Astrocytes, members of the glial family, interact through the exchange of soluble factors or by directly contacting neurons and other brain cells, such as microglia and endothelial cells. Astrocytic projections interact with vessels and act as additional elements of the Blood Brain Barrier (BBB). By mechanisms not fully understood, astrocytes can undergo oncogenic transformation and give rise to gliomas. The tumors take advantage of the BBB to ensure survival and continuous growth. A glioma can develop into a very aggressive tumor, the glioblastoma (GBM), characterized by a highly heterogeneous cell population (including tumor stem cells), extensive proliferation and migration. Nevertheless, gliomas can also give rise to slow growing tumors and in both cases, the afflux of blood, via BBB is crucial. Glioma cells migrate to different regions of the brain guided by the extension of blood vessels, colonizing the healthy adjacent tissue. In the clinical context, GBM can lead to tumor-derived seizures, which represent a challenge to patients and clinicians, since drugs used for its treatment must be able to cross the BBB. Uncontrolled and fast growth also leads to the disruption of the chimeric and fragile vessels in the tumor mass resulting in peritumoral edema. Although hormonal therapy is currently used to control the edema, it is not always efficient. In this review we comment the points cited above, considering the importance of the BBB and the concerns that arise when this barrier is affected. It has been one hundred years since Edwin E. Goldmann discovered the blood-brain barrier (BBB). Using trypan-blue dye intravenous injections in several animals, he observed that it spread throughout the body, except for the brain and spinal cord, which remained unstained. When trypan blue was injected into the subarachnoid space at the lumbar level, or in the cisterna magna in young rabbits, the Central Nervous System (CNS), including the choroid plexus (CP), were stained  and its major proteins, metabolites and a diversity of other molecules. Anatomically, the CP is a group of thin membranes located inside the lateral, third and fourth ventricles. It is basically composed of a monolayer of specialized epithelial cells derived from the ependyma that cover the ventricles walls. This monolayer coats a highly perfused stroma containing permeable fenestrated blood vessels, fibroblasts, dendritic cells and macrophages , which promotes neuronal proliferation and survival in the developing mouse cortex , microglia and neurons, all playing essential roles in CNS homeostasis , oligodendrocytes (oligodendrogliomas) and ependymal cells (ependymomas) . This classification is intended to improve the accuracy of prognosis, and thus the effectiveness of therapeutic management of patients. Gliomas classification has long been based solely on the histopathological features of the tumor tissues, obtained through neurosurgical resections, which are whenever possible, the first line of therapeutic intervention. The diagnostic criteria include the morphology of the tumor cells, tissue architecture, and the immunohistological marker profiles  tend to be well differentiated and have few cellular anomalies or atypias, but in general, they closely resemble their non-neoplastic cellular counterparts. These tumors contain specific genetic alterations  are anaplastic, showing signs of increased vessel density, cellular atypias, high mitotic activity and elevated cell density. Grade IV astrocytoma, better known under the name of glioblastoma (GBM) multiformis, or simply GBM, is the most common and aggressive form of glioma. Glioblastoma samples are characterized by a very high cell density, numerous atypias, areas of necrosis, and robust neoangiogenesis. Two forms of GBM have been identified: Glioblastoma growth is closely associated with the formation of new vessels and one of the most serious clinical complication is the development of vasogenic brain edema, which dramatically increases the intracranial pressure (ICP) by BBB leakage  present in the tumor mass  has been correlated to glioma progression. Aquaporins are a family of integral membrane transport proteins that facilitate water efflux across cell membranes in response to osmotic gradients. So far, 14 AQPs have been characterized in humans and rodents , using a contrast medium  acts as a tumor suppressor, decreasing the proliferation, self-renewal and tumor-initiation ability of GBM stem cells  to the plasma membrane and the consequent release of intraluminal vesicles (ILVs) of MVBs into the extracellular environment. At this point, ILVs are referred to as exosomes  that had been mentioned in the context of altered signaling pathways, whose expression could represent potential therapeutic targets in GBM  is caused by two main mechanisms, vasogenic and cytotoxic. While BTRE is classically associated with vasogenic edema, cytotoxic edema is being increasingly implicated in the pathophysiology of peri-tumoral swelling  analog, was studied in a phase I/II trial and seems to be well tolerated. Randomized phase III trials are now envisioned , because of the high risk of recurrence of seizures. The first-line AEDs, including valproate (VPA), phenytoin and carbamazepine, have demonstrated many major side effects and dangerous interactions with chemotherapeutic and antitumor drugs. The most commonly seen side effects with these drugs are cognitive impairment, liver dysfunction, dermatological reactions and bone-marrow suppression. Several studies have found that side effects are more common in patients with brain tumors than in the general epileptic population.Drug interactions between AEDs and commonly used tumor therapies can lead to inadequate control of the seizures or sub-therapeutic treatment of the tumor. Toxic effects have also been noted. Many of the common AEDs, especially carbamazepine, phenytoin and phenobarbital, induce cytochrome P450 enzymes, which accelerate the metabolism, reducing the half-life of corticosteroids and many other chemotherapy drugs that in this context exhibit decreased therapeutic effectiveness. Some chemotherapeutic agents also induce the P450 system, lowering concentrations of the AED.Among the older AEDs, valproate, in doses of 1–3 g per day, has been claimed to be the drug of choice because of its anticonvulsive efficacy. It has an intravenous formulation, which is very useful in perioperative management and also in cases of status epilepticus, which is seen in up to 26% of these patients, with the overall mortality rate reaching 30–40%. Also, its action as a histone deacetylase inhibitor may reduce proliferation rates and promote differentiation of cancer stem cells, the main mechanism responsible for tumor recidivism in the case of GBM , lacosamide and zonizamide have emerged that are without clinically important drug interactions. Most of them have shown similarly good efficacy, with better tolerability for oxcarbazepine, lacosamide and lamotrigine. Most data were based on uncontrolled retrospective studies. Of this group, only levetiracetam and lacosamide have intravenous formulations.Many studies have now proposed levetiracetam as the drug of choice in the treatment and prevention of peri- and post-operative seizures, in daily doses of 1.5–3 or 4 g. Levetiracetam has the advantages of excellent bioavailability , no protein binding, no liver-metabolism or enzyme-inducer effect, and a very good tolerability profile. It has a completely different mechanism of action, binding to the Synaptic Vesicle Protein 2 A, thus inhibiting neurotransmitter release. It also has shown histone deacetylase inhibitor, anti-inflammatory and antioxidant effects. Many recent trials have demonstrated its anticonvulsive efficacy and safety in the control of perioperative (88 to 100% seizure control) and long-term epileptic seizures (91% seizure freedom), as well as in the treatment of status epilepticus when combined with phenytoin  and much better tolerability for levetiracetam (long-term complication rates of 26.8% for VPA and 9.8% for LEV). Moreover, 38.5% in the VPA group changed to or added another AED, whereas only 17.6% in the LEV group did so. Another retrospective comparative study (Kerkhof et al., In a retrospective comparative study between valproate and levetiracetam with 282 patients with supratentorial brain tumors (Lee et al., The clinical management of seizures in this context is complex, requiring experts in this field, measurement of serum levels of the antiepileptic and oncologic drugs, serial blood analysis, monitoring of clinical side effects, and keeping in mind that the recurrence of seizures may represent a tumor recurrence.In this review, we have pointed out and discussed the implications of the interactions between normal glia or glioma cells and the BBB. Since its first discovery by Edwin E. Goldman, considerable progress has been made in the understanding of BBB. Although 100 years have passed since the BBB discovery, the mechanisms by which some drugs or parasites enter the brain are still unsolved. Furthermore, the real interactions of the endothelial cells with the extra-endothelial components of the brain microenvironment remain unclear. It is not completely understood how astrocytes endfeet interact with BBB and the astrocytoma derived signals that lead to an intense vasculogenesis into the tumor. Interestingly, GBM and BBB interactions may occur via extracellular proteins. For instance, the imbalance of tenascin and fibronectin in the tumor contributes to vessel formation as we have previously demonstrated (Alves 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.


