A biosensor can be defined as a compact analytical device or unit incorporating a biological or biologically derived sensitive recognition element immobilized on a physicochemical transducer to measure one or more analytes. Microfluidic systems, on the other hand, provide throughput processing, enhance transport for controlling the flow conditions, increase the mixing rate of different reagents, reduce sample and reagents volume (down to nanoliter), increase sensitivity of detection, and utilize the same platform for both sample preparation and detection. In view of these advantages, the integration of microfluidic and biosensor technologies provides the ability to merge chemical and biological components into a single platform and offers new opportunities for future biosensing applications including portability, disposability, real-time detection, unprecedented accuracies, and simultaneous analysis of different analytes in a single device. This review aims at representing advances and achievements in the field of microfluidic-based biosensing. The review also presents examples extracted from the literature to demonstrate the advantages of merging microfluidic and biosensing technologies and illustrate the versatility that such integration promises in the future biosensing for emerging areas of biological engineering, biomedical studies, point-of-care diagnostics, environmental monitoring, and precision agriculture. Biosensors are considered to be powerful analytical tools and are potentially useful for a wide range of applications ranging from drug discovery, to medical diagnostics, to food safety, to agricultural and environmental monitoring, and to security and defense . A bioseMicrofluidics is considered to be a multidisciplinary technology that links several different sciences including chemistry, biochemistry, engineering, physics, micro-technology, nano-technology and biotechnology . The larIn this paper, examples from the literature are presented to demonstrate the advantages of merging microfluidic and biosensor technology and illustrate the versatility that such a merging promises in the future biosensing for numerous areas of biological engineering, environmental monitoring, biomedical applications, agricultural monitoring, industrial monitoring, and point-of-care diagnostics. We have divided this review into three sections: (i) different types of biosensors ; (ii) different types of microfluidic platforms with their advantages and disadvantages; and (iii) examples of integrated biosensors in different microfluidic platforms.an independently integrated receptor transducer device, which is capable of providing selective quantitative or semi-quantitative analytical information using a biological recognition element”. This makes biosensing technology a powerful analytical tool capable of detecting biological or chemical molecules using electrical [In the past two decades, there has been a significant growth and interest in biosensor technology and research . Accordiectrical , opticalectrical , or massectrical . Figure The biological recognition sensing element dictates the selectivity and specificity that allows the biosensor to respond to a specific target or group of analytes, decreasing the possibility of interference with undesired substances . The selIn general, the biological recognition elements are immobilized using different methods such as adsorption, covalent binding, entrapment, and membrane confinement. 5 to 1017 greater than that of the uncatalyzed reactions [Enzymes are proteins that have the ability to catalyze chemical reactions with a rate enhancement ranging from 10eactions . Enzymeseactions , uses imOne of the advantages of using enzymes as a biological recognition element in biosensor technology is that they are highly selective to a specific substrate or a class of substrates. The second advantage is that during catalytic turnover, enzymes can produce ions, protons, heat, light, and/or electrons which are all measurable parameters .i.e., analyte of interest) does not need to be purified prior to detection [Recently, antibodies (Abs) have been applied widely  as recogetection ,24. Receetection .in vitro selection method called Systematic of Ligands by Exponential enrichment (SELEX) [In the past three decades, research in aptamers have grown dramatically and become one of the most important biological recognition elements competing with antibodies in the development of biosensors ,27. Apta (SELEX)  is used  (SELEX) .Aptamers have the ability to bind to their specific ligands with dissociation constants in the micromolar to picomolar range , and canBiosensors can be classified into several different kinds according to the kind of transducer used. The transducer transforms the biomolecule-analyte interaction into a measurable optical  or electA wide range of transducers has been developed and employed; however, the most popular methods are: (a) electrochemical; (b) optical (including colorimetric); (c) piezoelectric; and (d) magnetic . Table 1Recently, most of the biosensors used in the literature are mainly based on electrochemical detection ,39. It hOptical biosensing is a vital analytic and detection technique, which has a wider application in the field of medical diagnostics ,56 , fooMost of the colorimetric detection techniques involve monitoring the formed colored products as a result from the reaction between the analyte of interest and biological recognition element. The colored product, which can be identified by a naked eye or the use of an optical sensing instrument, is proportional to the analyte concentration. These types of detection are classified as a label-free detection method .One of the unique signal transduction methods in the configuration of biosensor is the fast mass transformation occurring due to the interaction between the analyte and the immobilized biological recognition element. When the mass of the crystal increases due to specific reaction or binding of an analyte to biological layer immobilized on the surface, a change in the oscillation frequency of the crystal occurs, identifying the concentration of the analyte . This tyThe use of magnetic sensing that relies on the use of micro/nanoparticles labelling has increased in recent years. Bio-molecules have virtually no magnetic properties; therefore, the addition of magnetic micro/nanobeads to bio-molecular samples could be used to separate, and quantify a known analyte within a given sample . A numbeNumerous microfluidic devices have been developed for different biological and chemical applications . ConventFabrication and designing of microfluidics platforms for biosensors need careful addressing and considerations such as: dimensions, materials, and the method used for fabrication to improve the biocompatibility and wettability of the fabricated device . Glass aRecently, a significant demand and effort in merging biosensors into lab-on-chip (LOC) technology using microfluidics systems has been demonstrated ,93 whichDue to the small size of micro-systems, a single microfluidic biosensor can perform full analysis  includinIn the next section, examples of microfluidics based biosensors will be explained based on the different kinds of microfluidics and the biological recognition element used as the sensing layer in the biosensor. In the last two decades, the continuous microfluidic system has improved. The huge advancement in microfluidic research has increased the study of biological systems ranging from molecules to small multicellular organisms. Microfluidics has added a great ability to sensing devises over conventional methods as it can sense small volumes of analytes, resulting in reducing reagent and energy consumption, less waste, reducing the cost and the integration of chemical and biological process on a single platform . Recentlet al. [2.A few examples on the application of the biosensor technology and its integration with the continuous microfluidic platforms have been reported and demonstrated in the past. For instance, an example for the integration of bienzyme functionalized nanocomposite with a microfluidic biosensor was demonstrated by Ali et al.   which provide a highly sensitive and selective, rapid and low-cost sensing platform for the detection of biomedical samples and protein-protein interactions.An example for the integration of a continuous microfluidic device and biosensor using antibodies as the biological recognition element is demonstrated by Lee et al.  for the et al. [−1).There are several advantages that make aptamers a preferable biological recognition element over antibodies. These advantages include simplicity in the structure of aptamers and the ability to design them in a way to yield an optical or electrical signal (upon binding with the target) without the need for secondary labeling and washing steps, reducing the cost and time ,112. An et al.  for the Droplet-based microfluidics is one of the most significant systems to be integrated with the biosensor technology. Recently, droplet-based microfluidics has extensively been used as a new platform for a wide range of applications including environmental, biomedical, security and defense with a better portability, and low energy . In the et al. [d-glucose, resulting in the byproduct of H2O2. The novel integrated microfluidic based electrochemical biosensor system provides a highly sensitive and low-cost glucose sensor with linear response up to 43.5 mM.An example of a droplet-based microfluidic electrochemical sensor using Pt-black microelectrode and enzymes for the detection of glucose was demonstrated by Gu et al.   has recently been developed by Golberg et al.  using mation see . The sysRecently, a “finger-powered” bead-in-droplet microfluidic system was developed using aptamer-beacons to detect the inflammatory cytokine interferon gamma. Droplets containing aptamer-functionalized microbeads were generated by a finger push-and-release sequence. When interferon gamma was present within the droplets, the fluorescence intensity of the aptamer-beacons was detectable . In anotDigital microfluidics (DMF), as another approach used for the manipulation of liquid samples as microdroplets, has offered reconfigurability and flexibility and limited contamination during sample preparation and analysis. This type of platform is compatible with a wide range of detection methods  and eliet al. [The integration of optical absorbance measurements system with digital microfluidics for the detection of body metabolites using glucose oxidase as the biological recognition element has been reported by Srinivasan et al.  .The integration of digital microfluidics with the FET-based sensor using antibodies as the biological recognition element has shown a great potential towards biomolecules transportation and detection without the need of pumps, bulky transduces or small fluidic channels.The integration of field effect transistor (FET) biosensor into the digital microfluidic technology using antibodies as the biological recognition element was demonstrated by Choi et al. . They deAptamers have been used in conjunction with antibodies in digital microfluidic biosensors. A droplet with fluorescently labeled IgE aptamer and magnetic nanoparticles coated with anti IgE was merged with a second droplet containing unlabeled IgE. The mixture was brought to a permanent magnet to hold the magnetic beads. The droplets of a washing buffer were passed over the beads to remove unbound IgE from the magnetic beads. The assay had a limit of detection (LOD) of 150 nM of IgE .i.e., continuous, droplet-based, and digital microfluidics) were presented. The most important part of this review includes examples of integrated biosensors into microfluidic platforms, illustrating the versatility of such as integration for emerging areas of biological engineering, biomedical studies, point-of-care diagnostics, and environmental monitoring.Standard methods for identification and detection of specific targets are expensive, time consuming, and suffer from the lack of portability. The integration of microfluidics and biosensors provides a powerful tool to replace the bulky traditional instruments with the ability to merge chemical and biological components into a single platform. Biosensors are considered to be a powerful analytical tools and are potentially useful for a wide range of applications ranging from drug discovery, to medical diagnostics, to food safety, to agricultural and environmental monitoring, and to security and defense . A biose


