Molecular Pathology of Rare Bleeding Disorders (RBDs) in India: A Systematic Review

Background

Though rare in occurrence, patients with rare bleeding disorders (RBDs) are highly heterogeneous and may manifest with severe bleeding diathesis. Due to the high rate of consanguinity in many caste groups, these autosomal recessive bleeding disorders which are of rare occurrence in populations across the world, may not be as rare in India.

Objectives

To comprehensively analyze the frequency and nature of mutations in Indian patients with RBDs.

Methods

Pubmed search was used (www.pubmed.com) to explore the published literature from India on RBDs using the key words “rare bleeding disorders”, “mutations”, “India”, “fibrinogen”, “afibrinogenemia”, “factor II deficiency”, “prothrombin” “factor VII deficiency”, “factor V deficiency”, “factor X deficiency”, “factor XI deficiency”, “combined factor V and VIII deficiency”, “factor XIII deficiency”, “Bernard Soulier syndrome” and “Glanzmanns thrombasthenia” in different combinations. A total of 60 relevant articles could be retrieved. The distribution of mutations from India was compared with that of the world literature by referring to the Human Gene Mutation Database (HGMD) (www.hgmd.org).

Results

Taken together, 181 mutations in 270 patients with different RBDs have been reported from India. Though the types of mutations reported from India and their percentage distribution with respect to the world data are largely similar, yet much higher percentage of small deletions, duplication mutations, insertions, indels were observed in this analysis. Besides the identification of novel mutations and polymorphisms, several common mutations have also been reported, which will allow to develop a strategy for mutation screening in Indian patients with RBDs.

Conclusion

There is a need for a consortium of Institutions working on the molecular pathology of RBDs in India. This will facilitate a quicker and cheaper diagnosis of RBDs besides its utility in first trimester prenatal diagnosis of the affected families.     

The 2015 Bioinformatics Open Source Conference (BOSC 2015)

The Bioinformatics Open Source Conference (BOSC) is organized by the Open Bioinformatics Foundation (OBF), a nonprofit group dedicated to promoting the practice and philosophy of open source software development and open science within the biological research community. Since its inception in 2000, BOSC has provided bioinformatics developers with a forum for communicating the results of their latest efforts to the wider research community. BOSC offers a focused environment for developers and users to interact and share ideas about standards; software development practices; practical techniques for solving bioinformatics problems; and approaches that promote open science and sharing of data, results, and software. BOSC is run as a two-day special interest group (SIG) before the annual Intelligent Systems in Molecular Biology (ISMB) conference. BOSC 2015 took place in Dublin, Ireland, and was attended by over 125 people, about half of whom were first-time attendees. Session topics included “Data Science;” “Standards and Interoperability;” “Open Science and Reproducibility;” “Translational Bioinformatics;” “Visualization;” and “Bioinformatics Open Source Project Updates”. In addition to two keynote talks and dozens of shorter talks chosen from submitted abstracts, BOSC 2015 included a panel, titled “Open Source, Open Door: Increasing Diversity in the Bioinformatics Open Source Community,” that provided an opportunity for open discussion about ways to increase the diversity of participants in BOSC in particular, and in open source bioinformatics in general. The complete program of BOSC 2015 is available online at http://www.open-bio.org/wiki/BOSC_2015_Schedule.Open in a separate window     

AntiAngioPred: A Server for Prediction of Anti-Angiogenic Peptides

The process of angiogenesis is a vital step towards the formation of malignant tumors. Anti-angiogenic peptides are therefore promising candidates in the treatment of cancer. In this study, we have collected anti-angiogenic peptides from the literature and analyzed the residue preference in these peptides. Residues like Cys, Pro, Ser, Arg, Trp, Thr and Gly are preferred while Ala, Asp, Ile, Leu, Val and Phe are not preferred in these peptides. There is a positional preference of Ser, Pro, Trp and Cys in the N terminal region and Cys, Gly and Arg in the C terminal region of anti-angiogenic peptides. Motif analysis suggests the motifs “CG-G”, “TC”, “SC”, “SP-S”, etc., which are highly prominent in anti-angiogenic peptides. Based on the primary analysis, we developed prediction models using different machine learning based methods. The maximum accuracy and MCC for amino acid composition based model is 80.9% and 0.62 respectively. The performance of the models on independent dataset is also reasonable. Based on the above study, we have developed a user-friendly web server named “AntiAngioPred” for the prediction of anti-angiogenic peptides. AntiAngioPred web server is freely accessible at http://clri.res.in/subramanian/tools/antiangiopred/index.html (mirror site: http://crdd.osdd.net/raghava/antiangiopred/).     