Arteriviridae, Genus Arterivirus) include an interesting group of enveloped positive stranded RNA viruses that infect domestic and wild animals and they share a strikingly similar genome organization and replication strategy to that of coronaviruses, but differ considerably in their genetic complexity and virion architecture. Until recently, arteriviruses included equine arteritis virus (EAV), porcine reproductive and respiratory syndrome virus (PRRSV), lactate dehydrogenase-elevating virus (LDV) of mice, and simian hemorrhagic fever virus (SHFV). Three of these viruses were first discovered and characterized a long time ago , whereas PRRSV emerged first in Europe and North America in the late 1980s and early 1990s. In 2012, arteriviruses were expanded to include the newly identified wobbly possum disease virus (WPDV) that causes neurologic disease among free-ranging Australian brushtail possums (Trichosurus vulpecula) in New Zealand. Similarly, four new genetically divergent SHFV variants were recently identified in a single male colobus monkey (Procolobus rufomitratus tephrosceles) and in African red-tailed (guenon) monkeys (Cercopithecus ascanius) from Kibale National Park, Uganda.The arteriviruses (FamilyErythrocebus patas), baboons (Papio anuibus), and African green monkeys (Ceropithecus aethiops). SHFVs of African origin are highly infectious and fatal in Asian rhesus monkeys . Although the human arterivirus is yet to be identified, there is significant concern about cross species transmission of some of these arteriviruses following xenotransplantation. There is no indication that any humans are infected with SHFV or any other arterivirus during disease outbreaks. However, it is obvious that transplantation of humans with tissues from SHFV-infected baboons or PRRSV-infected pigs would result in transfer of a considerable amount of these viruses, which could result in selection of a variant(s) that can replicate in humans. In this respect, PRRSV and SHFV should be seriously considered as a threat to xenotransplantation of tissues and organs from pigs and baboons.The arteriviruses are highly species specific, but share many biological and molecular properties, including virion morphology, a unique set of structural proteins, genome organization and replication strategy, and the ability to establish prolonged or true persistent infection in their natural hosts. However, the epidemiology and pathogenesis of the infections caused by each virus is distinct, as are the diseases they cause. EAV and PRRSV cause respiratory and reproductive disease in equids and swine, respectively. The natural hosts of LDV are field and the laboratory mice, but unlike other arteriviruses, LDV is generally non-pathogenic. Indeed this virus is a significant adventitious agent in the laboratory mice, and causes an increase of lactate dehydrogenase enzyme, contamination of transplantable tumors, clinically silent infection with mild pathology, and immunomodulation. SHFV establishes persistent, perhaps life long infections without disease in Patas monkeys (Of the five arteriviruses, EAV and PRRSV are the most economically important viruses and pose a significant threat to equine and swine industries, respectively. Both viruses are distributed throughout the world, but PRRSV infection has become the most serious infectious disease of the pigs costing over $560 million US dollars per year to the US swine producers. Genetic manipulation of full-length cDNA clones of these two viruses has become an especially important and widely used tool to study the biology, pathogenesis, and virulence determinants of both of these viruses. Due to the economic impact, there has been significant research interest in molecular characterization of PRRSV and its interaction with the host defense system. In conjunction with advances in PRRSV molecular virology, reverse genetics and immunology, more and more attention is being directed to development of safe and efficacious vaccines against PRRSV. However, development of effective vaccines against PRRSV infection has been a significant challenge due to the presence of two genotypes of the virus with rapid emergence of antigenic variants and delayed suboptimal neutralizing antibody responses to infection. Current modified live and killed vaccines have had limited success, prompting investigation of alternative approaches to design new and improved vaccines.In this special issue of Animal Arterivirus Infections by BioMed Research International dedicated to recent advances in arterivirus research, we have assembled seven manuscripts that describe original research data on PRRSV and EAV. The articles cover characterization of innate immune response to PRRSV and EAV, genetic diversity of PRRSV strains circulating in China, interaction between the structural and nonstructural proteins of PRRSV with the host cellular factors, vaccination-induced protection against homologous and heterologous challenge, and development of a recombinant DNA vaccine against PRRSV. These articles provide new insight into better understanding immune responses to PRRSV and EAV in the context of disease as well as vaccine-induced protection, which will help to develop better vaccines to control the growing worldwide burden of disease from these agents in the future. Clearly a comprehensive research approach combining molecular virology, immunology, viral pathogenesis, and host genetics is required to unravel the complexities of virus-host interactions of these viruses. We believe that we have assembled basic knowledge that encompasses these complexities, describes technologies that have contributed to this knowledge, and identifies at least some of the major problems faced in attempting to further understand the virus-host interactions that result in disease. We would like to thank all the authors and co-authors who contributed to this special issue of Animal Arterivirus Infections, as well as the numerous reviewers who reviewed each and every paper in a timely manner and provided their input to improve the quality and clarity of the final product. We would also like to acknowledge that there are many other colleagues and scientists who are active in the field of arteriviruses whose expertise has not been represented here.Denis ArchambaultUdeni B. R. BalasuriyaRaymond R. R. RowlandHanchun YangDongwan Yoo


In the last two decades, genetic and genomic studies have revealed the astonishing diversity and ubiquity of microorganisms. Emergence and expansion of the human microbiome project has reshaped our thinking about how microbes control host health—not only as pathogens, but also as symbionts. In coral reef environments, scientists have begun to examine the role that microorganisms play in coral life history. Herein, we review the current literature on coral-microbe interactions within the context of their role in evolution, development, and ecology. We ask the following questions, first posed by McFall-Ngai et al.  in their Symbiodinium  was isolated from water adjacent to the coral Galaxea fascicularis  isolated from an enrichment culture of a black band diseased coral  isolated from Montipora aequituberculata .The mainly oligotrophic habitat of corals, makes associations driven by metabolic complementation desirable, for instance association between diazotrophic bacteria and Nematostella due to proposed HGT    and horizontal transmission (uptake of symbionts from the environment) of Bacterial populations. Vertical transmission in Porites astreoides was recently demonstrated. Ectoderm-associated bacteria from the parent colony, mainly Roseobacter spp., were observed in both newly released, and in 4-day old planulae, with densities increasing after settlement  from parent colony to larvae after gamete release  from the Great Barrier reef , may contribute to the protection of spawn and larvae against uncontrolled bacterial colonization, as has been suggested by studies in adult corals and in the Cnidarian, Hydra  were found to form biofilms that were sufficient to increase metamorphosis rates of Acropora willisae and A. millepora by up to 50%  surfaces  —where settlement is triggered by larval contact with bacterial cell surface displays of phage-tail like proteins  must overcome competition with other benthic organisms to grow and form a robust adult colony. Bacteria may mediate the survival of coral recruits in the proximity of adult coral colonies or other coral competitors such as macro- and turf algae  of coral nubbins grown with algal competitors and appeared to be mediated by factors able to pass through a fine mesh  in the microbiota of three different coral species: Acropora tenuis, Pocillopora damicornis, and Tubastrea faulkneri, and subsequently documented increases of Roseobacter spp. and Alteromonas spp. in filtered seawater concurrent with release of gametes from A. tenuis, or larvae from P. damicornis  and/or tissue-death may result and lead to progressive death of the entire coral colony. The control and regulation of activities in the holobiont to maintain emergent function is termed homeostasis. In the coral holobiont, like any other animal holobiont, stressors that threaten homeostasis may be biological, chemical, or physical, and an initial stressor may lead to cascading interactions that trigger additional responses. To assess how homeostasis is maintained among components of the coral holobiont this section will focus on how physical and chemical gradients shape the microbiota in different habitats within the coral animal, and consider how perturbation of these gradients influence holobiont function. In particular, we will examine the following habitats: (1) the coral mucus and epidermal surface, (2) intracellular and interstitial spaces, (3) the gastrovascular cavity, and (4) the skeleton  and chemokinesis  guided by gradients of infochemicals such as DMSP that are produced by the holobiont and may increase during periods of temperature stress  cells is dominated by  Trench, , 1993. T Trench, . To date Trench, . Yet obtRitchie, . DespiteSymbiodinium may shape several intracellular and interstitial physicochemical gradients in the coral holobiont. Reactive oxygen production is associated with the activity of coral phagocytotic immune cells   with some evidence of genome-reduction associated with symbiosis  and the community composition included ribotypes observed in gut microbiomes of other marine and terrestrial taxa   and ecological interactions. In homeostasis, microbial populations are subject to the balance of resource limitation (bottom-up control) and antagonistic interactions that lead to population extinction (top-down control) Figure . DeviatiCyanothece , and organic carbon mediates the structure of the populations that persist and coexist. Perturbations that relieve resource limitation have both holobiont and ecosystem scale consequences.The microbial communities of several species of corals are hypothesized to harbor stable associates of the coral host, including strains of the nitrogen-fixing cyanobacteria Ritchie,  and popuSymbiodinium is quite apparent, photosynthetic microbes in the holobiont such as the cyanobacteria and anoxygenic phototrophs also need access to light energy for growth  , oxygenic photosynthesis renders most of the coral interior oxic during the day, however diurnal patterns of anoxia/hypoxia in the polyp gastrovascular cavity  was taken up more efficiently that the carbon-equivalent of bacterial cells  acroporid coral juveniles, promoting recruitment success during competition with benthic cyanobacteria  appears to be critical for the healthy function of most reefs dominated by coral. Perturbation of the reef environment by exposure to nutrients  or dissolved organic carbon (DOC) has been linked to changes in microbial community structure and adverse physiological consequences for corals Table . Nutrien Lesser, . Nutrien Lesser, . Other i Lesser, . NutrienAddition of DOC, directly or indirectly via nutrient-stimulated primary production, leads to high heterotrophic bacterial abundance and degradation of reef quality  contribute to this stable coexistence through a complex chemical ecology of antimicrobial compounds and signals that modulate microbial activity. Environmental stresses that disrupt coordinated activities of the coral holobiont pose a great risk to coral homeostasis