Multi-center epidemiological studies must ascertain that their measurements are accurate and reliable. For laboratory measurements, reliability can be assessed through investigation of reproducibility of measurements in the same individual. In this paper, we present results from the quality control analysis of the baseline laboratory measurements from the ELSA-Brasil study. The study enrolled 15,105 civil servants at 6 research centers in 3 regions of Brazil between 2008–2010, with multiple biochemical analytes being measured at a central laboratory. Quality control was ascertained through standard laboratory evaluation of intra- and inter-assay variability and test-retest analysis in a subset of randomly chosen participants. An additional sample of urine or blood was collected from these participants, and these samples were handled in the same manner as the original ones, locally and at the central laboratory. Reliability was assessed with the intraclass correlation coefficient (ICC), estimated through a random effects model. Coefficients of variation (CV) and Bland-Altman plots were additionally used to assess measurement variability. Laboratory intra and inter-assay CVs varied from 0.86% to 7.77%. From test-retest analyses, the ICCs were high for the majority of the analytes. Notably lower ICCs were observed for serum sodium  and serum potassium , due to the small biological range of these analytes. The CVs ranged from 1 to 14%. The Bland-Altman plots confirmed these results. The quality control analyses showed that the collection, processing and measurement protocols utilized in the ELSA-Brasil produced reliable biochemical measurements. The validity of inferences from clinical and epidemiological studies critically depends on the validity and reliability of measurements in the data collection process. Quality assurance and quality control measures are therefore needed throughout the stages of planning and data collection. For an individual patient in continued care, the use of quality specifications based on the intraperson biological variation of an analyte through a certain period of time is internationally accepted . For epiInvestigators in Brazil and in other low and middle income countries have increasingly been involved in multi-center studies with centralized laboratory analysis, but the necessary procedures for quality control have been infrequently reported. The ELSA-Brasil  study utilized a central laboratory for most of its analyses, offering an opportunity to describe the methodology and results of the quality assessment of these measurements.The resource of having a centralized laboratory in the ELSA-Brasil allowed each research center to have a team for the collection and processing of the biological material, which would be stored in cryotubes at -80°C for up to 30 days, and further transported to the central facility. The negative aspects of this strategy were the need for centralized training and certification of the teams from each center, and the increased risk of delay in the return of results to participants. The most important positive aspects were the facility for acquisition of the lab kits and the absence of inter-laboratory variability. The local processing of the samples decreased the volume of material transported, resulting in a reduction of costs.The objective of this report is to present the results of quality control analyses of baseline laboratory measurements of the ELSA-Brasil.The ELSA-Brasil study enrolled 15,105 participants in 6 field centers located in 3 different regions of the country from 2008 to 2010 , glycated hemoglobin (HbA1c), creatinine, sodium, potassium, uric acid, aspartate transaminase (AST), alanine transaminase (ALT), gamma-glutamyl transferase, total cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides, thyroid stimulating hormone (TSH), insulin (fasting and 2 h) and ultrasensitive C-reactive protein, as well as serology for Chagas disease were performed.Urine was collected over 12 h during the night, prior to the clinic visit, locally processed and stored for shipment in aliquots as described above. Determinations were performed centrally for sodium, potassium, calcium, creatinine and albumin.Given the large number of participants, multi-center nature and diversity of measurements, the ELSA-Brasil required effective and efficient mechanisms of quality assurance and control for laboratory determinations. As previously reported , the maiFor laboratory measurements, inter-assay CVs were calculated using the results of internal controls, and intra-assay CVs using data from several pilot studies on both fresh and frozen samples.In addition, we performed test-retest analyses of specific analytes in 10% of a randomly selected sample of study participants, with determination of intra- and inter-assay variability at the central laboratory. In selected participants, a single additional blood or urine sample was collected for duplication. Extra blood samples were collected at the end of the blood drawing process. These tubes ("blind replicates"), identified only by their bar code and thus, blind for determination, were handled identically to the original tubes and sent to the central laboratory at the same time as the original tubes.Laboratory results were transmitted to the study's data center, where the original and quality control (QC) results were matched through the bar codes to individual study participants.Reliability was estimated through the calculation of intraclass correlation coefficient (ICC) , using aWe calculated the CV as the ratio of the standard deviation of measurement error to the mean of the analyzed variable.Bland-Altman plots were also produced to graphically explore the agreement between measurements .Analyses were performed initially on all pairs. To minimize the effect of extreme outliers, we then reanalyzed the differences after removing pairs presenting a difference between measurements >3 standard deviations from the mean or for whom the pair's mean was >5 standard deviations from the overall sample mean. Analyses were performed using SAS software (USA), version 9.3 .The inter- and intra-assay CVs for the analytes are reported in The number of blind replicate QC samples obtained varied from 72 to 94 across analytes. Fewer QC samples were available for analytes obtained 2 h after the glucose load, as participants with diabetes were excluded from this test.In order to visualize the pairs and the influence of outliers, This study demonstrates the excellent quality of the sample collection and processing, and laboratory measurement of biochemical analytes of the ELSA-Brasil study, in a sample of free-living Brazilian adults. Laboratory intra- and inter-assay variability was almost always below recommended international standards. Between-person variability accounted for more than 90% of the total variability with few exceptions, and was usually greater than 98%. For epidemiological studies involving associations between exposures and diseases, the most inclusive and relevant of the QC measures is the ICC. The ICC, also called the reliability coefficient, can be interpreted as a ratio of the between-individual variance to the total variance. The existence of noise (underlying within-person variability) as a percentage of the signal  lowers the ICC values. Therefore, when testing epidemiological associations, the greater the noise the more difficult it is to detect real associations at a level of statistical significance. One important, and partially controllable source of this variability is the intraindividual variation, sometimes called "measurement error". The overall reliability coefficient evaluates the fraction of the population variability of a given variable that are not due to measurement errors . This measurement error could originate from biological variation or from either pre-analytical or analytical sources. Here, with samples taken from a single participant visit, the biological variation was not evaluated. Pre-analytical errors include those related to variability in collection , processing , storage  and transportation . Also to be considered is the possible mixing of tubes among participants during the collection processes. In this study, none of the outliers were clustered to a particular individual participant, indicating that the possibility of serious errors resulting from switching or erroneously labeling collection tubes or aliquots, or assigning laboratory results to the wrong participant, seem unlikely.Analytical errors include laboratory determination errors . The intra- and inter-assay CVs available from the ELSA central laboratory indicate very small error in this part of the measurement process, in general. With few exceptions, VCs of measured analytes were lower than the analytical quality specifications based on components of biological variation . Of noteThus, the bulk of the variability in reported measurements seems likely to be originated from a combination of smaller problems in analyte collection, processing and transportation, coupled with the specific analyte susceptibility to each of these problems.The markedly lower ICCs of the analytes serum sodium and potassium highlight an interesting phenomenon. As the CVs for these analytes were quite high, the low ICCs can be attributed to their low between-person variability, as illustrated by the narrow distribution of the values in the Bland-Altman plots for serum sodium  and potaThe CV, popular in reports of measurement error, is of utility in determining the extent to which the analyte value of a given participant will vary, in relation to the analyte's mean value, from one measurement to the next - a relevant issue in the determination of the analyte's clinical utility. However, as CV evaluation doesn't take other variabilities into consideration, it has a lesser role in evaluating the quality of measurements in epidemiologic studies of associations. In this study, the blind replicate CVs were higher than the intra- and inter-assay CVs, which was caused by the inclusion of not only laboratory measurement variability, but also pre-analytical sources of error usually present in multi-center studies with centralized measurements.The Bland-Altman plots provide a visual analysis of these QC results. One can see potentially systematic differences among replicates, such as that found for fasting insulin . This diThe decision to report results with and without outliers is not always easy. For example, it seems logical to use the ICC and CV obtained for alanine transaminase after exclusion of the outlier seen in the Bland-Altman plot, given that the mean of the outlier pair was more than 30 times the mean of the remaining pairs, leading to a potentially large weighting of this single value. The outlier value was apparently obtained from a participant with liver disease and, given the large difference within the pair (>50 U/L), likely outside the range of precision of the measurement technique. For precision evaluation in an epidemiological study in which the overwhelming majority of participants have values within or near the normal range, inclusion of such outliers makes interpretation of the findings difficult. As can be seen when comparing the Bland-Altman plots before and after the outlier exclusion, it obscures the analysis of measurement error of values within the relevant range. In contrast, outliers in other analytes, both for average values and those for differences between the QC sample and its pair, were close to the exclusion boundaries. As such, these values should probably be considered measurements within the relevant range of study, and their differences should be included in our final estimates of measurement variability.Two limitations of our analysis merit mention. First, as QC samples were collected, processed, and shipped simultaneously with their corresponding pair samples, some of the measurement variability occurring during these steps could have been underestimated. Additionally, an important aspect of variability - temporal biological variability - was not assessed, as participants were not recalled to repeat measurements on another day. Thus, for a thorough evaluation of the measurement reliability for epidemiological studies, consideration of this within-participant day-to-day variability would require incorporation of data from other studies.In conclusion, these analyses of measurement variability are important for QC documentation in multi-center studies during which strict QC measures are necessary to guarantee accurate results. The biochemical analytes of the ELSA-Brasil here reported were measured with high reliability. Based on this, they should serve well as exposure variables and co-variables for most studies of associations, especially given the large sample size of the cohort.pdf].Click here to view [


