Escherichia coli resistance to 3GC/carbapenems); (iii) screening travelers with a history of hospitalization and consumption of antibiotic drug returning from at risk areas  allowing an early warning detection of the emergence for quick control measures implementation in IOC.Antimicrobial resistance (AMR) is a major threat to human, animal health, and environment worldwide. For human, transmission occurred through a variety of routes both in health-care settings and community. In animals, AMR was reported in livestock, pets, and wildlife; transmission of AMR can be zoonotic with the probably most important route being foodborne transmission. The Indian Ocean Commission (IOC), composed of Comoros, Madagascar, Mauritius, Reunion (France), and Seychelles recognized the surveillance of AMR in both animal and human as a main public health priority for the region. Mayotte, French overseas territory, located in Comoros archipelago, was also included in this review. This review summarized our best epidemiological knowledge regarding AMR in Indian Ocean. We documented the prevalence, and phenotypic and genotypic profiles of prone to resistance Gram-positive and Gram-negative bacteria both in animals and humans. Our review clearly pointed out extended-spectrum β-lactamase and carbapenemase-producing Enterobacteriaceae as main human and animal health issue in IOC. However, publications on AMR are scarce, particularly in Comoros, Mayotte, and Seychelles. Thus, research and surveillance priorities were recommended (i) estimating the volume of antimicrobial drugs used in livestock and human medicine in the different territories [mainly third generation cephalosporin (3GC)]; (ii) developing a “One Health” surveillance approach with epidemiological indicators as zoonotic foodborne pathogen (i.e., couple  Increasing global antimicrobial resistance (AMR) is a major threat to human and animal endangering decades of improvements in health-care outcomes. It endangers modern human and veterinary medicine and undermines food safety .In humans, the global burden of AMR is longer duration of illness, higher lethality, increasing costs of treatment, and inability to cure infected patient . In animStaphylococcus aureus (MRSA)].Transmission of AMR to human can be zoonotic taking place through a variety of routes where the foodborne route is probably the most important . Direct The Indian Ocean Commission (IOC) is composed of five countries: Comoros, Madagascar, Mauritius, Reunion (France), and Seychelles. Mayotte, French overseas territory located in Comoros archipelago, was also included in the study. In 2015, the IOC identified AMR surveillance in both animal and human as a main priority for territories , and fecal–oral and foodborne bacterial species . We used available articles obtained through match searches using Google Scholar,The study performed from January to March 2017 included articles and conference abstracts published from 1990 to December 2016. Bacteria species included in the different searches were those prone to develop multidrug resistance as defined by Magiorakos et al. . Since 1998, S. aureus susceptibility profile changed tending to be more resistant to aminoglycosides and quinolones   was isolated during an uropathogenic survey  . In 2011c survey .Enterococcus spp. isolated in hospitalized patients were susceptible to vancomycin  use in livestock was correlated with VRE incidence in human populations .Pseudomonas aeruginosa is an opportunistic pathogen causing nosocomial infections (fections . Some stfections .P. aeruginosa isolates showed a moderate resistance to penicillin (piperacillin 12.8% and ticarcillin 31.9%) but were susceptible to ceftazidime and imipenem , 47% for ceftazidime, 73% for ciprofloxacin, and 40% for meropenem  and monolactams  . Outbreaare unit , 27, butoduction . The extolactams .P. aeruginosa in IOC are not clear, but acquisition of new gene of resistance to β-lactams is of concern. Rate of resistance to carbapenems identified in Mauritius are high even if decreasing. Identifying its burden in nosocomial infection is needed as well as aminoglycoside resistance. Recommended treatment for pseudomonas infection usually includes β-lactams and aminoglycosides.Trends regarding Acinetobacter baumannii is a troublesome pathogen for health-care institutions, and its ability to acquire resistance determinants is making it threatening the current antibiotic era  and imipenem was high (45.7%)  . Among s (45.7%) . No resi (45.7%) , but thinanarivo  could emnanarivo .Acinetobacter spp. isolated in hospitalized patients showed high rate of resistance to antibiotic tested with 86% for gentamicin and 50% for amikacin (both aminoglycosides), 95% for cefotaxime, 85% for ciprofloxacin, and 68% for meropenem , imipenem (12.9–8.3%), and ciprofloxacin (72.9–59.7%)] ] . In 2006-lactams . A wide entified .A. baumannii was identified in 2011 , Escherichia coli (37.0%), and Enterobacter cloacae (24.0%) with 75.0% of the CTX-M-15 type  and poultry (26%); main contaminated farms were located in Madagascar (40.5%), Mayotte (26.9%) followed by Mauritius (13.5%) and Comoros (6.7%) .E. coli, K. pneumoniae, and E. cloacae) in IOC seems increasing both in humans and animals. Broad spectrum antibiotics overuse is likely correlated with this evolution  .K. pneumoniae in IOC occurred in a Mauritius patient in 2009  gene in  in 2009 , in 2011 in 2009 , and in  in 2009 .E. cloacae of IMI-1 type occurred at the hospital , absent for quinolones, and moderated for ampicillin (35.7%) and ticarcillin (35.7%)  .Salmonella enterica serovar Enteritidis was isolated in humans without resistance identified for all antibiotic tested; transmission by chicken consumption was suspected  and nalidixic acid (1%)  .Salmonella spp. isolated in broiler chicken flocks were of 38.3% to streptomycin, 31.8% to tetracycline, and 16.8% to ampicillin (S. Hadar displayed reduced susceptibility to fluoroquinolones (80.8% to enrofloxacin)  .Salmonella spp. was identified between 1987 and 1988  (ctively) . Main inctively) .Campylobacter spp. was moderate in community children in 2008–2009, with overall resistance of 24.8% for amoxicillin, 2.2% for ciprofloxacin, 1.8% for erythromycin, and 1.1% for tetracycline , antibiogram panels, over different periods of time and targeting various bacteria species. Thus, results should be interpreted with caution but this attempt of review was not performed before and confirmed that AMR is threatening IOC. Main issue identified for IOC was ESBL and CPE which is in agreement with their increase worldwide over the past decade .Enterococcus spp., P. aeruginosa, Salmonella spp., Campylobacter spp., and Shigella spp.Literature was limited in Comoros, Mayotte, and Seychelles confirming needs to develop AMR surveillance and research in these territories and scarce for bacterial species: In IOC, the SEGA–One Health network was created in 2009 with the objective of monitoring outbreak-prone infections . It aims(i)A direct relationship between antibiotic consumption, emergence, and dissemination of AMR was demonstrated . Estimat(ii)A. baumannii in pets, humans, and livestock, and ESBLE in livestock and humans). Using standardized indicators (antibiotic drugs–bacteria species couple) for surveillance of AMR patterns across health-care settings, countries, and host species is essential. One relevant epidemiological indicator, for both animal and human, could be E. coli AMR, particularly 3GC and carbapenems. Surveillance should be accompanied by further investigations regarding genetic support of resistance between hosts for source of contamination and dynamics of propagation between reservoirs identification .Integrated AMR One Health approach including human, animal, and environment in both surveillance and research is clearly needed Spread of NDM-1-producing Enterobacteriaceae in IOC confirms needs for strengthening the early warning surveillance system of AMR emergences. The region is connected to hotspots of AMR as Asia (12.0% of traffic) characterized by high AMR prevalence in community [e.g., ESBL in China , importaThus, priorities should be established:Our article is the first attempt summarizing knowledge regarding AMR in both animal and human health sectors in IOC. This review clearly points out research and surveillance gaps and constitutes a tool for future activities to lead.NG performed the review, collected the data from literature, and wrote the first draft of the manuscript; EC, OB, and LFilleul revised and provided first feedback for the manuscript. All authors provided articles, have drafted, and revised the work critically for important intellectual contents and approved the final version. All authors agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We present VREX, a free open-source Unity toolbox for virtual reality research in the fields of experimental psychology and neuroscience.Different study protocols about perception, attention, cognition and memory can be constructed using the toolbox. VREX provides a procedural generation of (interconnected) rooms that can be automatically furnished with a click of a button. VREX includes a menu system for creating and storing experiments with different stages. Researchers can combine different rooms and environments to perform end-to-end experiments including different testing situations and data collection. For fine-tuned control VREX also comes with an editor where all the objects in the virtual room can be manually placed and adjusted in the 3D world.VREX simplifies the generation and setup of complicated VR scenes and experiments for researchers. VREX can be downloaded and easily installed from vrex.mozello.comThe online version of this article (doi:10.1186/s40359-017-0173-4) contains supplementary material, which is available to authorized users. Conducting both ecologically valid and at the same time highly controlled psychological and neuroscientific experiments is a notoriously difficult task, preventing it from widespread use . ControlHowever, 3D virtual reality environments are starting to provide a reasonable compromise between the expensive real-world and simplified computer screen experiments –4. Head Recently launched consumer grade VR headsets such as the Oculus Rift and HTC Vive allow 360° optical tracking and up to 16 square meters of movement space, while at the same time being affordable. These hardware breakthroughs in the last few years have made VR technology available for almost any lab studying experimental psychology. Also, major game engines such as Unity (Unity Technologies) and Unreal Engine  now have built-in VR support straight out of the box. However, this new research paradigm requires specialized knowledge in software and hardware technology in order to create immersive and presence-inducing virtual realities. In particular, knowledge of 3D modeling and texturing, game engine logic and scripting are all needed.Many psychology labs still lack these competences. The primary aim of the Virtual Reality Experiments (VREX) Toolbox is to help psychology researchers easily create experiments for virtual reality setups by providing an open-source software suites as an Unity add-on and standalone version  or incremental movement and turning. There is also an option to automatically move the participant on a previously defined path. This option can be set up in the 3D editor.Every time an experiment is run through VREX, a data log is created in the VREX system folder under Results. The following is logged by default: Participant ID, start time of the experiment, current environment, environment order, test type, score  and elapsed time. In case of change blindness and memory experiments additional parameters are stored, such as change count, changed object(s) and participants answer(s). Time is measured with millisecond precision.A more experienced user may want to modify or extend the functionalities of VREX. A collection of template scripts with basic elements in place to develop a custom experiment with new behaviours can be found under the assets folder in the Unity project. For example it is possible to create a script for tracking the user’s trajectory for spatial navigation tasks inside the environments, change the input scheme, add parameters that should be logged during an experiment or tweak the user interface for the participant.Different labs have different setups. The toolbox currently only works with the Oculus Rift DK2 and CV1 headsets due to the way the virtual camera is implemented. Other peripherals such as Leap Motion hand tracker or the HTC Vive VR headset can be introduced via the standard Unity interface.VREX is designed to allow easy creation of indoor VR environments and specific experiments. Although the software is far from being complete, it can already be used for practical research by novice programmers. Detailed examples of both change blindness and false memory experiment types are described below.Change blindness occurs when a person is unable to notice a big change in a scene, unless the scenario is presented numerous times or a hint is given . With VRFalse memory occurs when an individual remembers events that did not happen or were different in reality . StudyinVREX user interface is designed with very specific tasks in mind. This means that many options are limited and the user can not access the full potential of Unity through the toolbox menus. For general ease of use all key bindings are currently hardcoded and can not be easily remapped by the user, and this could be problematic for some types of experimental plans. Convenient access to such settings is where MazeSuite and WorldViz Vizard currently outperform VREX.Another limitation is that as VREX relies heavily on Unity 5, major future updates to the game engine may cause compatibility errors. This is common for all add-ons and recommended Unity version numbers should be noted when using the toolbox. For the standalone version of VREX this is not an issue.We have presented VREX, a Unity toolbox for VR research in psychology. By adding many powerful features and a simple user interface, the tool is designed to empower researchers with no or little prior knowledge in game engines to start working with VR. Together with the community we aim to develop the application further. The website of the VREX toolbox can be found on the following link: vrex.mozello.com.Project name: VREX: A UNITY® TOOLBOX FOR VR EXPERIMENTS.Project home page:http://vrex.mozello.com/.Archived version:https://drive.google.com/open?id=0B97-aac1_IQ5eEdiaW83TkZsa2c.Operating system: Microsoft Windows.Programming language: C#.Other requirements: Unity Engine 5 .License: Creative Commons BY-SA licence.Any restrictions to use by non-academics: none.


