Staphylococcus aureus is a major cause of nosocomial and community-acquired infections and represents a significant burden on the healthcare system. S. aureus attachment to medical implants and host tissue, and the establishment of a mature biofilm, play an important role in the persistence of chronic infections. The formation of a biofilm, and encasement of cells in a polymer-based matrix, decreases the susceptibility to antimicrobials and immune defenses, making these infections difficult to eradicate. During infection, dispersal of cells from the biofilm can result in spread to secondary sites and worsening of the infection. In this review, we discuss the current understanding of the pathways behind biofilm dispersal in S. aureus, with a focus on enzymatic and newly described broad-spectrum dispersal mechanisms. Additionally, we explore potential applications of dispersal in the treatment of biofilm-mediated infections. Staphylococcus aureus is a Gram-positive human commensal that persistently colonizes the anterior nares of approximately 20–25% of the healthy adult population, while as many as 60% are intermittently colonized , also known as polymeric N-acetyl-glucosamine (PNAG) O'Gara, . PIA is S. aureus surface protein (SasG), biofilm-associated protein (Bap), and clumping factor B (ClfB) , atl and lytM genes. Murein hydrolases degrade peptidoglycan and typically play an important role during cell wall rearrangements and cell division. Increased expression of these enzymes allows for autolysis in S. aureus. Autolysis is regulated through the activity of two operons, cidABC and lrgAB, that function together in a manner similar to bacteriophage holin/antiholin systems , two cysteine proteases (SspB and ScpA), and one metalloprotease (Aur)  that is encoded and produced by the agr operon. The AIP is detected by a two-component system that regulates virulence through the production of a regulatory RNA, RNAIII. The agr system regulates the virulence state of the cell by activating the production of secreted toxins and enzymes and the down-regulation of surface factors. The agr system induces the expression of both proteases and PSMs, which act as surfactants to disperse biofilms  and nuclease2 (Nuc2)  and modulating biofilm development. It has been shown that the expression of nuclease results in reduced biofilm formation nuc mutant, but the activity is very low , V8 serine protease (SspA), and nuclease (Nuc). The relative importance of each enzyme will depend on the strain-specific composition of the biofilm matrix. The proteases and surfactant molecules are under agr quorum-sensing control, and activation of this regulatory system is a known dispersing mechanism.The ability to form a biofilm is an important virulence determinant for the persistence of Going forward, additional studies are necessary to fill specific knowledge gaps. The targets of the major proteases  are still not fully described, although some candidate surface proteins, like the FnBPs and ClfB, have been identified. The function of Nuc in biofilm dispersal has not been examined in detail. It is likely other exo-enzymes, such as hyaluronidase and lipases, are also important in dispersal mechanisms, but have not been fully investigated in biofilm studies (Rosenthal et al., 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.


Animal colors play important roles in communication, ecological interactions and speciation. Carotenoid pigments are responsible for many yellow, orange and red hues in animals. Whereas extensive knowledge on the proximate mechanisms underlying carotenoid coloration in birds has led to testable hypotheses on avian color evolution and signaling, much less is known about the expression of carotenoid coloration in fishes. Here, we promote cichlid fishes (Perciformes: Cichlidae) as a system in which to study the physiological and evolutionary significance of carotenoids. Cichlids include some of the best examples of adaptive radiation and color pattern diversification in vertebrates. In this paper, we examine fitness correlates of carotenoid pigmentation in cichlids and review hypotheses regarding the signal content of carotenoid-based ornaments. Carotenoid-based coloration is influenced by diet and body condition and is positively related to mating success and social dominance. Gaps in our knowledge are discussed in the last part of this review, particularly in the understanding of carotenoid metabolism pathways and the genetics of carotenoid coloration. We suggest that carotenoid metabolism and transport are important proximate mechanisms responsible for individual and population-differences in cichlid coloration that may ultimately contribute to diversification and speciation. Vertebrates cannot synthesize carotenoids endogenously, but dietary carotenoids derived from photosynthetic organisms are responsible for red, orange and yellow hues of many species, including teleost fishes . In the Cichlid fishes (Perciformes: Cichlidae), a group of teleosts endemic to both the Old and New World, provide an excellent system in which to test hypotheses regarding coloration and speciation. Among the Old World cichlids of East Africa, which represent some of the most explosive adaptive radiations among vertebrates , body co2The display of carotenoid-based body coloration is costly. It requires the intake of sufficient amounts of carotenoids from the diet, diverts ingested carotenoids away from vital physiological processes to ornamentation, and makes its bearer conspicuous to predators . ConsequCarotenoids are part of the antioxidant arsenal of animals. Antioxidants quench the potentially harmful pro-oxidant molecules generated during normal metabolism. Oxidative stress arises when the balance between antioxidants and pro-oxidants is disturbed, which can be due to a deficiency of antioxidants or to a surplus of pro-oxidants produced by processes such as somatic growth or an immune response. Animals employ a variety of endogenously produced and food-derived antioxidant compounds, including pigments, vitamins, enzymes and other proteins, and the different compounds can interact with and compensate for each other. Apart from their role as antioxidants, carotenoids may also enhance the immune system through increased T-cell activation, macrophage capacity and lymphocyte proliferation .An influential hypothesis regarding carotenoids posits that trade-offs arise from the dual use of carotenoids for physiological functions and ornamentation . The “cain vivo antioxidants has been questioned  adopt a distinctive yellow or orange coloration  meeki. Fish fed a high-carotenoid diet were more likely to win in dyadic interactions against fish maintained on a low-carotenoid diet  meeki and Pundamilia, aggressive advantage may be affected by the perception of carotenoid-based coloration by potential rivals. Dominance of red males also occurs in staged contests between allopatric color morphs of the Lake Tanganyika endemic Tropheus moorii . Egg spots are yellow or orange spots on the anal fins, and are an important synapomorphy in the particularly species-rich haplochromines, a clade of maternal mouthbrooding cichlids comprising the species flocks of Lake Malawi and Lake Victoria . The conlticolor , while ie choice . Experime choice , which c44.1csf1ra in yellow-colored egg spots on the anal fins of Haplochromini cichlids (see below) and in the yellow tips of elongated ventral fins in Ectodini cichlids might be associated with xanthophore recruitment into these tissues  and for the ornamental fish trade. Several studies have shown that dietary supplementation increases the integument concentration of carotenoids in cichlids, but effects were not observed in all species and depended on the types of carotenoids that were added  and pigmentation (Pax7) as well as melanophore number and size (Pax3)  develop fewer but larger melanophores than the brown barred females. OB females vary in background color from white to orange depending on species and population, indicating that the identified allele affects the melanophore pattern but not the orange coloration in these cichlids .The orange blotch phenotype of certain Lake Malawi cichlid species, in which females display dark melanophore blotches on a background of xanthophores, was associated with up-regulation of the cichlid ubfamily . The orae (Pax3) . Cichlidcichlids . One posIn Midas cichlids and Lake Malawi cichlids with the orange-blotch phenotype, and likely in many other cichlid species, the extent of carotenoid-based coloration on the body is contingent on the distribution of melanophores. This interaction makes the processes controlling melanophore patterning relevant to understanding the processes underlying carotenoid coloration. Studies in zebrafish and medaka provided a wealth of information about the genetics of melanin coloration in these model organisms . Several5Carotenoids are important in many aspects of cichlid biology. The elaboration of carotenoid-pigmented structures via sexual selection is well documented. Much remains to be discovered, however, in areas such as carotenoid biochemistry, physiology and genetics. Basic data on the types and functions of carotenoids in cichlids, and their interactions with other metabolites, will allow the testing of hypotheses regarding the evolutionary significance of carotenoid coloration. Below, we specify some areas for future research that we believe will contribute significantly to our understanding of the evolution of carotenoid pigmentation in cichlid fishes, and in fishes more broadly.5.1The identification of esterified carotenoids – which are present in the integument of many cichlids – via high-performance liquid chromatography is more complicated than for non-esterified carotenoids, such as in bird feathers. The identification of integumentary carotenoids in cichlids e.g.  will all5.2P. nyererei: T. moorii: A. nigrofasciata: Poikilothermic vertebrates do not rely solely on dietary carotenoids to produce yellow and red coloration; they can also synthesize pterin pigments. Pterins have been detected in the integument of several fish species . The bal5.3Carotenoid-based coloration of cichlids responds to a number of factors including diet, health status and social stimulation . For exa5.4The conspicuous carotenoid-based coloration of many cichlids is likely to convey information on the bearer's condition, status and motivation. The use of this information in female choice  and male6Many of these questions outlined above can be addressed in other organisms, but we believe cichlids are a particularly important group for this kind of research for several reasons. On the one hand, understanding carotenoid physiology and biochemistry is important for our understanding of cichlid evolution. On the other hand, the extraordinary diversity of cichlids will help advance our understanding of the roles of carotenoids in organismal biology.A. burtoni, a model species in social neuroscience (First, the rich diversity of cichlid color patterns promises opportunities to uncover a variety of genetic, cellular and physiological processes involved in carotenoid coloration and color pattern differentiation . Variatioscience . Some cioscience . The relSecond, the aforementioned variation in coloration can help us to better understand the relationship between phenotypic plasticity and speciation . Cichlid