by weakening top-down control of microbiota structure and enabling growth of opportunistic microbes that proliferate at the expense of the coral host.Top-down forcing of holobiont structure through immune interactions, predation, and interference-based competition allows certain microbes to grow/persist in the holobiont while others are eliminated. Like all animal tissues, corals represent rich stores of nutrients kept out of bounds from potential microbial consumers by tissue barriers and immune functions. The epithelium and immune system of healthy corals are critical lines of defense to prevent indiscriminant microbial digestion of viable tissues and to allow stable coexistence of coral tissue and a microbiota. In addition, emerging research indicates that other components of the holobiont  and maternally provisioned antimicrobial peptides (AMP)  (Vega Thurber et al., The role of non-coral predators in the holobiont is not well-constrained although there is evidence that viruses and protozoa contribute to the mortality of coral and Bacteriophage sequences have been observed in all coral viral metagenomes (Wegley et al., Variation in physicochemical conditions can have distinct impacts on the coral holobiont community. Among healthy corals the bacterial community composition has been found to vary seasonally (Koren and Rosenberg, Scleractinian corals have survived dramatic environmental changes during their evolutionary history including changes in sea level, pH, and temperature—however current anthropogenically-induced changes are occurring at rates unprecedented in the geological record Glynn, . WhetherOur understanding of coral physiology and interaction as a holobiont is now enhanced by the expanded scientific focus on the diverse members of the coral microbiota. Access to whole genome data from the major players in a single holobiont, in particular, organisms that may be vertically transmitted from parent colony to offspring, will enable researchers to develop new, readily testable hypotheses regarding metabolic complementation, genome expansion/reduction, coevolutionary patterns, and rates of HGT across all genomes in a holobiont. Such information will yield insight into the adaptations that have enabled coral holobionts to be so successful in nutrient poor environments.In order to motivate future work to refine our mechanistic understanding of microorganisms as coral partners through evolution, development, and in ecological interactions within the holobiont we have selected eight questions to highlight based on recent literature Table . ContinuThe 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.


Neonatal chronic lung disease in the preterm infant, i.e. bronchopulmonary dysplasia (BPD) is characterized by impaired pulmonary development with its effects persisting into adulthood. Triggered in the immature lung by infectious complications, oxygen toxicity and the impact of mechanical ventilation, a sustained inflammatory response, extensive remodeling of the extracellular matrix, increased apoptosis as well as altered growth factor signaling characterize the disease. The current review focuses on selected pathophysiologic processes and their interplay in disease development. Furthermore, the potential of both, acute and long-term changes to the pulmonary scaffold and the cellular interface in concert with dysregulated growth factor signaling to affect aging and repair processes in the adult lung is discussed. With the increasing number of extremely premature preterm infants discharged from neonatal intensive care over the last decades, the development of neonatal chronic lung disease (nCLD), also known as bronchopulmonary dysplasia (BPD), accounts for a significant number of all pulmonary diseases in early infancy. Although major advances were made with respect to perinatal care, including surfactant replacement therapy, induction of lung maturation by antenatal corticosteroids, and improved invasive and non-invasive ventilation strategies, the incidence of BPD has not changed over the last decade . Due to The disease results in adverse long-term pulmonary outcome that may persist into adulthood and is associated with an increased risk for coexistent impairments in neurocognitive development underlining the importance of BPD on morbidity in this patient population .Clinically, BPD is defined by the need for supplemental oxygen and/or ventilator support for greater than 28 days (mild) or beyond 36 weeks PMA (moderate and severe) . SymptomHistopathological data from human tissue were mainly obtained from the lungs of late preterm infants in the pre-surfactant era, where the so called ‘old’ BPD was characterized by fibrotic changes and the side-by-side of over-inflation and atelectasis occurring after long-term ventilation and exposure to high oxygen concentrations. Although some of the characteristics are still seen in patients that now benefit from prenatal steroid treatment and surfactant therapy , the ‘nein utero and ex utero [Clinical studies, supported by findings from experimental models, have identified and confirmed both pre- and postnatal risk factors associated with the characteristic impairment in lung development observed in infants with BPD. Key pathophysiologic mechanisms are triggered by infections occurring both ex utero , oxygen ex utero  and the ex utero . These rex utero ,6,13.This review will focus on key pathophysiologic mechanisms leading to BPD, their interaction with one another, and the potential contribution to long-term lung pathology. Figure Intra-amniotic, congenital and nosocomial infections, oxygen toxicity, and MV contribute to the onset and perpetuation of an inflammatory response in the premature lung, characterizing the development of BPD -12. ThisBoth, the presence of infections as well as the impact of shear stress and oxygen toxicity, independently increase lung elastase and protease activity in the developing lung, due to the accentuated release from cellular and matrix sources. Interestingly, in the newborn rat, the impact of MV and hyperoxia leads to both common and distinct extracellular matrix (ECM) changes with respect to collagen deposition, increase in interstitial thickness, and elastin content . The , plays an important role in lung development by influencing the cellular composition of the lung via the regulation of endothelial and epithelial cell survival as well as influencing ECM production and remodeling . TGF-β sFrom the pseudoglandular to the canalicular stage of lung development, the appearance of primary and secondary crests marks an essential step in lung growth. Here, the epithelial-mesenchymal crosstalk is known to be critically involved in the gradual process. The resulting formation of new alveoli establishes the appropriate interface with the developing capillary bed allowing for sufficient gas-exchange . In the The (myo-) fibroblast, signaling through the platelet derived growth factor (PDGF), is a critical driver of secondary septation including its contribution to elastin production. Early abrogation of PDGF signaling therefore results in a failure of alveolarization indicated by an emphysematous lung phenotype .in vitro which might contribute to reduced and defective alveolar branching [Other growth factors orchestrating the cross-talk between the epithelial and mesenchymal cell compartment are members of the fibroblast growth factor family (FGF), essential mediators of alveolar morphogenesis and stimulators of elastin synthesis . Expressranching .On the transcriptional level, epithelial cell survival and differentiation is developmentally regulated by a multitude of different factors including FOX proteins and GATA6 . In a baThe vascular hypothesis establishes angiogenesis as a main driver of alveolarization and therefore a critical process for lung development . The pulThe VEGF pathway is of particular importance in promoting angiogenesis in the developing lung. This cell-specific endothelial growth factor is a critical agent for vascular development enabling formation and remodeling of pulmonary vessels and alveolar structures. Its expression peaks during the early stages of lung development and decreases to adult levels in the last stage of alveolarization . VEGF stWith the limited access to human tissue samples, the pathophysiologic understanding in BPD mainly relies on the findings generated by the use of these and other experimental models in lambs, baboons, rats and mice trying to mimic characteristic histopathological findings while still mirroring important clinical conditions . In the Nonetheless, although the histopathological findings in animal models resemble the changes seen in human lungs upon disease development, it has not been possible to fully imitate the pathological picture of BPD.Clinically, patients with BPD often suffer from respiratory exacerbations due to viral infections and asthma-like symptoms such as airway hyper-responsiveness and impaired exercise intolerance . In lineIncreasing evidence suggests that early pulmonary injury leading to the pathophysiologic changes characterizing BPD may contribute to premature aging of the lung . The susin vitro and in vivo studies have contributed to our understanding of molecular pathways involved in disease development and their critical interplay, leading to the identification of potential treatment targets. Most importantly, the use of experimental approaches and their  verification in clinical studies have connected critical pathways involved in the processes of angiogenesis, alveolarization and ECM remodeling in the context of BPD development [BPD is characterized by inflammation, apoptosis of various cell types and extensive ECM remodeling. Three main risk factors, i.e. pre- and postnatal infections, hyperoxia and MV lead to the complex interaction of pro- and anti-inflammatory proteins and the extensive alterations of signaling pathways associated with growth factor imbalance. Several elopment . Althougelopment . Further


We further demonstrated that these nucleases were well-tolerated and did not lead to off-target alterations of the exome in several corrected clonal cell populations. Collectively, these studies demonstrate the potential of engineered DNA-binding proteins to enable new approaches in medicine, science, and technology.The impact of reverse genetics, synthetic biology, and gene therapy has been restricted by the limitations of conventional genetic engineering technologies. To expand the capacity for genetic modification of mammalian cells, we are engineering artificial DNA-binding proteins, including zinc finger proteins, TAL effectors, and CRISPR/Cas9 to regulate and edit endogenous mammalian genes. For example, we have engineered both protein-based and RNA-guided transcriptional activators and repressors targeted to human genes relevant to medicine, science, and biotechnology. Delivery of combinations of transcription factors led to synergistic effects on gene activation and tunable expression levels. This approach recapitulates the previously intractable complexity of natural regulation of mammalian genes that is the product of cooperative actions of many transcription factors. We have also developed novel methods for controlling the activity of these proteins, such as optogenetic regulation of protein dimerization with blue light. Genome-wide analysis of the DNA-binding, gene regulation, and chromatin remodeling of these targeted epigenome modifiers has demonstrated their exceptional specificity. In other studies we have engineered synthetic nucleases to stimulate gene targeting to genomic safe harbor sites. This approach is particularly useful for generating isogenic cell lines. We showed that this method leads to a decrease in the variability of transgene expression within a clonal cell line and between multiple clones relative to conventional techniques. Finally, we have used similar methods to correct mutations causing genetic disease. We engineered synthetic nucleases targeted to the human dystrophin gene that is mutated in Duchenne muscular dystrophy patients. When we delivered these nucleases to cells from patients with this disease, the correct gene reading frame and expression of the functional dystrophin protein were restored Charles Gersbach is an inventor on patent applications and consultant in the area of genome editing.