DyCoNet: A Gephi Plugin for Community Detection in Dynamic Complex Networks

Community structure detection has proven to be important in revealing the underlying organisation of complex networks. While most current analyses focus on static networks, the detection of communities in dynamic data is both challenging and timely. An analysis and visualisation procedure for dynamic networks is presented here, which identifies communities and sub-communities that persist across multiple network snapshots. An existing method for community detection in dynamic networks is adapted, extended, and implemented. We demonstrate the applicability of this method to detect communities in networks where individuals tend not to change their community affiliation very frequently. When stability of communities cannot be assumed, we show that the sub-community model may be a better alternative. This is illustrated through test cases of social and biological networks. A plugin for Gephi, an open-source software program used for graph visualisation and manipulation, named “DyCoNet”, was created to execute the algorithm and is freely available from https://github.com/juliemkauffman/DyCoNet.     

Prediction and Analysis of Quorum Sensing Peptides Based on Sequence Features

Quorum sensing peptides (QSPs) are the signaling molecules used by the Gram-positive bacteria in orchestrating cell-to-cell communication. In spite of their enormous importance in signaling process, their detailed bioinformatics analysis is lacking. In this study, QSPs and non-QSPs were examined according to their amino acid composition, residues position, motifs and physicochemical properties. Compositional analysis concludes that QSPs are enriched with aromatic residues like Trp, Tyr and Phe. At the N-terminal, Ser was a dominant residue at maximum positions, namely, first, second, third and fifth while Phe was a preferred residue at first, third and fifth positions from the C-terminal. A few motifs from QSPs were also extracted. Physicochemical properties like aromaticity, molecular weight and secondary structure were found to be distinguishing features of QSPs. Exploiting above properties, we have developed a Support Vector Machine (SVM) based predictive model. During 10-fold cross-validation, SVM achieves maximum accuracy of 93.00%, Mathew’s correlation coefficient (MCC) of 0.86 and Receiver operating characteristic (ROC) of 0.98 on the training/testing dataset (T^200p+200n). Developed models performed equally well on the validation dataset (V^20p+20n). The server also integrates several useful analysis tools like “QSMotifScan”, “ProtFrag”, “MutGen” and “PhysicoProp”. Our analysis reveals important characteristics of QSPs and on the basis of these unique features, we have developed a prediction algorithm “QSPpred” (freely available at: http://crdd.osdd.net/servers/qsppred).     

Harmonization of the Practice of Independent Ethics Committees in Italy: Project E-Submission

Aim

The high variability of “centre-specific” documentation required by Independent Ethics Committee (IEC) plays a role in the time required for activation of participating centres of multicentre clinical trials. This study (a) provides a picture of the different activities, structural requirements and resources dedicated to the operation of the local IEC in Italy; (b) defines a detailed list of “centre-specific” documents considered as essential by the IEC for issuing its opinion and (c) suggests a “single document” to reduce the variability of the “centre-specific” documents required by the IEC.

Methodology

Two surveys were conducted through the portal of National Monitoring Centre of Clinical Trials (https://oss-sper-clin.agenziafarmaco.it/). The first survey focused on the local IEC resources and on the “centre-specific” documentation that local IEC required from the Sponsor and local Principal Investigator (PI). The second focused on “single document” required in the form of statements from the Sponsor and the PI. Answers were discussed and extended during regular scheduled teleconferences and plenary meeting.

Principal Findings

From 22/07/2009 to 15/12/2009, and from 19/04/2010 to 14/05/2010, 131 and 125 IECs responded to the first and the second surveys, respectively. 67% and 51% of IECs consider the structural requirements and the staff dedicated to the activity of the IECs as sufficient, respectively. Most of the IECs consider the “centre-specific” documentation as necessary for issuing the opinion, and a high percentage of IECs consider the proposed documentation as acceptable in substitution to any other “centre-specific” documentation already in use.