Poor adherence to antiretroviral therapy (ART) contributes to the development of drug resistance. HIV-infected children, especially those 5 years and under, are dependent on a caregiver to adhere to ART. However, characteristics of the caregiver, child, regimen, clinic and social context affect clinic attendance and medication-taking, both of which constitute adherent behaviour. We conducted nine interviews and three focus groups to determine how doctors, nurses, counsellors, traditional healers and caregivers understood the barriers and facilitators to ART adherence among children residing in rural South Africa. The data were transcribed, translated into English from isiZulu where necessary, and coded using Atlas.ti version 7. Results were interpreted through the lens of Bronfenbrenner's Ecological Systems Theory. We found that at the micro-level, palatability of medication and large volumes of medication were problematic for young children. Characteristics of the caregiver including absent mothers, grandmothers as caregivers and denial of HIV amongst fathers were themes related to the micro-system. Language barriers and inconsistent attendance of caregivers to monthly clinic visits were factors affecting adherence in the meso-system. Adherence counselling and training were the most problematic features in the exo-system. In the macro-system, the effects of food insecurity and the controversy surrounding the use of traditional medicines were most salient. Increased supervision and regular training amongst lay adherence counsellors are needed, as well as regular monitoring of the persons attending the clinic on the child's behalf. Excellent adherence (>95%) to antiretroviral therapy (ART) is required to achieve optimal outcomes of HIV treatment  and a nearby peri-urban Department of Health (DoH) clinic. The AC has supported the DoH Hlabisa HIV Treatment and Care Programme (ART programme) since its inception in 2004  South Africa (SA) (Doctors (IDI), nurses (IDI) and counsellors (FG) working within the ART programme;Caregivers (FG) of children ≤5 years attending a peri-urban clinic;Traditional healers (FG) consulting to people with HIV.Using open-ended questions we explored participants' understanding of the barriers and facilitators to ART adherence amongst children. Interviews and FGs were conducted under confidential conditions in the participants' first language (isiZulu) with the assistance of an isiZulu speaking translator, or otherwise in English, either at the AC, clinic or hospital.Semi-structured in-depth interviews (IDIs) and focus groups (FGs) were conducted with purposively sampled individuals:Examples of the questions used are shown in Ethical approval was obtained from Stellenbosch University, with reciprocity from the University of KwaZulu-Natal.www.atlasti.com) computer program  and entered into the Atlas.ti version 7 , 4 nurses , 12 counsellors , 10 traditional healers  and 11 caregivers . Doctors, nurses, counsellors and traditional healers were unanimous about the difficulty that children had with the palatability of Lopinavir/ritonavir. Caregivers indicated that mixing medicines with a sweetener such as a liquid multivitamin, juice or peanut butter improved the taste for children.Caregivers stated that their failure to disclose the child's status to household members disrupted their ability to administer medication. For example, respondents indicated that in some households the father refused to accept that his child was HIV-infected, which would force caregivers to hide the child's medication.Doctors, nurses and counsellors stated that language barriers created difficulties in communicating with caregivers about difficulties regarding medication administration. Although nurses and counsellors were able to communicate with caregivers in isiZulu, they reported that caregivers struggled to understand important information about their treatment such as changes in dose amounts. Further, although grandmothers were the primary caregiver, other household members also administered medication to the child, which became problematic when doses were changed. This problem was exacerbated when communication among these family members was poor.Doctors stated that the skill level of adherence counsellors was generally low, reflecting the poor quality of the training they received. They stated that counsellors were neglected in their role and received no debriefing, minimal supervision, and once-off training sessions that were in some instances poorly conducted. Regarding the pre-ART HIV sessions that caregivers received, doctors and counsellors agreed that caregivers were given too much information over a short period of time. They stated that it was emotionally and cognitively burdensome for a caregiver to accept the child's HIV diagnosis, receive counselling within two days and be expected to commence treatment immediately. Surprisingly, caregivers felt that the sessions were adequate, and that they were comfortable asking for help when needed.All the caregivers participating in the FG were unemployed, and some received a government grant see . RespondOur study is unique in its representation of each of the members involved in the treatment and care of a child on ART in this context. By including each of these role players we were able to demonstrate the relative influence of the micro-, meso-, exo- and macro-systems on paediatric adherence to ART. Vreeman et al.  demonstrDespite being largely confirmatory, our results indicate that children struggle with unpalatable syrups such as LPV/r  per day (D'Haese et al., Caregiver consistency is important and may take the form of regular monitoring of the person attending clinic, monitoring regular communication between caregivers and ensuring that treatment-related recommendations are effectively passed on to the person responsible. Finally, we recommend adequate training of adherence counsellors with regular supervision, training and debriefing.The study provides a broad range of perspectives expressed by the key role players in the treatment and care of children on ART. Due to the small sample size and nature of the data, the findings cannot be generalised, but may be used to inform future research.


The understanding of obsessive-compulsive disorder (OCD) has evolved with the knowledge of behavior, the brain, and their relationship. Modern views of OCD as a neuropsychiatric disorder originated from early lesion studies, with more recent models incorporating detailed neuropsychological findings, such as perseveration in set-shifting tasks, and findings of altered brain structure and function, namely of orbitofrontal corticostriatal circuits and their limbic connections. Interestingly, as neurobiological models of OCD evolved from cortical and cognitive to sub-cortical and behavioral, the focus of OCD phenomenology also moved from thought control and contents to new concepts rooted in animal models of action control. Most recently, the proposed analogy between habitual action control and compulsive behavior has led to the hypothesis that individuals suffering from OCD may be predisposed to rely excessively on habitual rather than on goal-directed behavioral strategies. Alternatively, compulsions have been proposed to result either from hyper-valuation of certain actions and/or their outcomes, or from excessive uncertainty in the monitoring of action performance, both leading to perseveration in prepotent actions such as washing or checking. In short, the last decades have witnessed a formidable renovation in the pathophysiology, phenomenology, and even semantics, of OCD. Nevertheless, such progress is challenged by several caveats, not least psychopathological oversimplification and overgeneralization of animal to human extrapolations. Here we present an historical overview of the understanding of OCD, highlighting converging studies and trends in neuroscience, psychiatry and neuropsychology, and how they influenced current perspectives on the nosology and phenomenology of this disorder. The firomething . It descomething . Compulsomething . Similaromething .Currently it is estimated that, in the USA, 2.3% of the adult population suffer from this condition . MoreoveFrom the early times of modern psychiatry, OCD has exerted a particular fascination on clinicians and researchers alike. Over more than a century of clinical and neurobiological research, OCD moved from being considered a typical neurosis  to beingfolie du doute (doubting madness) and délire du toucher , medial frontal gyrus, and anterior cingulate cortex (ACC), and increased volume of the ventral putamen . The mosMost of the evidence supporting a role for serotonin in the pathophysiology of OCD is based on the established therapeutic effects of serotonin reuptake inhibitors and the inefficacy of antidepressants with no effect on serotonin transmission or metabolism . Positroreduces impulsivity in animal models, which might partially explain the limited success of “serotonergic” treatments in OCD  or indirectly  in specific areas of the ventral striatum leads to OCD-like behavior changes (Gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the central nervous system, has been much less studied in OCD. Magnetic resonance spectroscopy studies found decreased levels of GABA in prefrontal cortical areas, including the ACC , with onhibition . Taken thibition . In line and OCD . Finally changes .hyperemesis gravidarum or dystocic labor, adverse life events, including childhood physical or sexual abuse, were all found to increase the risk of developing this disorder , and also to degradation or extinction of the contingency between performing the action and obtaining the outcome. Habitual behavior, on the other hand, persists even when the outcome is no longer rewarding or the action-outcome contingency is degraded or reversed. The phenomenological analogy between habitual action control and compulsive behavior has led to the hypothesis that OCD patients may be predisposed to rely excessively on habitual action control rather than on goal-directed behavioral strategies . EvidencHowever, the parallel between habitual action patterns and the elaborately ritualized, egodystonic behaviors that plague OCD patients may be an over-simplification. Importantly, the approaches used to date to study action control patterns in OCD generally fail to take into account the fundamental role of habits in the formation of chunked action-sequences, i.e., relatively invariant, rapidly deployed sequences of single action-units . Action The above-mentioned failure to develop adaptive and automatic action-sequences could be interpreted in the context of maladaptive goal-directed uncertainty monitoring, leading to repetitive behaviors, such as checking, in order to reduce uncertainty. This possibility is consistent with the evolving evidence on the role of the STN in the regulation of behavior, and more specifically in the neurobiology and psychopathology of OCD. As mentioned previously, the STN has been one of the targets for DBS in OCD. Linked to frontal cortical areas by the so-called hyperdirect pathway , the STNOne other unresolved question regarding the predominance of habitual action strategies in OCD is that such strategies result in highly adaptive action control patterns, allowing the subject to engage in alternate and cognitively demanding tasks, while performing a given well-learned action. This actually seems to be impaired in OCD patients, who not only engage explicit memory-linked medial temporal structures when performing implicit learning tasks , but alsIt is intuitive that the biological understanding of medical disorders should progress from semiology to etiology, rather than the opposite. For most medical disorders, understanding the physiology of signs and symptoms paved the way to the formulation of pathophysiological models and ultimately to the discovery of the disorder’s primary causation. Yet, in the case of OCD, it is clear that the evolution in our understanding of the disorder’s pathophysiology has had a profound impact on our conceptualization of the disorder’s phenomenology. Indeed we may say that the gradual dislocation of the main focus of pathophysiological models of OCD from cortical to subcortical structures has been paralleled by a metamorphosis of the conceptualisation of OCD phenomenology, whereby the initial emphasis laid on the control of explicit thought processes, thought contents, and thought-evoked anxiety, has been replaced by concepts rooted in behavioral neuroscience and animal models of action control. Contemporary literature on OCD is dominated by new phenomenological concepts, and even a new semantics for OCD-related concepts, now seen as characterized by a loss of inhibitory control over impulsive stimuli-response associations, by excessive harm-avoidance, or by a failure to flexibly shift between alternative action-sequences in response to changing environmental demands. Similarly, new insights into the role of dopamine and reward circuits in OCD led researchers in the field to emphasize the phenomenological analogies between addiction disorders and OCD, regarded here as a form of behavioral addiction, a disorder combining impulse control dysfunction with abnormal reward processing mechanisms. These phenomenological metamorphoses found echo in the very nosology of OCD, which in DSM-V was emancipated from anxiety disorders and raised to a new statute as an autonomous spectrum of disorders of behavior control.In very recent years, the phenomenological analogy between habitual action control and compulsive behavior brought yet another renewal of the semantics and the phenomenology of OCD. The idea that OCD patients may be predisposed to rely excessively on habitual rather than on goal-directed behavioral strategies now faces the field with a host of new concepts stemming from animal experimentation. To some degree, there seems to be a tendency to reduce the psychopathology of OCD to a deregulation of automatic behavior and pathological streams of non-functional stimuli-response associations, downplaying the role of cognition in the process. At the extreme of this evolution from a cognitivist toward a behavioral control centered conceptualisation of OCD, obsessions are eventually seen as secondary, retrospective cognitive constructions developed by the subject to assimilate these intrusive behavioral programs back into the self . Compulsin vivo neuronal recordings and non-invasive neuromodulation in humans, in order to more fully elucidate the mechanisms that may underlie the details of OCD psychopathology.Science has come a long way in understanding OCD. It has been a unique process, marked by dialectic interplay between the progress in neurobiological models of OCD and an increasingly sophisticated phenomenological characterization of this complex syndrome. Refined structural and functional imaging of the brain and technological progress in animal experimental modeling of OCD have opened the way to groundbreaking insights into the role of cortical and subcortical structures in the disorder’s pathophysiology, and revolutionized our perception and interpretation of obsessive-compulsive phenomena. Yet, in spite of much progress, we still lack an explanation for many of the disorder’s manifestations, and not least its extraordinary clinical variability. Extrapolations from animal models, themselves extrapolations of human phenomenology, while carrying extraordinary promise to further understanding of the disorder, necessarily ignore many complex aspects of human psychopathology. Care is thus needed to carefully cross-validate human-animal-human extrapolations, as well as analogies between psychopathology and constructs from experimental psychology and behavioral neuroscience literature, while avoiding impoverishing an extremely rich legacy of psychopathological detail and subtlety. In common with other psychiatric disorders such as bipolar disorder or schizophrenia, neuroimaging studies in OCD are often contradictory and difficult to interpret. This is due not only to phenotypic variability across subjects, but also to the far from clear meaning of volumetric and BOLD-signal differences between clinical subjects and healthy controls. Genetic studies have likewise failed to provide a much anticipated breakthrough in understanding and treating OCD and other mental disorders, probably because genetic factors lie too far upstream in the complex flow of factors and events that ultimately lead to the expression of mental illness in a given individual. A renewed interest for psychopathological refinement can certainly contribute toward the advancement of knowledge on the neurobiology of mental disorders in general, beyond explanation of each syndrome’s gross features and toward tackling of more subtle manifestations and of subtypes of each disorder. Nevertheless, the future also calls for more detailed studies using, for example, genetic animal models, improved neuroimaging technology, The Reviewer Sérgio Saraiva declares that, despite being affiliated to the same institution as the author Pedro Castro-Rodrigues, the review process was handled objectively and no conflict of interest exists. The other 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.