Previous studies have suggested that focusing an element can enhance the activation of the focused element and bring about a number of processing benefits. However, whether and how this local prominence of information interacts with global discourse organization remains unclear. In the present study, we addressed this issue in two experiments. Readers were presented with four-sentence discourses. The first sentence of each discourse contained a critical word that was either focused or unfocused in relation to a wh-question preceding the discourse. The second sentence either maintained or shifted the topic of the first sentence. Participants were told to read for comprehension and for a probe recognition task in which the memory of the critical words was tested. In Experiment 1, when the probe words were tested immediately after the point of topic shift, we found shorter response times for the focused critical words than the unfocused ones regardless of topic manipulation. However, in Experiment 2, when the probe words were tested two sentences away from the point of topic shift, we found the facilitation effect of focus only in the topic-maintained discourses, but not in the topic-shifted discourses. This suggests that the facilitation effect of focus was not immediately suppressed at the point of topic shifting, but when additional information was added to the new topic. Our findings provide evidence for the dynamic interplay between global topic structure and local salience of information and have important implications on how activation of information fluctuates in mental representation. The comprehension of a discourse involves constructing a mental representation of a discourse. As a discourse unfolds, the representation of the discourse is updated and keeping track of this information is essential for successful discourse comprehension. Given the limitation of cognitive resources, not all information in a discourse is fully attended to. Typically, salient or important information is more fully attended to than other information. One factor that marks the salience of information is linguistic focus.Who did Mary vote for?/Mary voted for [JOHN], JOHN supplies an answer to the preceding question. Hence, JOHN is focused (it was… who/that…), or by using focus-particles .Focus has often been defined as the most prominent information in a sentence . There a focused . BesidesIn the field of psycholinguistics, the effect of focus in online sentence/discourse comprehension has been an important issue that received considerable attention. Numerous studies have shown that focus can produce a number of processing benefits. Firstly, focused items receive more attention and are processed more quickly than unfocused items . For insThese studies on focus thus provide evidence that focusing an element can bring about a facilitation effect for the focused element. A general issue for these studies is that the effect of focus is always examined without the consideration of global discourse variables. In fact, one particular aspect for any natural discourse is that its subunits can form various structures, which can affect the activation of information in a discourse , 2014. GXiao Wang is a very kind man,” they may activate the memory nodes for Xiao Wang, kind, and man. This stage of laying mental foundations has been evidenced by the fact that comprehenders slowed down when reading the first sentence of a paragraph  throughout the passage. When there was a topic shift (shifted from the concert to the kitchen), this disadvantage disappeared. This was interpreted as suggesting that topic shifts suppressed the old discourse representation so that integration difficulty resulting from a change in the meaning of the ambiguous words disappeared.Apart from triggering readers to shift and build a new substructure, topic shifts can also produce a suppression effect on the old mental structure. Studies on anaphoric inferences and ambiguity resolution have suggested a suppression effect of topic shifts such that when encountering topic shifts, information related to the earlier topic is faded or is less available . For insThus, in light of the structure building framework and related evidence, topic shifts can be of particular relevance during discourse comprehension and most probably they would reduce the accessibility of information presented before the topic shifts. Given that focus produces a facilitation effect and topic shift produces a suppression effect, it is reasonable to speculate that these two internal components of discourse might interact with each other and affect discourse comprehension. Therefore, our first question was to examine whether there is interplay between these two factors during discourse comprehension.Focus structure and topic structure for Chinese has its peculiarity as well as general characteristics with other Western Germanic languages. For focus structure, Chinese uses more syntax and less phonology in focus realization compared with Western Germanic languages  . The praIf focus and topic shift interact to influence mental representations, a related question then arises as to at which point can the interaction be expected. As specified by the structure building framework, when encountering topic shifts, readers develop a new substructure and suppress the old one. Then readers move on pass the shifting stage and start to map incoming information onto the new substructure. Although topic shifts can produce a suppression effect on the content of the old topic , we do nTable 1 for an example stimulus).To address the issues mentioned above, we conducted two experiments in which a probe-recognition task was used to test the memory representation of the critical words. Four conditions were created depending on whether the critical word in the first sentence of each discourse was focused or unfocused and whether the second sentence of the discourse shifted or maintained the topic of the first sentence, namely: topic-shifted/focused, topic-shifted/unfocused, topic-maintained/focused, and topic-maintained/unfocused  to test whether an interaction between topic structure and information structure could be found when readers encountered a topic shift. In Experiment 2, the probe words were placed at the end of the fourth sentence of each discourse which was far away from the point of topic transition to test whether an interaction effect could be found when readers moved on past the shifting point.In Experiment 1, we aimed to test whether there would be a modulation effect of discourse topic on local information salience when readers encountered a topic shift. Thus, we test the accessibility of the critical words immediately after the presentation of the topic-shifted sentences. Based on previous findings suggesting that focusing an element leads to a facilitation effect on memory representation of the element , we expeSD = 2.2 years) volunteered to take part in the Experiment. All were native speakers of Chinese and were paid for their participation.Forty-eight college students  or maintained (topic-maintained condition) the topic of the first sentence. Then, two more sentences ended the discourse. These two sentences were identical across both conditions. Our definitions of topic were based on the commonly held beliefs about the features of topic in Chinese: (1) a topic is an entity that is introduced at the initial position of a sentence, (2) a topic is always followed by a comment . A topict(23) = -0.643, p > 0.05, mean ± SD = 3.35 ± 0.79; 3.42 ± 0.73; respectively for the topic-shifted and the topic- maintained condition].To control for discourse coherence as a confounding factor of topic manipulation, we conducted a rating study in which 24 participants who did not participate in the formal experiments reported here were asked to rate the coherence of the discourses on a five-point scale ranging from 1 (not coherent) to 5 (very coherent). Different conditions of the discourses were distributed to different lists so that each participant saw only one condition of a given discourse. On the basis of the rating scores, 40 sets of discourses were selected for the main study so that the coherence rating scores were matched between the topic-shifted and the topic- maintained condition . The trend was for the focused condition to yield shorter reaction times than the unfocused condition. Neither the main effect of topic shift nor the interaction between topic shift and focus was significant . A parallel analysis was conducted on the errors. A binomial family was used because of the binary nature of the responses. None of the fixed effects were significant .For reaction times, we found a significant main effect of focus . The trend was for the focused condition to yield shorter reaction times than the unfocused condition. More importantly, there was a significant focus by topic shift interaction . Planned comparisons showed that participants were faster to recognize a probe word when the word was in focus position than when it is outside the scope of focus in the topic-maintained discourses , but not in the topic-shifted discourses .For reaction times, there was a significant main effect of focus [Z = -2.42, p < 0.05). Participants were more able to make accurate responses when the probe word was focused than when it was unfocused. Neither the main effect nor the interaction between focus and topic shift were significant .A parallel analysis was conducted on the errors. A binomial family was used because of the binary nature of the responses. There was again a significant main effect of focus (These results again showed a processing advantage for the focused over unfocused words: readers were faster and more accurate at identifying the focused probe words. More importantly, an interaction effect between focus and topic shift was found, which clearly showed that the facilitation effect of focus was eliminated by the suppression effect of topic shift when more information was attached to the new topic. Specifically, focusing an item facilitated information retrieval only in the topic-maintained discourses, but not in the topic-shifted discourses. This provides clear evidence that topic shifts modulate the facilitation effect of focus during online construction of discourse representation.The goals of the present study were to determine whether and how the facilitation effect of focus was modulated by discourse structure. The results of Experiment 1 showed that the facilitation effect of focus was not immediately suppressed when upcoming sentences indicated a change in topic. However, in Experiment 2, we found that after reading additional sentences, the interaction between topic shift and focus appeared. These results suggest that the mental representation of a discourse involves dynamic interplay between the organization of discourse topic and the prominence of local information.Across experiments, we found that focusing an element produced a facilitation effect. Focused probe words were identified more quickly and more accurately than unfocused words. This agrees with previous studies that words were consistently better remembered when they had been focused . This faThe finding of the facilitation effect of focus in Experiment 2 is noteworthy. In this experiment, the focus effect was tested with three intervening sentences that separated the manipulation of focus and the probe word. This suggests