Although qualitative researches (QR) are invaluable in understanding complex healthcare situations, the quantitative systematic reviews could not treat them. To improve quality of healthcare services, results of QR should be considered in healthcare decision-making processes. Several methods and theories for synthesizing evidences of QR have been developed. In order to activate the narrative reviews and mixed methods reviews in Korean healthcare academies, I arranged the related nomenclatures and suggested some issues to conduct them. In providing healthcare services, the evidence-based medicine (EBM) paradigm -3—the idNot only the definition of qualitative research, but also the characteristic comparisons between qualitative and quantitative research are well documented in the papers by Murphy et al.  and DrapMeanwhile, because the synthesis of qualitative research results is inevitably different from a quantitative systematic reviews, there have been many attempts to overcome this gap , 37-40. Among the theories about qualitative research presented so far, works of Draper , BarnettAs can be seen in Searches of qualitative research to collect a body of related literature are more difficult than quantitative research -58. ThisBecause of the diversity of research methods and fields of application, qualitative research does not easily lend itself to standardizing the items of qualitative evaluation in the literature of interest ,59-64. NFor the synthesis of evaluation results, a best fit frameworks is established ,69-73 orAlso, a reporting guideline named RAMESES  has been developed for application when the research results of systematic reviews of qualitative research are to be reported in papers ,78. MakiThere are few systematic reviews on qualitative research in healthcare-related scholarship in South Korea. For the qualitative improvement of healthcare in this scarce situation, research in various healthcare fields should be conducted. Therefore, I suggest three things as below.First, establishing the nomenclature of systematic reviews on qualitative research is an urgent task, because systematic and coherent use of well-established terms is important when a multitude of suggested terms are in use, as shown in Second, the practical way to revitalize narrative reviews research when Korean researchers do not have much experience conducting narrative reviews is the critical reading of good research cases from various academic fields and their application to practical use. To facilitate this process, I organized useful research cases by academic field and presented them in Third, there is a need to establish a research-supporting organization and expand research human resources for qualitative research in the process of planning and conducting clinical studies . The bas


Aeromonas” . Aeromonas is a Gram-negative, motile, and facultative anaerobic bacterium. Some of the members are associated to infections in humans, fish, and other animals, and are widespread in aquatic environments  provides a more or less chronological history of the Aeromonas taxonomy and its major changes since 1891  with technological advances, from DNA–DNA hybridization coupled with physiological and biotechnological characterization to fingertprinting, gene sequencing, and whole-genome sequencing analyses. This has resulted in a great expansion of the genus Aeromonas although several taxonomic controversies persist for reliable species identification. General recommendations to meet such identification is provided by the author, which is a good point of this chapter.Chapter two (Aeromonas fish pathogens) and four (Aeromonas infections in humans) are dedicated to Aeromonas diseases. The first diseases of fish are perhaps known from Aeromonas pathogens, including the two oldest known fish pathogens: the motile A. hydrophila and the non-motile A. salmonicida. By reviewing data on taxonomy, diagnosis, ecology, virulence factors, and disease control, the author of chapter three provides evidence that the number of Aeromonas fish pathogen species increases with increasing research efforts, although major gaps persist, primarily on their ecology. Chapter four weakens the controversial role of Aeromonas in causing diseases in humans. The authors further extend the Aeromonas disease range in humans, from gastroenteritis to other infections .Chapters three  is short but original compared to other single-genus book dedicated to pathogens, because it provides examples of benign or even benefial associations of Aeromonas-animal pairings, primarily the case for the digestive tract symbiosis between A. veronii and medicinal leeches, a fascinating topic.Chapter five (Aeromonas antigenic structures), an overview of the plethora of surface structures that facilitate interactions between Aeromonas and diverse hosts and evironments are elegantly given to the readers, including flagella, pili and fimbriae, lipopolysaccharide (LPS), outer membrane proteins, capsule, and the S-layer. The chapter is specifically focused on the recent findings on those external antigenic structures thought to have roles in Aeromonas pathogenesis, which offer targets for the development of vaccines.In chapter six (New developments on the virulence mechanisms of Aeromonas hydrophila) covers new virulence factors and the related molecular requirements for causing diseases, focusing primarily on the type III and IV secretion systems and the role of quorum-sensing systems in modulating Aeromonas virulence. Emphasis is also given to the role of secreted effector proteins and other virulence factors.Chapter seven (Molecular diagnostics by genetic methods) deals with genetic methods used for classification and molecular diagnostics of Aeromonas species, including strategies designed for identifying strains at the species level from both pure cultures and environmental samples. Due to its complex historical taxonomy, Aeromonas appeared as one of the best examples to illustrate applications of molecular biology techniques in microbiology, integrating advantages and disadvantages, performance and limitations of molecular diagnostic microbial methods.Chapter eight  presents results from classical culture-dependent and 16S rRNA gene studies in the context of the ecology, prevalence and environmental factors, primarily temperature, influencing the proliferation and regrowth of Aeromonas in water, biofilm, and food.The nine and last chapter (Overall, I found the book timely because, as stated in chapter six, “since the previous book on Aeromonas over fifteen years ago there have been a number of aeromonad genomes sequenced that have been a great asset in helping laboratories to unravel the biology of this enigmatic pathogen.” Chapters are well-written, informative, well-referenced, and represent a valuable resource for scientist community in the Aeromonas field, including insights into relevant diagnostic methodologies. I particularly like the inclusion of chapter five on the beneficial Aeromonas-animal asssociations that opens the window to the evolutionary processes and genomic innovations which are the mother of the complex and generalized biological interactions in the modern world (Sime-Ngando et al., The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Host-microbe interactions have traditionally been viewed primarily from the perspective of pathogenesis and disease. The principal assumption is that a susceptible host will support microbial growth inside host tissues, leading eventually to the development of symptoms and disease. Inversely, non-susceptible (resistant) hosts will block pathogen infection and the expression of disease  beneficial co-existence between hosts and microbes  and related genera, which form obligate biotrophic associations with temperate grasses and play an important role in host physiology by enhancing growth and conferring abiotic and biotic stress tolerance. Due to their agronomic importance, Clavicipitaceous endophytes have been extensively reviewed elsewhere , for example, is a notorious pathogen of wheat, whose interaction with the host conforms to the gene-for-gene relationship (Kema et al., Z. tritici, which is known to have co-evolved as a specialized pathogen of wheat, the ability of some isolates to multiply asymptomatically inside the host as non-destructive colonists could reflect an evolutionary adaptation at a population level that maintains allelic diversity at virulence-associated loci, thereby enhancing the prospects for long-term reproductive success of the pathogen (Vanbaalen and Sabelis, Z. tritici, in particular, are wind transmitted and have been reported to travel for hundreds of kilometers, in contrast to the rain-splash, locally dispersed asexual pycnidiospores. Thus, maintaining a parasitic continuum in a pathogen population between non-destructive endophytic colonizers and virulent isolates may reflect an evolutionary adaptation that increases the long-term reproductive success of the organism. More importantly, the ability of some isolates to grow asymptomatically inside the host in a cryptic or latent manner could offer support to the idea that modern plant–pathogen interactions have arisen from the breakdown of past mutualistic interactions and a switch to a pathogenic lifestyle (Stukenbrock and Mcdonald, Whereas fungi such as The majority of studies in host-microbe interactions have thus far followed a rather reductionist approach, focusing mostly on aspects of pathogenesis and disease. However, it is possible that all of our hard-won knowledge about the activities of some species as pathogens, in fact represents a vanishing small part of the full spectrum of ecological activities that constitute their life history. Thus, for a holistic understanding of host-microbe interactions, it is vital to understand and appreciate the true breadth and dynamic nature of the parasitic, commensal or mutualistic continuum that exists between microbes and their hosts. Failing to do so may compromise efforts to manage species normally perceived as pathogens, and increase the risk of introducing invasive species into new areas. We consequently stress the importance of determining the extent to which fungi with a potentially pathogenic lifestyle exploit host plants that manifest no symptoms, and incorporating this information into epidemiological models and disease management strategies. Future studies should also focus on identifying the genetic and environmental factors that trigger a transition in the trophic interactions between hosts and parasites, and subsequently the phenotypic state of an infection. By studying in parallel the parasitic, mutualistic, and commensal lifestyles of traditional pathogens, key processes in the host-pathogen interplay and molecular communication can be identified that can be exploited for more sustainable disease control measures. Overall, we argue that a paradigm-shift in the study of host-microbe interactions is currently needed, to acknowledge the full extent of trophic interactions between parasites and hosts.The Associate Editor Benjamin Schwessinger declares that, despite being affiliated to the same institution as Ioannis Stergiopoulos and Thomas R. Gordon, the review process was handled objectively and no conflict of interest exists. 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.