Forty-three deaths were recorded among pigs fed boiled cassava meal at a private piggery over a period of two years. There were signs of sudden death in some cases with blood exuding from the external nares, vomiting, muscular weakness and pain or reluctance to move, emaciation, and stunted growth. The necropsy lesions included skeletal and cardiac muscle degeneration and necrosis, icterus, hepatic necrosis, and oedema of the dependent parts. The deaths and clinical signs are thought to be due to a non cyanide toxic principle in cassava, possibly the coumarins (scopoletin), which is found in high levels in cassava diet even after heat treatment. Therefore, the use of proper processing technology to obtain cassava products of high quality is recommended. Manihot utilisima) in form of the peels and pulp in the feeding of livestock has attracted renewed interest especially with the rising cost of commercially produced feeds [In Nigeria, commercially reared pigs under the intensive husbandry system are kept in concrete floors and fed balanced commercially produced feeds. The use of cassava  technique. The histopathological examination of the tissues revealed that, in the acute cases, the liver lobules showed spot-like degeneration and coagulative necrosis. In the chronic cases many liver lobules showed postnecrotic collapse and fibrosis, bile duct hyperplasia and scattered partially destroyed lobules, and mononuclear cellular infiltration  in the pulp of fresh material . BoilingScopoletin (6-methoxy-7-hydroxycoumarin) has been found in large quantities in cassava products including garri and cassava flour. The level of scopoletin is not affected by treatments such as sun drying, refrigeration, storage, and cooking . ScopoleScopoletin is absorbed by mitochondria and it uncouples oxidative phosphorylation at low concentrations and directly inhibits it terminally at the cytochrome oxidase level . This ef


It is known that patients with severe motor and intellectual disabilities (SMID) showed sudden unexplained death (SUD), in which autopsy failed to identify causes of death. Although the involvement of brainstem dysfunction is speculated, the detailed neuropathological analysis still remains to be performed. In order to clarify pathogenesis, we investigated the brainstem functions in autopsy cases of SMID showing SUD. We immunohistochemically examined expressions of tyrosine hydroxylase, tryptophan hydroxylase, substance P, methionine-enkephalin, and c-fos in the serial sections of the midbrain, pons, and medulla oblongata in eight SUD cases and seven controls, having neither unexplained death nor pathological changes in the brain. Expressions of tyrosine hydroxylase and tryptophan hydroxylase were reduced in two of eight cases, and those of substance P and/or methionine-enkephalin were augmented in the pons and medulla oblongata in seven of eight cases, including the aforementioned two cases, when compared with those in controls. The hypoglossal nucleus and/or the dorsal vagal nucleus demonstrated increased neuronal immunoreactivity for c-fos in seven of eight cases, although there was no neuronal loss or gliosis in both the nuclei. Controls rarely showed immunoreactivity for c-fos in the medulla oblongata. These data suggest the possible involvement of brainstem dysfunction in SUD in patients with SMID, and consecutive neurophysiological evaluation of brainstem functions, such as all-night polysomnography and blink reflex, may be useful for the prevention of SUD, because some parameters in the neurophysiological examination are known to be related to the brainstem catecholamine neurons and the spinal tract nucleus of trigeminal nerve. Severe motor and intellectual disabilities (SMID) describe a heterogeneous group of disorders with severe physical disabilities and profound mental retardation , 2. Patit-test for the comparison, and reported p values were shown in Table The clinical subjects comprised eight cases of SMID showing SUD (the SUD cases) between 1968 and 2001 at Tokyo Metropolitan Fuchu Medical Center for the Disabled and Tokyo Metropolitan Neurological Hospital and seven controls, aged from 7 to 55 years, showing neither SUD nor pathological changes in the central nervous system (Table A mild increase of the goblet cells was found in the SUD cases Table , but thaIn the SIDS victims, fine morphological changes have been noted in the brainstem respiratory network, including Kölliker–Fuse nucleus –9, althoSudden infant death syndrome victims were reported to show reduced immunoreactivity for tyrosine hydroxylase in the dorsal vagal nucleus and ventrolateral reticular formation in the medulla oblongata . Sudden It is controversial whether the immunohistochemical changes in this analysis are causes or results of SUD; however, the data suggest the possible involvement of brainstem dysfunction in SUD. Patients with SMID occasionally demonstrate abnormalities in both the sleep parameters in all-night polysomnography and the R2 components in blink reflex, which are related to the brainstem catecholamine neurons and the spinal tract nucleus of trigeminal nerve, respectively, both of which demonstrated changes in the SUD cases of SMID in this analysis . We beliSudden unexplained death remains to be one of the important causes of death in patients with SMID, although the exact pathomechanisms are not clarified fully. The data in this analysis suggest the possible involvement of brainstem dysfunction in SUD, and the detailed neurophysiological evaluation of brainstem functions is recommended in patients with SMID.Prof. MH planned this analysis and conducted all processes, such as selection of subjects, immunohistochemical staining, quantitative assessment of each specimen, taking photographs, and writing the manuscript. Prof. HS financially supported this analysis, supervised all processes, and discussed the context of 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.