The emergence of eusociality  in several insect lineages represents one of the most successful evolutionary adaptations in the animal kingdom in terms of species richness and global biomass. In contrast to solitary insects, eusocial insects evolved a set of unique behavioral and physiological traits such as reproductive division of labor and cooperative brood care, which likely played a major role in their ecological success. The molecular mechanisms that support the social regulation of behavior in eusocial insects, and their evolution, are mostly unknown. The recent whole-genome sequencing of several eusocial insect species set the stage for deciphering the molecular and genetic bases of eusociality, and the possible evolutionary modifications that led to it. Studies of mRNA expression patterns in the brains of diverse eusocial insect species have indicated that specific social behavioral states of individual workers and queens are often associated with particular tissue-specific transcriptional profiles. Here, we discuss recent findings that highlight the role of non-coding microRNAs (miRNAs) in modulating traits associated with reproductive and behavioral divisions of labor in eusocial insects. We provide bioinformatic and phylogenetic data, which suggest that some Hymenoptera-specific miRNA may have contributed to the evolution of traits important for the evolution of eusociality in this group. Most insect species are solitary, and behavioral interactions with conspecifics are primarily restricted to reproductive behaviors such as male–female courtship and male–male competition. This is in sharp contrast to social insects, where groups of genetically related individuals often live together in a colonial lifestyle. The size and stability of these colonies vary from a few individuals sharing a nest for a short period of time, to large perennial colonies composed of thousands of individuals . The mosThe recent sequencing of genomes from diverse social and solitary Hymenoptera clades offers a unique opportunity for identifying genome-level molecular events that may have supported the emergence of specific traits associated with the evolution of eusociality . The ability to compare whole-genome sequences, gene expression patterns, and other molecular properties of species with diverse forms of social lifestyles, has generated novel mechanistic and evolutionary insights into these complex behaviors. This approach has been used most successfully in studies of the division of labors in worker tasks  and reprDuring the early days of the molecular biology revolution, the majority of research on gene regulation was limited to transcriptional mechanisms of protein coding genes as originally defined by the “Central dogma of molecular biology” . HoweverFigure 1). miRNAs were first discovered in the nematode Caenorhabditis elegans, where the miRNA cel-lin-4 was shown to be necessary for the temporal timing of key developmental events (miRNAs are short (18–24 nucleotides) non-coding RNAs, which in animals seem to act primarily by repressing protein translation via interaction with the 3′UTR of mRNAs  throughout life. Although gamergates are morphologically worker-like, they reproduce and behave like a queen following the loss of the primary queen . This remarkable finding suggests that in honey bees, the consumption of exogenous miRNAs could play an important role in the differentiation of totipotent larvae into either sterile workers or reproductive queens. In this context, the conserved role of miR-184 in the regulation of neuroendocrine functions across different animal taxa .Three recent studies also examined the possible association between changes in brain miRNAs transcript levels and division of labor in honey bees . All thrame-miR-124 and ame-miR-275), which are differentially expressed in virgin and mated queens . Below we discuss two independent, non-mutually exclusive hypotheses for the possible involvement of miRNAs in the evolution of eusociality.Consistent with this premise, the evolution of bilateral animals from eumetazoans was associated with a great expansion in the number of miRNA genes . Other ede novo or via duplication events, facilitated the evolution of some eusocial traits in multiple independent clades that currently display eusociality. Under this hypothesis, novel miRNAs in current eusocial species act as essential nodes in genetic networks that support eusocial traits. If true, we expect that specific miRNAs would be more likely to be present in the genomes of eusocial species in comparison to related solitary species.Here, we hypothesize that, similarly to the evolution of novel transcription factors, the repeated evolution of specific new miRNAs, either Apis mellifera, the solitary wasp Nasonia vitripennis, and the fruit fly D. melanogaster (Table 2) using BLASTN . Consistent with data from other animal clades  seem to be associated with the expression of eusocial traits independent of phylogeny .As an initial test of this hypothesis, we searched for miRNA genes in the sequenced genomes of species in the Aculeata clade, which includes all living eusocial species in Hymenoptera. We first generated a list of all known annotated miRNA genes available in miRBase for the eusocial honey bee nogaster . Next, wg BLASTN . We onlyl clades , we founFigure 4A). Although the low sequence coverage for some of the analyzed ant genomes could lead to higher false-negative discovery rate, we reasoned that the likelihood that certain miRNAs will be falsely missing from all analyzed genomes is very low. Future miRNA sequencing data from many of the species studied here should further help reducing the possibility of false-negatives.To further refine our results, we subsequently extended the bioinformatic analyses to all available sequenced hymenopteran genomes, as well as several non-hymenopteran insect species, which served as outgroups , and one possibly unique to Aculeata (ame-miR-6065). The phylogenetic distribution of these five miRNAs indicated that multiple eusociality-associated miRNAs might have been gained and lost during the Hymenoptera radiation. In addition, we found that two eusociality-associated miRNA genes (miR-306 and miR-6065) were lost in the eusocial wasp Polistes dominula. Markedly, two of the eusocial-related miRNAs (miR-281 and miR-6065) were also present in the genome of the facultative eusocial bee Lasioglossum albipes. One possible explanation for this finding is that these specific miRNAs are important for traits associated with basal levels of sociality such as communal living, overlapping generations, and reproductive division of labor  belong to a single conserved miRNA-family , and possible functions in Drosophila. For example, members of the mir-279 family have been implicated in regulating neuronal development . However, two of these genes were were also present in the sawfly Athalia rosae (ame-miR-927b and ame-miR-3786), and therefore are not specific to Aculeata. Three additional genes  appear to have originated after the divergence of Vespidae and therefore did not fulfill the above criteria . Thus, our analysis revealed ame-miR-6001 as the single Aculeate-specific miRNA candidate gene that should be tested in the context of the above hypothesis. Similarly to Hypothesis 1 , the possible role of miR-6001 in the repeated evolution of eusocial traits in Aculeata is hypothetical. Directly testing the hypothesis we put forward here will require extensive molecular, biochemical, and phenotypic studies of its possible physiological and behavioral roles in eusocial traits.If true, we expect that specific miRNAs should be present in all Aculeate genomes, but absent from all other hymenopteran genomes, as eusociality has never been observed in hymenopteran species outside of the Aculeta. Our initial analysis revealed six Hymenoptera specific miRNA genes (A. mellifera. Thus, additional molecular and evolutionary analyses of non-hymenopteran eusocial insects as well as other eusocial and solitary clades in the Hymenoptera are required in order to better understand miRNA functions in the context of eusociality. Furthermore, to establish causation between the action of specific miRNAs and eusocial traits, new in vivo genetic and molecular techniques to manipulate social insects are required. Recent advances in molecular genetics of social insects (To date, the majority of data about the function of miRNAs in social insects come from studies of the European honey bee,  insects , and the insects , suggest insects .ES collected and analyzed genomic data. ES, GB, and YB-S gathered and synthesized relevant literature. ES, GB, and YB-S wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Numerous studies have documented increases in matrix metalloproteinases (MMPs), specifically MMP-9 levels following stroke, with such perturbations associated with disruption of the blood brain barrier (BBB), increased risk of hemorrhagic complications, and worsened outcome. Despite this, controversy remains as to which cells release MMP-9 at the normal and pathological BBB, with even less clarity in the context of stroke. This may be further complicated by the influence of tissue plasminogen activator (tPA) treatment. The aim of the present review is to examine the relationship between neutrophils, MMP-9 and tPA following ischemic stroke to elucidate which cells are responsible for the increases in MMP-9 and resultant barrier changes and hemorrhage observed following stroke. Over the last decade the matrix metalloproteinases (MMPs) have been widely investigated for their role in disruption of the blood-brain barrier (BBB), particularly the extracellular matrix (ECM), following stroke  1-knockout mice, a role for oxidative stress in the mediation of BBB disruption has been revealed  score, which is used to objectively quantify the impairment cause be stroke, has been observed , with MMP-9 levels correlating with larger infarct volume, increasing severity of stroke and poor functional outcome  and that these cells use MMP-9 to transmigrate into ischemic tissue, contribute to ischemic injury in the form of ECM degradation, BBB permeability changes, vasogenic edema, and HT, whilst also stimulating the release of MMP-9 from resident brain cells such as neurons and microglia. Clearly, the interaction of action of leukocytes at the BBB following stroke requires further investigation to elucidate the contribution of neutrophil-derived to ECM degradation, BBB disruption and subsequent vasogenic edema, hemorrhage and parenchymal injury.Many studies have focused on the protection of neurons as a therapeutic target. However, given that such a small percentage of all cells in the brain are neurons it seems more appropriate to consider protection across many cell types. Accordingly, investigation of the neurovascular unit and the complex interplay of endothelial cells, pericytes, astrocytes, and infiltrating leukocytes is likely to yield more appropriate therapeutic targets. Information regarding the relationship between MMP-9 and neutrophils may facilitate understanding of the mechanisms involved in BBB breakdown following stroke and ultimately guide therapeutic intervention.All authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Dear Editor,A 62-year-old male presented with a several-month history of headache and alteration inhis visual field. The diagnostic evaluation revealed a suprasellar mass that was causinghydrocephalus by extrinsic compression. We opted for ventricular shunt placement andsubsequent surgical excision of the mass. The surgical site was accessed through leftfrontal craniotomy. Histological analysis of the resected mass revealed that it was acraniopharyngioma.In the postoperative period, the patient evolved to significant worsening of theheadache, and a magnetic resonance imaging scan was requested in order to evaluate thecondition. The magnetic resonance imaging showed cerebellar hemorrhage affecting thevermis and the left cerebellar hemisphere , with st. The reported incidence of remote cerebellar hemorrhageafter supratentorial interventions ranges from 0.08% to 0.6%. However, it has been reported to occur after variousother surgical procedures involving the cranium or spinal cord-7.Remote cerebellar hemorrhage has been defined as bleeding within the cerebellarparenchyma, a rare complication that can occur after neurosurgical intervention. Theentity was first described in the 1970s, by Yasargil et al.. However, there isanother hypothesis that might explain the two findings in the case reported here. Thathypothesis is based on the supposition that opening the cisterns or the ventricularsystem promotes intracranial hypotension, triggering the process that culminates in thedistension and rupture of cerebellar veins, resulting in cerebellar hemorrhage.Several hypotheses have been suggested to explain the appearance of bleeding in thecerebellum away from the primary  surgical site. One suchhypothesis is that resection of a supratentorial lesion creates a pressure gradient,resulting in suction on the cerebellar veins, particularly in the upper portion of thevermis-7,9. In imagingexaminations, remote cerebellar hemorrhage has a characteristic presentation, with atendency for the blood to be distributed among the cerebellar folia with a curvilinearconfiguration. This aspect results in the pattern known as the zebra sign.Various neurosurgical procedures have been associated with the occurrence of remotecerebellar hemorrhage, including the clipping of aneurysms (ruptured or otherwise),tumor resection, drainage of parenchymal or extra-axial hematomas, and spinalsurgery, findings include dural thickening and diffuse duralenhancement; engorgement and dilatation of venous structures; subdural fluidcollections; downward displacement of the midbrain; and herniation of the cerebellartonsils.The symptoms of intracranial hypotension syndrome include headache that is orthostatic inpresentation, tending to improve in the recumbent position. In imaging studies of thebrains of patients with intracranial hypotensionThe case presented here demonstrates a chain of events that could have collectivelyresulted in the two central nervous system complications observed. The supratentorialsurgical manipulation and the placement of the ventricular shunt could have causedintracranial hypotension, resulting in the traction, distension, and consequent ruptureof cerebellar veins, as well as hemorrhage in the cerebellar parenchyma.Radiologist knowledge of these entities is relevant, because their proper, earlycharacterization can promote interventions aimed at their correction and at alleviatingthe associated symptoms.


Finally, when considering orchards grown in the same pedoclimatic conditions and of the same age, instead of the usual organic vs. conventional comparison, the effect of the management system on the apple fruit quality  was very limited to non-significant. The main factors of variation were the cultivar and the year of cropping rather than the cropping system. More generally, as each management system  encompasses a great variability of practices, this highlights the importance of accurately documenting orchard practices and design beside the generic type of management in such studies.Many biotic and abiotic parameters affect the metabolites involved in the organoleptic and health value of fruits. It is therefore important to understand how the growers' decisions for cultivar and orchard management can affect the fruit composition. Practices, cultivars and/or year all might participate to determine fruit composition. To hierarchize these factors, fruit weight, dry matter, soluble solids contents, titratable acidity, individual sugars and organics acids, and phenolics were measured in three apple cultivars  managed under organic, low-input and conventional management. Apples were harvested at commercial maturity in the orchards of the cropping system experiment BioREco at INRA Gotheron  over the course of three years . The main factors affecting primary and secondary metabolites, in both apple skin and flesh, were by far the cultivar and the yearly conditions, while the management system had a very limited effect. When considering the three cultivars and the year 2011 to investigate the effect of the management system  For apple , the effect of crop load on the fruit phenolic content has been demonstrated in some studies   per se apples by ; Table 1pples by . FurtherFinally, three management systems  instead of generally two systems were studied in a well-documented system experiment, displaying a range of cultural practices rather than chemical/non chemical archetypes, and eliminating the bias induced by uncontrolled environmental external factors. Under these controlled growing conditions, cultivar and year-to-year effects were by far the most effective in determining the quantitative primary and secondary metabolites composition of apples, while the management system  had little and no consistent influence on fruit composition. More broadly, all compounds of the apple contribute to its health value and its taste, and other quality criteria  can also influence the choices of consumers in buying ‘eco-friendly’ or organic apples  beside mS1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(DOCX)Click here for additional data file.


