Overall Abstract: In the past decade, rapid advances in the field of neuroscience resulted in a dramatic paradigm shift in the way we understand the role of glia in normal brain functions and brain disorder pathology. A growing body of evidence shows that diversified populations of astrocytes, microglia, oligodendrocyte precursors and mature oligodendrocytes play a critical role in the regulation of synaptic functions, blood-brain barrier, immune response regulation, myelination and axonal conduction, and in the synthesis of the extracellular matrix, a key regulator of neural plasticity. Building on this evidence, exciting new findings are beginning to emerge, shedding light on glia abnormalities in schizophrenia and their impact these functions. This symposium aims to discuss and integrate the current state of knowledge on direct evidence for glial abnormalities in schizophrenia and their underlying mechanisms.Dr. Juliana Nascimiento will present novel findings on the effects of NMDAr antagonists and antipsychotics influence glial cell lines and 3D cultures as neurospheres and cerebral organoids. Results from these studies point to the central role of glycolysis, EIF2 signaling and translational machinery in oligodendrocytes and astrocytes. Dr. Paul Klauser will report on elegant investigations on the implication of developmental redox imbalance inducing oxidative stress leading to impairments of oligodendrocytes, myelin formation and eventually to the disruption of white fibers integrity and conductivity, especially in brain regions where the metabolic demand is high. In patients, alterations of white matter were found to be inversely correlated with blood levels of GSH precursor cysteine and could be prevented by the early administration of the antioxidant N-acetyl cysteine. Dr. Sabina Berretta will discuss recent findings on novel modalities of interaction between glial cells, extracellular matrix and neurons, postulated to affect synaptic structural plasticity and axonal conductance. A growing body of evidence from her group shows disruption of such interactions in schizophrenia, potentially contributing to synaptic pathology and impacting neural connectivity. Dr. Dost Ongur will build on previous work showing abnormal diffusion of neuron-specific metabolite NAA in frontal white matter in patients with chronic schizophrenia in the absence of abnormalities in the diffusion of non-specific metabolites Cr and Cho. State-of-the-art recent studies on first episode psychosis patients and matched healthy controls show that NAA diffusion is normal in first episode patients but Cr and Cho diffusion is abnormal, suggesting that white matter abnormalities in non-neuronal elements in early phases of schizophrenia which are followed by neuronal damage in chronic disease.


Carbon nanotube field-effect transistors are used extensively in ultra-sensitive biomolecule sensing applications. Along with high sensitivity, the possibility of regeneration is highly desired in bio-sensors. An important constituent of such bio-sensing systems is the buffer used to maintain pH and provide an ionic conducting medium, among its other properties. In this work, we demonstrate highly-sensitive regenerative dopamine sensors and the impact of varying buffer composition and type on the electrolyte gated field effect sensors. The role of the buffer system is an often ignored condition in the electrical characterization of sensors. Non-enzymatic dopamine sensors are fabricated and regenerated in hydrochloric acid (HCl) solution. The sensors are finally measured against four different buffer solutions. The impact of the nature and chemical structure of buffer molecules on the dopamine sensors is shown, and the appropriate buffer systems are demonstrated. Dopamine is a chemical from the catecholamine and phenethylamine families, and performs crucial functions in the brain and the central nervous system (CNS) as a neurotransmitter and as a local chemical messenger that controls the immune system and digestive system, among others ,2,3. DysCarbon nanotube field effect transistor (CNTFET)-based biosensors have attracted a great deal of attention due to their unique electronic properties, special geometry (high surface area-to-volume ratio), high mechanical strength, and chemical stability ,17. In tIn addition to the highly sensitive experimental conditions like the effect of counterions, ionic strength of buffer also plays a role in deciding the performance of the biosensor . ResearcAnother big challenges in the commercial biosensor market is the cost of the sensors. Although expensive sensors are used extensively in research environments, low cost sensors have higher market potential. The high cost in sensor development is mainly attributed to high cost of instrumentation and materials. In addition to printed technologies and ambient processing techniques which are already leading to economical sensor development, the possibility of reusable sensors can be another way to further reduce the costs. Apart from economic reasons, the reusability of sensors is inevitable in applications where device-to-device variance can be a major source of error. Hence, the regeneration of sensors is of high interest among researchers . The finPatterns of source, drain, and gate contacts were formed by standard negative photo-lithography followed by lift-off process. On the patterned flexible polyimide substrate  has the ability to bind with dopamine and form a charged boronate ester which can then modulate the surface charge on the semiconducting channel ,31,32. TN-Ethyl-NN-hydroxysuccinimide) chemistry for the activation of carboxylic acids was exploited [N-morpholino)ethanesulfonic acid). Equal amounts  measurements were performed on the kapton sample before and after it was functionalized with CPBA using the Alpha spectrometer  controlled by OPUS software in the attenuated total reflection (ATR) mode. The spectral resolution was chosen as 2 cmironment . The secnic acid , hence cThe electrical characterizations of the CNTFETs was performed at ambient conditions using a Keithley Source Measuring Unit (SMU). For all electrical measurements, a PDMS chamber was mounted around the active area of the CNTFETs to serve as a compartment for around 60 To investigated the “unexpected” response of sensors to varying dopamine concentrations in HEPES and TRIS buffer, we look at the chemical structure of dopamine, TRIS, HEPES, and CPBA in THE ATR-FTIR spectra of sensors after they were measured in varying concentration of HEPES , detergents, glycine, and urea . For dopFor very high concentrations of dopamine (∼1 mM), the sensor saturates and cannot be turned on, as seen in A flexible dopamine sensor using a CNT field effect transistor has been demonstrated. Measurements in PBS buffer indicate a highly-sensitive sensor. Measurements were also performed for three other buffer solutions, namely HEPES, MES, and TRIS. The results clearly indicate that along with the most widely considered parameters like pH and ionic strength of the buffer, the chemical nature of buffer must also be considered. It is demonstrated that HEPES and TRIS buffers can themselves react to the functionalization, and the measurements in these buffer systems no longer hold correct in measuring dopamine. Finally, the regeneration of this dopamine sensor by treatment with acidic solutions is demonstrated.


Usage of statins is suggested to decrease the incidence of HCC. When it comes to different statin subtypes, the chemopreventive action remains controversial. We aim to compare the usage of different statins and reduction of HCC risk.Embase.com and Cochrane Library database up to August 10, 2015. Duplicated or overlapping reports were eliminated. We performed a traditional pair-wise meta-analysis and a Bayesian network meta-analysis to compare different treatments with a random-effects model.We searched PubMed, We reviewed five observational studies enrolling a total of 87127 patients who received at least two different treatment strategies including rosuvastatin, atorvastatin, simvastatin, pravastatin, fluvastatin, cerivastatin, and lovastatin or observation alone. Direct comparisons showed that usage of atorvastatin  and fluvastatin  could significantly cut the risk of liver cancer. The difference of indirect comparisons between the included regimens is not statistically significant. However, usage of all types of statins, such as fluvastatin , atorvastatin , simvastatin , cerivastatin , pravastatin , lovastatin  and rosuvastatin , appeared to be superior to observation alone. Notably, fluvastatin was hierarchically the best when compared with the six other statins.Our analyses indicate the superiority of usage of statins in reduction of liver cancer. Available evidence supports that fluvastatin is the most effective strategy for reducing HCC risk compared with other statin interventions. Hepatocellular carcinoma (HCC) is recognized as the sixth most prevalent malignancy and annually accounts for one of the most common causes of cancer deaths worldwide. HCC has a dismal 5-year survival rate, and to date there is no effective chemotherapy treatment , 2.in vitro and in vivo experimental studies have shown that statins have potential protective effects against primary and metastatic HCC via induction of apoptosis, arrest of cell proliferation, and inhibition of angiogenesis [Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, which is a key enzyme in the rate-limiting step in cholesterol synthesis. Statins are used as potent cholesterol-lowering medications in the primary and secondary prevention of cardiovascular and cerebrovascular disease , a growiogenesis .Results of human studies, however, remain controversial as to a beneficial role of statins in either preventing or curing patients presenting with HCC. Two randomized controlled trials showed that additional use of pravastatin prolonged the survival of patients with advanced HCC , 8. ObseTo date, there are no large randomized controlled trials assessing the potential effects of statin therapies in patients either susceptible to develop HCC or with advanced disease. Given the ambiguous results and limited epidemiological evidence for a link between statin use and risk of liver cancer, the current study sought to evaluate the association of statin use and the risk of HCC. Furthermore, we sought to define potential chemoprevention strategies for the treatment of HCC for respective statin subtypes. To attain a better understanding on this issue, we performed an updated Bayesian network-analysis of available observation studies to investigate the association between use of different types of statins and the risk of developing HCC.The PRISMA flowchart showing the electronic searching processes is shown in Figure Pairwise meta-analysis was accomplished for the eight different comparisons, with a lack of direct comparison between cerivastatin and lovastatin. The weighted RRs for the HCC occurrence were calculated for each comparison, the geometric distribution of which was displayed in Figure 2 statistic. Overall, statistical heterogeneity was moderate, although for some comparisons 95% CIs were wide and included values indicating very high or no heterogeneity. In the meta-analyses of direct comparisons for efficacy, I2 values higher than 