Fear and anxiety are adaptive responses but if left unregulated, or inappropriately regulated, they become biologically and socially maladaptive. Dysregulated emotions are manifest in a wide variety of psychiatric and neurological conditions but the external expression gives little indication of the underlying causes, which are inevitably multi-determined. To go beyond the overt phenotype and begin to understand the causal mechanisms leading to conditions characterized by anxiety and disorders of mood, it is necessary to identify the base psychological processes that have become dysregulated, and map them on to their associated neural substrates. So far, attention has been focused primarily on the medial regions of prefrontal cortex (PFC) and in particular their contribution to the expression and extinction of conditioned fear. However, functional neuroimaging studies have shown that the sphere of influence within the PFC is not restricted to its medial regions, but extends into dorsal, ventrolateral (vlPFC) and orbitofrontal (OFC) regions too; although the causal role of these other areas in the regulation of fear and anxiety remains to be determined and in the case of the OFC, existing findings are conflicting. Here, we review the evidence for the contribution of these other regions in negative emotion regulation in rodents and old world and new world monkeys. We consider a variety of different contexts, including conditioned and innate fear, learned and unlearned anxiety and cost-benefit decision-making, and a range of physiological and behavioral measures of emotion. It is proposed that both the OFC and vlPFC contribute to emotion regulation via their involvement, respectively, in the prediction of future outcomes and higher-order attentional control. The fractionation of these neurocognitive and neurobehavioral systems that regulate fear and anxiety opens up new opportunities for diagnostic stratification and personalized treatment strategies. Negative emotions such as fear and anxiety are highly adaptive and complex mental states that are the product of interactions between cognition, physiological responses and behaviors. The continuum of emotional responses stretches from unlearnt reflexes and fixed action patterns, through Pavlovian learning , to instrumental behavior, whereby the organism takes adaptive control of the environment. Via feedback loops between brain and body our fluctuating emotions influence attention, decision making, memory and social interactions. Once the source of fear or anxiety is dealt with or disappears, the emotional responses dissipate so that everyday activity can be resumed. Such effective regulation of negative emotional responses is critical for the physiological, psychological and social well-being of individuals. However, in some, these emotional responses become chronic or exceedingly recurrent even without an apparent external source. Failure to down-regulate or control one’s emotional responses, even when circumstances change from threatening to relatively safe, can have a devastating impact on a sufferer’s life.Emotional disturbance is a core symptom of mood and anxiety disorders and a prominent symptom of many other neuropsychiatric conditions, including schizophrenia  are considered as the first-line pharmacological treatment for anxiety disorders and depression, up to 40% of patients are estimated to be partially or completely resistant to the treatment  through repeated temporal pairings with an aversive event such as a foot shock . Sensory information of both CS and US are transmitted from peripheries to respective sensory processing areas and subsequently converge at the amygdala where the association of the two stimuli occurs. Through its connections to the brainstem, hypothalamus and motor areas, the amygdala is critically involved in the physiological and behavioral response to a learnt  or innate threat, especially when the threat is imminent and explicit LeDoux, . The conThe human PFC is located in the anterior portion of the frontal lobe and broadly includes cytoarchitectonically defined Brodmann areas (BA) 8, 9, 10, 11, 13, 14, 44, 45, 46 and 47/12  and infralimbic (ILc) cortices  played by ventral PFC in the generation and regulation of fear- and anxiety-like responses either between or within different species. Reductions in innate fear appear to be the most consistent effect of aspirative lesions of the OFC in macaques,  whilst in rodents, if anything, OFC lesions lead to heightened or generalized conditioned fear responses, alongside intact unconditioned anxiety responses. One obvious distinction between species is that conditioned fear responses have been the focus of rodent studies, whilst innate fear and experience-based (learned) anxiety-like responses have been more commonly used in macaques, components of negative emotion which may well involve distinct, although overlapping circuitry  marmosets develop discriminative conditioned heart rate (HR) and vigilant scanning responses to the auditory cue associated with the US+  lesioned and vlPFC lesioned groups on these fear- and anxiety-inducing tests, both lesioned groups displayed heightened emotionality to both the human intruder and the snake  already recognizes the different aspects of a psychological disorder in terms of dysfunctional emotions, maladaptive behaviors and cognitive processes. By understanding how they are parsed within associative neural systems will bring us one step closer to individualized treatment strategies. For example, cognitive re-appraisal, requiring a patient to shift attention from negative to positive aspects of an emotional situation may be more successful in a patient poor at predicting  than one deficient in attentional control . Thus, if we can fractionate the component cognitive processes that constitute the “top-down” regulatory mechanisms within ventral, dorsal and medial PFC, we will provide the necessary new insights into the varied causes of anxiety, which will allow for more precise stratification of the disorder, leading to individualized treatment strategies.YS and ACR: manuscript writing and idea building. AMS: conceptual discussion.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.


Nevertheless, whether the addition of the dopaminergic component produces more robust effects than single- or dual-acting agents, has yet to be demonstrated. This article reviews the main pathways regulating pain transmission in relation with the monoaminergic systems. It then focuses on the current knowledge regarding the in vivo pharmacological properties and mechanism of action of monoaminergic antidepressants including SSRIs, NRIs, SNRIs and TRIs. Finally, a synthesis of the preclinical studies supporting the efficacy of these antidepressants in analgesia is also addressed in order to highlight the relative contribution of 5-HT, NE and DA to nociception.Over 75% of depressed patients suffer from painful symptoms predicting a greater severity and a less favorable outcome of depression. Imaging, anatomical and functional studies have demonstrated the existence of common brain structures, neuronal pathways and neurotransmitters in depression and pain. In particular, the ascending serotonergic and noradrenergic pathways originating from the raphe nuclei and the locus coeruleus; respectively, send projections to the limbic system. Such pathways control many of the psychological functions that are disturbed in depression and in the perception of pain. On the other hand, the descending pathways, from monoaminergic nuclei to the spinal cord, are specifically implicated in the inhibition of nociception providing rationale for the use of serotonin (5-HT) and/or norepinephrine (NE) reuptake inhibitors , in the relief of pain. Compelling evidence suggests that dopamine (DA) is also involved in the pathophysiology and treatment of depression. Indeed, recent insights have demonstrated a central role for DA in analgesia through an action at both the spinal and suprasinal levels including brain regions such as the periaqueductal grey (PAG), the thalamus, the basal ganglia and the limbic system. In this context, dopaminergic antidepressants ( It also involves adaptation and modulation of the nociceptive messages by various neuromediators and related receptors. Since these neuromediators are present in the central network of brain structures that process or regulate nociceptive information, it is difficult to dissociate the affective dimension of pain from its sensory dimension . AnatomiAt the peripheral level, the pain comes from direct or indirect stimulation and sensitization of nociceptors by various endogenous signalling molecules, including ions, prostaglandins and/or leukotriens, histamine, bradykinines, but also monoamines . The act1.1.At the spinal level, the central terminal of the nociceptor forms synapses with neurons of the superficial dorsal horn of the spinal cord. Glutamate seems to be the main neurotransmitter released in response to a nociceptive stimulus. Glutamate then acts on post-synaptic receptors present in: (i) the projection of cells whose axons convey information to various parts of the brain; and (ii) interneurons (both excitatory and inhibitory) that all contribute to the local modulatory circuit in the spinal cord. Thus, the ascending pathways distribute spinal action potentials to brain areas related to the two dimensions of pain perception, sensory and affective: the somatosensory cortex, the periaqueductal grey (PAG), hypothalamus and basal ganglia. Spreading from central projections, corticolimbic pathways are also activated. These sites which process noxious activation include the thalamus, insular cortex, anterior and posterior cingulated cortex, prefrontal cortex -8 but alDescending inhibitory or facilitatory pathways from brain areas converge at the dorsal horn, controlling peripheral inputs from nociceptors. Monoaminergic fibers originating from various brainstem nuclei control pain perception through the release of 5-HT and NE in the superficial dorsal horn via the dorsolateral funiculus (DLF) . DLF fibCompared with the enormous literature devoted to 5-HT and NE, the spinal action of DA has received less attention. It has been proposed that the dopaminergic innervation of the spinal cord may originate in the substantia nigra and hypothalamus . Further1.2.The periaqueductal grey (PAG) is an important nociception modulation site where the emotional and cognitive sensations from thalamic or anterior cortical areas meet the vegetative aspects from the hypothalamus ,20. AlthThe dorsal raphe nucleus (DRN) is interconnected and functionally related to the PAG. Although both brain regions display strong anatomical interactions -28, the The LC and the A7 catecholamine cell groups, known to contain spinally noradrenergic neurons, are connected to the PAG through a monosynaptic pathway ,39. ThisDopaminergic neurons from the ventral tegmental area (VTA) have no clear anatomical and direct functional interactions with the PAG. Interestingly, several studies have localized the antinociceptive effects of morphine to the PAG ,44-46. TThe thalamus is the main relay site for nociceptive inputs to cortical and subcortical structures. It includes several nociceptive nuclei of the somatosensory and intralaminar thalamus. Thalamocortical networks that produce both sensory discriminative and affective components of the pain response generate conscious pain. Initial studies have indicated that the ablation of the parafascicular nucleus (PFN) selectively reduces the emotional suffering associated with acute and chronic pain in humans, and reduces responses to noxious stimulation in animals .The DRN projects axons to the thalamus including the PFN, to suppress the pain sensations. Stimulation of the DRN has been found to effectively inhibit the responses induced by noxious stimulation of neurons in the PFN . In contThe LC, but also the VTA, send projections to the somatosensory thalamus -59. The Apart from the well-known involvement in motoric circuitry of the basal ganglia, these brain nuclei are involved in many neuronal pathways having emotional, motivational, associative and cognitive functions as well. This brain region contains several nuclei including the putamen, caudate nucleus, globus pallidus, subthalamic nucleus, and nucleus accumbens that receive multimodal input from all sensory systems and thereby serve as a gating station for continuous sensory information, including pain. Several studies have suggested that basal ganglia may be involved in the sensory-discriminative aspect of pain, the affective and cognitive aspect of pain but also the modulation of nociceptive information and sensory gating of nociceptive information to higher motor areas, because they are the main link between the thalamus and the cerebral cortex . Data su2 receptor antagonist, attenuated the antinociceptive effect of morphine  as suggested by immunohistochemical and fluorescence histochemical studies . The CPuts of NE . The latts of NE .i.e., DA receptor agonists and antagonists) stimulation or electrolytic and chemical lesions of the CPu, GP and substantia nigra (SN) provide evidence that the basal ganglia can modify behavioral responses to noxious stimulation. For example, it has been shown that a unilateral lesion of the nigrostriatal pathways causes hyperalgesic responses to painful stimuli at contralateral side -5-HT or [3H]-NE reuptake in synaptosomes, is one of the most widespread method to assess the in vitro potency of reuptake inhibitors -naloxone demonstrating the lack of affinity of this SSRI for opioid receptors [The idea that the inactivation of SERT and the related increase in extracellular levels of 5-HT  could beeceptors . The siteceptors . Nevertheceptors  or studieceptors ,242.2C receptor for fluoxetine, sigma opioid receptor for fluvoxamine).It is thus possible that the antinociceptive effects of SSRIs in acute pain involves both spinal and supraspinal mechanisms. This hypothesis is supported by the fact that in “positive” studies comparing the effects of SSRIs in the tail flick and hot plate tests, these pharmacological agents produced analgesic responses in both tests . Indeed,In chronic neuropathic and inflammatory models of pain, citalopram reduced hyperalgesia . The obs3.2.Genetically modified mice lacking the NE transporter exhibit normal nociceptive threshold . This isIn various models of neuropathic and inflammatory pains, both desipramine and reboxetine displayed robust antinociceptive effects by reversing allodynia and/or hyperalgesia either after acute or chronic administration in mice ,272 and in vitro, may involve the noradrenergic property of paroxetine compared to fluoxetine  model of neuropathic pain has been examined. It was demonstrated that NRIs, desipramine and reboxetine, reversed allodynia, while the SSRI fluoxetine displayed a minimal activity suggesting that compounds with greater affinity for the NET are more effective in attenuating pain than compounds with a greater affinity for the SERT . Such reuoxetine . This is3.3.As observed with NRIs, duloxetine has no or modest effect in acute models of pain in mice  or rats Thus, although limited, studies comparing the analgesic effect of monoamines reuptake inhibitors in rodents suggest that dual acting antidepressants are more active in alleviating acute or chronic pain than selective NE reuptake inhibitors, which themselves appear more potent than SSRIs . The ran3.4.The role of DA reuptake inhibitor has been poorly studied in animal models of pain. This is likely due to the lack of selective DA reuptake inhibitors currently available. However, it was reported that the DA reuptake inhibitor, GBR12909 did not affect nociception in thermal or mechanical tests ,336 but in vitro and in vivo pharmacological property demonstrates that bupropion is a much weaker inhibitor of NE reuptake than reboxetine [The contribution of DA and the putative interest of DA reuptake inhibitors in relieving pain must be estimated also from the behavioral effects of agents displaying a blocking DAT activity such as bupropion or nomifensine. In rats, bupropion decreased allodynia induced by CCI and spinal nerve ligation models of neuropathic pain . The inaboxetine -340  neurokinin-1 (NK1) antagonist MK-869, which failed despite demonstrations of adequate exposure, penetration, and occupancy . There a4.This review emphasized the fact that pain and depression share common neurotransmitters pathways and that monoaminergic reuptake inhibitors may relieve chronic pain. If chronic pain may result in depression, pain is also a symptom frequently observed in depressed patients . If it iTo date, when comparative studies are available, clinical trials for pain invariably indicate a superiority of the dual-acting agents for pain treatment compared to single reuptake inhibitors. With the development of milnacipran, duloxetine and now the triple reuptake inhibitors such as bicifadine focusing in their ability to relieve pain, more extensive comparative data should be forthcoming to make a more evidence-based judgement on the superiority of the SNRIs and TRIs in these indications. In this context, preclinical studies are useful to unveil which antidepressants may display the best therapeutic profile and may determine whether doses that produce antidepressant effects also affect pain and reciprocally.One question that also should draw our attention is the management of side effects and particularly whether the simultaneous enhancement of serotonergic, noradrenergic and/or dopaminergic neurotransmission mitigate or accentuate these side effects. Despite the much improved side effect profile of newer antidepressants, all of them are still associated with adverse effects that vary between classes and within each class . SSRIs hRegarding TRIs, a concern with drugs that block DA transporters is their potential reinforcing property and abuse liability . NeverthIn conclusion, tolerability profiles of the antidepressant or combination of antidepressants should be considered when making treatment choices. The decision to use such combinations must be tempered by and weighed in conjunction with the knowledge that specific side effects are expected. It seems that the lack of research studies on the role of adjunctive treatments in the management of antidepressant-induced side effects favors monotherapy over polytherapies . In this