that the facilitation effect of focus is not transient, but can persist into memory as discourse unfolds. In previous studies, the effect of focus was mostly tested at the point of focus manipulation or immediately after it . Thus, tThe crucial hypothesis tested in the present study was whether and how focus interplayed with topic shift to influence mental representation. In Experiment 1, no interaction was observed when the probe words were placed immediately after the topic shift sentences. In Experiment 2, however, we found an interaction between focus and topic structure: The facilitation effect of focus was only found in topic-maintained condition, but not in topic-shifted condition. These results suggest that discourse structure can override local information salience and agree with previous findings that the processing of topic shifts can produce a suppression effect .Note that in Experiment 2 in which the interaction was found, the probe words were placed after two more sentences which elaborated on the new topic. According to the structure building framework, when encountering a topic shift, comprehenders shift to build a new substructure for the new topic and once the foundation for the new topic is laid, the mapping stage begins when readers gradually develop the representation by mapping on new information . Thus, oThe observed suppression effect of topic shifts on focus could be due to the allocation of cognitive resources. According to the structure building framework, the introduction of a new topic can trigger readers to shift and build a new substructure while at the same time suppress the structure of the old topic . HoweverThe presence and absence of the interaction effect observed in the current study have important theoretical implications: The mental construction of discourse representation involves dynamic interplay between discourse organization and local information structure. How the mental representation of a discourse is constructed has been a central issue for most theories of discourse comprehension. There are theoretical descriptions on the role of discourse organization such as topic shifts  and evena moment later vs. 1 year later as in in the classroom vs. went to the bar as in Shifts in topic is only one kind of shifts and there are also other shifts in a discourse that can trigger readers to shift from current representation and build a new substructure , such asTo conclude, in two experiments, we found that the facilitation effect of focus could still function when readers encountered topic shifts, but could be suppressed when additional information was added to the new topic. These results suggest that discourse organization can override local information structure. Our findings provide evidence for a dynamic relationship between discourse topic organization and local salience of information and highlight the need to consider their interaction for the model of discourse comprehension.This study was approved and carried out in accordance with the recommendations of the Institutional Review Board of the Institute of Psychology, Chinese Academy of Sciences with written informed consent from all subjects. All participants provided written, informed consent before taking part in our experiments.XY: experiment design, manuscript writing. XZ and RC: data collection, data analysis. CW: data analysis, manuscript writing. WL: manuscript writing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Rz1 is entirely embedded in the +1 reading frame of Rz, the genes were disembedded before using random mutagenesis to construct a library of lysis-defective alleles for both genes. Surprisingly, most of the lysis-defective missense mutants exhibited normal accumulation or localization in vivo, and also were found to be normal for complex formation in vitro. Analysis of the distribution and nature of single missense mutations revealed subdomains that resemble key motifs in established membrane-fusion systems, i.e., two coiled-coil domains in Rz, a proline-rich region of Rz1, and flexible linkers in both proteins. When coding sequences are aligned respective to the embedded genetic architecture of Rz1 within Rz, genetically silent domains of Rz1 correspond to mutationally sensitive domains in Rz, and vice versa, suggesting that the modular structure of the two subunits facilitated the evolutionary compression that resulted in the unique embedded gene architecture.Coliphage lambda proteins Rz and Rz1 are the inner membrane and outer membrane subunits of the spanin complex—a heterotetramer that bridges the periplasm and is essential for the disruption of the outer membrane during phage lysis. Recent evidence suggests the spanin complex functions by fusing the inner and outer membrane. Here, we use a genetics approach to investigate and characterize determinants of spanin function. Because  For Caudovirales, host lysis is essential for the release of phage progeny. For Gram-negative hosts, lysis is a three-step process requiring the function of three types of phage-encoded proteins: holins, endolysins, and spanins , and the endolysin degrades the peptidoglycan (PG), have been studied intensively for decades, and were considered necessary and sufficient for host lysis. However, recently we have found that lysis requires the disruption of the outer membrane (OM) by phage-encoded spanins  in Rz are involved in these bonds, as is the single Cys residue (C29) in Rz1  (Berry eRz1 was predicted to be largely unstructured due to its high proline content (10 prolines in the 40 aa mature Rz1). By contrast, the periplasmic domain of Rz was predicted to be highly structured, dominated by two coiled-coil helical domains  . When appropriate, ampicillin  and kanamycin  were also added. Growth and lysis of cultures were monitored by A550 as a function of time, as described previously  was added to the final concentration of 1 mM for plasmid induction. The pRE plasmid is a pBR322 derivative, which has the lambda late promoter pR’ located upstream of the Rz or Rz1 start site. To activate pR’, the antiterminator Q is supplied in trans by either the induced prophage or the pQ plasmid.The bacterial strains, bacteriophages, and plasmids used in this study are described in Rz  was used. The Rz1 gene is inactivated on the pRz plasmid by a nonsense mutation that is silent in Rz . OligonuKpnI and BamHI for Rz, and BamHI and HindIII for Rz1. The gel-purified, doubly digested, fragments were ligated into the pRE vector using T4 ligase, and transformed into XL-1 Blue cells. After overnight incubation at 37°, the transformants were pooled by slurrying, and plasmid DNA was extracted using the Qiagen spin miniprep kit. MC4100 (λ) lysogens carrying the nonsense alleles amQ100 Rz or amW38 Rz1 were transformed with the mutagenized plasmid pool of Rz or Rz1, respectively. To assess the frequency of mutation, 10 random colonies from each library were tested for lysis defects in liquid culture, and their spanin genes sequenced. Six Rz plasmids and four Rz1 plasmids did not complement the lysis-deficient phenotype. Of the remaining clones, three of the latter had missense changes that did not abrogate lysis phenotype. To enrich for lysis-defective mutants, a plasmid retention method was used. Colonies on the transformant plate were collected by slurring with LB, diluted and inoculated into 25 ml of LBM and appropriate antibiotics at an initial A550 ∼0.5, induced for lysis as described above. At 15 min past the normal lysis time (∼50 min), the culture was centrifuged at 4000 rpm for 5 min to harvest the nonlysed, Mg++-stabilized, spherical cells, the terminal phenotype of spanin-defective lysis. The harvested culture was carefully washed once with LBM before extracting plasmid DNA using a miniprep kit. Plasmids from the enriched mutant pools of Rz and Rz1 plasmids were used to transform λ lysogens of amRz and amRz1, respectively. Single colonies were picked and individually screened for a lysis defect by thermal induction in 5 ml LBM. Lysis-defective clones were sequenced by Eton Biosciences .Mutagenized PCR products were digested with Rz or Rz1 gene products was assessed by Western blotting of TCA precipitates as described previously . From this family, we selected six representatives of the i- and o-spanin C-terminal domains.https://cpt.tamu.edu/galaxy-pub/).Using Python and BioPython , we deveStrains and reagents are available upon request.amRz or lambda amRz1, respectively . The spanin genes were then subjected to random PCR-based mutagenesis in the context of these complementing plasmids, subcloned into a fresh vector, and transformed into a host carrying the amRz or amRz1 lysis-defective prophage. The pooled transformant libraries of pRz and pRz1 were selected for lysis-defective alleles by plasmid retention , and then screened individually for lysis after induction. A total of 266 pRz and 115 pRz1 lysis-defective clones were sequenced, of which 131 Rz and 79 Rz1 genes had single point mutations. The remaining lysis-defective Rz and Rz1 mutants had either multiple point mutations (48 Rz and 19 Rz1) or frameshifts, including both base insertions and deletions (87 Rz and 17 Rz1). Overall, the nucleotide changes were 42% transition and 58% transversion mutations, as expected for this type of mutagenesis, which indicates that the full range of possible mutations at each codon was equally accessible (Plasmids carrying Rz or Rz1 (pRz and pRz1) under the native pR’ late promoter were used to effect independent mutational analysis on each gene. Both pRz and pRz1 complemented the lysis defect of lambda cessible .For convenience and to emphasize structural features, the Rz sequence  is subdiRz alleles obtained by random mutagenesis, 82 alleles were nonsense changes in 34 different positions scattered throughout the periplasmic domain . Estimates of the Rz-Rz1 complex approximate the span of the periplasm: 170 residues (130 Rz and 40 Rz1) × 0.15 nm/residue, assuming α-helical structure, equals 25.5 nm . The nonsense mutations were distributed across the entire length of Rz, except for the extreme C-terminal region, suggesting that this domain of Rz is not essential  , which cause changes in the bulk of a hydrophobic side chain. This suggests that these residues are involved in interhelical packing of two parallel CC1 domains. One stretch of 10 residues, L64–A73, was the most mutationally sensitive region of Rz, with 13 missense alleles mapping to seven of the 10 positions, including five missense mutations at two “a” positions. Within this stretch, L64 and D65 were the most mutationally sensitive positions, with six missense alleles conferring a lysis defect. Although polar or charged residues are tolerated in a and d positions, it is unusual for Asp to be in the “a” position of a parallel helical interface  suggests this segment of CC2 interacts with Rz1.i.e., XCX or CXX) should be a good indicator of essential helical secondary structure. We used the One SNP Away tool  identified by the screen. Additionally, no prolines were identified in 22 such codons from positions 92–133. Conversely, proline substitutions were isolated in nine of 13 possible positions between 64 and 93. Similarly, there were two of five possible proline substitutions identified within an eight-residue stretch of CC2. Assuming proline substitutions obtained by this selection serve as an indicator of essential helical structure, the essential Rz