75% were recorded for seven comparisons. In addition, all P values for Begg's rank correlation test and Egger's test were difficult to assess due to the limited amount of research.With respect to statistical heterogeneity, it was assessed in two of the comparisons by the IWe summarized the results of the random-effects network meta-analysis for HCC rates in Figure The probabilities of best treatment for each intervention were ranked at each of the eight possible parameters Figure . ConsistFigure This present network meta-analysis, which was based on five observational studies involving 87127 people and 10993 liver cancer cases, was designed to investigate the association between different types of statin use and the risk of liver cancer. Our results showed that compared with observation alone, seven different types of statin conferred relative benefits to decrease HCC incidence. As presented in this network meta-analysis, fluvastatin specifically appeared to provide an overall advantage on reduction in HCC risk over the remaining interventions and was ranked first among the seven types of statins. Moreover, the likelihood of important selection or publication bias was small.Statins have been routinely used in patients with cardiovascular diseases thanks to their notable lipid-lowering effects. Despite the potential benefits of statin use in reduction of HCC, concerns about widespread adoption of this strategy are justified, because information on the balance between potential benefits and safety are scarce. Although there were concerns over carcinogenicity of statins raised by early animal studies, recent evidence has supported a potential chemopreventive role against liver cancer , 20, altBased on the evidence that the growth of HCC is critically dependent upon cholesterol, as well as statin selective localization to the liver and less than 5% reaches the circulatory system, statins are proposed to have a specific role in liver carcinogenesis. It is plausible that lipophilic statins  may be superior to hydrophilic statins  in liver cancer preventive qualities owing to better lipid solubility and membrane permeability , 22. Con2 values of comparisons, heterogeneity was high.There are several strengths to consider in our analysis. It is worth mentioning that our current study is the first Bayesian network meta-analysis performed on the relationship of subtypes of statin use and the reduced risk of HCC. Apart from direct active comparisons, our network meta-analysis figured out a comprehensive and complete picture to get precise effect estimates of respective type of statin, even when no head-to-head studies existed. First, this network meta-analysis compares all available statin therapies simultaneously and assesses each therapy individually, rather than only grouping them into statin use or not. Secondly, a rankogram of available types of stain for chemoprevention of HCC provides a formal rank order for treatment strategies. Application of a Bayesian network meta-analysis contributes to a general evaluation of multiple statin strategies , 25. In Our network meta-analysis has to be interpreted with caution in view of some limitations. First, most studies included in this analysis are observational but not prospective studies, which might affect the validity of overall findings. To our knowledge, by reason of the low incidence of liver cancer, it is difficult to complete a large randomized controlled trial within a finite time horizon. Secondly, owing to scant primary data, the size of each subtype group is particularly small after divided into different types of statins. The present debate about optimum statin treatments to prevent HCC will be assisted greatly by collection of robust data in future trials. Thirdly, the characteristics of populations were different, which mainly come from Asian, Europe, and North America, and yet few data was available from other countries or races. Fourth, more detailed data for duration or dosage of statin therapies is barely available, which may make a difference to the outcomes. Besides, there are some other confounders including different causes of HCC such as HBV/HCV infections or alcoholic liver disease, concomitant diseases, anti-viral therapies of HBV or HCV and so on, which might overestimate the preventive effect of statins as mentioned above. Lastly, statistical heterogeneity was moderate in the meta-analyses of direct comparisons, while no substantial inconsistency was found in the network meta-analysis.In summary, this network meta-analysis provides a useful and complete picture which convinced the associations between statin use and reduced HCC risk by using Bayesian analytical approach. Our analysis shows the superiority of usage of statins in reduction of liver cancer. Among all, fluvastatin is the most effective strategy for reducing HCC risk compared with other statin interventions. Our analysis of up-to-date evidence may provide new insights into controversies on this issue with valuable implications in clinical care and future research. Still, there is a strong need for well-designed randomized controlled trials for the chemoprevention of HCC.The protocol for the systematic review was performed in accordance with PRISMA  guideline . We searIn our trials for meta-analysis, included trials had to meet the following criteria: (1) RCTs, cohort studies or case-control studies; (2) original studies designed to assess the association between the use of statin and the risk of liver cancer with specific classification; (3) interventions: treatment with all kinds of statin treatments or observation alone; (4) population: no prior diagnosis of primary liver cancer or other cancers most likely to metastasize to the liver, such as stomach, colon, breast and so on; (5) risk estimates should be provided or could be calculated with sufficient usable outcomes data. Duplicated or overlap reports were eliminated according to the same title, author list or publication date. If more than one paper was derived from the same population, otherwise, only data from the most recent published report was included. Eligible studies should be published in the form of peer review articles in full length without language restrictions. Eventually, a total of five observational studies were included, which were all published in English though.Two investigators  independently reviewed the full manuscripts of eligible studies and extracted information into an electronic database: patients' characteristics study design, interventions, type of statins, adjusted confounders, the number of events of interest in each group. Any discrepancies regarding the extraction of data were resolved by an additional investigator (Zheng MH). When relevant information on design or outcomes was unclear, or when some needed data was unavailable directly from the study, the original authors were sought for eligible data by email.The quality of the methodology was independently assessed by two reviewers using the Newcastle-Ottawa Quality Assessment Scale with some modifications to match the needs of this study. The quality of the studies was evaluated by a ‘star system’ based on three items: patient selection, comparability of statin and observation group, and the ascertainment of either the exposure or outcome of interest for case-control or cohort studies respectively .2 statistic, which if >50% suggested statistically significant heterogeneity, if between 25% and 50% considered moderate heterogeneity levels, and if <25% implied low heterogeneity levels.First, we performed a traditional pair-wise meta-analysis which could synthesize studies that compared the same interventions directly by using STATA 12.0 . Then a Bayesian network meta-analysis to compare different treatments  to each other was followed respectively, with a random-effects model using Markov chain Monte Carlo methods in WinBUGS  as described in our previous work –30. The We also estimated the relative ranking probability that which treatment was the most efficacious therapy, the second best, the third best and so on. Treatment regimens were ranked in terms of HCC occurrence with the same methodology. Taken together, the multiple-treatments meta-analysis raised statistical power by incorporating evidence from both direct and indirect comparisons across all interventions.


In detail, the first class encompasses those methods based on the perturbation of the fluorescence signal, also known as ensemble-averaging methods, which average the behavior of many molecules (across many pores). The second class comprises those methods based on the localization of single fluorescently-labelled molecules and tracking of their position in space and time, potentially across single pores. Finally, the third class encompasses methods based on the statistical analysis of spontaneous fluorescence fluctuations out of the equilibrium or stationary state of the system. In this case, the behavior of single molecules is probed in presence of many similarly-labelled molecules, without dwelling on any of them. Here these three classes, with their respective pros and cons as well as their main applications to nucleocytoplasmic shuttling will be briefly reviewed and discussed.Molecules are continuously shuttling across the nuclear envelope barrier that separates the nucleus from the cytoplasm. Instead of being just a barrier to diffusion, the nuclear envelope is rather a complex filter that provides eukaryotes with an elaborate spatiotemporal regulation of fundamental molecular processes, such as gene expression and protein translation. Given the highly dynamic nature of nucleocytoplasmic transport, during the past few decades large efforts were devoted to the development and application of time resolved, fluorescence-based, biophysical methods to capture the details of molecular motion across the nuclear envelope. These methods are here divided into three major  Embedded in the NE are the nuclear pore complexes (NPCs), which allow the passage of ions and molecules across the NE and, at the same time, regulate the exchange of larger molecules, such as RNAs, proteins, or ribonucleoprotein (RNP) particles between nucleus and cytoplasm Dictyostelium discoideum by cryo-electron tomography (cryo-ET) The overall shape of the pore is known since pioneering studies, among others, were conducted yeast by electron microscopy (EM) At the molecular level, the whole NPC consists of about 30 different polypeptides designated nucleoporins (Nups), with a very controlled stoichiometry, and a total mass of ~ 125 MDa Given the highly dynamic nature of the overall process, a variety of time resolved biophysical strategies were applied to nucleocytoplasmic transport of molecules . By these methods, the characteristic time of molecular transport across the entire NE can be measured by averaging the behavior of many molecules, across many pores. A peculiarity of these methods is that they rely on the change of the optical properties within the sample only, whereas the characteristic dynamics and function of the molecules under study are not altered. Originally conceived by Peters in 1974 The second class comprises localization-based single particle tracking (SPT) methods. By these approaches, the trajectories of single molecules of interest are measured. By SPT, the transit times and