive suite of nest defenses they have evolved to protect their abundant stockpiles of food and the large colonies they sustain. In Asia, honey bees have evolved under tremendous predatory pressure from social wasps in the genusVespa, the most formidable of which are the giant hornets that attack colonies in groups, kill adult defenders, and prey on brood. We document for the first time an extraordinary collective defense used byApis ceranaagainst the giant hornetVespa soror. In response to attack byV.soror,A.ceranaworkers foraged for and applied spots of animal feces around their nest entrances. Fecal spotting increased after colonies were exposed either to naturally occurring attacks or to chemicals that scout hornets use to target colonies for mass attack. Spotting continued for days after attacks ceased and occurred in response toV.soror, which frequently landed at and chewed on entrances to breach nests, but notVespa velutina, a smaller hornet that rarely landed at entrances. Moderate to heavy fecal spotting suppressed attempts byV.sororto penetrate nests by lowering the incidence of multiple-hornet attacks and substantially reducing the likelihood of them approaching and chewing on entrances. We argue thatA.ceranaforages for animal feces because it has properties that repel this deadly predator from nest entrances, providing the first report of tool use by honey bees and the first evidence that they forage for solids that are not derived from plants. Our study describes a remarkable weapon in the already sophisticated portfolio of defenses that honey bees have evolved in response to the predatory threats they face. It also highlights the strong selective pressure honey bees will encounter if giant hornets, recently detected in western North America, become established.

Congenital cytomegalovirus (CMV) infection is a major cause of birth defects ranging from developmental disorders to stillbirth. Most newborns affected by CMV do not present with symptoms at birth but are at risk of sequelae at later stages of their childhood. Stored dried blood spots (DBS) taken at birth can be used for retrospective diagnosis of hereditary diseases, but detection of pathogens is challenged by potentially low pathogen concentrations in the small blood volume available in a DBS. Here we test four different extraction methods for optimal recovery of CMV DNA from DBS at low to high CMV titers. The recovery efficiencies varied widely between the different extractions (from 3% to 100%) with the most efficient method extracting up to 113-fold more CMV DNA than the least efficient and 8-fold more than the reference protocol. Furthermore, we amplified four immunomodulatory CMV genes from the extracted DNA: the UL40 and UL111A genes which occur as functional knockouts in some circulating CMV strains, and the highly variable UL146 and US28 genes. The PCRs specifically amplified the CMV genes at all tested titers with sufficient quality for sequencing and genotyping. In summary, we here report an extraction method for optimal recovery of CMV DNA from DBSs that can be used for both detection of CMV and for genotyping of polymorphic CMV genes in congenital CMV infection.

Water conservation and economics dictate that fresh produce processors reuse/recirculate the process water. However, the ensuing accumulation of organic matter in water depletes the chlorine sanitizer required for food safety. In this study, we comprehensively investigated chemical compounds that are responsible for water quality in relation to chemical oxygen demand (COD) and chlorine demand (CLD), the two most critical factors associated with water treatment and chlorine replenishment. Simulating commercial fresh-cut wash operations, multiple batches of diced cabbage (0.3 x 0.3 cm2) were washed in the same tank of water. The major components were isolated from the wash water and analyzed by HPLC. Sugars were the predominant compounds (82.7% dry weight) and the major contributor to COD (81.6%), followed by proteins/peptides (7.3% dry weight, 5.3% COD), organic acids (6.2% dry weight, 3.6% COD), and phenolics (0.5% dry weight, 0.5% COD). By repeated time course measures, the effect of these chemicals on CLD are dependent on the chemical structure, concentration in the wash water, and their rate of reaction. Proteins/peptides accounted for about 50% of the total CLD over a 120-min period and phenolics was 21% at 5 min, but diminished with time. The contribution by organic acids and sugars increased continuously, reaching 22% and 16% of total CLD at 120 min of chlorination, respectively. Collectively, these compounds represented 86% of the CLD in cabbage wash water at 5 min and greater than 94% CLD afterwards. This is the first systematic report on the source of COD and CLD during fresh produce washing. It provides essential information for the produce processors to develop safe, effective, and economical wash water treatment/reuse and chlorine replenishment strategies.