helices span from position ∼60 to ∼90 and ∼130 to ∼140. Using JPRED4, predictions based on primary structure find longer helices, from 27 to 87, and 121 to 150 . The only L2 lysis-defective mutants isolated, R91P and L93S, are within a predicted β-strand near CC1 , we hypoRz1 mutants, 79 had single point mutations; the rest had two or more mutations or frameshift mutations, and were excluded from analysis. Initially, the degree of saturation was thought to be less than that obtained for Rz, because the 36 nonsense mutations were found in only 10 of the 19 codons for which a single nucleotide change could yield a stop codon. However, four such codons were in the CTD of Rz1, beyond the last nonsense mutation (W46X) that was obtained in the lysis-defect selection. This raised the possibility that the extreme C-terminus of Rz1 is dispensable. This notion was confirmed when each of these six distal sites was converted to nonsense codon by site-directed mutagenesis, and tested for their function. None were found to have a lysis defect  (5) stretch  , especia stretch , residuefunction .5 stretch of Rz1, p15 has a proline stretch (PPAPPP). Like Rz1, the proline-rich motif in p15 is important for membrane fusion, and the fusion reaction is not sensitive to changes in the third position. Evidence has been presented that the role of polyproline helices in membrane fusion is to promote exposure of hydrophobic side chains of neighboring regions  protein . Most of the allele products accumulated to wild-type levels, indicating that the lysis defect is not due to protein synthesis or stability  or smears. Surprisingly, in the presence of Rz1, RzG143R is stabilized and RzL72F is unstable. We interpret this as evidence that Rz and Rz1 form a complex in vivo during the late gene expression period, and mutant products are stabilized, in the case of RzG143R, or destabilized (in the case of RzL72F) by conformational changes associated with complex formation. The accumulation of Rz1P33L, Rz1P35H, and Rz1P36Q appeared diminished, but these mutations fall within the epitope used for immunodetection, so it is unclear if accumulation is actually affected. Importantly, almost all Rz and Rz1 mutant products appear to accumulate exclusively as disulfide-linked homodimers (double asterisks), with the exception of Rz1W46C, which likely is largely blocked in an internal disulfide-bonded state , suggesting these mutants are defective in complex formation in vivo. Since abrogating negative charge at 150 blocked complex formation in vitro, we examined covariance at the C-termini of Rz and Rz1 equivalents in other lambdoid phages  identified by the screen, and three alleles created by site-directed mutagenesis . The only CC2 mutant that did not copurify with Rz1-His was Rzwith Rz1 . The RzQd phages . This ans at R59 . To addrin vivo . This stRz1-His. Similar to Rz CC1 mutants, all products tested from Rz1-His mutants exhibited parental coimmunoprecipitation with Rz .To screen Rz1 mutants for interaction defects, we created nine mutant alleles of  with Rz , suggestAs noted above, Rz1 could be truncated to position 55 without loss of function . This isRz and Rz1, which encode the subunits of the two-component spanin of phage lambda. The selection, based on a near-saturation selection for mutants that abrogated lysis, identified mutants that inactivate either Rz, the i-spanin, or Rz1, the o-spanin product encoded by the embedded gene. The selections were done on artificially disembedded genes, but, despite this architectural segregation, both genes exhibited mutational clustering in regions that corresponded to mutationally silent regions of the out-of-frame gene. These mutationally silent regions were tested by site-directed mutagenesis, and found to be replaceable by simple repeated linker sequences, thereby establishing that both Rz and Rz1 have flexible linker domains between the mutationally sensitive regions. Surprisingly, the mutants that were identified by the selection, despite the loss of lytic function, uniformly maintained the ability to form spanin complexes in vitro and in vivo, and most were not defective in the accumulation of gene products. The simplest interpretation is that these mutations blocked a step downstream of periplasm-spanning complex formation. We have proposed that the complex, once liberated from the constraints of the intact PG layer, undergoes oligomerization, and then causes fusion between the IM and OM (in vitro fusion system for the spanins.Here, we report the first genetic analysis of an embedded gene pair, of which both genes are required for the same biological function: in vivo by using high resolution cryo-EM, and super-resolution microscopy techniques.The Rz-Rz1 spanin system, with its powerful genetics, may be a useful platform for the study of membrane fusion in general. For example, because the spanin-mediated fusion event would have to occur within a 25 nm space between membranes at a precise time in the infection cycle, it may be possible to capture the hemifusion state www.g3journal.org/lookup/suppl/doi:10.1534/g3.116.037192/-/DC1.Supplemental material is available online at Click here for additional data file.Click here for additional data file.


There is an error in affiliation 1 for authors Jiaoyu Shan, Binbin Zhao, Zhao Shan, Jia Nie, Rong Deng, Andy Tsun, Kawing Lui, Rui Liang. Affiliation 1 should be:CAS Center for Excellence in Molecular Cell Science, CAS Key Laboratory of Molecular Virology and Immunology, Unit of Molecular Immunology, Institut Pasteur of Shanghai, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; University of Chinese Academy of Sciences, Shanghai, China


Phlebia radiata Cel 26 (a mutant with low cellulose degrading ability) were compared with pure pectinase treatment with regard to mechanical properties of the produced fibre/epoxy composites. For field retting a classification of the microbial evolution (by gene sequencing) and enzyme profiles were conducted. By phylogenetic frequency mapping, different types of fungi, many belonging to the Ascomycota phylum were found on the fibres during the first 2 weeks of field retting, and thereafter, different types of bacteria, notably Proteobacteria, also proliferated on the field retted fibres. Extracts from field retted fibres exhibited high glucanase activities, while extracts from P. radiata Cel 26 retted fibres showed high polygalacturonase and laccase activities. As a result, fungal retting gave a significantly higher glucan content in the fibres than field retting (77 vs. 67%) and caused a higher removal of pectin as indicated by lower galacturonan content of fibres (1.6%) after fibres were retted for 20 days with P. radiata Cel 26 compared to a galacturonan content of 3.6% for field retted fibres. Effective fibre stiffness increased slightly after retting with P. radiata Cel 26 from 65 to 67 GPa, while it decreased after field retting to 52 GPa. Effective fibre strength could not be determined similarly due to variations in fibre fracture strain and fibre-matrix adhesion. A maximum composite strength with 50 vol% fibres of 307 MPa was obtained using P. radiata Cel 26 compared to 248 MPa with field retting.Classical field retting and controlled fungal retting of hemp using  Cannabis sativa L.) are considered a particularly promising renewable raw material for production of high quality reinforcement of composite materials due to their high stiffness and strength to weight ratio  epidermis, (b) several layers of primary fiber cells , (c) layers of secondary cells that are smaller with a thinner secondary wall and (d) parenchyma cells. In the bast layer the primary and secondary fiber cells are bonded together by a pectinaceous matrix, the middle-lamella. The retting process is used to separate these cellulosic fibres by degrading the parenchyma cells, as well as in the middle lamella between fibres , processed into bales and finally separated into fibres and shives. The retting process is uncontrolled microbial wise, which can result in loss of fibre strength due to cellulase activity secreted by proliferation of native microorganisms on the hemp stems  and a pure enzymatic retting approach (pectinase enzymes) based on mechanical properties of the obtained hemp fibre reinforced composites. Genetic identification and environmental scanning electron microscopy (ESEM) observation of bacteria and fungi was carried out to identify their abundance on hemp fibre surfaces. Enzymatic and microbial characteristics were linked to changes in chemical composition of fibres and mechanical properties of the fibres and fibre/epoxy composites. Finally the changes in microbial community, fibre composition and fibre mechanical properties were followed versus the retting duration. The correlation between microbial retting duration, chemical composition of fibres, and mechanical properties of produced composites was thereby established.The objective of this study was to compare classical field retting with a pure microbial retting approach (Cannabis sativa L.), variety USO-31, was sown on May 5th 2013  by Bafa Neu GmbH and harvested on Sep 6th 2013  to avoid contamination with wild fungi. The fungal retting was conducted for 7, 14 and 20 days in 1 L Erlenmeyer flasks (15 g hemp stems of 15 cm in length per flask) at 28 °C. P. radiata Cel 26 was obtained from the Swedish Agricultural University, Uppsala, Sweden  placed into 2 mL Eppendorf tubes was extracted directly for obtaining genomic DNA using PowerBiofilm™ DNA Isolation Kit  according to the manufacturer’s instructions. Polymerase chain reaction (PCR) amplification for both bacterial and fungal DNA was carried out using a C1000™ thermo-cycler . Each DNA sample (1 μL) was used as template in the PCR reactions . Purified plasmids were sequenced using the 27F primer for bacteria and the ITS4 primer for fungi synthesized by the company Macrogen Europe . The identified sequences are published in the EMBL Nucleotide Sequence Database with the accession numbers LT622055-LT622085 outlined in Tables PCR products were purified using GFX PCR DNA and Gel Band Purification Kit . Cloning was performed using the pJet1.2/Blunt cloning vector (50 ng/µL) and T4 DNA ligase (5 U/µL). Ligation was carried out according to the manufacturer’s instructions  and the ligated product was used for transformation of electro-competent A GenBank nucleotide database search was conducted using the BLAST algorithm  to determine the closest relative of partial 16S gene sequences  supplemented with 0.1% (v/v) Tween 20 and 0.25 mM dithiothreitol in a glass tube at 0 °C and shaken at 120 rpm for 1 h. For each sample, the crude enzyme extracts were concentrated until the volume was reduced to 3.0 mL . Glucanase, polygalacturonase, galactanase and xyloglucan (XG)-specific endoglucanase activities were determined by measuring formation of reducing ends. The substrates used were for glucanase 10 g/L carboxymethyl cellulose  required to oxidize 1 µmol ABTS per minute under the assay conditions.Laccase activity was measured by monitoring the oxidation of Diammonium 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) by the enzyme extracts at 420 nm  through a 1-mm screen. Ground samples were extracted in a Soxhlet apparatus  operated at 10–15 kV (Fernando and Daniel Samples (5 mm long × 2 mm wide) were cut from bast fibre strips under a stereo microscope and fixed in 3% v/v glutaraldehyde, +2% paraformaldehyde