Throughout the first years of the twenty-first century, neurotechnologies such as motor cortex stimulation (MCS), transcranial magnetic stimulation (TMS), and transcranial direct current stimulation (tDCS) have attracted scientific attention and been considered as potential tools to centrally modulate chronic pain, especially for those conditions more difficult to manage and refractory to all types of available pharmacological therapies. Interestingly, although the role of the motor cortex in pain has not been fully clarified, it is one of the cortical areas most commonly targeted by invasive and non-invasive neuromodulation technologies. Recent studies have provided significant advances concerning the establishment of the clinical effectiveness of primary MCS to treat different chronic pain syndromes. Concurrently, the neuromechanisms related to each method of primary motor cortex (M1) modulation have been unveiled. In this respect, the most consistent scientific evidence originates from MCS studies, which indicate the activation of top-down controls driven by M1 stimulation. This concept has also been applied to explain M1-TMS mechanisms. Nevertheless, activation of remote areas in the brain, including cortical and subcortical structures, has been reported with both invasive and non-invasive methods and the participation of major neurotransmitters  as well as the release of endogenous opioids has been demonstrated. In this critical review, the putative mechanisms underlying the use of MCS to provide relief from chronic migraine and other types of chronic pain are discussed. Emphasis is placed on the most recent scientific evidence obtained from chronic pain research studies involving MCS and non-invasive neuromodulation methods , which are analyzed comparatively. Pain is clinically identified as an early and disabling symptom, extremely frequent and common to various diseases. However, rather than simply a sensory phenomenon, pain is better characterized as a complex experience extending beyond the sensory-discriminative component of pain, or the individual capacity to identify the nature  of a particular noxious stimuli. The affective-emotional aspect of pain , as well the involvement of attention, memory of previous experiences, and anticipation, termed the cognitive-evaluative pain dimension, are fundamental pieces of this still challenging and complex puzzle  and transcranial magnetic stimulation (TMS), when applied to the motor cortex for the treatment of chronic pains. To this purpose, the scientific evidence obtained with the invasive procedure, termed motor cortex stimulation (MCS), is always used as a reference.Given the clinical challenges that chronic pain management presents, scientific pain researchers have directed their focus toward the development of novel technologies and enhancement of known strategies that permit the modulation of cortical excitability in humans through non-invasive or minimally invasive procedures. Over the past years, several studies have investigated the analgesic effects of epidural/subdural MCS, especially in refractory or intractable neuropathic pain , the motor evoked potential (MEP) amplitude, the intracortical inhibition (ICI), the intracortical facilitation (ICF), and the electromyographic cortical silent period (CSP) , has been associated with a single session of anodal M1-tDCS in both healthy subjects  receptors in TMS-induced analgesia has also been explored. The establishment of this link has its origins in animal model studies  studies demonstrated increases in Glx, a combined marker of glutamine and glutamate, and N-acetylaspartate (NAA), which is considered to be a measure of neuronal integrity, in the parietal cortex underneath the anode (Clark et al., Recently, studies have also been explored the possible neurochemical actions of tDCS. Proton magnetic resonance spectroscopy (Placebo is a factor that must always be considered when analyzing the effects of chronic pain therapies. Although several clinical trials involving non-invasive brain stimulation for pain relief have found significant differences between active and sham stimulation (Fregni et al., Since the serendipitous observation that M1 stimulation produces significant clinical improvements in chronic neuropathic pain patients, this cortical region became the main target of several neuromodulatory techniques devoted to ameliorating chronic pain in human subjects. In fact, it has been reported that the stimulation of cortical regions adjacent to the primary motor cortex fail to produce similar analgesic effects, confirming the prominent role of M1 stimulation for pain control. Nevertheless, the intricate neurophysiological mechanisms that explain the clinical efficacy of M1 stimulation for pain relief are not completely understood. Evidence from MCS studies indicates that its analgesic mechanisms involve the activation of top-down controls related to the excitation of intracortical horizontal fibers. This concept can also be applied to TMS. However, results of neuroimaging studies also suggest that MCS and TMS act through modulation of deeper and remote brain structures related to pain, such as the insula, anterior, cingulate cortex, basal ganglia, thalamus, and brainstem. Interestingly, enhanced current flow in the same areas has also been demonstrated with tDCS. In addition, the neurochemical mediation driven by M1 stimulation has been recently unveiled in studies involving MCS, TMS, and tDCS. Opioidergic, glutamatergic, GABAergic and serotoninergic neurotransmissions are now considered components for the whole process. Nevertheless, there are still questions that must be answered, including those regarding the participation of other mechanisms of endogenous pain control, the clinical relevance of increasing anatomical and functional specificity in non-invasive procedures, and the presence and significance of a placebo effect. The answers to these questions are expected to be among the future perspectives of the field.MD, NF, RT, AC and AD drafted the manuscript. All authors read and approved the current version of this article.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.


Although digital angiography remains as the gold standard for imaging the celiacarterial trunk and hepatic arteries, multidetector computed tomography inassociation with digital images processing by software resources represents auseful tool particularly attractive for its non invasiveness. Knowledge ofnormal anatomy as well as of its variations is helpful in images interpretationand to address surgical planning on a case-by-case basis. The present essayillustrates several types of anatomical variations of celiac trunk, hepaticartery and its main branches, by means of digitally reconstructed computedtomography images, correlating their prevalence in the population with surgicalimplications. However, with the arrival ofmultidetector computed tomography (CT) angiography and modern image reconstructionprograms, this imaging method becomes an additional option for the detailedinvestigation of arteries with the significant advantage of itsnoninvasiveness.Multidetector CT angiography allows the visualization of small caliber shortarteries by means of maximum intensity projection (MIP) and three-dimensional volumerendering (VR) techniques.Angiography is the gold standard for CAT, and its branches, visualizationThe present essay is aimed at describing the normal anatomy and commonly found CATand HAS variations.The images shown in the present essay were collected from the personal files of theauthors and acquired in a Brilliance 64-channel multidetector CT apparatus .The scan protocol, with small sporadic variations, consisted in axial sections, slicethickness of 1 mm, pitch 0.8. The contrast agent Ultravist (Bayer) was utilized, ata concentration of 769 mg/mL, intravenously injected by means of an injection pumpat a rate of 5 mL/s, with bolus tracking time delay. A standard field of view (250mm) was utilized. The images reconstruction thickness was 2 mm. An ExtendedBrilliance Work Space workstation was utilized with the software Philips Brilliancefor computed tomography . In order to define the arterial pattern, analyses were performed in the axial planewith reconstruction techniques in the coronal and sagittal planes in multiplanarreconstructions (MPR), as well as three-dimensional reconstructions with the MIP andVR techniques. The normal patter and the main CAT and HAS variations weredemonstrated.. Song et al. have studied 5,002 cases of CAT and reported theoccurence of variations in 10.9% of cases. However, as concomitant CAT and HAS variations areconsidered, the rate increases to 55% of patients .CAT variations are not infrequent - is described as the trifurcation originating the left gastricartery, splenic artery and common hepatic artery,5 , wherethe left gastric artery originates from the splenic artery, forming a common trunkon trunk; hepatogon trunk. The absAs regards the HAS, it is described as normal in cases where the common hepaticartery originates the hepatic artery propria after the emergence of thegastroduodenal artery; and the hepatic artery propria, on its turn, divides intoright and left hepatic arteries within the hepatoduodenal ligament, at fewcentimeters from the liver surface.According to Koops et al., the frequencies of the normal HAS pattern as per theHiatt's classification , arecon .Amongst the most described variations, the following frequencies were found: 3-17%(type II), relocation of the left hepatic artery; 7-18% (type III), relocation ofthe right hepatic artery ; and 1.5.Normally, cases of Hiatt's type III are the most prevalent and play a relevant rolein procedures involving the liver, as after originating from the superior mesentericartery, the right hepatic artery runs posteriorly to the portal vein, which mightconfuse the surgeon, since in the normal pattern (type I) such artery is locatedanteriorly to the portal vein, within the hepatoduodenal ligament. Thus, one of thereasons for understanding those variations is avoiding iatrogenic eventsIn Hiatt's type II - left hepatic artery relocation -, procedures such as gastrectomyshould be cautiously performed, considering that in most of such cases, the lefthepatic artery emerges from the left hepatic artery; thus, in case of section of theleft gastric artery, a possible ischemia of the whole functional left hepatic lobemight occur..Understanding the pattern of variation of the hepatic arteries becomes imprescindiblefor the development of the liver transplant.With the introduction of laparoscopic surgeries with reduction of the surgical fieldview, it is necessary to understand the pattern of variations of the upperabdomen.The arterial patterns are relevant in the planning of the whole surgical andradiological procedure involving the upper abdomenConsidering the relevance of the mentioned variations, the authors suggest thatradiologists should investigate the arterial pattern and inform them in reports ofsurgeries and invasive examinations of the upper abdomen.Considering that the vascularization of a great part of the gastrointestinal systemoccurs from CAT and HAS branches, the knowledge about anatomical variations andrespective frequencies is of paramount relevance in the planning of upper abdomensurgeries to avoid procedural errors and medical iatrogenic events.