The entire pollen life span is driven by polyamine (PA) homeostasis, achieved through fine regulation of their biosynthesis, oxidation, conjugation, compartmentalization, uptake, and release. The critical role of PAs, from microsporogenesis to pollen–pistil interaction during fertilization, is suggested by high and dynamic transcript levels of PA biosynthetic genes, as well as by the activities of the corresponding enzymes. Moreover, exogenous supply of PAs strongly affects pollen maturation and pollen tube elongation. A reduction of endogenous free PAs impacts pollen viability both in the early stages of pollen development and during fertilization. A number of studies have demonstrated that PAs largely function by modulating transcription, by structuring pollen cell wall, by modulating protein  assembly as well as by modulating the level of reactive oxygen species. Both free low-molecular weight aliphatic PAs, and PAs conjugated to proteins and hydroxyl-cinnamic acids take part in these complex processes. Here, we review both historical and recent evidence regarding molecular events underlying the role of PAs during pollen development. In the concluding remarks, the outstanding issues and directions for future research that will further clarify our understanding of PA involvement during pollen life are outlined. Figure 1A); Put has an aliphatic tetramethylene backbone deriving directly from ornithine or indirectly from arginine or citrulline via N-carbamoylputrescine. The biosynthesis of higher PAs occurs by the addition of one or two aminopropyl groups to Put to form Spd and Spm, respectively. Whereas Put has positive charges on the primary amino groups, Spd and Spm also bear protonated internal iminic groups, at physiological pH. PAs are present in cells in both free and bound forms and their molecular mechanism of action is often associated with their polycationic groups able to establish hydrogen and ionic interactions with anionic groups of several biological molecules, among which proteins, nucleic acids, and membrane phospholipids. Moreover, they strongly bind in vitro to cell wall polysaccharides with a different binding capacity depending mainly upon the number of their positive charges. In addition, the covalent binding to some glutamyl residues of specific proteins, catalyzed by TGase, gives rise either to PA binding to proteins (mono-γ glutamyl-PAs) or to cross-links between proteins (bis-γ glutamyl-PAs) . These conjugates are components of the PCA-insoluble PA fraction , components of the PCA-soluble fractions. These are involved in the organization of the cell wall and are associated to fertility .In plant cells, PAs are mostly stored in the vacuole and in the cell wall, but Spm is present also in the nucleus . PAs plaPolyamines also control many aspects of pollen development, both under normal and stress conditions. Here, we summarize the involvement of PAs during the entire developmental program and functioning of pollen.Nicotiana tabacum pollen formation inside the anthers . At the stage of uninucleate microspore, transcripts for enzymes involved in the biosynthesis of PAs, mostly Put, have been found, namely transcripts for ADC and ODC . At the bicellular pollen stages, other transcripts are present for the oxidative metabolism of Put  ; additional transcripts for enzymes that participate in the urea cycle and metabolism of amino groups  are also present .Transcripts for enzymes involved in PA biosynthetic and oxidative metabolisms are present starting from the early pollen stages as observed during  and ODC  . These compounds were shown to crosslink different cell wall polymers via ester and ether linkages, suggesting a role in modulating the rigidity of the cell wall , catalyzing the conjugation of hydroxycinnamoyl CoA to Spd in anthers, was recently shown to take part in the organization of the cell wall. The sht mutant displayed irregularities, depressions and decreased auto-fluorescence of the pollen grain . Recently, the biosynthetic pathway of (HCA)-Spd based on the analysis of several Arabidopsis knock-out mutants was proposed   .2O2 is involved in cell wall stiffening. Pollen PAOs . In Oryza sativa seven PAO isoforms have been identified, and one of these, OsPAO7, is specifically expressed in anthers, with an expression peak at the bicellular pollen stage ; OsPAO7 produces H2O2 about 100 times more efficiently than other PAO isoforms  are active. The aborted pollen grains showed high SAMDC activity in wall residues, while functional pollen  showed low SAMDC activity, suggesting a possible regulatory role of Spd in the functionality of kiwifruit pollen , when the main PA biosynthetic enzymes  were present and active in vitro  and Spd synthase (SPDS) (cyclohexylamine), or D-arginine (inhibitor of Put synthesis) led to abnormal pollen grains in male-fertile plants with reduced viability and germination .Pollen can be stored for extended periods without loss of viability under dry and low-temperature conditions leading to reduced metabolism. PAs may contribute to maintaining viability during natural quiescence and/or storage . Moreover, the inhibition of PA biosynthetic enzymes by bis(guanylhydrazones) strongly affected pollen germination  across the plasma membranes, the apical pool of ROS . In fact, during pollen tube growth, the activity of cytoplasmic TGase was mainly detected in the tube apex and in the region closest to the grain. PA conjugation to actin and tubulin, catalyzed by TGase, affected their ability to assemble and their interaction with motor proteins both in vivo and in vitro . Moreover, pollen TGase secreted into the medium catalyzed the covalent linkage of PAs to released proteins and their cross-linking in vitro. This feature may contribute to regulating the pollen tube-style interaction .In addition, PAs might also control the assembly and properties of cell wall polysaccharides, such as pectins, which bind to PAs by ionic linkages . In cellArabidopsis thaliana pollen tubes, exogenously supplied Spd increased the concentration of cytosolic Ca2+; Spd oxidation by PAO generated H2O2, which activated Ca2+ channels, thus inducing Ca2+ influx beyond optimal levels and causing the inhibition of tube growth. Activation of Ca2+ currents by Spd was significantly disrupted in pao knock-out mutants, but Ca2+ channels could still be activated following application of H2O2  in Solanaceae, Rosaceae and Plantaginaceae; they enter the pollen and are degraded in compatible pollen while they are active in incompatible ones causing the degradation of pollen RNA (When pollens land on an incompatible stigma they may undergo the Self Incompatibility (SI) response. This is the most important evolutionary system of the Angiosperms to prevent inbreeding and requires a species-specific cell–cell recognition system. The female determinants can be either a cell membrane receptor as in llen RNA .Pyrus communis and in Citrus grandis. In Pyrus communis the content of free PAs (Put and Spm) was lower during incompatible as compared to compatible pollination . This could be related to the inhibitory effect of PAs on RNases; in fact, Put and Spd, and, even more, Spm, have been shown to halve the activity of RNase in Malus domestica pollen  was stimulated in Pyrus communis and Citrus grandis. This could have led to cross-links among cytoskeleton proteins, generating high-mass aggregates, similar to the actin foci observed in Papaver, and forming the tube tip plug (In contrast to compatible pollination, SI pollination in e growth . The dirllen PCD . Since etip plug .Figure  Progress in past decades has significantly advanced our understanding of how PAs exert multiple roles by different molecular mechanisms. However, further investigations on the physiological function of PAs and their molecular partners are still needed. In particular, knowledge would strongly benefit from a deeper understanding of PA transporters, which have been poorly studied. This could provide new insights on the interactions between the tapetal layer and the pollen grain during its development in the anther. It could likewise explain how HCAAs, PA biosynthetic and oxidative enzymes and other cell wall components are deposited during microsporogenesis. Moreover, despite recent findings on the composition and biosynthetic pathway of pollen HCAAs, clear evidence regarding their functions is still lacking. While some of the roles of PAs are rather evident, e.g., modulation of the cytoskeleton by TGase, others remain elusive, e.g., PA interactions with nucleic acids. Such information could explain their possible role in epigenetic control, the interconnection between PAs and ROS, and the role of free and conjugated PAs in the apoplast during the pollen–pistil interaction.Pollen development is a complex and well-coordinated process governed by genetic and enzymatic processes, some of which are modulated by PAs. Hence, these aliphatic polycations, drive pollen development throughout its life span, as summarized in IA, GC, DS-F, SD contributed to the design of the work as well as drafting the work and revising it critically for important intellectual content; then they made the final approval of the version to be published. They agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. In details, the idea to write a paper about the polyamines action during pollen life was proposed by IA and SD that is the supervisor of the entire work. Part of the data described in the text have been done in the labs of Bologna and Siena both with a solid expertise in polyamine and pollen. GC designed and realized the figure of the paper.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.