interactions at the level of isolated NPCs of several transport receptors and model cargo molecules were successfully probed. Contrary to other approaches, SPT experiments typically require complex experimental procedures: for instance, the molecule of interest must be purified, properly labelled , and introduced into the cell by microinjection or cell permeabilization. Also SPT measurements inherently require bright and isolated particles that must localized and tracked over time many times in order to acquire reasonable statistics.Finally, the third class encompasses spatiotemporal fluorescence correlation spectroscopy (FCS)-based approaches, which afford single molecule ‘sensitivity’ in presence of many similarly labelled molecules and in live, unperturbed cells. The recently developed spatial extension of FCS, named pair correlation function (pCF) approach, was proven to be particularly suited to study the shuttling of molecules across the NE In the following, the detailed contribution of these three classes of time-resolved methods to our understanding of nucleocytoplasmic transport of molecules will be summarized and discussed. A glimpse into the future directions in the field will be also provided.2Thanks both to the recent advancements in the optical microscopy technologies at our disposal and to the development of new fluorescent labels, several perturbation-based approaches were developed, including photobleaching methods , photoactivation , have been considered as a reference for the study of passive diffusion of other molecules through the NPC. In fact, GFP is nowadays used as an indispensable benchmark in any attempt to identify the NPC translocation mechanism of endogenous proteins.Concerning the available literature, in a pioneering work by Wei and colleagues For instance, Sunn and co-workers, based on the GFP reference, were able to show for the first time that the vitamin D receptor B1 (VDRB1) exploits a serum-dependent, active nuclear transport process, while no active nuclear export mechanism was found for the same protein Worthy of note, at this point, is the use of reversibly photoswitchable fluorescent proteins (RSFPs) (mentioned above in this section). In contrast to standard FRAP or photoactivation strategies, RSFPs enable the reversible optical highlighting of specific pools of molecules and thus the repeated measurements of protein dynamic behavior, definitely increasing the amount of information that can be retrieved from a single measurement. In the last 15 years, many of these FP variants have been proposed and new cell imaging applications discovered that exploit their properties The methods described so far, being inherently ensemble-averaging strategies, do not afford information on single molecules but averages over many similarly-labelled molecules. This inherent limitation, however, does not prevent perturbation-based methods from providing quantitative information on the molecular (kinetic and thermodynamic) details of the transport process. For instance, by combining FRAP with the calibration of intracellular protein (i.e. GFP) concentration, Cardarelli and co-workers were able to quantitatively define the nuclear transport saturability by the estimate of the effective dissociation constant of the NLS-Importin complex in the actual cellular environment 3As opposed to perturbation-based techniques, localization-based approaches potentially yield single molecule information, provided that the molecule of interest is properly labelled and introduced into the sample  at the desired concentration , fluorescence correlation spectroscopy techniques appear as ideal alternative strategies. Among others, the pair correlation function (pCF) method is particularly suited to study nucleocytoplasmic shuttling as it is able to measure the time needed for single molecules to migrate from one point to another within a living cell and in presence of many similarly labelled molecules  is a characteristic of the pCF-based analysis of nuceocytoplasmic transport. This is due to the fact that, each time that two points are correlated across the NE, the contribution of all the Worthy of mention, recent conjugation of the pCF approach to Number&Brightness analysis opens up new perspectives for the study nucleocytoplasmic exchange in live cells 5It is worth stressing here that perturbation-, localization-, and fluctuation-based approaches are here presented and discussed not in general, but strictly related to their application to nucleocytoplasmic transport. Within such a peculiar biological context each of them shows selected distinguishing benefits or limitations see . Ensembl6i) high spatial and temporal resolution, in order to properly describe the processes involved in nucleocytoplasmic transport; ii) single molecule sensitivity/resolution; iii) single pore resolution. In this regard, as already discussed, each of the presented strategies shows its own limitations. Perturbation-based methods, although informative, inherently fail to provide direct single-molecule and single-pore observations. SPT-based approaches are valuable as they potentially satisfy all the three requisites. Yet, it is not clear whether the experimental conditions required for SPT experiments  are compatible with unaltered pore function. Fluctuation-based analysis, on the other hand, is endowed with single molecule sensitivity and high spatiotemporal resolution under native conditions but are typically conducted across many microns within the cell, thus averaging the contribution of many pores at the same time. Worthy of note, high-speed atomic force microscopy (HS-AFM) has been recently proposed as a valuable innovative approach to the study of the spatiotemporal dynamics of the NPC transport barrier Xenopus laevis oocyte NPCs at a timescale of about 100 ms and conclude that the highly flexible, dynamically fluctuating FG Nups of the central channel rapidly elongate and retract, but do not cohere into a tightly crosslinked meshwork single NPC to probe the movement of single molecules with high accuracy As mentioned in the introductory section, intense debate is still ongoing about the molecular details of nucleocytoplasmic transport and the nature of the selective gating imposed by the NPC. In particular, the structural and functional spatiotemporal organization of the FG-rich nucleoporins of the central channel (and their role in regulating molecular transport under native conditions) remains largely obscure. Available models are mainly based on simplified in vitro attempts to reconstruct the organization of selected components of the pore and are largely contradictory As a proof of principle, Cardarelli et al. recently exploited feedback-based tracking of the pore position by using GFP-labelled Impβ1 as a dynamic marker of the NPC in live, unperturbed cells In summary, the orbital tracking method proved the potentiality to extract information on single-molecule events in a moving, nanoscopic reference system, in presence of many similarly-labelled molecules, and under physiological conditions. As such, this approach holds the promise to become a valuable technological platform to address the lingering questions at the level of single pores.


Introduction: Radiology plays a crucial role in the emergency care setting by delivering early and precise diagnoses under pressure of time, right at the beginning of patient treatment. Although there is a need for postgraduate education in emergency radiology, most of the national bodies responsible do not offer it in a uniform fashion and a general proof of qualification is missing in Europe. Therefore, the European Society of Radiology (ESR) has founded the (Sub-)Society of Emergency Radiology (ESER), prompting them to develop a European curriculum. This trend, which is currently also encouraged in many other non-radiological specialties which demand the highest professional qualifications, often lacks expertise in medical education.Goals: The goal of this article is the general description of the curricular planning process for a European postgraduate subspecialisation programme, using the example of Emergency Radiology , including the utilisation of TOOLS and recommendations derived from comparable projects.Project description: The project was divided into partial steps: the timeline displayed in a GANTT chart, and tasks and responsibilities assigned in a RASCI matrix. The curriculum was iteratively developed using the KERN approach and steps were prioritised using the PARETO principle. Furthermore, the following TOOLS were used: limitations and needs assessment, SWOT analysis, formulating learning objectives and categorising them after MILLER and SCLO, and using BLOOM’s taxonomy for cognitive learning objectives and operationalising them according to MAGER. Psychomotoric and affective learning objectives were assigned to CANMEDS roles, grouped by topic using CLUSTERING, and then mapped by MATRIX analysis to appropriate learning and evaluation methods. Striving for continuous improvement, the curriculum was finally embedded in curricular quality management.Results: The standardisation of the EDER access, considering the different national conditions, the minimisation of European learners’ attendance phases, restricting expenses by best possible use of existing structures, respecting the requirements and retaining the support of the European umbrella society ESR, finishing the project by a specific deadline and the demands of continuous improvement were particular challenges. A curriculum with the eligibility of five years’ speciality training in general radiology has evolved on schedule. The subspeciality training lasts at least one year and is divided into webinars, workshops during congresses  and one year practical training at the individual learner’s corresponding local hospitals, which adhere to a catalogue of requirements, comparable to national educational policies. The curriculum is completed by passing a written and oral exam (diploma) and re-accreditation every five years.Conclusions: Despite complex requirements, the TOOLS utilised allowed an almost seamless, resource-minimised, professional, location-independent distributed development of a European subspeciality curriculum within one year. The definitive implementation is still due. If any deviations from the draft presented should become necessary in the future, the embedment in the curricular quality management will lead to a redirection in the right way and, furthermore, secure a continuous improvement in the best way possible. CBL = Case-based learningECR = European Congress of RadiologyEDER = European Diploma in Emergency RadiologyEDiR = European Diploma in RadiologyESR = European Society of Radiology ESER = European Society of Emergency RadiologyETAP = European Training Assessment ProgrammeETC = European Training CurriculumL = LectureLO = Learning objectivesMC = Multiple choiceMini-CEX = Mini-clinical evaluation exerciseMSF = Multi-source feedbackNKLM = Nationaler Kompetenzbasierter Lernzielkatalog MedizinOSCE = Objective structured clinical examinationPBL = Problem-based learningPDCA = Plan–do–check–actQM = Quality managementRASCI = Responsible, Accountable, Support, Consulted, InformedSCLO = Swiss Catalogue of Learning ObjectivesSOE = Structured oral examinationSWOT = Strengths, Weaknesses, Opportunities, ThreatsUEMS = European Union of Medical SpecialistsWFME = World Federation for Medical EducationEmergency care is a general and permanent essential element in health care, with a very high proportion of emergency patients  and a fThe European countries have different training structures, durations and contents due to socio-economic, cultural and organisational reasons. The European Society of Radiology (ESR) is a radiological speciality society with over 63,000 members and currently integrates, as a European umbrella society, 47 national societies and 16 European subspeciality societies. One of the latter is the European Society of Emergency Radiology (ESER), which was founded in October 2011 , w, w13], wThe goal of this article is the general description of the curricular planning process for a European postgraduate subspecialisation programme, using the example of emergency radiology (European Diploma in Emergency Radiology: EDER), including the utilisation of TOOLS and recommendations derived from comparable projects.SW. as a member of the ESER Executive Board, was commissioned to develop a proposal for a curriculum for the subspecialisation in emergency radiology  to carry out cost-effective webinars, workshops and exams with a reduced organisational effort from the annual congresses. A sufficient web-based learning, teaching and administration platform for exam questions (i.e. the Item Management System . This platform allows the webinars to be streamed independent of time after recording.LEARNING , 39] an anWebinaLEARNING . The webLEARNING ) should Workshops: Five workshops per day, each of 90 minutes, will take place at the annual ESR and ESER congresses. The ten workshops are grouped by topic, similar to the webinars 1 to 9, and topics 10 and 11 are summarised in a tenth workshop  to (9), partially (2) and ungrouped contents.Scientific work: Within the scope of the ‘Scholar’ role, an understanding of research for decision-making on a scientific basis is a prerequisite for the participants or needs to be self-taught. In this particular case, ESER does not offer a concrete proposal, but requests a proof of performance (see 4.2.6)Teaching hospitals: In addition to all the psychomotor skills and competencies, almost all affective learning objectives need the environment of an acute care hospital, which is why the teaching of those learning objectives needs to be transferred to decentralized institutions. Consequently, learning objectives were separately categorised, grouped and listed, and a relevant logbook conceptualised. The institutions will be bound by contract to fulfil a given framework of content and time: lectures, a catalogue of autonomously conducted diagnostics and interpretations, intermediate exams, feedback discussions, attitudes and on-call duties.The evaluation takes place on four levels regarding the learners and the programme; training at the teaching hospital will be evaluated semi-annually. Webinars/workshops will be evaluated formatively after each lecture (L) and summatively at the end of the programme. All formative evaluations use the same Likert scale from 1-6 (strongly agree – strongly disagree) and contain free text questions (“What was good?” and “What could be better?”).Self-directed learning: The summative evaluation of the obligatory literature occurs during the final exams.Webinars: Preparation and follow-up each include five multiple choice questions (MC) chosen randomly from a pool of questions. The lecture itself will be evaluated formally. Questions are, for example, “The knowledge offered was new to me?”, “The number of contents was appropriate for the duration of the lecture?” and questions about the quality of the teacher. Proof of performance for the 11 webinars are the MC tests passed and certificates of attendance.Workshops: Questions in the KEY-FEATURE format , e, eWorkshScientific work: Proof of performance is the individual presentation of a paper or poster about an emergency radiological topic at an ESER-approved congress, or the acceptance as a first or last author of a manuscript for review in a indexed, peer-reviewed journal. Alternatively, three co-authored manuscripts will be accepted. This scientific work may be carried before the beginning of the programme and emergency radiological relevance will be verified by ESER.Teaching hospital: Eleven radiology-adapted mini-clinical evaluation exercises (Mini-CEX)  an anTeachiini-CEX) , 48] (r (rTeachiini-CEX)  from 1-6Final exam: Admission follows the submission of all proofs of performance named above. An application for the recognition of alternative proofs of performance can be filed. A successful certificate of training for national speciality training or a European Diploma in Radiology (EDiR) must be presented before admission. The summative evaluation will be provided, at least, during each ESR and ESER congress and includes a structured oral examination (SOE) , , . As suchThe approach at the beginning of the project was unusual and occurred because of internal considerations to increase the necessary, but not ensured, support of the ESER board. Therefore, e-mail communication was not considered to be expedient and instead, a private meeting, during the few in-person occasions was chosen. The initial BRAINSTORMING at the board meeting concerning strengths, weaknesses, opportunities and threats promoted, in addition to an early estimation regarding the feasibility of the project, the communication between curricular staff and revealed individual ideas. The following SWOT analysis  derived An early commitment to possible educational methods was helpful during the concrete implementation. Webinars, for instance, are already being held and only need to be restructured. It should also be easy to maintain a majority of the teachers currently active. Additionally, we chose the requirement to fulfil the learning objective in research and science since Seaburg et al. (2016) in a recently published study, found a significant association between the number of publications and ratings in clinical evaluations of speciality trainees , and we Our first estimation of the curriculum development under the situation given, using the approach and TOOLS above, is subjectively positive. An objective opinion will be possible after the concrete implementation, arrival of the evaluation results and comparison with the target values. The possibility of an external assessment is present with the European Training Assessment Programme (ETAP), an initiative founded by the ESR Education Committee in 2001 and part of the European Union of Medical Specialists (UEMS) . The ETAThe TOOLS presented and the general approach of curriculum development have already been published multiple times. The originality of this article lies in the condensing of information for the increasingly frequent case of comparable projects initiated by professional societies, particularly on a supranational level, often planned and implemented with the same challenges. This happens frequently with established specialist competencies, but without added comparable competencies in medical education and without the possibility of secondary consultations. The article presented should raise awareness and serve as a compendium for a professional approach.The informal targeted needs analysis of the learners (Step 2 of the KERN cycle ), carrieEvery institution operating as a teaching hospital in the future, should be charged for certification by ESER, auditioned annually and recertified every three years. The ESER board will seek professional consultation to decide on the specific standards used. We have to proceed as is common at the German Medical Association (and others) for reasons of feasibility: A simple written confirmation about the fulfilment of performances/requirements catalogued is produced which, in principle, is not verifiable by ESER itself.The summative evaluations at the end of the curriculum are limited by the time available at the annual congresses and the number of teachers and examiners. Should the number of candidates interested in EDER rise, as was the case with EDiR (less than 100 at the beginning of 2011 to more than 1000 candidates at the end of 2015), the SOE could reach its limitations fast. The European Board of Radiology, which conducts exams for EDiR, implemented a new assessment format successfully in 2016, which replaced the oral exam and simultaneously improved the assessment methodology . In addiThe preparation of a European subspeciality qualification in Emergency Radiology has led to a curriculum development with the following limits and requirements which may apply to many other European curricula for postgraduate medical education:Limited time is confronted with the pressure to succeedPeople responsible do not always have a high expertise in medical educationEligibility needs to be unified, despite various national conditionsMinimal synchronous attendance time for learners, teachers and people responsibleNo separate financial or personnel resourcesWe recommend a close co-operation with experts of medical education and QM for any comparable projects. The initial needs analysis with the determination on a subsequent framework, followed by a stepwise PARETO-prioritised development, protected by a QM for continuous improvement exists as a working model. From our perspective, the following project TOOLS were shown to be helpful: BRAINSTORMING, CHECKLISTS, SWOT analysis, GANTT chart, RASCI matrix, MATRIX analysis and CLUSTERING of topics. We suggest NKLM, Revised BLOOM’s Taxonomy and CANMEDS roles for categorising learning objectives. Webinars and workshops during speciality congresses seem beneficial as teaching formats. The inclusion of teaching hospitals is inevitable for practical training with real patients. We recommend the limitation of a few educational methods, since it is easier to mediate those to the teachers on a high level and this, consequently, yields better handling: keynote presentations and mainly SCAFFOLDING in SANDWICH technique, FBL, POL, FEEDBACK BUZZ groups and the extensive use of E-Learning for online preparation and follow-ups. We suggest SMP for oral and a mix of MC with KEY-FEATURE as assessment formats for written exams. By implementing special expertise and utilising the methods and TOOLS above, an almost seamless, resource-efficient, professional and location-independent development of a European subspeciality training curriculum succeeded, including the integration of a QM, within one year.Although the text was phrased gender neutrally in principle, the generic masculine form was used in some passages for reasons of simple legibility.The project is not financially supported. ESER, as a society, has received funds from sponsors for congress events for an adequate value in the past. The total amount in the last three years was below 100,000 Euro and these funds had no influence on the manuscript presented here.Parts of this work originated within the framework of the dissertation project of Martin G. Wagner.ESER thanks the ESER office in Vienna, represented by the deputy, Wolfgang Duchek, and the project secretary, Sabine Grab, Department of Radiology at the Ludwig-Maximilians-University in Munich, for their great support.The authors declare that they have no competing interests.