in 0.1 M Na-cacodylate buffer (pH 7.2). After fixation, the samples were dehydrated in aqueous ethanol using: 20, 40, 60, 80, 90 and 100% for 15 min in each solution. Subsequent dehydration was performed in 33, 66 and 100% acetone in ethanol before samples were critical point dried using an Agar E3000 critical point dryer  with liquid CO®, Buckinghamshire, United Kingdom). The gauge length was 10 mm and the displacement rate 0.5 mm/min (corresponding to a strain rate of 5% min−1). Tensile testing was performed on 20 specimens for each treatment. The cross-sectional area (Af) was determined from measured fibre mass, length and density [1.50 kg/dm3  of mass in the range of 5–20 mg were used for tensile testing  and its amine hardener (Aradur® 3489) both supplied by Huntsman Corporation, Houston, USA, were mixed at a 100/28 mass ratio and degassed in a vacuum oven. The setup for vacuum infusion and mould processing has been described  in the range 0–0.70 g/g composite. When Wf was below 0.30, the composite specimens had irregular surfaces and their density (ρc) was determined by the buoyancy method (Archimedes principle) using water as displacement medium. Otherwise the composite specimens had flat surfaces and ρc could be calculated based on their dimensions . The volumetric composition of composite samples was determined as previously described  was assumed to increase linearly versus the fibre volume content (Vf) based on a proportionality constant (porosity factor) αpf (Eq. pm) was assumed to be zero since no air bubbles were observed in the cured epoxy matrix by ESEM  with a load cell of 10 kN was used. Strain measurements were conducted using two extensometers and the displacement rate was 1 mm/min (corresponding to a strain rate of 2.5% min−1). Based on measured stress–strain curves, composite stiffness (Ec)  and UTSc were determined. For each treatment, at least ten specimens with varied fibre content were tested. The UTS and stiffness curves of the composites were compared by assessing the responses by linear regression comparison within the tested range of Vf. Effective fibre stiffness (Ef) in the composites were thereby determined from Eq. m).k is the slope of the regression line, i.e. of Ec vs. Vf. The effective fibre strength cannot be calculated similarly due to effects of fibre-matrix adhesion and Weibull distribution of the fibre strength  , particularly during field retting was noted presumably due to a lack of lignin degrading enzymes produced by the microbial retting flora. The increase can also be due to microbial biomass formation and that the degradation rate of lignin is much lower than that of other components i.e. pectin.The changes in chemical composition of fibres during field retting Table  were accESEM microscopy observation showed that neither bacteria nor fungal hyphae were present on the hemp fibre surface before field retting Fig. a. After α-ß-γ-proteobacteria and Bacteroidetes  and that of γ-proteobacteria decreased to 38%. Bacteroidetes were identified after 14 and 20 days of retting with 8 and 25% of the bacteria, respectively.16S rRNA from samples with different field retting durations was analyzed for bacterial diversity resulting in a total of 81 different sequences Table . Sequences Table . In the es Table . From viMassilia aurea after 7 and 14 days retting with frequencies of 14 and 25%, respectively, decreasing to 16% after 20 days. Rhizobium soli constituted 14–17% of the bacterial diversity during the whole retting period. The Pseudomonas fraction of the bacteria  decreased from 28% after 7 days to 16% after 14–20 days. The Shigella sonnei part of the bacteria decreased from 29 to 0% after 20 days. In addition minor but increasing percentages (0 → 3–10% of bacteria) were found for Erwinia aphidicola, Chryseobacterium scophthalmum, Hymenobacter sp. and Pedobacter sp.The most frequent phylotype was Figure Basidiomycota phylum and to six genera within the Ascomycota phylum  and Alternaria infectoria (9 → 20%). However C. uredinicola dominated initially contributing 50% of the diversity in the fungal community after 7 days. Low and decreasing frequency percentages of Cryptococcus  and Sphaerulina amelanchier were observed with 10 and 7% of the fungal community after 0 days, respectively.Sequences of the fungal community derived from the ITS gene corresponded to five genera distributed within the um Table . BetweenBasidiomycota after 14 days of retting, presumably as a result of faster growth of Ascomycota fungi. The analysis also indicates that many well-known fungal species were present initially with 14 blue labels showing a high biodiversity. With time, the diversity of the fungal community decreased as evident from the decrease to only four orange labels after 20 days represented by Alternaria, Stemphylium and Cladosporium  up to 20 days . Thereby the more efficient pectin degradation by fungal retting  of 2.7 GPa. By comparison of the slopes (k) of the lines of Ec versus Vf, of composites with differently treated fibres, it can be seen that composites with P. radiata Cel 26 retted fibres, irrespective of retting duration, had much higher stiffness and strength than with field retted fibres  and strength (UTSf) were estimated by linear regression within the Vf range tested  for 7-, 14-, and 20-days retted samples using P. radiata Cel 26, respectively. In contrast, 20 days field retted fibres showed that Ef was only 51 GPa. If P. radiata Cel 26 retting was replaced with pectinase treatment higher stiffness of 74 GPa was obtained. The results demonstrate thereby that P. radiata Cel 26 retting produced better fibres than field retting.The effective fibre stiffness (Eed Table . It was pf) of composites with differently treated fibres were determined by Eq. . For composites made with P. radiata Cel 26 retted fibres, the composite porosity was found to decrease vs. the retting duration due to smaller fibre bundles in general obtained with more defibration   than composites made with field retted hemp fibres (248 MPa). However a pure pectinase approach resulted in even higher composite strength (325 MPa) (Liu et al. Field retting and


The emerging understanding of molecular changes in a wide range of brain tumours has led to a significant shift in how these tumours are diagnosed, managed and treated. This article will provide a hands-on overview of the relevant biomarkers and their association with newly defined biological tumour entities. The traditional approach to diagnose brain tumours is the examination of a histological specimen. Conventionally, pathologists make the histological diagnosis by assessing morphological features of cellular atypia, variation of nuclear size (anisonucleosis), shape (pleomorphism), mitotic activity, cell density, characteristic architectural patterns, vascular properties, and cell necrosis. With the exception of histone-mutant diffuse midline gliomas, these cytoarchitectural characteristics are taken into account when assigning a malignancy grade to the tumour according to the WHO classification scheme . This isThe focus of this review article is limited to primary, intrinsic brain tumours occurring in adults. Each molecularly defined diagnostic group, i.e. IDH-mutant astrocytomas or oligodendrogliomas, histone-mutant gliomas, BRAF-mutant gliomas and ependymomas will be discussed separately with an emphasis on the specific molecular alterations in each group and their clinical relevance.IDH1, IDH2) in 2008 was a major breakthrough towards molecular biomarker-driven diagnosis of adult gliomas  show a comparatively good prognosis [Ependymomas (and their variants subependymoma and myxopapillary ependymoma) are glial neoplasms predominantly arising within, or in the vicinity of, the ventricles and the spinal cord. The molecular classification of ependymomas has proven more meaningful for prognostication than the histological grading , 58, andrognosis . As all rognosis . The ST-rognosis , the latrognosis .Fig. 3DiIt is recommended that outside clinical trials, the WHO grading of ependymal tumours should not be used for the treatment decision . Insteadhttp://www.molecularneuropathology.org). In those entities, e.g. Histone H3-, or IDH-wildtype glioblastomas, where no driver mutations are found, identification of molecular heterogeneity and characterisation of molecular subgroups in histologically similar tumours will help in the design of novel, more effective targeted therapies. The main clinical relevance of identifying specific (epi) genetic alterations in each tumour is in their potential to serve as target for inhibitor drugs, or for the development of therapies, such as immunotherapy with vaccines aiming at the destruction of cells expressing a mutant protein [Many of the tumour entities described in this review are now defined by the presence of a mutation that serves as a biomarker, and there is now a consensus that certain brain tumour types should be diagnosed according to distinct biomarker profiles rather than histological features alone. It is striking, that the histological grade in some entities has become subordinate to the molecular profile, e.g. in ependymomas  and poss protein .


Introduction. Retrograde pedal access has been well described in the literature as a secondary approach for limb salvage in critical limb ischemia (CLI) patients. In this manuscript we are presenting a case where retrograde tibiopedal access has been used as a bail-out procedure for the management of superficial femoral artery (SFA) intervention complications. Procedure/Technique. After development of a perforation while trying to cross the totally occluded mid SFA using the conventional CFA access, we were able to cross the mid SFA lesion after accessing the posterior tibial artery in a retrograde fashion and delivered a self-expanding stent which created a flap that sealed the perforation without the need for covered stent. Conclusion. Retrograde tibiopedal access is a safe and effective approach for delivery of stents from the distal approach and so can be used as a bail-out technique for SFA perforation. Retrograde pedal access has been well described in the literature as a secondary approach for limb salvage either in critical limb ischemia (CLI) patients after failure of antegrade femoral approach or in patients who are not candidates for the antegrade approach. In this manuscript we are presenting a case where retrograde tibiopedal access has been used as a bail-out procedure for the management of superficial femoral artery (SFA) intervention complications.A 76-year-old female with history of diabetes mellitus, hypertension, coronary artery disease, and smoking was complaining of intermittent severe claudication in her left leg with a Rutherford classification of 3. Left leg ABI was 0.61.Preintervention angiogram showed totally occluded mid SFA that reconstitutes to the distal SFA, which was patent as shown in Peripheral arterial occlusive disease (PAD) is a common and debilitating disease that affects an estimated 10–12 million people in the United States . HoweverRetrograde tibiopedal access is a safe and effective approach for delivery of stents from the distal approach and so can be used as a bail-out technique for SFA perforation. Physicians who are involved in peripheral intervention will require having a full understanding of this approach and acquiring the necessary skills to perform it safely and effectively.