In 1943, Austin Moore developed the first endoprosthesis fashioned from Vitallium, providing the first alternative to traditional amputation as primary treatment of bone tumors. The success of the Vitallium endoprosthesis has since then led to the development of new materials and designs further advancing limb salvage and reconstructive surgery. Combined with the advent of chemotherapy use and imaging advances, conservative treatment of musculoskeletal tumors has expanded greatly. As the implantable options increased with the development of the Lewis expandable adjustable prosthesis and the noninvasive Phenix Growing prosthesis, receiving the diagnosis of a bone tumor no longer equates to automatic limb loss. Our review details the history and development of endoprostheses throughout orthopedic oncology in the treatment of musculoskeletal tumors. During the early 1900s, multiple unsuccessful attempts were made to incorporate the use of metal equipment into the body. These failures were primarily attributed to the researchers’ inabilities to find a suitable metal that could withstand corrosion from bodily fluid without causing an unfavorable reaction in soft tissue  began toIn 1943, Vitallium was used for the first metallic endoprosthesis in orthopedic oncology and, possibly, in the entire field of orthopedics. Using the alloy, Austin Moore  created It was not until the 1970s that endoprosthetic implants began to emerge as a valuable treatment option in orthopedic oncology. This was largely due to the various advances in medical technology, specifically in the fields of radiology, pathology, chemotherapy, and radiotherapy, that allowed staging and assessing tumoral local extension and led to an increase in survival in musculoskeletal tumor patients.With the development of antineoplastic drugs, orthopedic physicians began to use chemotherapy for primary musculoskeletal tumors starting in the early 1970s. The use of chemotherapy in orthopedic oncology began as adjuvant therapy following removal of the primary tumor. This followed the hypothesis that chemotherapy would destroy any microscopic residual disease missed during surgery. The results were promising. Studies showed increased survival rates in patients who received adjuvant chemotherapy following bone resection , 14. PhyImaging studies have always played a vital role in diagnosing musculoskeletal tumors. Although plain radiograph is an essential component in early diagnosis of bone tumors, it was not until the early 1970s, with the developments of computed tomography (CT) , 19, andTwo important advancements in materials is the use of titanium alloys, specifically Ti-6Al-4, and silver coating. Pure titanium has a high affinity for oxygen, but Ti-6Al-4V undergoes a process called self-passivation to create a protective film that can resist corrosion. This quality has created many uses for this alloy in orthopedic implants . AdditioThe use of endoprostheses in limb-salvaging procedures started to gain popularity in the 1970s as chemotherapy improved and total joint replacements were increasingly used. To further improve the development of custom endoprostheses in musculoskeletal oncologic limb-salvaging procedures, researchers began thorough analyses of failure modes for these prostheses, thus encouraging continuous redesign and improvement of endoprostheses.One of the common modes of failure in early custom endoprostheses was failure of the stem component. Suboptimal fabrication during the manufacturing process and the use of alloys that could not withstand high stress were two of the early causes of stem failure , 31. ImpAnother challenge in implant use is attaching the tendon directly to a metallic implant. A tendon directly attached to a metallic implant, with no scaffold, is held together by fibrous ingrowth, which is a weak interface. This type of attachment has <20 % of the strength of a normal tendon insertion . NormallDue to advances in the field of chemotherapy, musculoskeletal oncologists began to see an increase in the number of available candidates for limb-salvaging procedures. With this increase came the birth of current modular endoprosthetic systems that seek to create a single system, thereby simplifying bone and joint replacement procedures . Prior tTwo of the most crucial characteristics essential in modular systems are reliability and simplicity. To provide a straightforward and dependable system, developers of modular endoprostheses began to use the Morse taper between joining components . This taAlthough the original groundwork on modular systems remains, biomechanical engineers continue to develop new methods to advance the system with more reliable components and fewer complications.As limb-salvage procedures became a popular method for treating malignant bone disease in adults, physicians found it difficult to treat bone tumors using standard endoprostheses in growing children. With many bone tumors occurring around the growth plates of immature patients, the use of standard endoprostheses caused limb discrepancies when these patients reached adulthood. To address this issue, researchers began to explore the idea of expandable prostheses.The first expandable prosthesis, the Lewis expandable, adjustable prosthesis, consisted of a screw extension mechanism that was turned by making a small incision and using a chuck key to turn the screw , 47. ThiOne of the earliest internally expandable prostheses was the Phenix growing prosthesis  . This syThe evolution of machining, adjuvant therapies, and imaging techniques has contributed greatly to the advances in limb-salvage surgery. As we move further into the twenty-first century, the possibilities for advancements in limb-salvage reconstruction seem bright. Improved techniques of stem fixation and soft tissue attachment will further increase implant survivorship, improve quality of life, and continue to improve the prognosis and hope for those diagnosed with a musculoskeletal tumor .


Plasmodium vivax, persistence of Plasmodium falciparum and clinically suspected malaria, continuous import of malaria cases, underreporting of mixed infections , and climate change. In this study, we report, in addition, on altitudinal shifts of malaria vectors in Nepal.Malaria elimination is a goal of many endemic countries with the ultimate goal of global malaria eradication. Based on the achievements the country has made over the last decade, Nepal has been preparing for a malaria pre-elimination phase since 2011 with the ambitious vision of a malaria-free Nepal by 2026. However, a number of challenges have been identified in recent studies including high rates of relapse/re-infection with To obtain data on the present altitudinal distribution of malaria vectors, entomological surveys were carried out in eastern Nepal between 60 and 2,500 m above sea level. The surveys were conducted from September to October 2012 and April to May 2013 using aspirator and CDC light trap collection of adult mosquitoes in houses, cattle sheds and natural outdoor shelters. In addition, Anopheles larvae were collected from potential breeding sites, and geographical position and environmental data were recorded for all collection sites. The collected mosquitoes were morphologically identified in the field and then deep frozen for future molecular analyses.Anopheles fluviatilis, Anopheles annularis and Anopheles maculatus complex members) were recorded up to 1,820 m above sea level. This is the highest record of malaria vectors in eastern Nepal so far. Moreover, larvae of Anopheles spp. were recorded up to 2,310 m above sea level; their species-level identification is in progress. The densities of these vectors were significantly higher in the post-monsoon season (end of September to October) compared to the pre-monsoon (April-May) season.Based on the findings of the morphological identification, known malaria vectors in Nepal (Taking into account the evidence from climate change studies and previous entomological surveys in Nepal, it can be concluded that malaria vectors have already shifted to higher altitudes of Nepal due to climatic and other environmental changes, indicating a serious health risk for mountain people and tourists. The presence of potential malaria vectors in the highland areas, lack of vector-control interventions, higher rates of warming in the mountains compared to the lowlands and the continuous introduction of malaria cases from abroad may pose challenges for achieving the goal of malaria elimination in Nepal by 2026.


Together with new genetic and molecular techniques to optically trigger and monitor neural activity, a new generation of optical neural interfaces is being developed, mainly thanks to the exploitation of both bottom-up and top-down nanofabrication approaches. This review highlights the role of nanotechnologies for optical neural interfaces, with particular emphasis on new devices and methodologies for optogenetic control of neural activity and unconventional methods for detection and triggering of action potentials using optically-active colloidal nanoparticles.In last decade, the possibility to optically interface with the mammalian brain  Understanding, monitoring and manipulating neural activity with high spatial and temporal resolution New strategies and technologies to systematically monitor thousands of functional links that each neuron forms with other neurons are being developed. Nanoscience and nanotechnology can play a key role in developing new ideas and experimental approaches to create detailed maps of the human and mammalian brain  implanted directly in the target area. A straightforward and multi-purpose implementation of that was presented in 2013 in Kim et al. . The so-2; McAlinden et al., 2 . These probes respond to a variation of neural activity by changing their fluorescence intensity and are widely adopted in microscopy techniques in vivo to monitor electrical activity of cortical neural circuits , the authors of João et al.  suggesteAnother promising, but still only theoretical, application of colloidal nanoparticles is the readout of neural activity by using semiconductor nanocrystals (NCs). NCs are nanometer-sized semiconductors that present quantized levels in both valence and conduction band, rather than the quasi-continuum of states allowed in bulk semiconductors. A photon absorbed by a NC generates a bound electron-hole (e-h) pair, which can recombine following radiative  or non-radiative channels. For NCs made of II-VI semiconductors, emitted light is in the visible spectral range and, at physiological conditions, it has a well-defined single-peak emission spectrum with a ~30 nm-large Gaussian distribution around the peak wavelength. When specific types of nanoparticle are inserted into an electric field, however, the e-h pairs polarize along the direction of the applied field, and both the emission wavelength and the lifetime of the excited state change accordingly (Galland et al., Nanotechnology, exploiting surface and bulk nano and micromachining and self-organized nanochemistry, has dramatically improved the capability to produce devices that deliver, shape, and collect light with high spatial and temporal resolution. In the past decade all those technologies have been applied to ICT applications and for biosensing assays, with many materials and methods derived from the telecom and nanoelectronic industry.in vivo biophotonic methods, such as optogenetics, and new nanobiosensing approaches and clinical applications, there has been an increasing need of optical tools and convergence of integrated nanophotonic technologies toward lifescience. However, biological tissue are dispersive media and they do not allow a straightforward propagation and control of light inside organs and, specifically, in the brain.Since the advent of different In this paper we have shown how the scientific community is facing this challenge. So far light delivery to deep brain regions cannot be achieved without being extremely close or in contact with the region of interest, and the proposed approaches always rely on waveguides, optical fibers or wired LEDs mounted on rigid or flexible supports. At the current stage, optogenetics allow to target genetically defined classes of neurons, while the recently developed multipoint devices add the feature of controlling closely spaced neurons belonging to the same class also in deep brain regions, complementing the high spatial resolution obtained with microscopy in the firsts cortical layers. Moreover, the integration of multipoint extracellular recording of neural activity is allowing for unprecedented spatial resolved stimulation/readout in free-moving animals (Grosenick et al., All authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.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.