Bronchial asthma is one of the most common chronic inflammatory diseases of the airways. In the pathogenesis of this disease, the interplay among the genes, intrinsic, and extrinsic factors are crucial. Various combinations of the involved factors determine and modify the final clinical phenotype/endotype of asthma. Oxidative stress results from an imbalance between the production of reactive oxygen species and reactive nitrogen species and the capacity of antioxidant defense mechanisms. It was shown that oxidative damage of biomolecules is strongly involved in the asthmatic inflammation. It is evident that asthma is accompanied by oxidative stress in the airways and in the systemic circulation. The oxidative stress is more pronounced during the acute exacerbation or allergen challenge. On the other hand, the genetic variations in the genes for anti-oxidative and pro-oxidative enzymes are variably associated with various asthmatic subtypes. Whether oxidative stress is the consequence of, or the cause for, chronic changes in asthmatic airways is still being discussed. Contribution of oxidative stress to asthma pathology remains at least partially controversial, since antioxidant interventions have proven rather unsuccessful. According to current knowledge, the relationship between oxidative stress and asthmatic inflammation is bidirectional, and genetic predisposition could modify the balance between these two positions—oxidative stress as a cause for or consequence of asthmatic inflammation. Bronchial asthma (BA) represents the most common chronic respiratory disease in children, and its prevalence is constantly increasing especially in the developing countries . The disRecently, many attempts were made to classify and stratify the diseases into different phenotypes or endotypes, which are characterized by specific patterns and aspects with respect to the development of inflammation. Phenotype means a cluster of characteristics that define asthma and its subsets. Till date, several asthmatic phenotypes have been determined and characterized based on the triggers  or clinical presentation  . Another450, NADPH oxidases, nitric oxide synthase, xanthine oxidase), immune and non-immune cells , and others . On the other hand, there are also many exogenous sources of ROS, such as cigarette smoke, ultraviolet light, ionizing radiation, pollutants, ozone, organic solvents, metals, and some medicaments . Both endogenous and exogenous sources of ROS can play an important role in the pathogenesis and worsening of various inflammatory conditions, especially through continual accumulation of the oxidative changes in biomolecules  or extrinsic  antioxidant substances –17.Decreased activity or dysfunction of antioxidant enzymes , 19.Over-activity of pro-oxidative enzymes –22.Many authors showed that BA is significantly associated with increased oxidative stress expressed by the increased markers of oxidative damage. Based on the development and persistence of oxidative stress, several crucial aspects and mechanisms can be identified :OverproProduction of highly reactive oxygen species leads to the progressive damage of various biomolecules  with the functional and structural consequences. Under physiological condition, antioxidant defense mechanisms are able to eliminate and repair these changes. However, unregulated overproduction of ROS, during inflammation of various origins, leads to accumulation of the changes without sufficient repair. Oxidative damage of the biomolecules influences the signaling pathways, enzymatic functions, gene expression, and many other essential biological processes. Structural changes are contributing to the chronic remodeling of the tissues. Another possible contribution to the oxidative damage of biomolecules in addition to ROS is RNS. These are formed from the reaction of nitric oxide with oxygen or ROS. RNS could modify the thiol groups  or change the tyrosyl residues to nitrotyrosine. This process usually leads to the inactivation or dysfunction of the modified proteins , 23.Oxidative stress represents a substantial component of BA development and persistence. Chronic inflammation leads to the overproduction of ROS and RNS. During the allergen challenge or other situations , the pronouncement of inflammatory cascades is accompanied by the accelerated production of ROS/RNS. This could contribute to the structural changes in the airways, support the remodeling processes, decrease the sensitivity to the anti-inflammatory and anti-asthmatic treatment, and worsen the clinical course of the diseases. Uncontrolled asthma with the sinusoidal acceleration of ROS/RNS production pronounces the pathophysiological aspects of the inflammation and dysfunction of the biomolecules and is clinically associated with remitting–relapsing course of the disease Figure  8, 12, , 12, 23.Many studies showed that the cells formatting airway inflammation in BA are an important source of ROS in these patients . OxidatiExposure to environmental allergens has been shown to stimulate overproduction of ROS/RNS, resulting in the damage of DNA , proteins, and lipids with decrease or even loss of their physiological functions. Moreover, accumulation of oxidative damage causes the structural changes in the airways tissues . It was Due to low concentrations of ROS, their extreme reactivity, and very short lifetime, direct measurement of ROS is very complicated and, therefore, various indirect markers of oxidative damage, such as markers of lipid peroxidation , acrolein, hexanal, heptanal, nonanal, 4-hydroxyhexanal, 4-hydroxynonenal), protein oxidative damage , DNA damage , or general total antioxidant capacity of plasma could be used . OxidatiIt was shown that oxidative stress markers in BA are associated with worse clinical control, worse disease severity, or reduced lung function . The amoInterestingly, umbilical cord blood-derived basophils from the neonates born to mother with atopic asthma showed increased markers of oxidative stress, decreased activity of glutathione peroxidase, and increased production of interleukin 4. This could contribute to the development of allergic hyperreactivity in children at risk of asthma .Based on the observation from the previous studies that BA is associated with decreased antioxidant protection due to changes in the functions of antioxidant enzymes of decreased concentrations of non-enzymatic antioxidants, the use of different antioxidants could be an interesting mode of supportive and complementary anti-asthmatic therapy , 28.Epidemiological data suggest that antioxidants have a significant effect on the incidence and severity of BA. It was shown that BA and airflow limitation are associated with deficiency of various antioxidants, such as carotenoids, retinol, coenzyme Q10, and vitamin C, D, and E , 40–45. In several studies, the supplementation with a nutraceutical of antioxidants and anti-inflammatory compounds  was associated with reduction of airway inflammation, as documented by a decrease in fractional exhaled nitric oxide (FENO)  or proloTobacco smoke exposure was confirmed to be an important risk factor for bronchial hyperreactivity development. It increases the oxidative stress with its further consequences. Moreover, it was shown that toxic chemicals in tobacco smoke are able to induce epigenetic changes with increased expression of various pro-inflammatory genes . TherefoOxidative stress is a dynamic process with very tiny border between protective and harmful effects and with problematic predictability of the threshold after which disease ensues and, therefore, this could explain a general inconsistence in the clinical trials with antioxidant supplementation in asthma management. Although the supplementation of various antioxidants appears to be a promising adjuvant therapy for asthma, various studies did not confirm the significant benefits over standard therapy. The potential of antioxidant therapy could be improved by taking into consideration individual characteristics of each particular asthmatic  and environmental risk factors, instead of treating oxidative stress in the airways broadly , 41.GST-T1) and catalase (gene CAT).In our research, we studied and analyzed the possible role of oxidative damage in the development of BA in children from several aspects. We studied the markers of oxidative stress in exhaled breath  and in the peripheral blood (concentration of free thiol groups as a marker of protein oxidation and concentration of TBARS as a marker of lipid peroxidation) and also analyzed the selected single nucleotide polymorphisms in the genes for two important antioxidant enzymes, such as polymorphisms of two important antioxidant enzymes, theta isoform of glutathione transferase  and catalase. These two enzymes represent essential mechanisms in the protection against oxidative damage, because they utilize a wide range of products of oxidative damage as substrates. Children with asthma had higher prevalence of the GST-T1 null genotype compared with healthy controls and these genotypes increased the risk for asthma development by 3.17 folds. Interestingly, atopic asthmatics had a lower prevalence of GST-T1 gene null genotype than non-atopics (CAT), the TT genotype was more frequent in asthmatics than in healthy children (OR = 5.63). This genotype correlated positively with the concentration of –SH groups and negatively with the content of TBARS  in asthmatic children compared to healthy subjects. Atopics showed significantly decreased concentration of –SH groups compared to non-atopic asthmatics. We also evaluated a marker of lipid peroxidation—concentration of TBARS. Asthmatics yielded higher concentration of TBARS than healthy controls, especially of atopic phenotype and during acute asthmatic exacerbation , 62. Besof TBARS .Oxidative stress represents an important part in the pathogenesis of asthma, but on the other hand, it is only a part in the complex mosaic of BA development. Contribution of oxidative stress to asthma pathology remains at least partially controversial, since antioxidant interventions have proven rather unsuccessful. According to current knowledge, it is not possible to definitely resolve whether oxidative stress is the reason or a consequence of chronic inflammation in asthmatic airways. Since the therapeutic use of antioxidants was not generally proven in clinical studies for asthma, the appropriate selection of asthmatic patients with the potential to benefit from antioxidant therapy needs further investigation. Future research should be focused on the detection of the individual asthmatics, in which application of a particular antioxidant strategy could modify the clinical course and support the standard medication. Moreover, the development of effective antioxidant therapy (new antioxidants or modification in the existing anti-asthmatic molecules with increased antioxidant properties) with complex biological effects could improve the clinical management of asthmatic and allergic patients. Another issue in the context of preventive allergology would be the development of preventive antioxidant strategies that would help in the global burden of allergic diseases.All the authors equally contributed to the concept of the work, performed literature search, written the text, revised it critically, and approved the final version of the manuscript to be published.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.