LSCM is capable of showing MGs modifications during aging and in the most diffuse ocular surface diseases such as dry eye, allergy, and autoimmune conditions and in the drug-induced ocular surface disease. On the other hand, LSCM may help clinicians in monitoring the tissue response to therapy. In this review, we summarized the current knowledge about the role of in vivo LSCM in the assessment of MGs during aging and in the most diffuse ocular surface diseases.Meibomian glands (MGs) play a crucial role in the ocular surface homeostasis by providing lipids to the superficial tear film. Their dysfunction destabilizes the tear film leading to a progressive loss of the ocular surface equilibrium and increasing the risk for dry eye. In fact, nowadays, the meibomian gland dysfunction is one of the leading causes of dry eye. Over the past decades, MGs have been mainly studied by using meibography, which, however, cannot image the glandular structure at a cellular level. The diffusion of the Meibomian glands (MGs) are holocrine glands embedded in the tarsal plate of the eyelids. Each gland comprises multiple acini connected by a long common central duct running throughout the entire length of the gland . The fun in vivo . In VKC, extensive periglandular Langerhans cell infiltration, with blurred lumen contours and hyperreflective solid matter in the lumen, was described , 39. IbrIn conclusion, in patients with dry eye, confocal microscopy revealed that the inflammation is the first and most important pathogenetic mechanism involved in the development of the MG alterations. The main findings were the increased acinar density and the reduced orifice diameter, which differentiated DES from primary or secondary MGD . In thisAKC and VKC are two of the most common and important ocular allergies. AKC is a bilateral chronic hypersensitivity disease characterized by conjunctival papillary hypertrophy, acute and chronic conjunctivitis, keratitis and corneal ulceration, eyelid eczema, and blepharitis –53. TearVKC is a recurring seasonally inflammatory condition of the cornea and conjunctiva, characterized by giant tarsal papillae and pathological changes of the conjunctival epithelium and MGs. LSCM documented intense infiltration of eosinophils and multinucleated granulocytes in the superficial conjunctival epithelium, extensive periglandular lymphocytic cells infiltration. These inflammatory alterations lead to macroscopic modifications of MGs, represented by glandular atrophy, and ductal dilation . The samIn allergic patients, the fibrotic involution of MG acinar units is more extensive than that observed in obstructive MGD, where the pressure induced by the stagnating meibum induces enlargement of MG acini .In summary, MG modifications in patients with ocular allergy diseases are in part different from those observed in MGD and dry eye, since the periacinar fibrosis, along with the inflammatory infiltration and the meibum stagnation, represents the main finding . In thesIn the last years, LSCM was also used to evaluate the impact of glaucoma therapy on ocular surface structures –7, 55, iMGs presented a reduction of glandular density and area and increased reflectivity of the acinar secretion, more evident in patients treated with two or more drugs compared to patients in monotherapy. The reduced density and area were expressions of glandular loss and reduced meibum production, respectively. Higher secretion reflectivity indicated increased secretion viscosity.These modifications were similar to those observed in patients with MGD, with the exception of the glandular area that appeared to be reduced in glaucoma . Higher Other aspects characterizing MGs in glaucoma are the ductal orifice dilation and the inflammation of the glandular wall and interstice, which confocal microscopy documented with the presence of punctate hyperreflective elements. The ductal orifice dilation was probably an adaptive mechanism to overcome the high secretion density and duct blockage induced by treatment.The potential induction mechanisms are the toxicity and an inflammatory or immune-mediated response, with the inflammation probably being the first step in the cascade of glandular modifications , 12.β-blockers. Therefore, BAK and PGA, with their toxic and inflammatory stimuli, play the most crucial role in the induction of glaucoma therapy-related MGD.In patients in monotherapy LSCM contributed to clarifying the role of preservatives ) and active compounds in the acinar modifications: preserved drugs were more toxic than preservative-free (PF) formulations, with preserved prostaglandin analogues (PGA) being more toxic than preserved Very interestingly, all the microscopic modifications of MGs significantly correlated with the ocular surface disease index (OSDI) score, break-up time (BUT), and Schirmer test I (STI). These correlations indicated that MGs alterations, as assessed with LSCM, were indicators of dry eye and have a main role in development of the therapy-related ocular surface disease in patients with glaucoma. Thus, the preservation of the structural and functional integrity of MGs represents a critical challenge during medical management of glaucoma.In conclusion, the long-term antiglaucoma therapy has a strong negative impact on MGs functionality: the iatrogenic changes of MGs, in fact, lead to meibomian gland dysfunction very similar to primary MGD, with the exception of the reduced acinar area . It is hThese modifications present evident clinical implications since they could strongly affect adherence and persistence of treatment.Until recently, the microanatomy evaluation of the ocular surface structures was limited to the impression cytology, which is based on sampling the superficial epithelial layers. This methodology, despite being highly reproducible, does not allow exploring deep tissues and induces discomfort to patients. in vivo biopsy of all ocular surface tissues, at different depths, and the tissue analysis at cellular and subcellular level. The meibomian gland is one of the ocular surface structures that mostly benefited from the confocal assessment, since the standard meibography can only give a macroscopic analysis of the entire gland. The possibility of imaging MGs at cellular level, in vivo, and in a noninvasive way permit studying these structures in the most diffuse and important ocular surface diseases such as dry eye, ocular allergy, and drug toxicity.The rapid diffusion of LSCM in the last decades permitted anIn this way, LSCM may allow a preclinical diagnosis of MG conditions, potentially before glands begin to malfunction. In addition, since confocal microscopy does not produce significant discomfort for patients, this technique offers the advantage to strictly monitor the disease over time, to anticipate the treatment when required, to follow the response to therapy, and to modify the therapy regimen accordingly.Moreover, the fine microscopic assessment could also clarify the pathophysiology of these conditions. This is even more important considering that the incidence of ocular surface diseases is exponentially growing in the last years.


In the last decade, the availability of the cardiac biomarkers N terminal pro-natriuretic peptide type-B (NT-proBNP) and troponins has radically changed the approach to diagnosis, staging, and response assessment of cardiac amyloidosis. Despite similar appearance at standard imaging, the main types of amyloidosis involving the heart are characterized by different rates of progression and different outcomes. In general, patients with ATTRm amyloidosis have lower concentrations of cardiac biomarkers and usually a better prognosis than subjects suffering from AL amyloidosis. Nevertheless, the clinical manifestations of ATTRm amyloidosis are heterogeneous. Different amyloidogenic TTR mutations give rise to different cardiac phenotypes, ranging from exclusively neuropathic diseases, through mixed phenotypes, to mutation characterized by severe cardiac dysfunction. For instance, one of the mutations associated with cardiac involvement in ATTRm amyloidosis, Ile68Leu, presents with high NT-proBNP concentrations, comparable to those observed in patients with cardiac AL amyloidosis. In our series, survival of patients with Ile68Leu ATTRm is not different from that of cardiac AL patients. Thus, it is possible that NT-proBNP represents a viable marker for early diagnosis of cardiac involvement in ATTR patients with “aggressive” mutations. This marker could be considered in the screening of carriers of these mutations. Despite the lack of systematic studies of cardiac biomarkers in ATTR amyloidosis, the results of several small published series and the analysis of the Pavia patient population, indicate that NT-proBNP and cardiac troponins have prognostic value in this disease, correlating to amyloid burden and adding to known prognostic factors. The availability of several novel therapeutic options for ATTRm, require objective means for the assessment of treatment efficacy in routine clinical practice and in clinical trials. Early results from the Pavia series indicate that NT-proBNP progression portends a poor prognosis, and cardiac biomarkers will probably find their place in the evaluation of response to therapy in ATTRm amyloidosis. Cardiac biomarkers will most likely become increasingly useful in the management of patients with ATTRm amyloidosis in the near future. However, studies of biomarkers in this disease are hindered by small numbers and disease heterogeneity. Prospective studies and international collaboration are warranted to define the role of cardiac biomarkers in the diagnosis and follow-up of cardiac ATTRm amyloidosis.