In order to ensure their physiological and cellular functions, plasma membrane (PM) proteins must be properly conveyed from their site of synthesis, i.e., the endoplasmic reticulum, to their final destination, the PM, through the secretory pathway. PM protein homeostasis also relies on recycling and/or degradation, two processes that are initiated by endocytosis. Vesicular membrane trafficking events to and from the PM have been shown to be altered when plant cells are exposed to mutualistic or pathogenic microbes. In this review, we will describe the fine-tune regulation of such alterations, and their consequence in PM protein activity. We will consider the formation of intracellular perimicrobial compartments, the PM protein trafficking machinery of the host, and the delivery or retrieval of signaling and transport proteins such as pattern-recognition receptors, producers of reactive oxygen species, and sugar transporters. Plants co-exist with a vast diversity of mutualistic or pathogenic microbes such as bacteria, fungi, and oomycetes. Most of the land plants form a mutualistic relationship with symbiotic fungi such as mycorrhizal fungi, and legumes with rhizobia bacteria. Only a small fraction of plant–microbe encounters results in pathogenic relationship.Indeed, the plants have developed intricate defense mechanisms, referred to as innate immunity, to identify and defend themselves against a wide range of pathogens . The plaMany plant defense responses to infection are shared by PTI and ETI resistance such as protein phosphorylation, changes in ion fluxes, production of reactive oxygen species (ROS), synthesis of antimicrobial compounds, and pathogenesis-related proteins associated with cell wall reinforcement. ETI elicits more prolonged and robust immune responses than PTI, and is more often associated with the hypersensitive cell death that inhibits pathogen growth .Accordingly, the plant cell PM contains integral or peripheral proteins that regulate plant biotic interactions by recognizing microbes, activating subsequent signaling cascades, and controlling cellular entrance and exit of molecules.trans-Golgi network (TGN) to be delivered to the PM by exocytosis  and then transported along the secretory pathway through the Golgi apparatus and the ocytosis . Once atocytosis . A membrocytosis .Protein trafficking through the secretory and endocytic pathways relies on membrane-bound vesicles and a complex set of proteins involved in vesicle formation, transport, docking, and fusion with the respective target membrane . By adjuPlants can establish intracellular associations with mutualistic as well as pathogenic microbes. The intracellular accommodation of microbes allows them to gain access to host nutrients. In a mutualistic relationship, the host cell also benefits from specific metabolites originating from the microbes. Microbes penetrate the cell wall and differentiate specialized intracellular feeding compartments by invagination and extension of the host cell PM to colonize the intracellular space Figure . The perMutualistic association of legume plants with rhizobia culminates in the organogenesis of nitrogen-fixing root nodules. Rhizobia are first hosted in an intracellular infection thread (IT) that guides them through the root cell layers. The developing IT results from the local hydrolysis of the plant cell walls and invagination of the PM . When thMedicago root upon AM symbiosis evidenced flotillin-like proteins and proteins involved in vesicle trafficking supporting the importance of membrane trafficking events in mycorrhiza establishment and functioning  associated with plant roots. Intracellular fungal hyphae are enveloped by the periarbuscular membrane  in PM microdomains  that catalyze the exchange of bound GDP with cytosolic GTP, and GTPase activating proteins (GAPs) that enhance GTP hydrolysis .Pseudomonas syringae destabilizes the Arabidopsis ARF-GEF AtMIN7 through the host 26S proteasome that leads to suppression of immunity , which is the most studied PM-resident SNARE, plays a pivotal role in cell wall-based resistance against the powdery mildew agent, bidopsis , barley bidopsis , and grabidopsis . The potnfestans . A loss-nfestans  suggestiArabidopsis  group , has bee tritici . TaNPSN1bidopsis . The worA. thaliana plants expressing GFP-tagged SYP/SNAREs, has identified several disease resistance proteins of the TIR-NBS class as potential interactants . Moreover, accumulation of the exocyst subunit EXO84b was revealed at the site of PAM biogenesis in AM , and sugar transporters.The plant cell PM contains a plethora of receptors (PRRs) that sense changes in the biotic environment and trigger specific signaling cascades and physiological outputs inside the cell. PRRs do this by recognizing evolutionarily conserved MAMPs/elicitors or molecules released from damaged plant cells. PRRs are divided into two classes: leucine-rich repeat receptor-like proteins (LRR-RLPs) and receptor-like kinases (LRR-RLKs). Increasing evidence supports the claim that PRRs are part of multi-protein complexes associating regulatory proteins .N-glycosylation pathway, which permit a correct trimming of oligosaccharides on nascent proteins, are major actors in the ER-quality control.The ER is the first compartment of the secretory pathway. It ensures the quality-control process (ER-QC) for folding and maturation of client secretory proteins during and after their biosynthesis . Many ERArabidopsis. A recent review suggests that, owing to the diversity of ER-proteins engaged, the anterograde trafficking of PRR may be a potential point of regulation to generate appropriate defense responses , which take part of ER network structuration , have beRecently, ACD6 (ACCELERATED CELL DEATH6), a multipass membrane protein with an ankyrin domain, has been involved in the formation of a complex with FLS2 at the ER to promote FLS2 targeting to the PM and immune response under the signaling defense hormone salicylic acid SA .Environmental changes could alter the abundance of plant PM proteins by the process of endocytosis . Once reInternalization of cell surface receptors, via the clathrin-mediated pathway or any other endocytic pathway, would serve to terminate signaling through degradation, sustain signaling through recycling, or relay signaling inside the cell through the formation of signaling endosomes .Arabidopsis  in tomato  upregulates the expression of two SWEET genes in rice, a plant response that results from the direct binding of TAL  effectors to the SWEET promoters  at the PM are controlled by endocytosis during plant development . On the In AM, the symbiotic fungus offers nutrients, such as phosphorus and nitrogen, to the plant and get, in return, carbohydrates produced by photosynthesis . The tomInterestingly, an RNA-seq transcriptomic approach has reported, in addition to host transporter genes such as SUTs and SWEET1, a highly significant upregulation of remorin genes in ectomycorrhizal interaction . It may in vivo. Alternatively, protein clustering in microdomains could promote internalization by endocytosis and thus extents signaling intracellularly or ensures its termination. Future studies will have to characterize the dynamics of association of proteins with microdomains at the PM but also in intracellular compartments to figure out the precise role of lateral compartmentalization of membranes in pathogenic or mutualistic interactions.Interestingly, all PM proteins discussed above, and whose intracellular trafficking is modulated upon exposure to microorganisms, have been found in detergent-insoluble membranes or to co-localize with microdomain-associated proteins. Membrane microdomains are thought to temporally and spatially organize proteins and lipids into dynamic signaling complexes . Such a Pseudomonas syringae on Arabidopsis plants through the disruption of the actin cytoskeleton and the inhibition of endocytosis  trigger a massive remodeling of the host cell PM that leads to the formation, and maintenance, of a perimicrobial membrane with a unique identity. Perimicrobial membranes are characterized by the presence of microdomain-associated proteins of the remorin and flotillin families. These proteins are believed to play a role as molecular scaffolds for the building of signaling platforms. Future should reveal whether they are involved in the polarized delivery of specific PM proteins and/or the endocytic retrieval of others.Recent studies have identified vesicle trafficking regulators as modulators of the plant response to pathogenic or mutualistic microorganisms. A challenge that lies ahead will be to identify commonalities and differences in the vesicle pathways implicated in the different types of interactions. It will also be important to determine to what extent pathogen effectors affect the accumulation of PM proteins for subversion of plant defense. Deciphering new mechanisms of pathogenicity as well as novel aspects of plant immunity would benefit from the identification of the whole repertoire of protein trafficking components that are targeted by pathogen effectors. The identification of new R-genes should help to design novel strategies for manipulating crop plants toward resistance to ravaging pathogens.The Guest Associate Editor Daniel Wipf declares that, despite being affiliated to the same institution as the authors, 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.