ERAS approach (Enhanced Recovery After Surgery) is a multimodal, perioperative pathway designed to achieve early recovery after surgery. ERAS has shown documented efficacy in elective surgery, and the concept of “multimodal” and “multidisciplinary” approach seems still to be of higher importance than each single item within ERAS protocols. New perspectives include the use of ERAS in emergency surgery, where efficacy and safety on outcome have been documented, and flexibility of traditional items may add benefits for traditionally high-risk patients. Obstetric surgery, as well, may open wide horizons for future research, since extremely poor data are currently available, and ERAS benefits may translate even on the baby. Finally, the concept of “outcome” may be extended when considering the specific setting of cancer surgery, in which variables like cancer recurrence, early access to adjuvant therapies, and, finally, long-term survival are as important as the reduced perioperative complications. In this perspective, different items within ERAS protocols should be reinterpreted and eventually integrated towards “protective” techniques, to develop cancer-specific ERAS approaches keeping pace with the specific aims of oncologic surgery. ERAS approach (Enhanced Recovery After Surgery) is a multimodal, perioperative care pathway designed to achieve early recovery after surgery . The conSurgical stress response (SSR) is represented by hormonal and metabolic changes resulting in immune and endocrine responses, proportionally with the extent of surgical tissue lesion. Local changes impact the inflammatory reaction in the whole body, leading to widespread effects on organ functions and on the development of complications. SSR plays a major role in respiratory, cardiac, and thromboembolic complications, resulting in a general catabolic state and in strong immune-depressant effects [ effects . Prevent effects .Because of evidence on the positive effects of this approach, ERAS has now extended to several types of surgeries. In this narrative review, we want to present the newest field of applications of this philosophy. Despite the fact that several possible topics may be discussed and much progress is still possible within ERAS, we decided to focus only on the most innovative ones (even when few studies exist), that is, those that are supposed to have the higher development in the future years and produce some changes in the philosophy of ERAS towards a patient-specific (and not only surgery-specific) approach. The latest applications in emergency and obstetric surgery are presented, as well as implications in the specific subset of cancer patients, in which other “outcomes” may add to the traditional ones and may promote the development of cancer-specific ERAS protocols in the future.The benefit and safety of ERAS protocol have been validated in different studies, and the validity of ERAS approach is no more questioned. A specific society entitled to the ERAS approach exists, including the most important experts in the field and producing procedure specific guidelines easily accessible on the web and able to guide any perioperative physician who wants to develop an ERAS approach within his/her institution . GuideliDespite the amount of strategies included in ERAS protocols, introduction in clinical practice resulted to be feasible in different realities, and a recent study reported that the rate of compliance to each ERAS item within a protocol correlates with the length of the hospital stay  .Authors who have historically conceived and validated ERAS protocols have already suggested key points for the development of successful ERAS programs, which are based on knowledge of institutional reality, on continuous communication within the multidisciplinary team, and on application of common protocols for patients care .In contrast to what happened for elective surgery, there has been little improvement in outcomes or morbidity for patients undergoing emergency procedures; emergency surgical operations carry a mortality rate at least ten times higher than many similar elective procedures  and ERASUnfortunately, few studies investigated the use of ERAS model in emergency surgery. The setting of emergency obviously limits compliance with the entire protocol, since this was initially designed for elective situations. Moreover, emergency patients might be considered at higher risk . Howevercolonic resection performed during an unplanned hospital admission”; they have demonstrated that many of the ERAS items can be applied for such situation, and no significant adverse effect was observed despite higher American Society of Anesthesiologists (ASA) status, Portsmouth-POSSUM (P-POSSUM) scores, and more stressful procedures in urgent patients [Despite this, Roulin et al. have studied the application of ERAS in urgent colectomy, defined as “patients .Another study evaluated the safety and efficacy of ERAS approach on patients undergoing laparoscopic repair for perforated peptic ulcer. ERAS resulted in improved surgical outcome by reducing the occurrence of postoperative ileus; the authors suggested this result to be associated with the lack of routine nasogastric decompression and with early oral feeding. The use of nonsteroidal anti-inflammatory drugs for postoperative pain seemed to provide beneficial effects, as well .In conclusion, although studies on ERAS in emergency surgery are rare, early results account for the flexibility, effectiveness, and safety of this approach. Further studies are needed to confirm such evidence in wider range of surgical procedures, in the perspective of developing patient-related, surgery-specific ERAS approaches for emergency procedures (as previously done for elective pathways).Cesarean section (CS) is an interesting field of research for ERAS, whereas the return to normal function is more precious than ever in this type of patients. The need to standardize ERAS protocols for parturient undergoing CS is perceived by the research and clinical community. First experiences in this field have been starting in the last years. A recent study has shown that the application of the ERAS protocol resulted in a substantial increase in the proportion of women leaving hospital the day after elective CS, without increasing the rates of readmission . OtherwiEven with the best surgical technique, surgery for cancer is associated with release of tumor cells .Several perioperative factors contribute to the development of remote metastasis , but sinAdequate pain control for the entire perioperative period is mandatory in ERAS protocols and is even more required in the perspective of oncological outcome. Indeed, pain activates the hypothalamic-pituitary axis and the sympathetic system, giving immunosuppression . AuthorsThus, pain control is the main target, but the choice in the analgesic strategy is also crucial, since the benefit of pain control may be overcome by tumor promoting effect of some drugs. The opioid mu-receptor agonists commonly used for anesthesia and perioperative analgesia are immune-suppressive: morphine and fentanyl suppress the innate and acquired immunity , and opiNevertheless, in vitro studies gave conflicting results on the topic. Long-term morphine treatment reduces leukocyte migration and recruitment to reduce angiogenesis and tumor growth in long-term cultures of mouse Lewis lung carcinoma cells . In anotAlthough there is no definitive evidence that opioids worsen outcomes in cancer patients , opioid-Despite this evidence, the role of epidural anesthesia (EA) has been questioned  because Otherwise, no studies have investigated any protective effect on SSR using such techniques instead of epidural. EA has the advantage of reducing the sympathetic stimulation and opioid consumption; EA reduces cortisol  and epinRegarding the potentially limited effect on SSR and considering the concomitant use of rescue opiates in the postoperative period, the efficacy of such new techniques may be questioned in the specific setting of cancer patients, where main outcome variables include cancer spread and recurrence, access to adjuvant therapies, and, finally, long-term survival. In this perspective, specific cancer-oriented, epidural-based ERAS approaches may be adopted, differently from noncancer patients .In this perspective, some authors have already suggested a protective role of regional anesthesia on cancer recurrence . The use Persistent Postsurgical Pain (PPSP), which may condition long-term opioid therapies and more difficult access to life-saving therapies (radiotherapy and chemotherapy).Finally, a limiting factor in cancer outcome isAgain, key elements of ERAS protocols are part of the so-called “protective anesthesia,” an innovative anesthetic regimen  that relPrevention and treatment of postoperative nausea and vomiting (PONV) are another important component of ERAS protocols, as PONV may result in retardation of enteral feeding. Early nutrition is a major component of ERAS protocol and adds benefits in maintaining metabolic homeostasis : it has Dexamethasone is one of the most widely prescribed drugs against PONV, also considering its low cost; there is plenty of evidence on the efficacy of dexamethasone, which is supposed to act through central and peripheral mechanisms: inhibition of prostaglandins, production of anti-inflammatory factors, and decreased production of endogenous opioids . The effOtherwise, concerns still exist on wound healing and on the immune-depressant effect of dexamethasone in the context of cancer surgery; a recent study recommends prudence in pharmacological decisions, despite the fact that any association was demonstrated between the use of dexamethasone and the increase in surgical site infections .Few studies have addressed the topic of cancer outcome and use of perioperative dexamethasone, and results are conflicting. In patients affected by rectal cancer, a better three-year survival was noticed in the untreated group compared with patients given dexamethasone perioperatively . In anotA key element in ERAS approaches is the use of minimally invasive surgery. Video-Laparoscopic Surgery (VLS) has been shown to determine lower stress response and fewer complications and to reduce hospital stay and mortality  and is cA meta-analysis of the six studies concluded that VLS + ERAS is a more reliable treatment for colorectal malignancy, compared with laparoscopic conventional care treatment. VLS + ERAS reduces complications while carrying similar risks of anastomotic leakage, wound infection, obstruction, and readmission . Other sJoining ERAS and VLS seems even more crucial in cancer patients. As mentioned above, VLS results in a lower stress response , reIn conclusion, the ERAS approach in abdominal surgery has shown documented efficacy in elective surgery, and the concept of “multimodal” and “multidisciplinary” approach seems still to be of higher importance than the single items themselves. New perspectives include the use of ERAS in emergency surgery, where the first studies were able to demonstrate flexibility and to bring benefits in outcome, and obstetric surgery, where a whole world of possibilities to be explored exists, since extremely poor data are currently available. Finally, it would be important to extend the ERAS outcomes beyond the perioperative period in oncological surgery, analyzing the different components of the protocol to develop cancer-specific ERAS approaches.