The hypothalamic-pituitary-adrenal (HPA) axis is critical for life. It has a circadian rhythm that anticipates the metabolic, immunoregulatory and cognitive needs of the active portion of the day, and retains an ability to react rapidly to perceived stressful stimuli. The circadian variation in glucocorticoids is very ‘noisy’ because it is made up from an underlying approximately hourly ultradian rhythm of glucocorticoid pulses, which increase in amplitude at the peak of circadian secretion. We have shown that these pulses emerge as a consequence of the feedforward–feedback relationship between the actions of corticotrophin hormone (ACTH) on the adrenal cortex and of endogenous glucocorticoids on pituitary corticotrophs. The adrenal gland itself has adapted to respond preferentially to a digital signal of ACTH and has its own feedforward–feedback system that effectively amplifies the pulsatile characteristics of the incoming signal. Glucocorticoid receptor signalling in the body is also adapted to respond in a tissue-specific manner to oscillating signals of glucocorticoids, and gene transcriptional and behavioural responses depend on the pattern (i.e. constant or pulsatile) of glucocorticoid presentation. During major stressful activation of the HPA, there is a marked remodelling of the pituitary–adrenal interaction. The link between ACTH and glucocorticoid pulses is maintained, although there is a massive increase in the adrenal responsiveness to the ACTH signals. It is 65 years since Hans Selye  demonstrThe principal endogenous glucocorticoids are cortisol  and corticosterone (the majority of rodents and birds), which we shall collectively refer to as CORT. CORT is characterised by its circadian pattern of basal secretion from the adrenal glands, with the highest levels being seen just prior to the start of the active cycle (i.e. just prior to awakening), followed by a gradual decline, reaching nadir levels during the inactive phase. This daily rhythm is under central regulation by corticotrophin-releasing hormone (CRH) and arginine vasopressin (AVP) from the paraventricular nucleus (PVN) of the hypothalamus. The PVN receives a powerful input from the suprachiasmatic nucleus (SCN) ,11, predde novo synthesis and release into the systemic circulation, CORT is able to act at target tissues to exert metabolic, cardiovascular, immunological and cognitive responses in response to the relevant inputs , which in turn is released into the general systemic circulation and stimulates the adrenal cortex to produce CORT. After t inputs . At physt inputs ,16.Splanchnic nerve innervation of the adrenal glands also contributes to the circadian rhythm. The adrenal glands receive autonomic (sympathetic) innervation that lies under the influence of neuronal projections of the autonomic portion of the PVN and, consequently, the SCN to the spinal cord . The splThe diurnal pattern of peak and trough CORT secretion is considerably more pronounced than the circadian pattern of ACTH. Studies examining 24-h cortisol and ACTH profiles of healthy individuals reveal a four- to six-fold difference between the circadian peak and trough amplitudes of CORT, whereas the corresponding ACTH difference is approximately two- to three-fold. The characteristics of these 24-h profiles alter with age (trough CORT and ACTH levels are higher in elderly people)  or in reThe circadian variation of cortisol also interacts with several other important biological oscillations, such as activity, body temperature  and the The classic transcriptional (genomic) effects of CORT are relatively slow and are mediated by activation of its cognate intracellular nuclear receptors, the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR) ,28. MRs The distribution of these receptors is tissue-specific, providing an additional mechanism for regulatory control, with GRs being present throughout the brain and periphery and MRs having a more limited localisation; they are mainly present in cardiovascular tissues, liver and kidneys, as well as in most corticolimbic regions of the brain, including those involved in HPA axis regulation . HippocaThe physiology underlying nongenomic, rapid glucocorticoid effects has been linked to various second messenger cellular systems under different conditions. For example, the binding of pharmacological doses of CORT to GR stimulated phosphatidylinositol 3-kinase and protein kinase Akt, leading to acute, cardioprotective endothelial NO synthase activation and nitric oxide-dependent vasorelaxation in the heart of rat models of myocardial infarction . At the The circadian variation seen in CORT's natural rhythm is made up of underlying discrete pulses of HPA activity, a rapid and dynamic ultradian rhythm Fig. 43). Th. Th43). The origin of the ultradian rhythm has been subject to many hypotheses, although it has always been assumed to result from some elusive hypothalamic pulse generator. Recently, however, it has become clear that, unlike the circadian rhythm, ultradian pulsatility is not under central regulation from the SCN because, as in SCN-lesioned rodents, the ultradian rhythm persists despite a loss in circadian rhythmicity . The cenin vivo experimental work. This modelling was based on the previously accepted understanding that (i) there is a delay in CORT secretion because of the requirement for its de novo synthesis and (ii) this CORT can induce a rapid, nongenomic negative-feedback at the level of the pituitary corticotrophs to inhibit CRH induced ACTH secretion  suppressed rodents, pulsatile ACTH infusions result in pulsatile corticosterone secretion. When a constant infusion of ACTH is given at the same total dose, there is no adrenal response at all . We havein vivo microdialysis studies that confirmed synchronous total blood and hippocampal free CORT pulses  only. Indeed, oscillations in CORT induce a phenomenon known as ‘gene pulsing’  and, to a lesser extent, to albumin. It is only the free fraction of CORT that is active, and this comprises only approximately 5% of total CORT under basal (nonstressed) trough levels . The bioT pulses ,77. The ing’ Fig. 78,79);,79;in viing’ Fig., as welling’ Fig.,82. GR ting’ Fig..fos mRNA response to the neural activation by noise stress is markedly altered in the amygdala, hippocampus and hypothalamic PVN depending not only on the pattern of presentation (constant versus pulsatile), but also on the phase within each pulse. During the rising phase of the pulse, an exaggerated noise stress response was observed in comparison to the falling phase  for which the neuroanatomical bases lie in the corticolimbic areas and primarily the hippocampus. Within these systems, glucocorticoids interact at multiple levels, resulting in structural and frequently opposing or bidirectional functional consequences . At the A receptors, whereas basal levels reduce neurotransmission  populations in hippocampus and PFC by increasing the number or the probability of vesicular exocytosis at the presynaptic level via a rapid nongenomic MR effect . This issmission .In relation to the wide spectrum of glucocorticoid actions, and under different physiological or pathological circumstances, pulsatility offers a means of temporally dissociating or combining the MR- from the GR-dependent actions Fig.. This teReturning to the effects of glucocorticoids on synaptic activity and neurotransmission, a recent elegant study  provideset al.  . The aim


In many European estuaries, extensive areas of intertidal habitats consist of bare mudflats and sandflats that harbour a very high abundance and biomass of macrobenthic invertebrates. The high stocks of macrobenthos in turn provide important food sources for the higher trophic levels such as fish and shorebirds. Climate change and associated sea-level rise will have potential to cause changes in coastal and estuarine physical properties in a number of ways and thereby influence the ecology of estuarine dependent organisms. Although the mechanisms involved in biological responses resulting from such environmental changes are complex, the ecological effects are likely to be significant for the estuarine benthic macrofauna and hence the consumers they support. This paper reviews the utilisation patterns of estuarine intertidal habitats by shorebirds, fish and crustaceans, as well as factors affecting the distribution, abundance and biomass of estuarine macrobenthos that is known to be important food source for these estuarine predators. This study also provides simple conceptual models of the likely impacts of sea-level rise on the physical and biological elements of estuarine intertidal habitats, and implications of these results are discussed in the context of sustainable long term flood and coastal management in estuarine environments. Estuaries are ecologically important coastal environments situated between freshwater rivers and the sea, characterized by highly varying physicochemical, morphological and hydrological conditions ,2, whichThis paper considers the implications of climate change and associated sea level-rise for estuarine and coastal management on time scales of decades up to a century (medium- to long-term) with particular reference to the ecology of macrofauna on an estuarine intertidal habitat. This study first reviews the utilisation patterns of estuaries by shorebirds, fish and crustaceans, as well as factors affecting the distribution, abundance and biomass of estuarine intertidal macrobenthos that is an important food source for those predators. This is followed by a brief account of the most likely sea-level rise scenarios and an account of the likely impacts on the physical elements of coastal and estuarine ecosystems. Simple conceptual models are then presented to consider the likely impacts of climate change and sea-level rise on the physical and biological properties of estuarine intertidal habitats with reference to a case study of macrobenthic biomass in relation to changes in sediment particle size, beach morphology and salinity regime in the Humber estuary, UK. How changes in other environmental parameters such as ambient temperatures or nutrient loading may interact with the effects of sea-level rise to shape the structure and function of invertebrate assemblages is also considered. Finally, the adaptive strategies likely to be adopted to cope with sea-level rise are discussed and implication is given as to how to identify appropriate ecological objectives for sustainable shoreline schemes in estuarine environment in the future.In coastal and estuarine ecosystems, birds are often regarded as top predators and fish occupy intermediate trophic levels, both of which are supported by a large amount of intertidal benthic macrofaunal prey. The high benthic biomass is a reflection of the presence of extensive intertidal flats and fine sedimentary deposits created through processes such as tidal asymmetry and flocculation . TypicalSpartina anglica, over the mud flats in southern England resulted in reductions in the numbers of Dunlin using affected areas [Much of the interest from conservation and management agencies in evaluating the effects of sea-level