Most ribosomal antibiotics obstruct distinct ribosomal functions. In selected cases, in addition to paralyzing vital ribosomal tasks, some ribosomal antibiotics are involved in cellular regulation. Owing to the global rapid increase in the appearance of multi-drug resistance in pathogenic bacterial strains, and to the extremely slow progress in developing new antibiotics worldwide, it seems that, in addition to the traditional attempts at improving current antibiotics and the intensive screening for additional natural compounds, this field should undergo substantial conceptual revision. Here, we highlight several contemporary issues, including challenging the common preference of broad-range antibiotics; the marginal attention to alterations in the microbiome population resulting from antibiotics usage, and the insufficient awareness of ecological and environmental aspects of antibiotics usage. We also highlight recent advances in the identification of species-specific structural motifs that may be exploited for the design and the creation of novel, environmental friendly, degradable, antibiotic types, with a better distinction between pathogens and useful bacterial species in the microbiome. Thus, these studies are leading towards the design of “pathogen-specific antibiotics,” in contrast to the current preference of broad range antibiotics, partially because it requires significant efforts in speeding up the discovery of the unique species motifs as well as the clinical pathogen identification. The severe increase in antibiotic resistance and cross-resistance among pathogenic bacterial strains presents a significant health threat. Hence, focusing on the specific properties of antibiotics targets in pathogenic bacteria, and on the molecular mechanisms acquiring resistance to them, are of prime importance. So far, the main efforts to combat antibiotic resistance are based on attempts at the production of new antibiotics by various approaches, such as mining underexplored microbial niches or designing new chemical probes for improving the antibiotic performance of known molecular scaffolds, and, to a lesser extent, the development of novel therapeutic agents . PresenProtein biosynthesis is a key life process in all organisms; hence, it is targeted by many antibiotics. This process, namely, the translation of the genetic code, involves decoding the genetic information and the creation of nascent proteins. Ribosomes, the universal flexible and dynamic giant multi-protein-RNA assemblies, perform both tasks in all living cells, including pathogenic bacteria. They are built of two structurally independent riboprotein subunits that associate upon initiation of protein biosynthesis. In all organisms, the small subunit accommodates the mRNA and provides the site for the decoding of the genetic information, and the large subunit contains the site for peptide bond formation, called the peptidyl transferase center (PTC), and the tunnel along which the nascent proteins progress until they emerge from the ribosome . Owing tConcurrent with the determination of the high-resolution structures of bacterial ribosomes, intensive efforts have been made to identify molecular modes of antimicrobial action, in order to reveal the principles of their selectivity, to shed light on their synergism, and to elucidate the mechanisms of acquiring resistance. Consequently, the target site and the mode of action of at least one member of each family of ribosomal antibiotics have been located and described in detail, showing that they inhibit protein biosynthesis by targeting functional regions in the ribosomes . ExampleEscherichia coli ..S. aureuStaphylococcus aureus (MSRA) and vancomycin-resistant Enterococcus faecium (VREF). This combined drug is made of a pair of semi-synthetic streptogramins, called dalfopristin (component A of Synercid™) and quinupristin (component B), which bind simultaneously to the PTC and to the macrolide binding pocket at the entrance to the exit tunnel, and exploit the inherent flexibility of these sites [A common resistance-acquiring mechanism is mutating the anchors between the antibiotic agent and its pocket. Hence, a potential approach to partially overcome (or reduce) resistance is to increase the number of anchors. This can be achieved by the use of pairs of antibiotics that bind to two different functional adjacent sites that can interfere synergistically with ribosomal function, although their synergy may occur independently of their individual effects on the translation . Synercise sites . The synStreptomyces rochei, which uses dual genes harbored in a large plasmid. This dual production indicates a synergistic mode of action, which was found to display modest ability to inhibit cell growth as well as cell-free translation. In accordance with the biological results [Although Synercid™ has been in use for a rather short period, resistance has already been reported. Hence, searches for additional synergetic pairs are being pursued. One of them is the lankamycin (LM)/lankacidin C (LC) pair, produced by a single organism,  results , the cry results ,53. BothOverall, the binding and functional modes of action of antibiotic drug are common to all eubacteria, including most of those comprising the microbiome. Therefore, an unintentional outcome of the currently preferred broad-spectrum antibiotic treatments is modifying the delicate composition of the microbiome, which was found to be exceedingly influential in issues related to several significant diseases. Since species specificity in antibiotic susceptibility of various pathogenic bacteria targeting the large ribosomal subunit has been reported ,55, it iS. aureus (SA) led to the identification of unique structural motifs that may be exploited for the design of advanced species-specific antibiotics [S. aureus were detected in the stem loops of several rRNA helices . Hence, their non-degradable metabolites, which are rather toxic, are reaching the environment, contaminating it, and thus increasing antibiotic resistance. Some of the newly identified potential sites, such as the extended and exposed rRNA helices, can, in principle, be exploited for the design of species-specific potent degradable antibiotics. For pursuing this idea, preliminary studies showed that compounds such as complementary DNA or PNA can interact with these sites and hamper in vitro translation. The ongoing research is based on previous experiments that showed that, oligonucleotides could be used as ribosomal inhibitors as well as tools for structural and functional studies. For example, short DNA oligonucleotides were used as “antisense DNA” to probe rRNA accessibility and for locating specific functional regions , before -Minimal inhibitory concentration (MIC) values of known antibiotics can be optimized. For example, the additional of a single hydrogen bond between an antibiotic and its pocket improves the MIC dramatically . Flexibi-It is suggested that species-specific structural motifs should be exploited for the creation of novel antibiotics with a better distinction between pathogens and useful bacterial species in the microbiome. In fact, the next generation antibiotics should be degradable and species-specific. Thus, the aim of immediate research should be to minimize resistance to antibiotics while preserving the microbiome as well as reducing the contamination of the environment.-The proposed design of “pathogen-specific antibiotics,” which is a revolution in the current concepts of antibiotics, is of immediate need. “Pathogen-specific antibiotics” means antibiotic drugs specific for each and every pathogen. This strategy requires the clinically fast identification of pathogens that is already being addressed .-The practical application of “pathogen-specific antibiotics” requires the swift determination of the structures of antibiotics targets  of all or most pathogens. For this aim, the recent exciting development of single particle 3D cryo-electron microscopy should be more suitable than X-ray crystallography, since it can be performed by the use of relatively small amounts and does not require crystals.The studies reported here highlighted the interplay between internal flexibility, motion complementarity, cellular regulation, and antibiotic resistance. Furthermore, although so far based on a single, albeit important, example, focusing on selected acute issues in current antibiotics usage led to several unexpected and less obvious results, some of which are briefly mentioned below.


Danio rerio) has proven to be a highly useful tool, mostly because of its fast growth and development, as well as the ability to absorb the molecules diluted in water through its skin and gills. In this review, we focus on the reports concerning the application of zebrafish as a model for assessing the impact of toxicants on skeletal muscles, which share many structural and functional similarities among vertebrates, including zebrafish and humans.The rapid progress in medicine, agriculture, and allied sciences has enabled the development of a large amount of potentially useful bioactive compounds, such as drugs and pesticides. However, there is another side of this phenomenon, which includes side effects and environmental pollution. To avoid or minimize the uncontrollable consequences of using the newly developed compounds, researchers seek a quick and effective means of their evaluation. In achieving this goal, the zebrafish ( Many different lines of evidence have revealed that the zebrafish is an excellent model to study various aspects of developmental biology and molecular genetics. In the recent past, the zebrafish was also used as a model for toxicological and drug discovery research which were described in excellent reviews ,2,3. Howsilent heart, polka, breakdance, and hiphop  dimethylethylammonium bromide), which were applied from the zebrafish 5-somite stage onwards. Only one of them (GAL) provided an identical image of phenotypic changes to that observed for ache mutants. In the case of both the ache mutant and GAL-treated embryos, three common characteristics of the phenotype were observed: motility impairment induced by myopathy, no effect on the differentiation of neurons of the spinal cord, no changes in nACh-receptor (nicotinic acetylcholine receptor) activity. The other inhibitors exhibited additional impacts that were not seen in ache mutants. Two inhibitors, TAC and EDRO, caused a disturbance in the development of spinal cord neurons. Exposure to the inhibitor ESE gives similar effects to the genetic removal of nAChE-R in ache mutants—suppression of muscle defects acetate) is commonly used to relieve pain and inflammation. It is a member of non-steroidal anti-inflammatory drugs (NSAIDs), acting as an inhibitor of cyclooxygenase, an enzyme responsible for the synthesis of prostanoids. The exposure of zebrafish embryos to diclofenac results in various developmental defects including cardiovascular and muscle degeneration. The mechanism of diclofenac toxicity is based on interactions with macromolecules such as DNA, and induction of alteration of genes expression or the regulation of downstream genes . MoleculAmong the substances designed for beauty enhancement and protection containing ultraviolet (UV) filters are sun creams. They protect human skin from UVA and UVB radiation, but they also exert several side effects. Active compounds in UV filters can bind to hormone receptors, show hormonal activity and might have endocrine disruptive effects  see Sec. Due to A zebrafish model was used to test teratogenicity of 4-methylbenzylidene camphor (4-MBC) during early development . 4-MBC iRecently Balázs’  group exEthanol has a severe influence on early human development, including cardiac and central nervous system abnormalities, abnormal craniofacial features, and general developmental and intellectual delays. The spectrum of these disorders is known as fetal alcohol syndrome (FAS) . Seriousv/v) from 8 to 24 hpf. General morphological changes include deformities, higher mortality rate, delays in hatching and fewer bouts of swimming in response to touch. Morphological effects tend to increase in severity and frequency in a dose-dependent manner. The microscopic analysis showed that slow muscles displayed abnormalities such as a lack of segment division, altered angles between dorsal and ventral hemi-segments and smaller muscle fibers. Fish exposed to alcohol had shorter and narrower muscle fibers.Zebrafish has been used as an animal model to study different aspects of alcohol influence on embryonic development ,151,152.According to data published by the International Organization of Coffee, the popularity of caffeine products is on the increase, and the consumption of coffee in 2015 was more than 9.1 million tons per year, about 7% higher than in 2012. Caffeine can have both positive and negative health effects. On the one hand, many studies have shown positive health effects such as lowering the risk of type 2 diabetes mellitus and obesity  and reduImmunohistochemical methods were used to better understand the toxic effects of caffeine on the development of the neuromuscular system in zebrafish. Treated animals displayed disruption in neuromuscular junction development and abnormal neurotransmitter secretion. Skeletal muscles specific antibody (MF20) in caffeine-treated embryos exhibited an unequal staining pattern of MF20 in the whole somites and rounded somitic edges. In addition, shortening and misguided axons of motoneurons with accumulation of secretory vesicles in the terminal part of the axon were observed .The World Health Organization (WHO) defines nicotine addiction as one of the biggest public health threats. It is estimated that the tobacco epidemic kills six million people annually, of which more than 600,000 deaths concern non-smokers who are exposed to the harmful effects of tobacco smoke .Nicotine has been shown to affect fetal development, leading finally to spontaneous abortion, low birth weight, and even sudden infant death or significant cognitive, intellectual, and behavioral deficiencies in offspring ,163.The toxic effects of nicotine exposure on the motor system are connected with the over-activation of skeletal muscle-specific AChRs leading to motor skill impairment, including paralysis.Svoboda et al.  revealedsofa potato) which lacks muscle-specific AChRs [In addition, immunohistochemical studies on a non-transgenic zebrafish strain confirmed that nicotine delayed development of the dorsal axons arising from secondary motoneurons and impaired innervation of the musculature ,81. Of iic AChRs ,166.Besides drugs, cosmetics and other man-made artificial compounds designed for direct application, there is an urgent need for monitoring and assessing the effects of various bioactive chemicals which can affect the environment and thereby also human health, e.g., through bioaccumulation. These substances are either intentionally synthesized to find application in, e.g., agriculture as pesticides, or an unwanted waste of human activity such as farm effluents. Regardless of their origin, they need to be treated as potential pollutants, and their use if possible should be preceded by careful evaluation of the risks and benefits.The studies presented in this review demonstrate the possibility of zebrafish application in toxicological research. The availability of various mutant lines creates the possibility of advanced phenotype analysis and enables deep insight into the toxic effects and biological activity of exogenously delivered compounds. The studies conducted in this field improve our understanding of the toxic effects of chemical compounds on vertebrate neuromuscular system development and function and provide an opportunity for further studies, which will allow the development of toxicant antidotes to neutralize the whole or different aspects of their toxicity. Moreover, they determine the direction for the development of modern biosensors based on zebrafish transgenic lines with improved specificity and sensitivity of different chemicals’ detection.