Understanding how major organismal lineages originated is fundamental for understanding processes by which life evolved. Major evolutionary transitions, like eukaryogenesis, merging genetic material from distantly related organisms, are rare events, hence difficult ones to explain causally. If most archaeal lineages emerged after massive acquisitions of bacterial genes, a rule however arises: metabolic bacterial genes contributed to all major evolutionary transitions. Making sense of the origins of major lineages of life, and therefore of the ways by which novel physiologies, ecological systems, and classes of organisms evolved is possibly as important as understanding the origin of species The contribution of bacterial genes to eukaryotic evolution is well-acknowledged et al. A major evolutionary transition such as eukaryogenesis, giving birth to a high-level lineage, by the merging of genetic material from distantly related organisms, is usually assumed to be rare (no more than a few events per billion years). Being rare, however, does not entail that such events do not obey rules, but simply that causal rules are difficult to discover. In a recent series of original works, Nelson-Sathi et al. encourages strengthening the research program on evolutionary transitions, by specifically tracking the motion and the transformative role of metabolic genes from bacterial origin in the web of life. It could provide novel and broader angles to address the issue of the origins of lineages, and the diversity of life on Earth.This unparalleled role of metabolic bacterial genes in shaping lineages has also been observed in several endosymbiotic events, which turned non-photosynthetic eukaryotes into photosynthetic ones


Hypertrophic cardiomyopathy (HCM) is an autosomal dominant disease characterized by variable expressivity, age penetrance, and a high heterogeneity. The transcriptional profile , epigenetic modifications, and posttranslational modifications seem to be highly relevant for the onset of the disease. miRNAs, small noncoding RNAs with 22 nucleotides, have been implicated in the regulation of cardiomyocyte function, being differentially expressed in several heart diseases, including HCM. Moreover, a different miRNA expression profile in the various stages of HCM development is also observed. This review summarizes the current knowledge of the profile of miRNAs characteristic of asymptomatic to overt HCM patients, discussing alongside their potential use for diagnosis and therapy. Indeed, the stability and specificity of miRNAs make them suitable targets for use as biomarkers for diagnosis and prognosis and as therapeutical targets. Mut(MYBPC3) , 4. The (MYBPC3) . A high ickening . Due to ickening . This ischain MYH and cardOne of the most striking features of HCM is the inexistence of a correlation between genotype and phenotype, has family members carrying the same mutation developed distinct symptoms , 8, 9. TSince their discovery in 1993, miRNAs have been increasingly recognized as an important class of regulatory small noncoding RNAs that function as negative regulators of gene expression , 21. App MCM7 gene, or in exons of protein coding genes, as miR-985 located in the last exon of CACNG8 gene [ DLEU2 gene and from exons of noncoding genes such as miR-155 coded by an exon of BIC gene [miRNAs are transcribed by RNA polymerases II and III from different genomic locations –27 and cBIC gene , 29 , which are double stranded oligonucleotides including the mature miRNA sequence and the complementary passenger strand , 65. TheHCM therapeutics based on miRNAs involves the modulation of expression of these noncoding RNAs in cell, by inactivating the function of prohypertrophic miRNAs using synthetic miRNAs or by using miRNAs mimics oligonucleotides of antihypertrophic miRNAs . in vivo studies [The possibility of the use of miRNAs based therapy in treatment of HCM is highlighted by several studies , 66, 67  studies  successf studies . HoweverDespite the promising use of therapy based miRNAs, modulation of the function of miRNAs raises several concerns. The uptake of miRNA-mimics can result not only in the restoration of miRNA function in affected cells but also in the overexpression of the miRNA in other cells. In the same way, the inhibition of miRNA activity by antimiRs can be done on off-target locations. Therefore, targeting would be important in order to meliorate miRNA therapy, which can be accomplished with nanotherapies . FurtherThe study of miRNAs expression profiles that will allow its use for diagnosis, prognosis, and therapeutics of HCM is still in its infancy. The high heterogeneity of HCM pathology renders the detection of a disease specific miRNA profile difficult. Furthermore, due to the invasive procedures required for human cardiomyocytes analysis, the analysis of the cardiac miRNA profile is only possible when the disease reached a terminal state. The use of murine models has proven to be fruitful to study the several stages of the disease. Nanobiotechnology allows the simultaneous detection and silencing of miRNAs directly from blood samples will speed up early diagnosis, prognosis, and therapy in hypertrophic cardiomyopathy.


Nanoscale vesicles have become a popular tool in life sciences. Besides liposomes that are generated from phospholipids of natural origin, polymersomes fabricated of synthetic block copolymers enjoy increasing popularity, as they represent more versatile membrane building blocks that can be selected based on their specific physicochemical properties, such as permeability, stability, or chemical reactivity. In this review, we focus on the application of simple and nested artificial vesicles in synthetic biology. First, we provide an introduction into the utilization of multicompartmented vesosomes as compartmentalized nanoscale bioreactors. In the bottom-up development of protocells from vesicular nanoreactors, the specific exchange of pathway intermediates across compartment boundaries represents a bottleneck for future studies. To date, most compartmented bioreactors rely on unspecific exchange of substrates and products. This is either based on changes in permeability of the coblock polymer shell by physicochemical triggers or by the incorporation of unspecific porin proteins into the vesicle membrane. Since the incorporation of membrane transport proteins into simple and nested artificial vesicles offers the potential for specific exchange of substances between subcompartments, it opens new vistas in the design of protocells. Therefore, we devote the main part of the review to summarize the technical advances in the use of phospholipids and block copolymers for the reconstitution of membrane proteins. Compartmentalization is a key feature of eukaryotic cells to spatially separate distinct biochemical processes from each other. Lipid bilayer membranes serve as impermeable barriers that effectively separate subcellular compartments. This (i) enables the simultaneous operation of metabolic pathways that utilize the same intermediates and (ii) allows for the adjustment of specific reaction conditions inside individual organelles. In order to translate this natural principle of biological organization, the use of membranes to encapsulate chemical reactions has attracted interest in the design of synthetic systems. Lipids or block copolymers are commonly used to build membranes in synthetic systems, in so-called liposomes or polymersomes, respectively.Liposomes consist of a shell of amphiphilic lipid species, such as phospholipids, that encapsulate an aqueous solution. The lipids are arranged in a bilayer with the polar head groups of the two leaflets facing toward the inside and the outside aqueous phase and the hydrophobic tails of the phospholipids facing toward each other Figure B. Based Both liposomes and polymersomes have become popular as vectors for targeted and tailored drug delivery and for the application in biochemical microreactors. Due to the amphiphilic nature of the lipid or polymer building blocks, a spontaneous assembly into vesicles occurs in aqueous environments  and alcohol dehydrogenase (ADH) were encapsulated separately into intrinsically porous sub-micrometer-sized PS-b–PIAT polymersome subcompartments, which, in turn, were engulfed by PB-b-PEO GUVs  (PVFc-b-P2VP) or the pH- and temperature-responsive polymer polystyrene-b-poly (PS-b-PDMAEMA). On the other hand, the cargos dimethyldodecylamine (DDA) and diphenyl disulfide (DPDS) were selectively switchable from the water-insoluble to the soluble form by pH change or H2O2 redox trigger, respectively. The substances were stored in the water-insoluble form inside the polymersomes and were then specifically released by an external stimulus that allowed the passage of DPDS (by oxidation) or DDA (by pH change) across the membrane.In addition, stimuli- and cargo-selective content release was achieved in dual stimuli-responsive polymersomes with two kinds of cargo , which is a trimeric integral membrane protein that enhances the passive diffusion of small hydrophilic molecules  identify the best expression vector compatible with the cell-free expression system , other, non-detergent-mediated fusion techniques have been elaborated, including the work of Kahya et al. . Here, tIn other reconstitution techniques, solubilized proteins are added to the already formed liposomes. Defects in the liposome membrane are either induced by sonification pulses (Rigaud and Lévy, 0F1-ATPases, and porins (Elani et al., Some classes of proteins, e.g., cytochromes, bacteriorhodopsin and FArtificial vesicles are a versatile resource to establish compartmentation in synthetic biochemical nanoreactors. To this end, compartmentation can either be achieved by immobilizing artificial liposome or polymersome vesicles to the matrix of microfluidic reactors or by generating nested or concatenated vesicular systems such as vesosomes.In order to maximize yield of biochemical processes inside vesicular nanoreactors, it is indispensable to control the exchange of substrates and products across the membranes between the reactor compartments. Technically, it is possible to reconstitute entire metabolic pathways into liposomes or polymersomes, and by the use of vesosomes or immobilized vesicles, it seems even feasible to mimic the natural compartmentation of these pathways in synthetic systems. However, the exchange of substances across membrane boundaries needs to be highly specific to make compartmented reconstituted biochemical pathways work.Exchange of substances across membranes can either be accomplished by utilizing lipid and coblock polymers that change permeability in response to physicochemical triggers such as heat, redox potential, or pH. However, only unspecific mixing of contents can be achieved by altering the permeability of the membrane boundary. The incorporation of unspecific protein pores into the membranes of vesicle compartmented reactors, e.g., porins or α-hemolysin, allows for a more selective exchange of low molecular weight substances but still permits the passage of a variety of intermediates that are similar in charge and/or structure. Ultimately, the reconstitution of specific membrane transporters that only allow the passage of individual substrates is necessary for the functional reconstitution of compartmented biochemical pathways. As outlined in this review, diverse approaches can be undertaken to achieve successful reconstitution of membrane transport proteins into artificial membranes. Since the optimal conditions for successful reconstitution are quite specific for each individual transport protein and can hardly be transferred to other candidates, we have taken care to summarize the available techniques for membrane transporter reconstitution. We believe that the integration of metabolite transporters into vesicle-based nanoreactors will largely advance bottom-up approaches in the development of compartmented protocells in the future.CS, AL, and LV wrote the article. NB, VS, and LV conceptualized the 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.