Conclusions

The harmonization of IECs practice in Italy is the first step to facilitate multicentre clinical trials. Similar efforts should be directed to reduce the total number of IECs and to standardize clinical trials approval procedures, focusing on administrative procedures as well.     

A Preliminary Investigation of Individual Differences in Subjective Responses to D-Amphetamine,Alcohol, and Delta-9-Tetrahydrocannabinol Using a Within-Subjects Randomized Trial

Polydrug use is common, and might occur because certain individuals experience positive effects from several different drugs during early stages of use. This study examined individual differences in subjective responses to single oral doses of d-amphetamine, alcohol, and delta-9-tetrahydrocannabinol (THC) in healthy social drinkers. Each of these drugs produces feelings of well-being in at least some individuals, and we hypothesized that subjective responses to these drugs would be positively correlated. We also examined participants’ drug responses in relation to personality traits associated with drug use. In this initial, exploratory study, 24 healthy, light drug users (12 male, 12 female), aged 21–31 years, participated in a fully within-subject, randomized, counterbalanced design with six 5.5-hour sessions in which they received d-amphetamine (20mg), alcohol (0.8 g/kg), or THC (7.5 mg), each paired with a placebo session. Participants rated the drugs’ effects on both global measures (e.g. feeling a drug effect at all) and drug-specific measures. In general, participants’ responses to the three drugs were unrelated. Unexpectedly, “wanting more” alcohol was inversely correlated with “wanting more” THC. Additionally, in women, but not in men, “disliking” alcohol was negatively correlated with “disliking” THC. Positive alcohol and amphetamine responses were related, but only in individuals who experienced a stimulant effect of alcohol. Finally, high trait constraint (or lack of impulsivity) was associated with lower reports of liking alcohol. No personality traits predicted responses across multiple drug types. Generally, these findings do not support the idea that certain individuals experience greater positive effects across multiple drug classes, but instead provide some evidence for a “drug of choice” model, in which individuals respond positively to certain classes of drugs that share similar subjective effects, and dislike other types of drugs.

Trial Registration

ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT02485158","term_id":"NCT02485158"}}NCT02485158     

A statistical model for describing and simulating microbial community profiles

Many methods have been developed for statistical analysis of microbial community profiles, but due to the complex nature of typical microbiome measurements (e.g. sparsity, zero-inflation, non-independence, and compositionality) and of the associated underlying biology, it is difficult to compare or evaluate such methods within a single systematic framework. To address this challenge, we developed SparseDOSSA (Sparse Data Observations for the Simulation of Synthetic Abundances): a statistical model of microbial ecological population structure, which can be used to parameterize real-world microbial community profiles and to simulate new, realistic profiles of known structure for methods evaluation. Specifically, SparseDOSSA’s model captures marginal microbial feature abundances as a zero-inflated log-normal distribution, with additional model components for absolute cell counts and the sequence read generation process, microbe-microbe, and microbe-environment interactions. Together, these allow fully known covariance structure between synthetic features (i.e. “taxa”) or between features and “phenotypes” to be simulated for method benchmarking. Here, we demonstrate SparseDOSSA’s performance for 1) accurately modeling human-associated microbial population profiles; 2) generating synthetic communities with controlled population and ecological structures; 3) spiking-in true positive synthetic associations to benchmark analysis methods; and 4) recapitulating an end-to-end mouse microbiome feeding experiment. Together, these represent the most common analysis types in assessment of real microbial community environmental and epidemiological statistics, thus demonstrating SparseDOSSA’s utility as a general-purpose aid for modeling communities and evaluating quantitative methods. An open-source implementation is available at http://huttenhower.sph.harvard.edu/sparsedossa2.     

Functional Genetics in the Post-Genomics Era: Building a Better Roadmap in Drosophila

In this commentary, Rob Kulathinal describes two articles from the Perrimon lab, each describing a new online resource that can assist geneticists with the design of their RNA interference (RNAi) experiments. Hu et al.’s “UP-TORR: online tool for accurate and up-to-date annotation of RNAi reagents” and “FlyPrimerBank: An online database for Drosophila melanogaster gene expression analysis and knockdown evaluation of RNAi reagents” are published, respectively, in this month’s issues of GENETICS and G3.     