Midbrain Ventral Tegmental Area (VTA) has long been known for its pivotal role in mammalian reward system. A growing body of literature is now focusing on identification of the dominant element modulating neural populations of VTA, leading to Operant conditioning, a well-known mechanism by which a reward following a given stimulus can underscore a specific behavior, commonly termed as reinforcement learning.We focus on the recent findings from Yoo et al. , who impTo identify the presumed glutamate neurons projections to the VTA, they sterotactically infused an Adeno-associated viral vector expressing Channelrhodopsin-2 (ChR2) fused with a red fluorescent reporter. After pretest control measures addressing whether the target identifier was properly injected into dopaminergic regions in the PPTg, they put mice whose brain's VTA areas were inserted with fluorescent ChR2 and those inserted with fluorescent marker alone, for behavioral measurements on the nose poke assay and two bottle tests  increase dopamine release in VTA  to activate the reward pathway, possible simultaneous increase in the other neurotransmitter was not measured. Besides, both dopamine and glutamate release were shown to mediate activation of the reward system. Moreover, the study mainly focuses on the dominancy of afferent glutamate projections to the VTA, however, taking these combinatorial neural populations into account, these afferent projections might essentially activate glutamate-dopamine populations, instead of mere dopamine neurons, therefore, activating the reinforcement learning through both dopamine and glutamate release.In conclusion, although a great number of former studies has emphasized on the dominancy of one specific neurotransmitter underling operant conditioning process, accumulated evidence considering these combinatorial neural populations is now suggesting that further illustrations on compartmentalization of neural populations of VTA and an evaluation on their level of involvement in reinforcement-based behavior is required.MM drafted the manuscript and proposed the idea. FR made critical review of the subject and helped drafting the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AJR and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


In the original publication  two Goog3. Sherstyuk VP, Sarapulova OO, Shvalagin VV. Luminescent hybrid nanocomposites and prospects of molecular and nanophotonic systems in modern packaging and printing. Repino, St. Peterburg, Russia: Book of Abstracts of the 3-International Symposium “Molecular Photonics”; 2012. p. 79.https://scholar.google.com.ua/scholar?hl=uk&q=Luminescent+hybrid+nanocomposites+and+prospects+of+molecular+and+nanophotonic+systems+in+modern+packaging+and+printing&btnG=http://onlinereg.ru/mph2012/Molecular_photonics2012_Abstracts.pdf#page=7913. Sarapulova O Kyrychok T, Sherstiuk V, Orlov A. Modern printing technologies for micro- and nanoelectronics. Proceedings of IEEE XXXIII International Scientific Conference Electronics and Nanotechnology (ELNANO) April 16–19, 2013 Kyiv, Ukraine 2013: 151–155.https://scholar.google.com.ua/scholar?q=Sarapulova+O+Kyrychok+T%2C+Sherstiuk+V%2C+Orlov+A.+Modern+printing+technologies+for+micro-+and+nanoelectronics.&btnG=&hl=uk&as_sdt=0%2C5


Traditional imaging techniques are quite limited for the study of the relationship between blood vessels and lymphatic vessels. Therefore, a new imaging technique is required based on blood vessel and lymphatic endothelial-specific molecular markers. In this short report, vascular molecular markers are reviewed and a new molecular imaging technique for blood vessel and lymphatic co-staining is introduced.


The microstructure of the lead-free piezoelectric nanofibers was observed by field emission scanning electron microscope (FE-SEM) and the orientation was analyzed by fast Fourier transform (FFT) images. X-ray diffraction (XRD) analysis confirmed that the phase was not changed by the electrospinning process and maintained a perovskite phase. Polarization-electric field (P-E) loops and piezoresponse force microscopy (PFM) were used to investigate the piezoelectric properties of the piezoelectric nanofibers, according to the degree of alignment—the well aligned piezoelectric nanofibers had higher piezoelectric properties. Furthermore, the output voltage of the aligned lead-free piezoelectric nanofibers was measured according to the vibration frequency and the bending motion and the aligned piezoelectric nanofibers with a collector rotation speed of 1500 rpm performed the best.Flexible lead-free piezoelectric nanofibers, based on BNT-ST (0.78Bi On the other hand, flexible piezoelectric polymers such as polyvinylidenefluoride (PVDF) and P(VDF-trifluoroethylene(TrFE)), are poor in piezoelectricity and some other ways to improve the piezoelectric properties of PVDF nanofibers were studied [In the field of wearable electronic devices, there is a strong demand for portable smart devices that are not bulky and do not have to be too frequently charged. Thus, producers of wearable electronic devices are striving to create electronic devices that meet this demand. Piezoelectric modules are attractive as an emerging solution for this issue because they convert mechanical energy to electrical energy and harvest energy from motions of the human body ,2. For t studied ,5,6.A study on improving piezoelectric properties of piezoelectric nanofibers was also done using the alignment process of electrospinning ,8,9. Par0.5Na0.5TiO3-0.22SrTiO3)/PVDF-TrFE composites were fabricated by the electrospinning method and the effects of the alignment degree of the piezoelectric nanofiber were investigated according to the rotation speeds of the collector during electrospinning. The microstructure of the collected nanofibers was analyzed by field emission scanning electron microscope (FE-SEM) and the orientation was analyzed by fast Fourier transform (FFT) images. The crystal phase of the piezoelectric nanofibers was analyzed by X-ray diffraction (XRD) analysis and the piezoelectric properties were analyzed by Polarization-electric field (P-E) loops and piezoresponse force microscopy (PFM). The output voltage of the piezoelectric nanofiber modules according to the bending motion and vibration frequency was measured by an oscilloscope and the aligned piezoelectric performed the best.In this study, piezoelectric nanofibers of BNT-ST. The BNT-ST ceramic particles were controlled to less than 100 μm by mash control. To prepare an electrospinning precursor solution, PVDF-TrFE copolymers with mole fractions of 75% VDF and 25% TrFE , N,N’-dimethylformide  and acetone  were vigorously mixed at room temperature for 24 h. The weight ratio of each substance in the precursor solution was as follows: PVDF-TrFE:acetone:DFM = 2:5:5. After stirring, the BNT-ST ceramics powders were added to the precursor solutions in an amount of 60 wt % and the solutions were vigorously stirred for 24 h at room temperature to obtain a homogeneous solution.Lead-free BNT-ST piezoelectric ceramics powder were fabricated by a traditional solid-state reaction method. High purity BiThe homogeneous precursor solution was loaded to a 10 mL plastic syringe with a metallic needle of 21 G and the needle was connected to a voltage power supplier. A positive high voltage of 10–15 kV was applied and the feeding rate of the precursor solutions was 1 mL/h. The nanofibers were electro-spun on the aluminum foil attached to a cylindrical drum, which was placed at a distance of 10 cm from the tip of the needle. To align the nanofibers, the rotation speed of the cylindrical drum was adjusted to 0 rpm, 500 rpm, 1000 rpm and 1500 rpm. All the experiments were conducted at room temperature under a relatively low humidity of 20–40%. After the electrospinning, the as-spun BNT-ST nanofibers with different rotation speeds were fabricated as piezoelectric modules with interdigitated copper electrodes between the bottom and top of the nanofibers by a Warm Isotropic press (WIP) process at 70 °C and 80 bar and then high voltages of 1 kV were applied to the fabricated piezoelectric modules at room temperature for 1 h for the poling process to improve of the piezoelectric performance . A FE-SEM  was used for microstructure characterization of the BNT-ST nanofiber according to the rotation speed of the collector. The crystal structures of the BNT-ST nanofibers collected at the cylindrical drum with different rotational speeds were analyzed by XRD . The local piezoelectric response was characterized using PFM . The output voltages as a function of frequency for the nanofibers with different collector rotation speeds were measured using a vibration exciter , a function waveform generator , a bipolar amplifier  and an oscilloscope . The output voltages according to the bending motion of 1 Hz using a bending machine  were measured and recorded by an oscilloscope .The SEM images and fast Fourier transform (FFT) images of the BNT-ST nanofibers according to the rotation speed of the collector are shown in max) and the longitudinal piezoelectric coefficient (d33) were calculated based on the amplitude hysteresis loops. For the BNT-ST nanofibers with a 0 rpm rotation speed, which has a random array in the SEM images in max and 13 pm/V of d33 were observed in the P1 area and 0.92 pm of Smax and 0.7 pm/V of d33 in the P2 area. On the other hand, the BNT-ST nanofibers at a rotation speed of 1500 rpm, which showed the aligned nanofibers in the SEM images in max and 148.1 pm/V of d33 were observed in the P1 area and 11.04 pm of Smax and 8.4 pm/V of d33 in the P2 area. These results mean that the BNT-ST ceramic phase had greater piezoelectricity than the P(VDF–TrFE) polymer phase and as the rotation speed of the collector increased, the BNT-ST nanofibers became aligned in the advancing direction and had a better piezoelectricity. Furthermore, the piezoelectric coefficient d33 (148.1 pm/V) of the aligned BNT-ST nanofibers at P2 was considerably larger than that of the reported values for the PVDF nanofibers (−58.5 pm/V) and PZT nanofibers (83.4 pm/V) [To evaluate the nanoscale electromechanical response of the BNT-ST nanofibers with different collector rotation speeds, 3D topography images, phase hysteresis loops and amplitude hysteresis loops were analyzed by PFM as a function of applied DC voltage from −8 V to 8 V, as shown in max (maximum polarization) values calculated based on the P-E hysteresis loops of the BNT-ST nanofiber modules according to the rotation speed of the collector. As the rotation speed of the collector increased, the Pmax values increased. Particularly, in the BNT-ST nanofiber modules with a rotation speed of 1500 rpm, which were the aligned nanofibers, the Pmax value was 1.62 μC/cm2. This value was about 2.7 times higher than the Pmax value (0.6 μC/cm2) of the randomly arrayed BNT-ST nanofiber modules with a 0 rpm rotation speed. On the other hand, it was confirmed that the Pmax value of the BNT-ST nanofiber modules with the rotation speed of the collector from 0 rpm to 1000 rpm was slightly increased to from 0.6 μC/cm2 to 0.8 μC/cm2. These results confirmed that a slight increase in the alignment degree of the BNT-ST nanofibers does not contribute significantly to the piezoelectric performance and there is the sufficient alignment degree for the improved piezoelectric performance. In other words, as the rotation speed of the collector increased until over 1500 rpm, the aligned BNT-ST nanofiber modules showed a higher degree of polarization and better piezoelectric characteristics.pp value at 2000 Hz was 0.99 V, which is about a 23.7% increase over the 0.80 V of the saturated Vpp of the randomly arranged BNT-ST nanofiber modules with a 0 rpm rotation speed. As the rotation speed of the collector increased, the saturated Vpp values at 2000 Hz increased. The output voltages of the BNT-ST nanofiber modules with different collector rotation speeds according to bending motion with 1 Hz are shown in pp value of 0.31 V, the well-aligned BNT-ST nanofiber modules with a rotation speed of 1500 rpm had a Vpp value of 0.56 V. As the rotation speed of the collector increased, the Vpp value according to the bending motion increased. These results indicate that when the piezoelectric nanofibers were well aligned in a certain direction by the increase in the rotation speed of the collector and the well-aligned BNT-ST nanofiber modules had better piezoelectric characteristics. The electric displacement in the BNT-ST nanofiber modules between adjacent electrode fingers of the interdigitated electrodes was parallel to aligned direction and as the polarized direction and the applied voltage direction match, we considered that the piezoelectric characteristics are maximized.pp value of 0.99 V at 2000 Hz in the vibration frequency test and a Vpp value of 0.56 V in the bending motion. In other words, as the piezoelectric nanofibers became more aligned in a certain direction by the increase in the rotation speed of the collector, the better piezoelectric characteristics were achieved.Flexible lead-free piezoelectric nanofibers based on BNT-ST ceramic and PVDF-TrFE copolymers were fabricated by an electrospinning method and the lead-free piezoelectric nanofibers were well aligned with a higher collector rotation speed. The microstructure, orientation and crystal structure of the lead-free piezoelectric nanofibers were observed by FE-SEM images, FFT images and XRD analysis. Compared to randomly arranged BNT-ST nanofiber modules, the well-aligned BNT-ST nanofiber modules demonstrated higher piezoelectric characteristics in the P-E hysteresis loop analysis and in the output voltage analysis according to vibration frequency and bending motion. In particular, the well-aligned BNT-ST nanofiber modules with a rotation speed of 1500 rpm had a saturated V