Many studies of the immune system in human and animals have been focused on how it protects the body from parasitic and infectious agents , while, Many parasitic diseases are characterized by the long-term persistence of parasites in the host, due to the not fully effective host immunity or inadequate therapy. These conditions have created a particular and somewhat complicated host-parasite relationship supported by mechanisms of immune evasion by parasites, such as parasites covered by self-substances of the host, antigenic variations, and others .Moreover, the first contact between the host and the parasite stimulates the innate immunity, the first immunological, nonspecific mechanism for fighting against infections. This immune response is rapid and is mediated by numerous cells including phagocytes, T-cells, mast cells, basophils, and eosinophils, as well as the complement system .However the infection is established when innate immunity failed in eliminating parasitic and infectious agents and in that case adaptive immunity develops. The primary functions of the adaptive immune response are the recognition of specific “non-self”-antigens, the generation of immunologic pathways for specific pathogens, and the development of an immunologic memory . The celImmune mechanisms are involved in the pathology of many parasitic infections. The main factors responsible for the tissue injury during parasitic infections are chronicity of the infections, release of parasites or host cells in tissues and within the blood, alteration and destruction of the host tissue, presence of antigenic components shared by the host and the parasite, and relative inefficiency of the host in eliminating the antigens or cross-reacting antibodies. The three main pathogenic mechanisms proposed to explain the role of infectious factors as triggers of autoimmune diseases during parasitic infections are (1) polyclonal B- or T-cell activation, (2) increased molecular mimicry, or (3) immunogenicity of self-antigens secondary to the infection-mediated inflammation –7.Even though both immune mechanisms contribute to the development of immunopathological lesions in parasitic diseases, in the past more attention has been given to the humoral response, especially immune complexes and complement.Immune complexes (ICs) are formed when antibodies bind to relevant antigens: in parasitic diseases, this can occur at the site of parasite penetration, in the circulation, and within tissues or organs. Normally, during the course of the infection, the excess of ICs formed is removed by specific cells, while in pathological conditions the release of massive amounts of antigens from dying parasites which can substantially influence the ratio between antigen and antibody in favour of an antigen excess leads to high amount of ICs that can be detrimental. Furthermore, failure of the reticuloendothelial system to remove ICs from the blood can be another reason of the excess of circulating ICs .Circulating ICs may localize in the walls of blood vessels or in the organs as described in the kidney during leishmaniasis, malaria, trypanosomiasis, schistosomiasis, and other parasitic infections. The local formation of ICs and related diseases has been also observed in several organs such as heart and skeletal muscle lesions in canine leishmaniasis and others .Increased levels of IgE in the host infected with different parasites are a well-known process and they have been used as helpful diagnostic criteria. Their significance in pathology was usually attributed to a type I hypersensitivity reaction, while more recent data have shown that the biological importance of IgE includes other mechanisms .Cell-mediated mechanisms of immunopathological lesions in parasitic diseases are mainly represented by delayed hypersensitivity reactions. A typical example is the granulomatous reaction around eggs or parasites entrapped in the liver, lung, gut, or intestine .It is evident that immune-reactive cells including T-cells, B-cells, and macrophages play important roles in the pathogenesis of these immune-mediated diseases . RecentlOne example is interleukin- (IL-) 6 that is considered one of the major proinflammatory cytokines. It acts on a variety of cells, including immune-competent cells and hematopoietic cells, causing their proliferation and differentiation . The oveBased on these findings, IL-6 was thought to be a valuable and attractive therapeutic target for drug discovery and some research groups have developed anti-IL-6 receptor antibodies for the treatment of patients with inflammatory autoimmune diseases.Based on these considerations, much attention has to be given to the response of the immune system during parasitic infections considering immune-mediated associated diseases in the host and considering new targets for combined therapeutic approaches in such conditions.Orlando PacielloChiara PalmieriIwona Otrocka-DomagalaLaura RinaldiJorge Morales-MontorPeter Geldhof


The main purpose is to generate patient-specific iPS cell lines from the somatic cells that carry mutations or genetic instabilities for the aim of studying their differentiation potential and behavior. This new technology will pave the way for future development in the field of stem cell research anticipating its use in clinical settings and in regenerative medicine in order to treat various human diseases, including neurological and neurodegenerative diseases.With the help of several inducing factors, somatic cells can be reprogrammed to become induced pluripotent stem cell (iPSCs) lines. The success is in obtaining iPSCs almost identical to embryonic stem cells (ESCs), therefore various approaches have been tested and ultimately several ones have succeeded. The importance of these cells is in how they serve as models to unveil the molecular pathways and mechanisms underlying several human diseases, and also in its potential roles in the development of regenerative medicine. They further aid in the development of regenerative medicine, autologous cell therapy and drug or toxicity screening. Here, we provide a comprehensive overview of the recent development in the field of iPSCs research, specifically for modeling human neurological and neurodegenerative diseases, and its applications in neurotrauma. These are mainly characterized by progressive functional or structural neuronal loss rendering them extremely challenging to manage. Many of these diseases, including Parkinson's disease (PD), Huntington's disease (HD), Amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD) have been explored  Stem cell research is considered one of the most captivating areas of cell biology mainly due to the unique properties of stem cells and their potential use in cell-based therapies to treat a variety of diseases, including Parkinson's disease (PD), Alzheimer's diseases (AD), Diabetes Mellitus (DM), and many others , which are derived from the inner cell mass (ICM) of blastocysts, are pluripotent cells that have the ability to proliferate indefinitely. But still they maintain their pluripotency with the capability to differentiate into cells of all three germ layers: ectoderm, mesoderm and endoderm  and the nervous system . The development of the vertebrate nervous system is shown to be regulated temporally and spatially by gradients of signaling molecules that may have either inhibitory or activating roles. These molecules are important for neuronal migration  endonuclease is considered the less cumbersome and the most flexible system to execute precise genome editing in human pluripotent stem cells , Huntington's disease (HD), Amyotrophic lateral sclerosis (ALS) and Alzheimer's disease (AD), and other applications in neurotrauma, through summarizing the different protocols that are used in each for the reprogramming and differentiation processes Figure .The study of Parkinson's disease (PD) is challenging due to the inaccessibility of affected human midbrain dopaminergic (mDA) neurons on which to base experimental research, and the rarity of animal models that follow the major disease characteristics. Despite this, it has been uncertain whether the involved mechanisms would also occur in human neurons affected by the disease. Consequently, molecular pathways underlying this pathology are still not well-defined. Although the majority of PD cases are sporadic, some rare familial forms of this disease have led to the discovery of PD-linked genes. This discovery was imperative for deciphering the cellular and molecular mechanisms of PD Gasser,  and for DA neurons that were first generated from mouse iPSCs in 2008 and transplanted into the striatum of a rat PD model, have shown to ameliorate functional deficits  or four (OKSM) lentiviral factors were allowed to commence differentiation into DA neurons via dual SMAD inhibition for 1 day and feeder-free floor plate induction  in the differentiation of human ESCs into mDA precursor cells Human iPSC-derived PD-cell models were later used to have a mechanistic insight into the gene-environmental interaction involved in the pathogenesis of PD, such as the use of small-molecule high-throughput screening to identify new pathways (like MEF2C-PGC1α pathway) as therapeutic targets to combat PD  is an autosomal dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin gene that generates long polyglutamine stretches in the encoded huntingtin protein (HTT). This leads to a massive loss of medium spiny neurons in the striatum and loss of neurons in the cortex as the disease progresses. Personality changes, weight loss, involuntary movements and dementia are the principal changes mostly developed among the people carrying the huntingtin gene mutation.Transgenic mouse models of HD expressing exon 1 of the human HD gene were developed to mimic the features of the human HD. The R6/2 mouse model has represented the most rapid symptoms with widespread huntingtin inclusions in the brain  is an incurable neurodegenerative disorder that leads to the loss of upper and lower motor neurons. It has a genetic background in which 10% of the cases have a positive family history of mutations in the superoxide dismutase 1 (SOD1) gene which is associated with the most common familial form of ALS. In addition, there is an estimate of 30 genes directly linked to the pathophysiology of the disease, including TARDBP, FUS, OPTN, VCP, UBQLN2, C9ORF72, PFN1…etc., and over 120 other genes indirectly associated with ALS  proteinopathy, such as mislocalization of TDP-43, increased total cellular levels of TDP-43, and decreased cell survival. However, upon co-culture of the derived astroglia with derived motor neurons from the same iPSCs of the ALS patient or from control normal patients, there were no effects of any kind on the neurons in culture suggesting other involved mechanisms as previously described for SOD1 ALS  is the most common form of age-related dementia . A recent clinical trial on humans, the SciStar study, has been using oligodendrocyte progenitors to treat people with recent SCIs. This study not only has proven to be effective so far but has lately passed major safety issues and therefore was approved to expand by increasing the number of both enrolled patients and the transplanted cells per patient  2016). Although this study aims at evaluating the safety and effectiveness of AST-OPC1 agent, which consists of oligodendrocyte