rise on estuaries lies in the potential impact on these higher trophic levels, especially migrating shorebirds ,19. In Ned areas . Similared areas . Sea leved areas .Playtchthys flesus, Pleuronectes platessa), gobies (Pomatoschistus spp.), crabs (Carcinus maenas) and shrimp (Crangon spp.), move onto the flats to feed on mobile epifauna and sedentary infauna [Arenicola and Heteromastus, the feeding tentacular crowns of fanworms and siphons of bivalve molluscs [Gadus morhua or whiting Merlangius merlangus are heavily dependent upon estuarine macrobenthos through direct feeding on benthic infauna, or through feeding on other fish species, crab C. maenas, shrimp C. crangon which have themselves fed on the benthic invertebrates [Many fish and crustaceans also utilise the intertidal flats with the flood tide to feed ,7. A div infauna . They almolluscs ,31. Eventebrates . Migratotebrates . With retebrates . In troptebrates . In viewtebrates  and extetebrates . Some letebrates .Corophium voltator, on the Ythan estuary, Scotland [Arenicola marina [−2 yr−1, which accounts for 30% of the estimates of total benthic production, in the Oosterschelde in the Netherlands, whereas in Ythan estuary in Scotland, the fish are estimated to consume three times the amount of food consumed by the birds [Degree of dependence on estuarine benthic macrofauna can also be measured by predation pressure from those fish, crustaceans and shorebirds. For instance, redshank are estimated to remove 16%–38% of intertidal amphipod crustacean, Scotland , bar-taia marina , and oysa marina . Food cohe birds . Given tEurydice pulchra, dominated, but with moderating exposure, the proportion of polychaetes increased. The highest biomass was found on sheltered beaches dominated by molluscs, mainly the bivalve Tellina tennuis. Similarly, McLachlan [Mytilus edulis) can form extensive biogenic reefs with substantial biomass on at lower shore levels, particularly at the mouths of meso- and macro-tidal estuaries where there is favourable tidal flow [Exposed shores and sandy intertidal flats are often located at the outer region of an estuary. The overall morphology and dynamics of the beach, captured by the concept of “dissipative”  and “reflective”  spectrum, is one of the predominant factors in controlling the distribution and abundance of intertidal faunal assemblages on these exposed sandy intertidal flats . For inscLachlan  reviewedcLachlan . In NW Edal flow . Except Nerine and Ophelia become more frequent in area of moderate disturbance, and with more sheltered conditions, burrowing bivalve molluscs, such as tellinids and large clams, and epibenthic mussels become abundant [Neomysis integer, Melita obtusata, Dexamine spinosa, Stenothoe marina, Idotea spp.).Sheltered shores are found in areas of low energy and they have poorly sorted sediments with high levels of organic matter and a high silt content . Fine paabundant . In addiNereis diversicolor; a mid-tide zone of the bivalve molluscs Scrobicularia plana, Cerastoderma edule and Macoma balthica, and where some sand is present, the lugworm Arenicola marina; a low-shore zone with the bivalves Mya arenaria and possibly Tellina fabula, depending on the sediment particle size. The relationships between tidal level and abundance or biomass of macrofauna tend to show similar patterns between tidal flats in different locations. For instance, in different parts of the Dutch Wadden Sea, Beukema [The nature of the substratum and tidal level can markedly affect the distribution patterns of benthic macrofauna within an intertidal area . In Brit Beukema  showed t Beukema . It is cArenicola marina also diminishes with distance from the sea [In contrast to studies of macrofaunal biomass and production, many publications on the distribution range of species and the diversity of benthic assemblages in estuaries have focused on salinity gradient ,48,49. D the sea . It has  the sea , and Lit the sea  points o the sea ,50,56,57 the sea  showed tCerastoderma edule, Nephtys hombergii, and Lanice conchilega, showed minimal biomass values due to low over-winter survival during severe winters. In addition, bivalve species such as Macoma balthica show a significant negative correlation between winter temperatures and subsequent recruitment, with higher M. balthica densities in the year following a cold winter, probably due to an increase in fecundity [Cerastoderma edule is unusually high in the summers following widespread mortality of the consumers of their spat [Inter-annual variability in climatic conditions will have strong influence on temporal variability of estuarine macrobenthic assemblages through changes in individual growth rates, fecundity and recruitment success. In the long-term study in the Wadden Sea in the Netherlands, Beukema and colleagues  noted thecundity ,59,60. Peir spat . Changeseir spat  investigeir spat . Indireceir spat  and the There is an increasing body of evidence which to confirm food supply regulates benthic biomass and secondary production by affecting individual growth and fecundity of adult invertebrates at the estuarine system scales ,17,65. FThe Earth’s climate has warmed by 0.6 ± 0.2 °C during the last century (1901–2000)  and reseSea-level changes reflect climate change mainly through thermal expansion of the upper layer of the oceans and release of water from glacier or ice sheet melt ,70. Sea-At the global scale, models predict that the annual global mean surface air temperature will rise 1.1–6.4 °C by 2100 based on increases of greenhouse gases , and theHowever, there are likely to be marked regional and local-scale variations relating to sea-level rise ,75,76, aIn addition to global warming and increased sea level, there are predicted changes in the frequency of severe droughts, excessive precipitation and extreme events together with changes in seasonal and diurnal temperatures at local and regional scales ,77. ThesThere are several estuarine physical processes in response to sea-level rise that are of great relevance for the management of estuarine ecosystems. Rising sea level will result in changes in coastal and estuarine geomorphology as a consequence of increased water depth and enhanced wave and tidal energy . Such phIn addition to the loss of habitat due to coastal squeeze, change in sedimentation and geomorphological processes over intertidal flats in response to sea-level rise will have important implications for the estuarine ecosystem. Estuarine intertidal mudflats with fine muddy sediment and a rich organic matter content typically support high numbers of benthic invertebrates, which in turn plays a central role in maintaining estuarine food webs ,55. ThroFinally, there will be effects of sea-level rise on the estuarine salinity gradient from freshwater to marine conditions. The structure of this gradient varies according to estuarine morphology, freshwater run-off and turbulence or mixing ,15. ManyThese general processes will be modified by local geology, the land use of drainage basin, the size, shape and human usage of the estuary , and it The above review indicates that benthic macrofauna plays a pivotal role in estuarine trophic interactions and the standing stock of macrobenthos is likely to be affected by a number of environmental parameters. Based on this, Given uncertainties and the stochastic nature of some of the environmental stressors which primarily affect temporal aspect of estuarine macrobenthos, construction of deterministic models based on all the inter-relationships presented in Along the longitudinal gradient, present macrobenthic biomass per unit area is controlled mainly by salinity gradient and sediment particle size distribution (the black solid line in Arenicola marina and Macoma balthica which normally establish at higher shore levels, may be adversely affected. These scenarios indicate that some intertidal habitats are likely to experience loss of macrobenthos not only through the quantitative reduction in spatial area of intertidal habitats but also their qualitative reduction due to adverse shifts in local physical conditions in response to sea-level rise, and combination of such effects could lead to a disproportionate reduction of macrobenthic biomass than a straightforward loss of intertidal area would suggest in an estuarine ecosystem.Along the vertical gradient from mean high to low water levels, present macrobenthic biomass per unit area is affected by the tidal height (the black solid line in The relations in In the case of estuaries with narrow, restricted entrances or with a bar, it is possible that a general increase in sediment supply could lead to accretion of material on intertidal flats as the suspended matter enters a less hydrodynamically energetic environment , as seen−2, whereas other extensive mudflat of Spurn Bight situated at the outer region of the Humber showed an average biomass of 17.1 g AFDW m−2. This indicates that creation of 400 ha intertidal habitat in the upper region by implementing managed realignment could not even compensate for the loss of 1 ha of mudflat in Spurn Bight in the Humber in terms of securing total macrobenthic biomass available for higher trophic levels at estuarine system scale. Replacement sites must therefore be identified and their appropriate size must be calculated on the basis of a sound ecological understanding of estuarine ecosystems if we are to counteract the future loss of intertidal habitats and to maintain the current levels of macrobenthic biomass through the implementation of such restoration schemes. In typical European estuaries, 90%–95% of the estuarine intertidal habitats are bare mudflats and sandflats  that are2 of wetland area were attributable to the increased rate of relative sea-level rise during the 20th century [A further consideration is that any delay of implementing appropriate adaptive strategies in the coastal region will reduce their effectiveness in reducing ecological and socio-economic impacts over the medium and long term, when the effects of sea-level rise could be catastrophic. On a regional scale, there is already increasing evidence that intertidal habitats is being lost at alarming rate as a result of rising sea-levels coupled with coastal subsidence. For example, almost 60% of wetland loss observed along the northern Gulf of Mexico was due to the net effect of sea-level rise and subsidence , and alo century .Finally, sea-level rise is just one of many stressors which will affect estuarine environments in the future and it is by no means clear whether the combined impacts of different stressors can be simply treated as additive. If they are non-additive in their impacts, then a small change in sea-level rise may exert an impact greater than expected. Further studies are needed to increase certainty in predicting how estuaries are likely to change and interact with contemporary climate change and associated rising sea levels in order to prevent any one of the stressors from triggering catastrophic impacts on the ecology of intertidal habitats, which may in turn cause widespread impacts on heavily estuarine dependent higher trophic levels including humans.