The limited success of cancer vaccines in clinical could be owing to their low efficacy that can be largely affected by their formulation. Vaccine formulation usually consists of three components, antigen, delivery system, and adjuvant. Recently, long peptide antigen (LPA) has been attracting interest, because (1) one LPA can provide antigen presenting cells (APCs) with multiple T cell epitopes and (2) it avoids undesired presentation by non-professional APCs that can lead to the exhaustion of antigen-specific T cells. While a LPA can include multiple epitopes, the prerequisite design of junction of epitopes  for robust immunogenicity is not yet established, and we explored it in this study. On the other hand, accumulating data has been suggesting the importance of delivery system for cancer vaccines. By employing a novel nanogel-based delivery system, we tried to improve the immunogenicity of LPA vaccine.LPA was designed to include three mouse cytotoxic T cell (CTL) epitopes that were linked with the IES of oligotyrosine, oligothreonine, oligoglycine or oligoproline. The complex of LPA and cholesteryl pullulan (CHP) nanogel was fabricated, and subcutaneously injected to mice with TLR agonist such as CpG oligoDNA or poly-IC RNA as an adjuvant. Frequency of vaccine-induced CTLs specific to each epitope was then determined by intracellular IFN-γ staining assay. Using similar mouse model, the effect of CHP nanogel delivery system on transportation of LPA to APCs in the draining lymph node (DLN) as well as T cell response to vaccination was also examined, comparing with a conventional delivery system, incomplete freund adjuvant (IFA).The IES candidates were selected based on in silico prediction of the sensitivity to proteasomal cleavage. The sensitivity of oligotyrosine and oligothreonine was predicted high, while that of oligoglycine and oligoproline was low. In the vaccinated mice, the LPA including the oligotyrosine IES most robustly stimulated specific CTLs towards all three epitopes included in LPA. The LPA including the oligothreonine IES was second best. In contrast, LPA including the oligoglycine or oligoproline IES often induced very limited CTL response. In the evaluation of delivery system, the CHP nanogel transported LPA to APCs in the DLN much more efficiently than IFA did. The kinetics of the CHP nanogel-mediated LPA transportation was closely similar to that of APC activation by TLR agonists, suggesting that the CHP nanogel harmonises the action of LPA and adjuvant. Indeed, in the presence of TLR agonists, the CHP nanogel greatly augmented the immunogenicity and anti-tumour effect of LPA vaccine.We succeeded in the enhancement of immunogenicity of LPA vaccine by the rational design of IES in LPA and the CHP nanogel-mediated efficient antigen transportation to the DLN. These technologies will bring a remarkable improvement of vaccine efficacy.


Solanum lycopersicum) affects the metabolism of ripening fruit at several levels. Most strikingly, recent experiments have shown that some of the molecular mechanisms originally devoted to sense and respond to environmental light cues have been re-adapted during evolution to provide plants with useful information on fruit ripening progression. In particular, the presence of chlorophylls in green fruit can strongly influence the spectral composition of the light filtered through the fruit pericarp. The concomitant changes in light quality can be perceived and transduced by phytochromes (PHYs) and PHY-interacting factors, respectively, to regulate gene expression and in turn modulate the production of carotenoids, a family of metabolites that are relevant for the final pigmentation of ripe fruits. We raise the hypothesis that the evolutionary recycling of light-signaling components to finely adjust pigmentation to the actual ripening stage of the fruit may have represented a selective advantage for primeval fleshy-fruited plants even before the extinction of dinosaurs.Besides an essential source of energy, light provides environmental information to plants. Photosensory pathways are thought to have occurred early in plant evolution, probably at the time of the Archaeplastida ancestor, or perhaps even earlier. Manipulation of individual components of light perception and signaling networks in tomato ( Light has a dual role in plants as an essential source of energy for driving photosynthesis and, on the other hand, as an environmental cue that modulates many aspects of plant biology such as photomorphogenesis, germination, phototropism, and entrainment of circadian rhythms . The abiArabidopsis thaliana, shade decreases the production of carotenoids in photosynthetic tissues  or perhaps even earlier, before the occurrence of the endosymbiotic event that gave rise to photosynthetic eukaryotes over more than a billion years ago . Through tissues  in part  tissues . De-reprse light .Solanum lycopersicum), which is the main model system for fleshy fruits, that is, fruits containing a juicy fruit pulp. Further, we will discuss potential selection pressures that might account for the evolutionary recycling of light-signaling components in fleshy fruits.Light signals in general and PHYs in particular also modulate the genetic programs associated to fruit development and ripening. Here we will revise current and emerging knowledge on this area based on work carried out in tomato , i.e., relatively recently in the history of plants , as an aUpon fertilization, the development of fleshy fruits such as tomato can be divided into three distinct phases: cell division, cell expansion, and ripening . These dhp) mutants hp1 and hp2 are two of the best characterized. These mutants owe their name to a deep fruit pigmentation derived from an increment in the number and size of plastids, which in turn result in elevated levels of carotenoids such as lycopene  and DEETIOLATED1 (DET1), respectively . Other components that participate in the same light-signaling pathway that HP1 and HP2 have also been shown to impact tomato fruit metabolism. For instance, silencing the tomato E3 ubiquitin-ligase CUL4, which directly interacts with HP1, also produces highly pigmented fruits . Transgenic plants with downregulated transcripts of COP1 and HY5 produce tomato fruits with increased and reduced levels of carotenoids, respectively  has also shed light on the subject. Tomato plants overexpressing the blue light photoreceptor cryptochrome 2 (CRY2) produce fruits with increased levels of flavonoids and carotenoids . Another key difference between tomato leaves and fruits is the cuticle, which is far more pronounced in the fruit. While a potential role of the cuticle in altering the spectral properties of the light that reaches the pericarp cells remains to be investigated, it is now well established that the occurrence of chlorophyll in fruit chloroplasts significantly reduces the R/FR ratio of the light filtered through the fruit fresh . Chlorophyll breakdown at the onset of ripening reduces the self-shading effect, consequently promoting PHY activation, degradation of PIF1a, derepression of PSY1, and eventually carotenoid biosynthesis . In this manner, the genetically controlled expression of PSY1 (and hence the production of carotenoid pigments) is fine-tuned to the actual progression of ripening  , at a tiGinkgo biloba, Taxus baccata, and Ephedra distachya) produce fleshy colorful tissues around their seeds and, similar to that occurring in angiosperms, these fruit-like structures undergo a ripening process that also serves as a visual advertisement for animals to eat them and disperse their seeds. Recent evidence supports the hypothesis that the main molecular networks underlying the formation of the fleshy fruit were originally established in gymnosperms  might have evolved by co-option of components from the preexisting shade-avoidance responses  by altering the behavior of the receivers  . Importaesponses . This evBL, LA, and MR-C searched and discussed the literature and wrote the 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.


The first and the sixth National Iranian Stroke Congress were held on 27-29 November 2013 in the Petrochemical Recreation Complex, Tabriz, Iran. The congress was supported by World Stroke Organization and Turkey Cerebrovascular Association and almost 100 physicians and researchers across Iran and other countries participated in the congress.Dr. Afshin Divani from USA, Dr. Manouchehr Seyyedi Vafaei from Denmark, Prof. Olsen Niles from Denmark and Dr. Ozcan Ozdemir from Turkey were the international guests of the congress. The congress was managed by Prof. Mehdi Farhoudi as Chief and scientific secretary and Dr. Kaveh Mehrvar as executive secretary. This congress was supported nicely by the president of Iran Stroke Association Dr. Mohammad Reza Gheini.The main goal of this congress was to gather stroke-related specialists and masters  as well as paramedic staff  to contribute updating stroke-related practical knowledge with the latest scientific achievements and recent developments. The other aim of the congress was recognition of related problems and obtaining their solutions in prevention and treatment fields.Totally, 220 abstracts were submitted to the congress, 30 selected for oral presentation and 171 for poster presentation. Furthermore, 25 invited lectures were also presented.The milestone of this scientific meeting comprised four predominant sections.Oral PresentationsThe main focus of the congress was discussing the acute stroke management. In the mornings main lectures presented and other presentations planned afternoons. In this congress, domestic and foreign prominent masters and specialists pointed the main ideas about acute phase stoke management. New medical findings in the field of stroke management including diagnostic methods, the usual treatments of stroke and related protocols, importance of teamwork in stroke management were also discussed in detail.PanelsIn addition, besides the scientific works and presentations two panels were provided in the congress. The first panel was about “difficulties of thrombolytic therapy in Iran” and the second one discussed the “importance of the medical team working in acute stroke” to help stroke patients.WorkshopsDuring the 3-day congress, eight major workshops were held in various areas of acute stroke. The title of these workshops, include nursing care in stroke, intravenous thrombolytic therapy, intra-arterial thrombolytic therapy, how to check National Institutes of Health Stroke Scale, neurosonology, imaging in acute stroke, rehabilitation after acute stroke, and empirical models of stroke. The workshops were regarded with acceptable number of attending participants considering practical discussion of experts and specialists.ExhibitionAs a peripheral program of the congress, an exhibition of pharmacological companies active in the field of stroke treatment and management was held. In this exhibition, the pharmacological companies in both traditional and modern medicine, research companies in the field of dealing and management of stroke-related patients’ education presented their latest products.Finally, this international annual congress not only provides new fields to discuss stroke-related causalities, treatments or management, but also attendance and participating foreign neurology masters and specialist contributes to establish an international relationship and connects Iran to science world and international neurology centers across the world.As the main goals of this congress, including updating target group information, discuss the problems and suggestion practical approaches through analyzing the oral presentations of the urban and foreign masters and ideas suggested as posters, the scheduled panels were investigated as above-mentioned.Finally, as a consensus conclusion, the contents of the statement were listed as followings:th coincides with October 29 - world stroke day - as the stroke day in the national calendar to improve society health and lower stroke prevalence by correcting the food habits, society culture, and lifestyles.According to the increase of stroke incidence, considering more prevalence among young aged groups, and significant casualties thus to stroke, according to WHO warning, prophylactic assessments and related management should be considered through improving the point of view of all society and officials. As mentioned, the second cause of vascular disease mortality and the most prevalent fact of disability among adult age group is stroke. So that as a proposal, it is been expected to coin Aban 7According to either the proved effect of thrombolytic therapy in the acute phase of stroke in choice group patients and prominent role of this therapy in after stroke disability rehabilitation or considering the usefulness of the therapy due to its golden time, these points should be regarded:Stoke should be considered as one of the medical emergencies like trauma and myocardial infarction.Tissue plasminogen activator medication should be concluded in the conservative portfolio of health insurance.