In epidemiological surveys and surveillance the application of molecular tools is essential in detecting submicroscopic malaria. A genus-specific conventional cytochrome b (cytb) PCR has shown high sensitivity in field studies, detecting 70% submicroscopic malaria. The main objective of this study was to assess the conversion from conventional to real-time PCR testing both SYBR and probe protocols, and including quantitative (q) PCR. The protocols were assessed applying well-defined clinical patient material consisting of 33 positive and 80 negative samples. Sequencing of positive PCR products was performed. In addition, a sensitivity comparison of real-time PCR methods was done by including five relevant assays investigating the effect of amplification target and platform. Sensitivity was further examined using field material consisting of 111 P.falciparum positive samples from Tanzanian children (< 5 years), as well as using related patient data to assess the application of q-PCR with focus on low-level parasitaemia. Both the cytb SYBR and probe PCR protocols showed as high sensitivity and specificity as their conventional counterpart, except missing one P. malariae sample. The SYBR protocol was more sensitive and specific than using probe. Overall, choice of amplification target applied is relevant for achieving ultra-sensitivity, and using intercalating fluorescence dye rather than labelled hydrolysis probes is favourable. Application of q-PCR analysis in field projects is important for the awareness and understanding of low-level parasitaemia. For use in clinical diagnosis and epidemiological studies the highly sensitive and user-friendly cytb SYBR q-PCR method is a relevant tool. The genus-specific method has the advantage that species identification by sequencing can be performed as an alternative to species-specific PCR. publishedVersion

Moving Bed Biofilm Reactors (MBBRs) can efficiently treat wastewater by incorporating suspended biocarriers that provide attachment surfaces for active microorganisms. The performance of MBBRs for wastewater treatment is, among other factors, contingent upon the characteristics of the surface area of the biocarriers. Thus, novel biocarrier topology designs can potentially increase MBBR performance in a significant manner. The goal of this work is to assess the performance of 3-D-printed biofilter media biocarriers with varying surface area designs for use in nitrifying MBBRs for wastewater treatment. Mathematical models, rendering, and 3D printing were used to design and fabricate gyroid-shaped biocarriers with a high degree of complexity at three different levels of specific surface area (SSA), generally providing greater specific surface areas than currently available commercial designs. The biocarriers were inoculated with a nitrifying bacteria community, and tested in a series of batch reactors for ammonia conversion to nitrate, in three different experimental configurations: constant fill ratio, constant total surface area, and constant biocarrier media count. Results showed that large and medium SSA gyroid biocarriers delivered the best ammonia conversion performance of all designs, and significantly better than that of a standard commercial design. The percentage of ammonia nitrogen conversion at 8 hours for the best performing biocarrier design was: 99.33% (large SSA gyroid, constant fill ratio), 94.74% (medium SSA gyroid, constant total surface area), and 92.73% (large SSA gyroid, constant biocarrier media count). Additionally, it is shown that the ammonia conversion performance was correlated to the specific surface area of the biocarrier, with the greatest rates of ammonia conversion (99.33%) and nitrate production (2.7 mg/L) for manufactured gyroid biocarriers with a specific surface area greater than 1980.5 m2/m3. The results suggest that the performance of commercial MBBRs for wastewater treatment can be greatly improved by manipulation of media design through topology optimization.

When clothes are worn and washed, they emit fibres into the ecosystem via discharges of sewage that have been linked to the global dispersion of clothing fibres. Facilities that treat sewage divert some fibres from sewage effluent to sludge, but no current methods of filtration eliminate their environmental release. While filters for washing-machines are sold to consumers with the argument they will reduce the emissions of fibres from clothes to the environment, there is insufficient scientific peer-reviewed evidence assessing their ability to retain fibres from washed clothes and reduce environmental contamination. To improve our understanding and develop more realistic methods to assess the efficiency of filters, we washed replicate cotton and polyester garments in replicate domestic front-loaded washing-machines with and without replicate filters (micro- and milli-meter-sized pores), and then quantified the masses of the fibres retained by the filters and those released in the effluent. Here we show micrometer-sized filters significantly reduced the mass of cotton by 67% (F-2,F-6= 11.69, P

The self-cleaning screen filter is one of the most common types used in drip irrigation systems. Backwashing pressure difference and backwashing time for two screen filters with one geometry and two different screens (178 μm and 124 μm) using two water qualities (tap water and sand-water mixture) were studied in a total of 88 runs (42 runs for tap water, 22 and 24 filtration cycles for sand-water mixture and backwashing, respectively). The backwashing pressure difference and backwashing time were calculated using the experimental data, and the results were largely in the range of measured values. Three constraint conditions (flowrate, sand condition and filtration time) of backwashing pressure difference were analysed, and the optimal values of backwashing pressure difference were given as 60.0 and 70.0 kPa for 178 μm and 124 μm filters, respectively. The backwashing time of the screen filter should be an optimal value that ensures that the pressure difference between the internal and external surfaces of the screen decreased to the initial value, and the sand concentration of the backwashed outlet decreased to a small, stable value. Based on the results of the backwashing experiment and prototype observation, the optimal backwashing time was given as 30 to 45 s for both screen filters.