The United Nations General Assembly debate on antimicrobial resistance (AMR) recognizes the global significance of AMR. Much work needs to be done on technology capability and capacity to convert the strategic intent of the debate into operational plans and tangible outcomes. Enhancement of the biomedical science–clinician interface requires better exploitation of systems biology tools for in-laboratory and point of care methods that detect sepsis and characterize AMR. These need to link sepsis and AMR data with responsive, real-time surveillance. We propose an AMR sepsis register, similar in concept to a cancer registry, to aid coordination of AMR countermeasures. Biocompstrategy . So, to strategy . A robusstrategy , 12. An strategy . Both spstrategy . New anastrategy . The comstrategy . Howeverstrategy . Practicstrategy . The O’Nstrategy :A massi…incredible that doctors must still prescribe antibiotics based only on their immediate assessment of a patient’s symptoms, just like they used to when antibiotics first entered common use in the 1950s.The O’Neill Review recognizes that no single measure will solve the problem of AMR and only seeks to lay out a broad agenda. The review’s introduction emphasizes the inability of current diagnostic procedures to provide rapid and comprehensive answers, noting that it isAntibiotic prescribers face three major obstacles: (a) AMR is an abstract concept for all but its victims and their physicians; (b) detection of specific forms of AMR does not conclusively determine the best choice of anti-infective therapy; and (c) in severe infections, the wait for laboratory evidence on which to base a choice of antibiotic can have fatal consequences. This last consideration remains a key promoter of emerging AMR and could be described as poorly targeted personal medicine; the antithesis of precision public health. Half a millennium ago, Machiavelli observed that the increase in diagnostic certainty with the passage of time leads to reduced treatment success . This maKlebsiella pneumoniae (in vivo expression and an unpredictable impact on clinical outcome. A carbapenem-resistant K. pneumoniae septicemia cannot be treated with a carbapenem, whether the mechanism of resistance is NDM-1, OXA-48, VIM, or IMP. The antimicrobial susceptibility phenotype is, therefore, a critical decider in the sepsis management continuum, even if the laboratory result comes 24–48 h after the initial choice of presumptive antimicrobial therapy. The susceptibility phenotype currently determines definitive therapy and ultimately informs the wider public health community. At present, surveillance data on antimicrobial susceptibility vary with laboratory capability, capacity, and locally determined public health priorities. These are all under-resourced, particularly in remote regional settings and in low-income countries (The mechanisms of AMR are numerous, increasing in variety, prevalence, and geographic distribution , but theeumoniae  happenedeumoniae . The widountries . Neverthountries –31. The ountries .Rapid, culture-independent phenotypic tests are needed that improve precision in antimicrobial prescribing , 18. In Countermeasures need purpose, intent, direction, and evidence for their efficacy. An understanding of the complex intersection of laboratory, clinical, and public health insights will improve their beneficial effect . AMR-speAntimicrobial resistance has become a global tragedy of the commons, driven by a complex bacterial survival trade-off at a cellular level . Now thaThe authors are working together on culture-independent pathology test development. TI prepared the initial draft. NU reviewed, edited, and supplemented the first draft with an emphasis on sepsis. Subsequent versions of the manuscript were exchanged between the authors who both approved the final version.The authors are supported by a Grand Challenges award from the Bill and Melinda Gates Foundation, as stated in the acknowledgments above. This and research translation grants from the Government of Western Australia are being used to develop culture-independent pathology tests for sepsis and AMR countermeasures. Thermo Fisher Scientific and Biomerieux have provided in-kind support to the authors’ research group, under supervision of the WA Health Department’s Research Development Unit. Neither author has received funding from these companies for any purpose. No supporting organization or its members had any role in the preparation of this manuscript, which is the opinion of the two authors.


In the second study (study 2), we used a within-subjects design to test effects of dopaminergic stimulation on dopamine synthesis capacity. In study 1, eight volunteers received two 18F-DOPA scans on separate days, both at rest. In study 2, 12 healthy male volunteers received two 18F-DOPA positron emission tomographic (PET) scans after treatment with either the dopamine partial agonist apomorphine (0.03 or 0.005 mg kg−1) or placebo. In study 1, no significant correlation was found between baseline and change in dopamine synthesis capacity between scans . In study 2, a significant negative correlation was found between baseline dopamine synthesis capacity and percentage change in dopamine synthesis capacity after apomorphine challenge . This correlation was significantly different (P<0.01) from the correlation between baseline and change in dopamine synthesis capacity under unstimulated conditions. One-way repeated-measures analysis of variance showed a significant group (study 1/study 2) × time interaction =11.5, P=0.003). Our findings suggest that regulation of dopamine synthesis capacity by apomorphine depends on baseline dopamine function, consistent with dopamine stimulation stabilizing dopaminergic function. Loss of this autoregulation may contribute to dopaminergic dysfunction in brain disorders such as schizophrenia, substance dependence, and Parkinson's disease.Dopaminergic function has a key role in normal brain function, dopaminergic dysfunction being implicated in numerous neuropsychiatric disorders. Animal studies show that dopaminergic stimulation regulates dopaminergic function, but it is not known whether this exists in humans. In the first study (study 1), we measured dopamine synthesis capacity (indexed as  Supporting this, a study in rhesus monkeys showed that apomorphine reduces DSC, and that this effect was greater in animals with greater baseline DSC.18F-DOPA PET, in healthy humans. In study 1, we determined the reliability of 18F-DOPA imaging and the relationship between baseline DSC and change on re-scanning in untreated healthy volunteers to provide reference data for comparison. In study 2, we determined the effect of apomorphine on DSC using doses designed to preferentially act on presynaptic autoreceptors39 to test the hypothesis that apomorphine would alter DSC, and that this would depend on baseline DSC.Therefore, although these findings are consistent with an autoregulatory effect of dopamine agonism in disease, the regulation of DSC in healthy humans remains to be determined. In view of this, we aimed to determine the relationship between baseline dopamine synthesis capacity and the effect of dopamine stimulation, using apomorphine, on dopamine synthesis capacity, indexed using 18F]-DOPA for both the studies.Study 1 was approved by the South London and Maudsley/Institute of Psychiatry NHS Trust. Study 2 was approved by the Hammersmith Research Ethics Committee. The Administration of Radioactive Substances Advisory Committee (ARSAC) granted permission to administer GTPγS binding both alone and in combination with dopamine points to it having partial agonist effects, as noted above.29 Nevertheless, it is important to note that partial agonism has not been shown in intact preparations, where apomorphine is generally found to behave as a full agonist, so this interpretation of our findings should be considered tentative at this stage.Conversely, in healthy volunteers with relatively low DSC and presumed relatively high tonic auto-inhibition by dopamine, apomorphine will compete with dopamine to occupy the autoreceptor. As apomorphine lacks the full agonist action, it will decrease the net functional agonist effect when it displaces dopamine, thereby causing increased DSC. This is in line with competitive binding studies.Our study was not intended to examine dose effects. Consequently, the sample size for the higher dose of apomorphine was small, which means that our analysis of dose effects lacks power and should be considered preliminary. It would be useful to use a wider dose range in future work to definitively test the dose effects. Our study design does not prove a causal relationship between baseline DSC and response to dopamine agonism. This could be done in preclinical experiments using optogenetic or other techniques to alter baseline DSC.The test–retest comparison group had a longer duration of time between scans than the apomorphine challenge group. Despite this length of time, however, there was good test–retest reliability between scans in this group, and no evidence of regression to the mean. Nevertheless, it would be useful for future studies to test this over the same duration as used in the challenge group. It should also be noted that a few of the participants in the test–retest study showed large absolute differences, of −10% to 20% although at the group level, the variability was much less. This likely reflects subject and scan acquisition-related variability, such as small movements of the subject during one scan session. However, this individual level variability would reduce our power to detect the relationship between baseline dopamine synthesis capacity and change under apomorphine. As such, it does not account for our findings, and could reduce the strength of the relationship.18F-DOPA activity in cerebellum at baseline and after apomorphine that we found would suggest that apomorphine is not having an effect on blood flow (please see Other considerations include possible effects of apomorphine on blood flow in the reference region (cerebellum), and the possibility that apomorphine could have effects on the peripheral elimination or metabolism of DOPA. However, these are unlikely to explain our results for the following reasons. First, we did not find differences in DOPA uptake in the reference region pre- or post-apomorphine. The consistency of 35 Assuming an effect of apomorphine on blood flow, as shown in preclinical studies,66 this would have been uni-directional (either an increase or decrease compared with baseline conditions) and in contrast to our findings, which suggest bidirectional