Furthermore, it has been recognized recently that defects in neuronal migration lead to brain malformations and diverse psychiatric and neurological disorders including schizophrenia, epilepsy, and autism. Accordingly, it is important to elucidate the molecular mechanism of neuronal migration in the neocortex, in order to understand not only the basic principles of brain development but also the pathological processes of these disorders. In this special issue, we attempt to cover topics ranging from the basic mechanisms of neocortical development to the malformation and evolution of the neocortex, with a special focus on neuronal migration.The mammalian neocortex shows an extremely well-organized structure that underlies higher brain functions such as cognition, language, and memory. The neocortex consists of a six-layered structure, in which excitatory and inhibitory neurons form complex neural circuits in concert with glial cells. As a result of recent technological innovations in live imaging and Toma and Hanashima reviewed the mechanisms that regulate the changes in RGC competency and neuronal subtype transitions, focusing on the regulatory networks of various transcription factors including Foxg1. At the earlier stage of neocortical development, RGCs predominantly produce a large number of neurons, but later they change into glia-restricted progenitors. After the discovery of the importance of astrocytes in synaptic plasticity and blood flow, the mechanisms of glial development have attracted increasing interest for many neuroscientists. Tabata reviewed the mechanism controlling the production of diverse types of astrocytes and their migration behavior, demonstrating the multiple origins of glial cells in the neocortex.Radial glial cells (RGCs) are primary progenitors capable of generating various types of neurons and glial cells, which include Cajal-Retzius cells, subplate neurons, pyramidal neurons, interneurons, oligodendrocytes, and astrocytes. Thus, it is important to know how these diverse types of cells are generated from RGCs and integrated into complex neocortical circuits. Sakakibara and Hatanaka reviewed the sequential events in polarization processes of both excitatory and inhibitory neurons, and they discussed the underlying molecular mechanisms.Neocortical circuits consist of highly interconnected excitatory glutamatergic and inhibitory GABAergic neurons, which are generated from distinct pools of RGCs. The excitatory neurons are generated from RGCs localized in the ventricular zone of the dorsal telencephalon and migrate radially toward the pial surface in an inside-out manner . On the other hand, inhibitory neurons mainly originate from the ventral telencephalon and migrate tangentially into the neocortex . In spite of such different developmental origins, both excitatory and inhibitory neurons go through the multipolar stage with several minor processes in the neocortex before axon extension. Then, they undergo dramatic morphological changes to initiate axon formation, namely, neuronal polarization. Azzarelli et al. reviewed the roles of Rnd proteins, “atypical” Rho family members, in neuronal migration and discussed its upstream and downstream pathways. The functions of many cytoplasmic proteins including cytoskeletal components are regulated by phosphorylation and dephosphorylation processes. Ohshima focused on protein kinases, including CDK5 and JNKs, and reviewed their regulatory roles in cytoskeletal organization during multipolar-bipolar transition and radial migration. Ohtaka-Maruyama and Okado comprehensively summarized the molecular pathways involved in these developmental processes, emphasizing the importance of subplate neurons in the development and evolution of the six-layered neocortical structure.At the multipolar stage, the excitatory neurons transiently use a multipolar migration mode, namely migration with no fixed direction, in the subventricular and intermediate zones. Then, they adopt a bipolar shape during neuronal polarization and migrate quickly toward the pial surface along RGC processes, which is called locomotion mode. Many kinds of molecules are involved in these dynamic changes in the morphology and behavior of neurons. Small GTP binding proteins belonging to the Rho family play critical roles in cytoskeletal regulation during such dynamic processes. Kondo et al. demonstrated that subplate neurons transiently express high levels of secretary proteins such as connective tissue growth factor, neuroserpin, and insulin-like growth factor binding protein 5, which may be involved in cortical circuit formation. Greenman et al. reported a novel finding that autotaxin (ENPP2), a secretary enzyme bearing lysophospholipase D activity, regulates the localization and adhesion of neural progenitor cells independent of its catalytic activity. Maeda reviewed the roles of proteoglycans in neuronal polarization and migration and discussed the possibility that extracellular matrix regulates the distribution and activity of multiple secreted factors in the developing neocortex. In addition to the long-range gradient of secreted factors, axon pathfinding is also regulated by short-range guidance cues and direct cell-cell contacts mediated by guidepost cells. Squarzoni et al. reviewed the roles of already known guideposts such as Cajal-Retzius cells for entorhinal-hippocampal axons and corridor cells for thalamocortical axons, and further proposed a new class of guidepost cells, microglia, in the cortex.It is apparent that neuronal migration and wiring are regulated by various secreted factors such as growth factors, chemokines, and extracellular matrix molecules, although their mechanisms are poorly understood. Hayashi et al. reviewed the differences in the migratory behaviors of neocortical and hippocampal neurons, which lead to the formation of distinct layered structures in these two cortical regions. Defects in the migration of excitatory and inhibitory neurons can lead to the various neurological and psychiatric disorders. Kato reviewed recent development in the understanding of the genetic bases of neuronal migration disorders in terms of genotype-phenotype correlations, focusing mainly on lissencepahaly. Muraki and Tanigaki discussed the possible relationship between neuronal migration defects and behavioral abnormalities relevant to schizophrenia based on studies using genetically defined animal models. The evolutionary approaches should greatly deepen our understanding of the mechanisms underlying neocortical development. Nomura et al. established the method of in ovo electroporation and ex ovo culture of reptilian embryos. Comparative studies using this method will provide significant insights into the origin of the mammalian neocortex.Hippocampal formation has a close relationship with the neocortex both functionally and structurally, but it shows a distinct arrangement of pyramidal neurons from that of the neocortex. It is hoped that the special issue entitled “Mechanisms of Neuronal Migration during Corticogenesis” will serve as a valuable resource for many neuroscientists to promote their research perspectives. Finally, as topic editors, we would like to express our sincere appreciation to all the authors for their outstanding contributions and to all the reviewers for their insightful comments on the papers. We also thank the editorial office and the production staff for their unceasing efforts and dedication.CO wrote the manuscript. KN, AP, and NM revised the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


A 16-year-old boy with an unremarkablemedical history presented with complaints of chest asymmetry present since birth. Patient has no other complaints, including pain, decreased strength, or restricted range of motion.What is Poland syndrome?How does this condition present and how is it diagnosed?What are the treatment options?Poland syndrome is a sporadic, congenital condition with a wide spectrum of presentation. Absence of the sternocostal head of the pectoralis major muscle is considered pathognomonic for the condition. There is also an association with ipsilateral chest wall and limb derangements, the most common being syndactyly. The severity of Poland syndrome differs from person to person and is often not diagnosed or reported. The causes are unknown, although an interruption of the embryonic blood supply to the subclavian arteries around the 46th day of development is the prevailing theory.Diagnosis of Poland syndrome is clinical, as the deficiencies presented are largely cosmetic. The most common (simple) form presents with a unilateral absence of the sternocostal head of the pectoralis majorwith subclavicular hollowing and a deficient axillary fold. Other common findings include axillary webbing that can be seen on arm abduction and elevation, smaller breast/chest with the nipple-areola complex displaced toward the axilla, shortening of the hemithorax, upper extremity shortening, aberrant chest hair distribution in males on the affected side, and syndactyly. Systemic findings may be present, especially in the more severe form of Poland syndrome. The simple form may be known to the patient but remain unannounced to family members until early adolescence when the patient becomes more conscious of the asymmetry. The severe form, however, is usually noted during infancy by the parents because the chest wall is quite visibly asymmetrical and a brachysyndactyly may also be present.The surgical options for chest wall asymmetry depend on anatomical severity, gender, associated anomalies, and patient preferences. The understanding of the intervention options are complicated by the lack of detailed, long-term studies. Treatment options include autologous fat injection, pedicled latissimus dorsi muscle transfer, transverse rectus abdominus musculocutaneous flaps, deep inferior epigastric artery perforator flaps, custom-made chest wall prosthesis, nipple-areola complex repositioning, mammary prosthesis, sternal/rib reconstruction, contralateral operations (to ameliorate asymmetry), or a combination of these techniques.In the case described earlier, left chest fat transposition was performed with approximately 100 cc of autologous fat infiltrated into the left chest with reasonable contour at the completion of the procedure. Three months later, however, much of the fat graft resorbed and the asymmetry again became pronounced. A left latissimus dorsi pedicled flap was then performed. The ipsilateral latissimus muscle was mobilized and tunneled through an incision along the anterior axillary line . Three sComplications of this procedure may include bleeding, infection, seroma formation, and wound dehiscence. This procedure leads to a lack of volume of the posterior axillary fold and ipsilateral back due to the absence of the latissimus dorsi. There is also a risk of postoperativeweakening of the shoulder girdle. The simple deformity of Polands syndrome in males is effectively effaced with a latissimus dorsi transfer, in accordance with Seyfur et al. It is important to note that the complex deformity may require furthermusculoskeletal repair.


Bacteria, fungi, and oomycetes that colonize the living plant tissues are encased in this narrow region in the initial step of infection. Therefore, the apoplastic region is believed to be an interface which mediates the first crosstalk between host and pathogen. The secreted proteins and other metabolites, derived from both host and pathogen, interact in this apoplastic region and govern the final relationship between them. Hence, investigation of protein secretion and apoplastic interaction could provide a better understanding of plant-microbe interaction. Here, we are briefly discussing the methods available for the isolation and normalization of the apoplastic proteins, as well as the current state of secretome studies focused on the  Magnoporthe oryzae and TALEs of Xanthomonas oryzae  causes rice blast disease which results in huge loss of productivity of this most important cereal grain worldwide  infection led to the identification of 501 proteins, of which only 31 (6.2%) were secreted from C. miyabeanus, whereas 470 were secreted from rice. These results suggest that the host-secreted proteins are more abundant during the C. miyabeanus infection. Proteins with decreased abundance were mainly related to the Calvin cycle and glycolysis, whereas abundance of the proteins involved in the TCA cycle, amino acids, and ethylene biosynthesis was increased  differs significantly, and likely their functions. However, it is worth mentioning here that the ratio of pathogen secreted proteins also varies in accordance with the compatible or incompatible interaction with the host. During the incompatible interactions, the growth of pathogens is very much reduced with much less secretion of protein while during compatible interactions the growth of pathogen is highly pronounced which results in secretion of more proteins   and extra-invasive hyphal membrane (EIHM), to separate the plant cytoplasm from the haustorium of oomycetes and invasive hypha of filamentous fungus respectively (Yi and Valent, in-planta secreted proteins. However, as gel based approaches have several limitations including identification of low-abundance proteins, it is possible that some of the key proteins of both plant as well as microbe, would be missed during the analysis. As an example, analysis of secreted proteins from V. longisporum-Arabidopsis or Brassica did not identify any fungal protein, may be due their low-abundance.All the studies conducted so far to investigate the plant-microbe interactions have utilized a gel-based proteomics approach for the identification of With the advancement in the proteomics technologies, new and more precise gel-free quantitative proteomics approaches with improved sensitivity are being developed which could be applied for protein identification, dynamic regulation, and analyzing the post-translational modifications (Picotti et al., in-planta protein extraction techniques at multiple times post-infection will reveal the “real” composition and dynamic changes of the apoplast and can result in the identification of more components of PTI. Furthermore, unraveling complete proteome of apoplastic region in multiple plants, pathogens, and their interactions would be highly fruitful for understanding the biology of plant-pathogen interactions, and that will help in designing new strategies for generating the next-generation crops resistance to multiple pathogens and environmental stresses.Upon infection, both plants as well as pathogens secrete molecules including proteins which determine the fate of their interaction. While pathogen secreted proteins are involved in infection and pathogenicity, plant secreted proteins play crucial roles in its resistance. Therefore, identification and functional analyses of apoplastic proteins will open new horizons in our understanding of plant-pathogen interactions. Moreover, application of the 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.