Assimilable organic carbon (AOC) is known to correlate with microbial growth, which can consequently degrade drinking water quality. Despite this, there is no standardised AOC test that can be applied to drinking water distribution systems (DWDS). Herein we report the development of a quick, robust AOC that incorporates known strains Pseudomonas fluorescens strain P-17 and Spirillum strain NOX, a higher inoculum volume and enumeration using flow cytometry to generate a quicker (total test time reduced from 14 to 8 days), robust method. We apply the developed AOC test to twenty drinking water treatment works (WTW) to validate the method reproducibility and resolution across a wide range of AOC concentrations. Subsequently, AOC was quantified at 32 sample points, over four DWDS, for a year in order to identify sinks and sources of AOC in operative networks. Application of the developed AOC protocol provided a previously unavailable insight and novel evidence of pipes and service reservoirs exhibiting different AOC and regrowth behaviour. Observed correlations between AOC and microbial growth highlight the importance of monitoring AOC as an integral part of managing drinking water quality at the consumers tap.

The current technologies of algal biofuels production and wastewater treatment (e.g., aerobic) process are still in question, due to the significant amount of fresh water and nutrients requirements for microalgae cultivation, and negative energy balance in both processes, especially when considered in the context of developing counties around the world. In this research, a simplistic sustainable approach of algal biofuels production from wastewater was proposed using a Hybrid Anaerobic Baffled Reactor (HABR) and Photobioreactor (PBR) system. The study suggests that the HABR was capable of removing most of the organic and solid (>90% COD and TSS removal) from wastewater, and produced a healthy feedstock (high N: P = 3:1) for microalgae cultivation in PBRs for biofuels production. A co-culture of Chlorella vulgaris, Chlorella sorokiniana, and Scenedesmus simris002 showed high lipid content up to 44.1%; and the dominant FAMEs composition (C16-C18) of 87.9% in produced biofuels. Perhaps, this proposed low-cost technological approach (e.g., HABR-PBR system) would connect the currently broken link of sustainable bioenergy generation and wastewater treatment pathway for developing countries.

Coal gangue has the shortcomings of low calorific value and refractory burnout, while polyvinyl chloride has the advantages of a long combustion process and high calorific value. In order to make up for these shortcomings of coal gangue, the possibility of a treatment method based on co-combustion of coal gangue with polyvinyl chloride, which can be centrally recovered from municipal solid waste, is proposed. In order to analyze the combustion effect of a mixture of these two substances, experimental samples were prepared by mixing these two substances in three different ratios, and they were tested by thermogravimetric analysis. The experimental results were compared, analyzed and evaluated. The effects of the proportion of polyvinyl chloride in the mixture on the temperature parameters, activation energy, and interaction during co-combustion were analyzed. In order to analyze the interaction during co-combustion of the two, a coupling analysis method for mixed combustion is presented, and the effectiveness of this method is verified by comparing with the correlation analysis results of co-combustion. The results show that co-combustion can mitigate the ignition difficulty and burnout of coal gangue. When the proportion of polyvinyl chloride in the mixture was increased from 20% to 80%, the maximum weightlessness rate of the first stage rapidly increased from 4.5%/min to 15.6%/min; however, that of the second stage slowly increased from 3.7%/min to 4.2%/min. A 20% proportion of polyvinyl chloride showed the most significant promotion of co-combustion, with a maximum coupling coefficient of 0.00318, which was 1.11 and 1.35 times greater than that of 50% and 80% proportions, respectively. Co-combustion can reduce the activation energy of coal gangue during the initial and end stages. Therefore, co-combustion is helpful to improve the problems of low calorific value and refractory burnout of coal gangue.

Organic materials in fresh-cut produce wash water deplete free chlorine that is required to prevent pathogen survival and cross-contamination. This research evaluated water quality parameters frequently used to describe organic load for their fitness to predict chlorine demand (CLD) and chemical oxygen demand (COD), which are major needs identified by the industry-led produce food safety taskforce. Batches of romaine lettuce, iceberg lettuce, or carrot of different cut sizes and shapes were washed in 40 liters of water. Physicochemical properties of wash water including CLD, COD, total organic carbon (TOC), total suspended solids (TSS), total dissolved solids (TDS), turbidity, total sugar content, and pH, were monitored. Results indicate that pH is primarily commodity dependent, while organic load is additionally impacted by cutting and washing conditions. Significant linear increases in COD, TOC, CLD, TDS, and turbidity resulted from increasing product-to-water ratio, and decreasing cut size. Physicochemical parameters, excluding pH, showed significant positive correlation across different cut sizes within a commodity. High correlations were obtained between CLD and COD and between COD and TOC for pooled products. The convenient measurement of TDS, along with its strong correlation with COD and CLD, suggests the potential of TDS for predicting organic load and chlorine reactivity. Finally, the potential application and limitation of the proposed models in practical produce processing procedures are discussed extensively.