modulation of dopamine synthesis. Two subjects did not receive placebo during baseline scan, which may introduce an expectancy effect. However, taking these two subjects out of the analysis did not significantly change magnitude of effect size or P-value, as shown above. We accept that, in the subjects who reported side effects with apomorphine, breaking of the blind could have occurred. This could have potentially affected dopamine release in these individuals.67 It should be noted, however, that in those people receiving apomorphine at the lower dose (with no side effects) the results were unchanged, and magnitude of correlation was larger.Although we were unable to test whether apomorphine would affect metabolism of DOPA in our sample, previous animal literature has failed to find any effect.iK is calculated from iK=K1 × k3(k2+k3), which is a rectangular hyperbola, if iK is higher, a change in k3 will have different effects on iK than when Ki is lower. We therefore ran a simulation using standard values for K1, k2 and k3.68 We do not know the real change in k3 but around control values the relationship between iK and k3 is not exactly linear. Assuming a +30% change in k3, this would translate into a +33% change for subjects with the largest iK and in a +40% change for those with the lowest iK. Therefore, it is unlikely that our results reflect a statistical artefact. It could also be hypothesized that the conversion rate, k3, may be saturable, and therefore one would not see a change in those with higher baseline iKcer. However our results argue against this. In our study, similar changes were seen in those with both high and low baseline iKcer, in keeping with other pharmacological studies using the F-DOPA ligand, which have shown a decrease in iKcer in those with relatively higher baseline iKcer.69 Furthermore, in normal conditions, AADC is not the rate-limiting step in dopamine synthesis and apomorphine at the doses used in the study is not known to directly influence AADC levels, but modifies it functionally via dopamine receptors.70 In these conditions, tracer quantities of 18F-DOPA will not saturate AADC. However, it should be noted that AADC may be saturable when high doses of L-DOPA are given, for example, as a treatment for Parkinson’s disease. Finally, it is worth examining the possible acute and chronic effects of apomorphine. Animal work suggests differential effects of apomorphine acutely .71 In humans, chronic treatment with D2 partial agonists with high intrinsic activity, aimed at selectively engaging the autoreceptors, has not been effective in treating schizophrenia.72 The main reason attributed has been desensitization of D2 autoreceptors. However, animal literature is mixed in this regard.74It is conceivable that, as 14 our findings indicate that a low level of dopamine agonism may act to reduce elevated DSC. The low dose used suggests there will not be significant postsynaptic effects, reducing the risk of exacerbating psychosis or other side effects. Indeed, there is evidence of efficacy of apomorphine in schizophrenia, although initial positive results75 were not replicated,72 possibly because doses used in some studies were higher (up to 6 mg) than those we used, as well as desensitization, as noted above. Dopamine agonism could also have a role in substance dependence (particularly stimulant dependence) to stabilize dopaminergic neurotransmission by potentially providing tonic dopaminergic stimulation and reducing surges in dopamine associated with drug-related cues. Our findings support studies in these conditions to determine the effect of dopaminergic stimulation on dopamine function.Our finding that low-dose apomorphine has a stabilizing effect on dopaminergic function has implications for the treatment of a number of disorders in addition to Parkinson’s disease. In schizophrenia, where DSC is predominantly elevated,


The promise for real precision medicine is contingent on innovative technological solutions to diagnosis and therapy. In the post‐genomic era, rational and systematic approaches to biological design could provide new ways to dynamically probe, monitor, and interface human pathophysiology. Emerging as a mature field increasingly transitioning to the clinics, synthetic biology integrates engineering principles to build sensors, control circuits, and actuators within the biological substrate according to clinical specifications. A particularly tantalizing goal is to develop novel versatile, programmable and autonomous diagnostic devices intertwined with therapy and personalized for the patient to get closest, finest, and most comprehensive diagnostic information and medical procedures. Here, we discuss how synthetic biology could be preparing the future of medicine, supporting and speeding up the development of diagnostics with novel capabilities to bring direct improvement from the clinical laboratory to the patient, while addressing healthcare evolution and global health concerns. Developing high clinical value diagnostics remains a major technological challenge of medical sciences. Conventional diagnostic technologies are facing evolving economical and resource imperatives, while still mostly relying on analytical chemistry or antibody‐based platforms centralized in clinical laboratories to match standards of specificity, sensitivity, throughput, and robustness. However, these approaches are often not clinically informative enough to accommodate precision medicine, and confront complex syndromes, rare or rapidly emerging diseases.Obtaining a comprehensive view of patient pathophysiology, its exposure to the environment and its trajectory over time require new modes of detection for complex and dynamic biomarker signatures still mostly inaccessible to measurement. For instance, most microbiological diagnostics cannot provide timely infectious etiology before tedious laboratory manipulations, thereby imposing important mortality burdens. Likewise, achieving dynamic, real‐time, and non‐invasive monitoring of endocrine function, exposure to toxics, metabolic parameters such as glycemia, or early molecular onset of cancer would constitute medical breakthroughs.This would require autonomous diagnostics capable of direct analysis of new types of biological parameters in complex matrices. Ultimately supported by portable and low‐cost devices, implantable diagnostics could prove extremely valuable in many clinical situations that would benefit from precise processing of localized cellular and molecular information. Interestingly, integration of medical reasoning within diagnostics in the form of programmable signal processing could allow complex measurements while considerably speeding up and improving diagnostic to treatment efficiency. Engineering the next‐generation diagnostic devices to meet modern medical needs remains a critical challenge, which necessitates innovative approaches and is therefore increasingly drawing the interest of synthetic biologists.et al, The last decades can be regarded as the descriptive phase of molecular biology that permitted the advent of synthetic biology. Synthetic biology has become a science of structuring biological matter in a rational and systematic manner to achieve control on energy and information processing through classical engineering strategies: standardization, hierarchical abstraction of complexity, and modularity . This strategy involves synthetic gene circuits acting as intracellular molecular prosthesis that autonomously monitor disease‐associated biomarkers and coordinate adjusted and timely therapeutic responses. This approach could prove extremely valuable in many diseases with asymptomatic onset, complex dynamics, and clinical situations where therapeutic benefit depends on the delay in analytical methods, information management, interpretations, and effective patient care, for instance, in the long‐term surveillance of cancer, or chronic autoimmune, infectious and inflammatory diseases. For example, mammalian cells were engineered to detect clinical levels of psoriasis‐specific TNF and IL‐22 cytokines and deliver systemically IL‐4 and IL‐10 immunomodulatory cytokines in real time. When implanted in vivo, these devices prevented psoriatic onset, stopped acute events, and restored physiological tissue morphology. Similarly, engineered autologous T cells have been tested in clinical trials for cancer theranostics, and a variety of adoptive cell therapeutics approaches could benefit from constantly increasing precision and capabilities. Mammalian cells have also been designed to perform metabolic disorders, urate homeostasis, obesity, and diabetic ketoacidosis management (Kojima et al, in vivo triggering selective and robust apoptotic responses to malignancy. Using gene expression levels that are commonly used to diagnose prostate and small‐cell lung cancer, synthetic genes circuits could be programmed to multiplex molecular biomarkers in vivo and compute appropriate therapeutic outputs. Alternatively, devices made of networked synthetic nucleic acids have been developed, such as autonomous DNA nanomachines capable of multiplexed analysis of cancer cell surface markers and logic‐gated delivery of medical molecular payloads in live animals.Synthetic cell‐based biosensors also hold promises as autonomous devices for continuous monitoring of pathophysiology et al, Since the lack of specific and predictive endogenous biomarkers limits the diagnosis of many complex diseases, attention has recently been given to the engineering of synthetic biomarkers. Administered in the circulatory system to record molecular events associated with pathological states, they could enable the non‐invasive monitoring of non‐classical parameters by producing disease‐specific molecular signatures. The potential for early disease stage detection and monitoring has been reported with experimental models of liver fibrosis, cancer and solid tumors, or cardiovascular diseases. For instance, synthetic biomarkers scoring pathological enzymatic activities at disease sites release reporters in circulation that concentrate in urines to be measured user paper devices (Warren in vivo synthetic devices to act as self‐contained theranostics is almost established and could evolve toward multipurpose implantable systems, potentially introducing a new era for clinical practices and precision medicine. Realizing these novel approaches could benefit individuals as well as society, improving therapeutic outcome and reducing healthcare costs, while also benefiting regions with poor infrastructure.Synthetic biology has advanced enormously in the past few years. Engineered molecular and cellular devices with biosensing and information‐processing capabilities proved to be clinically compliant and are now transitioning to trials. It provides with an opportunity to decrease the size and resource requirements of diagnostic devices, increase portability to get closer to the patient, and give access to real‐time, personalized, and physiologically meaningful diagnostic measurements Table . The capSynthetic biology could serve as a technological support for medical biology, simplifying decision rules for clinicians through more precise and sophisticated diagnostic tools. However, uncertainty remains about the extent to which clinicians will to adopt these diagnostic tools. Although recent technological biosafety and risk assessment advances have been made, regulatory concerns remain to be addressed. In this perspective, abiotic synthetic biology is likely to provide the next medical tools and paper‐based diagnostic platforms appear extremely promising.in vivo implementation of quantitative models to test molecular hypotheses and probe biological networks. In that perspective, synthetic biology represents an unprecedented approach to exploring pathophenotypes, discovering new biomarkers and augmenting molecular definition of syndromes.Last, we envision that the field also holds considerable value to explore human pathophysiology. The reverse engineering of biosynthetic pathways, genes, and networks constitutes resources for the multi‐level screening of disease mechanisms. It allows iterative design and The authors declare that they have no conflict of interest.