progenitor cells produced from human ESCs, in patients with recent SCI, results can be promising as regards potential use of iPSCs instead of human ESCs in this context.Oki et al.  providedin vivo (Okita et al., While the iPSCs technology holds the promise to becoming an efficient therapy for many neurodegenerative diseases that currently have no cure, there remains the risk of encountering multiple unanticipated outcomes when applying them on humans. The risks range from unwanted biological effects and immune response, toxicity, neoplasm formation, disease transmission, reactivation of latent viruses, to rejection of the cells by the body. It is difficult to determine and pinpoint the risks as they depend on several factors, including the cells that are used to achieve pluripotency, the status of the differentiated cells, their proliferation capacity, the technique of administration of the pluripotency genes, the level of manipulation, the growth factors used, the dilemma of retaining epigenetic memory, the intended site of injection, the reversibility or even irreversibility of the applied treatment, the susceptibility of the administered cells for disease, the incomplete suppression of the four transgenes after differentiation, the persistence of undifferentiated cells, and the survival of the transplanted cells An important risk of iPSC therapy is associated with using lentiviruses or retroviruses. These viruses, although are genetically tailored to hold the genes required for an iPSC state transformation, will integrate into the host cell genome and consequently add multiple viral integration sites and be the cause for several safety issues (Howe et al., in vivo, plus these cells show similar functionality to cells derived from iPSCs (Lopez-Leon et al., Trans-differentiated neurons offer a way to bypass the tumorigenicity risk associated with passing through a pluripotency state. This method allows lineage reprogramming as in directly converting a somatic cell type into another through transgenic expression of transcription factors or miRNAs. On the other hand, the procedure of re-differentiating iPSCs into specific cell types is considered lengthy, costly and arduous. In addition to that, having multiple stages before reaching the intended outcome may lower the efficiency of the generated cell type. So far, in this context, trans-differentiation seems to be also an interesting path in research that also offers the same concept of therapeutic approaches and disease modeling as iPSCs. Trans-differentiated cells show no tumorigenicity when transplanted Although, iPSCs research has been a revolution in the scientific field as it provides new hope for the treatment of many diseases, protocols describing the differentiation of iPSCs into neural cells in neurodegenerative diseases and in the context of neurotrauma are still being modified and studied to assess the effect of this process on gene mutations in these cells and vice versa. Advances have been made recently to uncover the underlying molecular pathways of several neurodegenerative diseases, yet more work has to be done before one can say complete cure from such disorders is possible.in vitro disease models and investigating in particular the human polygenic disorders where animal models are not sufficient (Lindborg et al., A recent approach of generating 3D brain tissues, namely “cerebral organoids”, closely reproduces the endogenous developmental program. This approach can give rise to retinal identities, ventral telencephalon, developing cerebral cortex, and choroid plexus, within 1–2 months (Lancaster and Knoblich, in vitro and in vivo disease modeling and drug and genetic screening (Figure Yet, many hitches like mutations, incomplete epigenetic reprogramming and tumors formation, which accompany the use of iPSCs, should be solved as well. Therefore, further understanding of iPSCs, including a genome-wide epigenetic characterization of those cells and further studying of their genomic stability, is needed before beginning their clinical applications in the area of regenerative medicine for treating human diseases, mainly the intractable ones Table . Indeed,g Figure .HB, OH, FC, and WA worked on study conception and design. HB, OH, FC, KC, BD, and AM screened titles for relevance and abstracted the data from the eligible full text articles. HB, OH, and WA analyzed and interpreted the data. HB, OH, FC, and WA drafted the manuscript. HB, OH, and WA critically revised the manuscript with input from the entire team. All authors have read and approved the final draft.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.


Trees are then rooted a posteriori by identifying the ancestral and derived transformational homologs. These multiple interrelationships make the entire retrodiction enterprise a challenging endeavor of reciprocal fulfillment.Since Hennig , cladistDespite the usefulness of phylogenetic reconstruction and over 170 years of conceptual advances following Richard Owen's structural interpretation of homology, many continue to make evolutionary inferences with definitions of homology that are independent of history. Recently, a group of supporters of the ancient “RNA world” theory devised an algorithm that subjectively guarantees the structural origin of the ribosome in its biosynthetic RNA heart, the peptidyl transferase center (PTC) , weaken their theory of ribosomal history. Recently, we examined putative IFs in small and large rRNA subunits  trees describing RNA structural evolution, (ii) a model of evolutionary change for optimization of those changes on the trees, and (iii) a process-free rooting criterion. More importantly, the algorithm cannot confirm nor deny the historical validity of IF evidence, since IFs are not homologies testable on trees. Thus, the work of Petrov et al. , 2015 ilAll authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.Computational biology in the Evolutionary Bioinformatics laboratory is supported by grants from NSF (OISE-1172791) and USDA (ILLU-802-909). DC is recipient of NSF postdoctoral fellowship award 1523549.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.


Deep brain stimulation (DBS) of the subthalamic nucleus (STN) and globus pallidus interna (GPi) is considered an essential therapy in the management paradigm of Parkinson's disease (PD). Its success stems essentially from its remarkable efficacy and, compared to the lesions created with thalamotomy and pallidotomy, its flexibility through programming that allows modification of the stimulation delivered to the precise brain targets, thereby obtaining maximal benefit with minimal side effects. In light of the impressive pace of advances in DBS technology, relentless exploration of new targets, development of programming paradigms, and continued controversy on patient selection and timing of DBS surgery, a special issue on this fascinating topic is pertinent and timely.The choice of the DBS target between STN and GPi is driven by the constellation of motor and nonmotor symptoms which are key determinants to quality of life. U. Yazdani et al. discuss the pros and cons of each target and emphasize the need for these considerations while determining the final choice in a given individual. A widely accepted notion is that GPi DBS causes less cognitive decline than STN DBS and can be considered as the preferred target in PD patients with preoperative cognitive impairment. R. Mehanna et al. reevaluate this issue in a review of 72 studies totaling 2,410 STN DBS patients and 702 GPi DBS patients and draw a set of recommendations regarding the cognitive impact of DBS in PD patients.While STN and GPi are the main targets of DBS in PD patients as they alleviate a broader spectrum of motor symptoms, in contrast to Vim which helps primarily with tremors , the benIn addition to appropriate patient screening and target selection, careful programming is critical for a positive clinical outcome. Although general guidelines for DBS programming are available, a systematic protocol is lacking. Programming can thus be challenging, time-consuming, and labor-intensive. Nevertheless, with the advent of technological improvements, programming algorithms are expected to become more effective and less frustrating. A. Wagle Shukla et al. review the current approaches to DBS programming and summarize the most recent advances in the DBS field, including interleaving of DBS pulses, fractionated current, directional steering of current, use of biphasic DBS pulses, and closed-loop stimulation. The authors also discuss the role of computer-guided programming and the possibility of remote Internet-based programming which are promising approaches to impact access to DBS care in the near future.Although the clinical efficacy of DBS in PD is well established, its mechanism of action is still partially understood but is being actively explored, especially in animal studies . There aIn 2013, the EARLY-STIM trial provided Class I evidence for the use of DBS earlier in PD . This fiHowever, in a survey of 23 Swedish patients interviewed at a mean of 8 years after diagnosis, M. Sperens et al. report that patients in moderate stages of PD seem to be resistant to an earlier intervention. Despite the EARLY-STIM trial, a significant subset of patients still consider DBS as a last resort procedure. According to the authors, patients in their cohort had a reasonable perception of DBS, expressing caution “and well considered attitudes towards its outcome.” Their resistance to early intervention is thus not stemming from inappropriate concern or relative ignorance.very adequate, an additional 46% rate it as adequate, while 3.6% find it to be completely inadequate. This study underscores the need for a better preoperative education in order to insure realistic expectations and successful DBS outcomes [On the other hand, in a retrospective study of 29 patients, K. LaFaver et al. report that while all patients are at least partially content to have undergone DBS and will recommend it, only one-third feel that their preoperative education was Finally, determining predictors of functional and quality of life outcomes after DBS in PD would be useful to better tailor treatment to the individual patient's needs. H. Abboud et al. report on a retrospective review of pre- and postoperative data in 130 patients, suggesting that postural instability and worse pre-DBS motor score are the strongest predictors of poorer functional and QOL outcomes, while age at surgery and duration of the disease did not seem to influence the outcome. On the other hand, the presence of tremors and the absence of dyskinesia and of freezing of gait are reported as the greatest predictors of global improvement, confirming prior reports , 8. ThisIn summary, this special issue provides an updated overview of the ever-expanding field of DBS for PD, while probing into emerging and controversial themes. It compares and reevaluates effectiveness of STN versus GPi DBS targets on motor and nonmotor symptoms, objectively analyzes the available data on the impact of other potential targets, and challenges the conventional wisdom on factors that predict better outcome, optimal timing of surgery, and the appropriateness of patients, including their perceptions of the procedure prior to DBS surgery.Raja MehannaHubert H. FernandezAparna Wagle ShuklaJawad A. Bajwa