Stormwater biofilters are used to attenuate the flow and volume of runoff and reduce pollutant loading to aquatic systems. However, the capacity of biofilters to remove microbial contaminants remains inadequate. While biochar has demonstrated promise as an amendment to improve microbial removal in laboratory-scale biofilters, it is uncertain if the results are generalizable to the field. To assess biochar performance in a simulated field setting, sand and biochar-amended sand biofilters were periodically dosed with natural stormwater over a 61-week conditioning phase. Impact of media saturation was assessed by maintaining biofilters with and without a saturated zone. Biochar-amended biofilters demonstrated improved Escherichia coli removal over sand biofilters during the first 31 weeks of conditioning though media type did not impact E. coli removal during the last 30 weeks of conditioning. Presence of a saturated zone was not a significant factor influencing E. coli removal across the entire conditioning phase. Following conditioning, biofilters underwent challenge tests using stormwater spiked with wastewater to assess their capacity to remove wastewater-derived E. coli, enterococci, and male-specific (F+) coliphage. In challenge tests, biochar-amended biofilters demonstrated enhanced removal of all fecal indicators relative to sand biofilters. Additionally, saturated biofilters demonstrated greater removal of fecal indicators than unsaturated biofilters for both media types. Discrepant conclusions from the conditioning phase and challenge tests may be due to variable influent chemistry, dissimilar transport of E. coli indigenous to stormwater and those indigenous to wastewater, and differences in E. coli removal mechanisms between tests. Mobilization tests conducted following challenge tests showed minimal (

Cerato-platanin (CP) is a non-catalytic, cysteine-rich protein, the first member of the cerato-platanin family. It is a single-domain protein with a double Ψ/β barrel domain resembling the D1 domain of plant and bacterial expansins. Similarly to expansins, CP shows a cell wall-loosening activity on cellulose and can be defined as an expanisin-like protein, in spite of the missing D2 domain, normally present in plant expansins. The weakening activity shown on cellulose may facilitate the CP-host interaction, corroborating the role of CP in eliciting plant defence response. Indeed, CP is an elicitor of primary defences acting as a Pathogen-Associated Molecular Patterns (PAMP). So far, structure-function relationship study has been mainly performed on the bacterial BsEXLX1 expansin, probably due to difficulties in expressing plant expansins in heterologous systems. Here, we report a subcloning and purification method of CP in the engineered E. coli SHuffle cells, which proved to be suitable to obtain the properly folded and biologically active protein. The method also enabled the production of the mutant D77A, rationally designed to be inactive. The wild-type and the mutated CP were characterized for cellulose weakening activity and for PAMP activity (i.e. induction of Reactive Oxygen Species synthesis and phytoalexins production). Our analysis reveals that the carboxyl group of D77 is crucial for expansin-like and PAMP activities, thus permitting to establish a correlation between the ability to weaken cellulose and the capacity to induce defence responses in plants. Our results enable the structural and functional characterization of a mono-domain eukaryotic expansin and identify the essential role of a specific aspartic residue in cellulose weakening.

The use of Polymerase Chain Reaction (PCR) assays for pathogen detection in travelers' diarrhea (TD) field studies is limited by the on-site processing and storage requirements for fecal specimens. The objectives of this investigation were to i) characterize the pathogen distribution in deployed military personnel with TD using the TaqMan® Array Card PCR (TAC) on frozen stool and diarrheal smears on Whatman FTA Elute cards (FTA cards), and to ii) compare TAC detection of enteropathogen targets using smeared FTA cards and frozen stool, using TAC on frozen stool as the 'reference standard'. Stool samples, obtained from active duty personnel with acute TD enrolled in a field trial, were smeared onto FTA cards and stored at room temperature. A corresponding aliquot of stool was frozen in a cryovial. FTA cards and frozen stool samples were tested at a central lab, using a customized TAC for detection of TD pathogens. 187 paired frozen stool samples and smeared FTA cards were stored for a median of 712 days (IQR 396-750) before testing. Overall detection rates were 78.6% for frozen stool and 73.2% for FTA cards. Diarrheagenic Escherichia coli were the most common bacteria identified. Using the TAC results on frozen stool as the reference, the overall sensitivity and specificity of TAC on FTA cards was 72.9% and 98.0% respectively. TAC on FTA cards demonstrated a decrease in sensitivity with increasing frozen stool quantification cycle (Cq) (90.0% in FTA cards with a corresponding frozen stool Cq < 30, and 72.9% in samples with a corresponding frozen stool Cq < 35). Our findings support the use and further development of FTA cards in combination with a quantitative PCR assay for enteropathogen detection in TD field studies.