GeneComps and ChemComps: a new CTD metric to identify genes and chemicals with shared toxicogenomic profiles

The Comparative Toxicogenomics Database is a public resource that promotes understanding about the effects of environmental chemicals on human health. Currently, CTD describes over 184,000 molecular interactions for more than 5,100 chemicals and 16,300 genes/proteins. We have leveraged this dataset of chemical-gene relationships to compute similarity indices following the statistical method of the Jaccard index. These scores are used to produce lists of comparable genes (“GeneComps”) or chemicals (“ChemComps”) based on shared toxicogenomic profiles. GeneComps and ChemComps are now provided for every curated gene and chemical in CTD. ChemComps are particularly significant because they provide a way to group chemicals based upon their biological effects, instead of their physical or structural properties. These metrics provide a novel way to view and classify genes and chemicals and will help advance testable hypotheses about environmental chemical-genedisease networks.

Availability

CTD is freely available at http://ctd.mdibl.org/     

Genome sequence of the Lebeckia ambigua-nodulating “Burkholderia sprentiae” strain WSM5005T

“Burkholderia sprentiae” strain WSM5005^T is an aerobic, motile, Gram-negative, non-spore-forming rod that was isolated in Australia from an effective N₂-fixing root nodule of Lebeckia ambigua collected in Klawer, Western Cape of South Africa, in October 2007. Here we describe the features of “Burkholderia sprentiae” strain WSM5005^T, together with the genome sequence and its annotation. The 7,761,063 bp high-quality-draft genome is arranged in 8 scaffolds of 236 contigs, contains 7,147 protein-coding genes and 76 RNA-only encoding genes, and is one of 20 rhizobial genomes sequenced as part of the DOE Joint Genome Institute 2010 Community Sequencing Program.     

Rainy weather and medical school admission interviews

Mood can influence behaviour and consumer choice in diverse settings. We found that such cognitive influences extend to candidate admission interviews at a Canadian medical school. We suggest that an awareness of this fallibility might lead to more reasonable medical school admission practices.Admission offers to medical school are competitive and sometimes based on an interview. Psychology research suggests, however, that interviews are prone to subconscious biases from extraneous factors unrelated to the candidate.¹ One of the most fundamental observations is that people interviewed on rainy days tend to receive lower ratings than people interviewed on sunny days.² We studied whether this bias also extends to admission interviews at a large Canadian medical school.We analyzed the results of consecutive medical school interviews at the University of Toronto between 2004 and 2009. We included all data available with no exclusions. Almost all interviews occurred in the early spring. Scores for each interview were obtained from the admissions office as recorded from 0 to 20.³ This Likert scale was anchored with integer values where 10 denoted “unsuitable,” 12 denoted “marginal,” 14 denoted “fair,” 16 denoted “good,” 18 denoted “excellent” and 20 denoted “outstanding.”We obtained weather data from the official government archive and defined a priori the day as “rainy” if precipitation (including freezing rain, snow and hail) occurred in the morning or afternoon.⁴ Otherwise, we defined the day as “sunny.” We did not examine more complex combinations with time lags, such as when a sunny day followed multiple rainy days.A total of 2926 candidates were interviewed over the 6-year period. As expected, their demographic characteristics were unrelated to the weather (Appendix 1, available online at www.cmaj.ca/cgi/content/full/cmaj.091546/DC1). Overall, those interviewed on rainy days received about a 1% lower score than those interviewed on sunny days (average score 16.31 v. 16.49, p = 0.042). This pattern was consistent for both senior interviewers (16.39 v. 16.55, p = 0.08) and junior interviewers (16.23 v. 16.42, p = 0.041). We next used logistic regression to analyze subsequent admission decisions. The difference in scores was equivalent to about a 10% lower total mark on the Medical College Admission Test.Open in a separate windowWe suggest that cognitive patterns evident in controlled psychology laboratories can also occur in regular medical settings. The magnitude of the specific influence may be modest, but such small differences can be important in some cases because each year there are about 100 candidates who receive a score within 1% of the admission threshold.⁵ In this study, we examined only one extraneous influence on mood. Many additional factors may also affect mood (e.g., ambiance, deportment, humour and scent).² Calling attention to these issues may diminish their impact on judgment.¹     

A Biotic Game Design Project for Integrated Life Science and Engineering Education

Engaging, hands-on design experiences are key for formal and informal Science, Technology, Engineering, and Mathematics (STEM) education. Robotic and video game design challenges have been particularly effective in stimulating student interest, but equivalent experiences for the life sciences are not as developed. Here we present the concept of a "biotic game design project" to motivate student learning at the interface of life sciences and device engineering (as part of a cornerstone bioengineering devices course). We provide all course material and also present efforts in adapting the project''s complexity to serve other time frames, age groups, learning focuses, and budgets. Students self-reported that they found the biotic game project fun and motivating, resulting in increased effort. Hence this type of design project could generate excitement and educational impact similar to robotics and video games.

This Education article is part of the Education Series.

Hands-on robotic and video game design projects and competitions are widespread and have proven particularly effective at sparking interest and teaching K–12 and college students in mechatronics, computer science, and Science, Technology, Engineering, and Mathematics (STEM). Furthermore, these projects foster teamwork, self-learning, design, and presentation skills [1,2]. Such playful and interactive media that provide fun, creative, open-ended learning experiences for all ages are arguably underdeveloped in the life sciences. Most hands-on education occurs in traditionally structured laboratory courses with a few exceptions like the International Genetically Engineered Machine (iGEM) competition [3]. Furthermore, there is an increasing need to bring the traditional engineering and life science disciplines together. In order to fill these gaps, we present the concept of a biotic game design project to foster student development in a broad set of engineering and life science skills in an integrated manner (Fig. 1). Though we primarily discuss our specific implementation as a cornerstone project-based class [4], alternative implementations are possible to motivate a variety of learning goals under various constraints such as student age and cost (see supplements for all course material).Open in a separate windowFig 1We developed a bioengineering devices course that employed biotic game design as a motivating project scheme. A: Biotic games enable human players to interact with cells. B: Conceptual overview of a biotic game setup. C: Students built and played biotic games. Image credits: A C64 joystick by Speed-link, 1984 (http://commons.wikimedia.org/wiki/File:Joystick_black_red_petri_01.svg); Euglena viridis by C. G. Ehrenberg, 1838; C Photo, N. J. C.Biotic games are games that operate on biological processes (Fig. 1) [5]. The biotic games we present here involve the single-celled phototactic eukaryote, Euglena gracilis. These microscopic organisms are housed in a microfluidic chip and are displayed in a magnified image on a video screen. Players interact with these cells by modulating the intensity and direction of light perpendicular to the microfluidic chip via a joystick, thereby influencing the cells’ phototactic motion. Software tracks the position of individual euglena with respect to virtual objects overlaid on the screen, creating myriad opportunities for creative game design and play. For example, in a simple game, points might be scored when a cell hits a virtual box (see S1 Video).The biotic game design project we developed was intended to motivate all the broad categories of theoretical and hands-on skills for creating any integrated instrument intended to house and to interface with biological materials, i.e., optics, electronics, sensing, actuation, microfluidics, fabrication, image processing, programming, and creative design. We termed the synthesis of these skills “biotics” in analogy to mechatronics. Our intended audience for this course was bioengineering undergraduate students at Stanford University who already had some programming experience but little to no experience in device design, fabrication, and integration. We also incorporated bioethics into the curriculum to emphasize the social responsibility of every engineer and demonstrate the potential for the biotic game project to motivate multiple fields. The course we taught spanned ten weeks, divided roughly equally into a set of technical units and the biotic game project, with two 4-hour lab sections and a single 1.5-hour lecture each week. For details and all course documents, please refer to the supplemental material.The technical section of the course focused on developing hands-on skills and theoretical understanding related to devices in a conventionally structured laboratory setting. We introduced students to fundamental electronics concepts and components such as voltage, current, resistors, capacitors, LEDs, filters, operational amplifiers, motors, microcontrollers (Arduino Uno), and breadboards. We followed a similar traditional approach in introducing optics, presenting the thin lens equation, ray tracing, conjugate planes, basic optical system design, and Köhler illumination. We covered additional topics in less detail: MATLAB programming, particle tracking, computer-aided design (CAD), fabrication, and microfluidics (learning objectives are provided at the beginning of each unit in the supplemental material).During the project-based section, students built their own biotic games. We left specific choices of implementation, architecture, and design to the students to encourage creativity and exploration but required students to revisit the technical skills they learned in the first section by integrating some specific requirements into their games (Fig. 2). Students built a bright field microscope with Köhler illumination and projected their images onto a webcam (optics). Glass and polydimethylsiloxane (PDMS) components comprised the microfluidic chip (microfluidics) and housed the euglena (microbiology). The holder for the chip and euglena-steering LEDs was designed in Solidworks (CAD) and 3-D printed (fabrication). The students constructed a polycarbonate housing for the game controller using a band saw and drill press (fabrication). The students revisited electronic breadboarding and soldering when creating the electronic circuits to communicate between the LEDs, joystick, microcontroller, and computer. Finally, they used MATLAB to program the microcontroller, implement real time image recognition, and provide the user interface for the game experience (image processing and programming).Open in a separate windowFig 2Biotic game-based courses encourage students to integrate a versatile set of relevant STEM topics.Image credits: Taken by N. J. C. (credit for the work and artifacts to the students who took the course).We challenged students to consider the ethical implications [6] of manipulating life in a game context before building their projects. Although phototaxis experiments with euglena are commonplace in education, and have hitherto raised no ethical concerns, the equivalent manipulation in the form of a game warrants its own ethical analysis as provided by Harvey et al. [7]. The students read and discussed this paper, then wrote a 200-word essay on whether they found it permissible or not to make and play biotic games. Students had the choice to switch to a nongame project of equivalent complexity. All students found euglena-based games permissible, pointing out that “they are nonsentient and cannot feel pain,” followed by a diverse range of considerations such as “the euglena are still free to act as they please,” “there needs to be an educational intention,” or “a pet…provides a way…to work on responsibility and caring.” Based on further student-initiated discussions that spontaneously emerged throughout the course, we believe that biotic games are effective in providing a stimulating, student-relevant, in-class context for bioethics.We motivated the game design project to the students as having educational potential at two levels, i.e., learning by building and learning by playing; we lectured them about the needs and opportunities for new approaches to K–12 STEM education [8,9]. The students were then asked to consider building a game that had educational value for the player. Educational value has many aspects, which was reflected in students’ statements regarding their intended educational outcomes for their games on their course project websites. These ranged from more factual learning objectives (“learn about…” “…inner working,” “…structural detail,” “… light responses,” “…euglena behavior”) to objectives affecting attitude (“spark interest,” “generate fascination,” “encourage to explore,” “respect for life”). We also had a game designer give a guest lecture to the students. For pragmatic reasons, we requested the students keep games very simple (ideally having just a single in-game objective) and cap game duration at one minute. Before, during, and after their projects, students received feedback from instructors as well as from their peers on their games from technical and user perspectives.The games that the students ultimately produced were diverse and creative (Fig. 2 and S1 Video), including single and multiplayer scenarios, games where euglena hit virtual targets, and games where euglena pushed virtual objects. Games that involved pushing objects across the screen (relying on collective motion of many organisms) were generally more consistent at correlating player strategy to scored points than those that involved hitting target objects. The quality and robustness of these integrated projects naturally varied, and individual groups placed more or less emphasis on different aspects based on personal preferences and learning goals (for example, fabricating a more elaborate housing for the game controller versus programming more complex game mechanics). A key point was that the students did not rely on prepared materials or platforms to develop their games but rather had to design, build, and test their game setups from scratch, thereby revisiting and deepening the primary learning goals of the course with some freedom to follow their own learning aspirations (Fig. 2). The final project deliverables were a two-minute project demonstration video, a website describing the elements of the project, and a game that all instructors and students played on the final day (Fig. 1B), which led to lots of laughter as well as in-depth discussions on technical details.Many students self-reported that they enjoyed the project and that it led to increased motivation and effort during the course. In response to the question “Do you think you were motivated to try harder or had more fun (and thereby learned more) during your final project because you were making a game (rather than just building a technical instrument, for example)? If so—please give some examples:” 15 out of 17 students responded “Very/definitely” on a five point scale. As examples, students listed: “wanted to make the best game,” “want to make it clever and cool in the eyes of classmates who are play testing,” “motivated during final push,” “willing to put in more time,” “was fun”/”made it fun,” “create a game that actually works,” “reinforced what was learned before,” and “provided room for creativity.” These comments reflect the overall excitement we saw for the biotic game project. While these responses do not constitute rigorous proof regarding course effectiveness (which will require more detailed and controlled assessments in the future), we consider this course a success based on our teaching experiences.45 students have now taken this class over the past three years, with 18 students in our most recent offering. We used each year to iterate and improve our implementation. For example, we changed the organism and stimulus from Paramecia galvanotaxis [5] to Euglena phototaxis, which gave more reliable long-term responses. We also added a simple microfluidics unit enabling students to build more robust organism housing chambers. We changed the microscope structure from LEGO to Thorlabs parts (essentially trading the emphasis on 3-D structural design, flexibility, and cost for a more in-depth focus on high-end optics and their alignment). Finally, we explicitly asked the students to design and fabricate a housing for the game controller to better incorporate fabrication skills like using a band saw and tapping screw threads. So far, we primarily used MATLAB as the programming component given its widespread use in education and research and the available Arduino interface. However, MATLAB is not particularly well-suited to support game design and is also not free, making translation into lower resource settings challenging. For the future, we are considering moving to smartphone-based control (such as Android) given that these mobile environments are very flexible and increasingly used for control of scientific and consumer instruments and are becoming more widespread in education. We also see the opportunity to better emphasize and teach the approach of iterative design; for example, by letting students prototype and test their game ideas on paper [10] and simple programming environments like Scratch [11] first, before attempting the full implementation. It would likely also be very rewarding for the students to be able to take their project home at the end of the course. In summary, many different course design decisions can be made based on specific intended educational outcomes. Not all of these can be fit into one course at the same time, and clear decisions should be made on how to balance covering a breadth of topics with depth on a selected few.As a preliminary test of another age range, time frame, and budget, we taught a greatly simplified 3-hour workshop where high school and middle school students assembled a low-cost microscope and microfluidics chamber, attached it to a smartphone, and stimulated euglena using a preprogrammed Arduino-based controller (see supplements). We had no game interface implemented yet on the phone, but the students could observe the euglena responses to the light stimuli. All students were able to complete the project and take their microscopes home. Over half of our undergraduate student teams also volunteered to present their game projects for this outreach event which took place multiple weeks after their class had ended. This separate experience suggests that the biotic game concept holds promise for reaching a wider age range in a shortened timespan and at a greatly reduced budget, and that completed games can be used in outreach activities. We are currently developing a kit modeled after this unit.In conclusion, we consider biotic games promising in motivating integrated, hands-on learning at the interface of life science and engineering. Our efforts so far indicate that this concept could be adapted to various age groups and learning goals with the potential for wider future impacts on education. We draw upon the analogy to robotics, where microcontrollers went from initially unfathomable as an educational tool to the vision of Papert and collaborators and their use of programmable robotics with children [12], eventually leading to multiple commercial realizations (LEGO mindstorm, Arduino, etc.), a large public following, and a major role in education both in the classroom and through competitions such as First Robotics [1]. We also see additional potential for integrating more creative and artistic aspects into STEM, i.e., leading to generalized Science, Technology, Engineering, Arts, and Mathematics (STEAM) disciplines [13]. We invite others to join us in these endeavors—all instructional materials are available in the appendix for further adaptations and educational use.     

Phyletic Profiling with Cliques of Orthologs Is Enhanced by Signatures of Paralogy Relationships

New microbial genomes are sequenced at a high pace, allowing insight into the genetics of not only cultured microbes, but a wide range of metagenomic collections such as the human microbiome. To understand the deluge of genomic data we face, computational approaches for gene functional annotation are invaluable. We introduce a novel model for computational annotation that refines two established concepts: annotation based on homology and annotation based on phyletic profiling. The phyletic profiling-based model that includes both inferred orthologs and paralogs—homologs separated by a speciation and a duplication event, respectively—provides more annotations at the same average Precision than the model that includes only inferred orthologs. For experimental validation, we selected 38 poorly annotated Escherichia coli genes for which the model assigned one of three GO terms with high confidence: involvement in DNA repair, protein translation, or cell wall synthesis. Results of antibiotic stress survival assays on E. coli knockout mutants showed high agreement with our model''s estimates of accuracy: out of 38 predictions obtained at the reported Precision of 60%, we confirmed 25 predictions, indicating that our confidence estimates can be used to make informed decisions on experimental validation. Our work will contribute to making experimental validation of computational predictions more approachable, both in cost and time. Our predictions for 998 prokaryotic genomes include ∼400000 specific annotations with the estimated Precision of 90%, ∼19000 of which are highly specific—e.g. “penicillin binding,” “tRNA aminoacylation for protein translation,” or “pathogenesis”—and are freely available at http://gorbi.irb.hr/.     

Effect of a musical intervention on tolerance and efficacy of non-invasive ventilation in the ICU: study protocol for a randomized controlled trial (MUSique pour l’Insuffisance Respiratoire Aigue - Mus-IRA)

BackgroundNon-invasive ventilation (NIV) tolerance is a key factor of NIV success. Hence, numerous sedative pharmacological or non-pharmacological strategies have been assessed to improve NIV tolerance. Music therapy in various health care settings has shown beneficial effects. In invasively ventilated critical care patients, encouraging results of music therapy on physiological parameters, anxiety, and agitation have been reported. We hypothesize that a musical intervention improves NIV tolerance in comparison to conventional care. We therefore question the potential benefit of a receptive music session administered to patients by trained caregivers (“musical intervention”) to enhance acceptance and tolerance of NIV.Methods/designWe conduct a prospective, three-center, open-label, three-arm randomized trial involving patients in the intensive care unit (ICU) who require NIV, as assessed by the treating physician. Participants are allocated to a “musical intervention” arm (“musical intervention” applied during all NIV sessions), to a “sensory deprivation” arm (sight and hearing isolation during all NIV sessions), or to the control group. The primary endpoint is the change in respiratory comfort (measured with a digital visual scale) before the initiation and after 30 minutes of the first NIV session. The evaluation of the primary endpoint is performed blindly from the treatment group. Secondary endpoints include changes in respiratory and cardiovascular parameters during NIV sessions, the percentage of patients requiring endotracheal intubation, day-90 anxiety/depression and health-related quality of life, post-trauma stress induced by NIV, and the overall assessment of NIV.The follow-up for each participant is 90 days. We expect to randomize a total of 99 participants.DiscussionAs music intervention is a simple and easy-to-implement non-pharmacological technique, efficacious in reducing anxiety in critically ill patients, it appeared logical to assess its efficacy in NIV, one of the most stressful techniques used in the ICU. Patient centeredness was crucial in choosing the outcomes assessed.

Trial registration

ClinicalTrials.gov: {"type":"clinical-trial","attrs":{"text":"NCT02265458","term_id":"NCT02265458"}}NCT02265458. Registered on 25 August 2014.

Electronic supplementary material

The online version of this article (doi:10.1186/s13063-016-1574-z) contains supplementary material, which is available to authorized users.     

Prospective Antiretroviral Treatment of Asymptomatic,HIV-1 Infected Controllers

The study of HIV-infected “controllers” who are able to maintain low levels of plasma HIV RNA in the absence of antiretroviral therapy (ART) may provide insights for HIV cure and vaccine strategies. Despite maintaining very low levels of plasma viremia, controllers have elevated immune activation and accelerated atherosclerosis. However, the degree to which low-level replication contributes to these phenomena is not known. Sixteen asymptomatic controllers were prospectively treated with ART for 24 weeks. Controllers had a statistically significant decrease in ultrasensitive plasma and rectal HIV RNA levels with ART. Markers of T cell activation/dysfunction in blood and gut mucosa also decreased substantially with ART. Similar reductions were observed in the subset of “elite” controllers with pre-ART plasma HIV RNA levels below conventional assays (<40 copies/mL). These data confirm that HIV replication persists in controllers and contributes to a chronic inflammatory state. ART should be considered for these individuals (ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01025427","term_id":"NCT01025427"}}NCT01025427).