Emerging evidence indicates that the signaling activity of these receptors, as for other GPCR, is influenced by allosteric modulators, which interact with the receptor in a binding site distinct from the binding site of the agonist and modulate the receptor signaling activity in response to the orthosteric ligand. Allosteric modulators have a number of potential advantages over orthosteric agonists/antagonists as therapeutic agents and offer unprecedented opportunities to identify extremely selective drug leads. Here, we resume evidence of allosterism in the context of chemoattractant receptors, discussing in particular its functional impact on functional selectivity and probe/concentration dependence of orthosteric ligands activities.Chemoattractants control selective leukocyte homing  Physiologically, when the extracellular signal interacts with the so-called “orthosteric binding site” of a GPCR, a conformational change occurs that conveys the signal through the plasma membrane, thus triggering intracellular signaling cascades via heterotrimeric G proteins and other signal transducers (G protein-coupled receptors (GPCRs) are the largest family of cell-surface receptors encoded by the human genome and are involved in most pathophysiological aspects , 2. GPCRnsducers . Becausensducers , 4, 5, tvia chemoattractant/receptor interactions is pivotal for the organization of the immune system and for protection against infectious diseases. Chemoattractants are also key players in the development and exacerbation of immunomediated pathological conditions, such as allergic responses, autoimmune diseases, and other acute and chronic inflammatory disorders, and their fine regulation plays a crucial role for the development of an appropriate immune response  profiles to be in parallel evaluated in relevant in vivo models.While offering unprecedented opportunities for the design of highly selective pharmacological tools, the allosterism concept implies a profound revision of the entire drug discovery process having impact on the design and characterization of novel lead candidates targeting the GPCR family. From the structural point of view, biased allosteric modulation, probe dependence, and ligand cooperativity require the ability to model multiple conformational states in the presence of different ligands that still represents a major hurdle for the rational design of drugs. Several independent studies have shown how subtle structural and electronic modifications in a class of allosteric GPCRs modulators may result in dramatic changes of the biological activity, thus limiting the possibilities to conduct large and efficient lead optimization programs –41. In tvia conformational coupling between the orthosteric and allosteric binding sites, or modulate efficacy by altering the functional response of the receptor to orthosteric ligand binding, thus resulting in positive, negative, or neutral effects on receptor activation .The most relevant efforts to develop chemokine receptor inhibitors have been focused on drugs blocking HIV infection  that is able to block in a probe-independent manner the receptor activation process. Interestingly, mechanistic studies support the rationally derived binding mode hypothesis, thus confirming that the allosteric site is conserved among the two receptor subtypes. The binding mode of the molecule with CXCR1 and CXCR2 is in keeping with the concept that allosteric sites in the TM domains of GPCRs could represent valuable targets for selective allosteric inhibitors able to finely modulate receptor signaling, and suggests their therapeutic investigation in inflammatory disorders. Pharmacological studies were conducted to investigate the potency of CXCR1/CXCR2 inhibition for the prevention of inflammation- and autoimmunity-mediated damage of pancreatic islets. Blockade of CXCR1/CXCR2 was associated with inhibition of insulitis and modification of leukocytes distribution in blood, spleen, bone marrow, and lymph nodes, and was effective in preventing diabetes in an inflammation-mediated model based on multiple low dose injections of streptozotocin and in preventing diabetes in NOD mice (Reparixin (formerly known as repertaxin) and ladarixin represent the first examples of non-competitive allosteric modulators of chemokine receptors, showing the ability to behave as NAMs of CXCR1/CXCR2 without affecting the cognate ligand binding affinity. Interleukin-8  and related ELRnditions –49. The nditions –53. Modunditions . Reparixnditions . The effnditions , 56 and nditions , thus panditions . LadarixNOD mice . Pharmachttps://cortellis.thomsonreuterslifesciences.com).The complement has long been recognized as a potentially useful therapeutic target, and a number of strategic approaches and therapeutic agents have been developed during the last years  and chemokine GPCRs  entered clinical trials, and in some cases, successfully reached the market . The pharmacological implications potentially deriving from the availability of multiplicity of molecules that, acting through a single receptor, differentially regulate its signaling activity are still far from being fully exploited and will offer opportunities for the development of new drugs targeting chemoattractant receptors in the near future.MA, MC, and ML contributed equally to writing and critically revised 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.


The control of diseases shared with wildlife requires the development of strategies that will reduce pathogen transmission between wildlife and both domestic animals and human beings. This review describes and criticizes the options currently applied and attempts to forecast wildlife disease control in the coming decades. Establishing a proper surveillance and monitoring scheme (disease and population wise) is the absolute priority before even making the decision as to whether or not to intervene. Disease control can be achieved by different means, including: (1) preventive actions, (2) arthropod vector control, (3) host population control through random or selective culling, habitat management or reproductive control, and (4) vaccination. The alternative options of zoning or no-action should also be considered, particularly in view of a cost/benefit assessment. Ideally, tools from several fields should be combined in an integrated control strategy. The success of disease control in wildlife depends on many factors, including disease ecology, natural history, and the characteristics of the pathogen, the availability of suitable diagnostic tools, the characteristics of the domestic and wildlife host(s) and vectors, the geographical spread of the problem, the scale of the control effort and stakeholders’ attitudes. Diseases shared with wildlife are multi-host infections with an impact on human health, economy, and wildlife management or conservation were wildlife itself plays a significant role on infection maintenance. Shared diseases represent a significant burden that affects public health, global economies, and the conservation of biodiversity –3. It haDisease emergence in wildlife , and difficulties in the eradication of endemic shared diseases such as classical swine fever (CSF) and tuberculosis (TB), have, over the last few decades, prompted a growing interest in disease control in wildlife reservoirs , 11–14. M. bovis but feral hogs in Australia are considered spill over hosts .Ixodes tick control  has been shown to reduce Ixodes scapularis populations by up to 94%, and acaricide application to deer decreased nymphal I. scapularis populations by up to 83%. However, the effect of these strategies on the incidence of Lyme disease in human beings remains unknown , reduced the number of engorging female ticks, their weight and their reproductive capacity, thus resulting in the reduction of tick infestations and in the prevalence of some tick-borne pathogens (Cervus elaphus) and white-tailed deer with an overall vaccine efficacy of approximately 80% for the control of R. microplus infestations in white-tailed deer  numbers increased, as well as grazing pressure and habitat damage, when carnivore culling was conducted in Canada  culling, similar to test and cull schemes applied to domestic animals. Such actions can be very expensive, and their feasibility depends on access to the animals, the availability of convenient, sensitive and specific tests, the prevalence of the infection, and the spatial distribution of the target population Table .Random and selective culling strategies are more likely to succeed in isolated populations than on large geographical scales, and the results will probably consist of a certain reduction of disease prevalence in the wildlife host and in the domestic host targeted, rather than in the total eradication of the infectious agent . The sucIn this context, wildlife vaccination to reduce infection prevalence emerges as a valuable alternative or complementary tool in disease control. Disease control through the vaccination of wildlife reservoirs may potentially have advantages over other approaches. As opposed to culling, vaccination may be more acceptable to the general public  since itThe best vaccination method for wildlife populations spread over a wide geographical area is oral vaccination using baits. The oral vaccination of wildlife is the only disease management tool with proven efficacy on large spatial scales. This has been shown most clearly in the case of fox rabies control in Western Europe . Table 2However, wildlife disease control can eventually interfere with wildlife ecology. In diseases where vaccination significantly reduces target host mortality, effects on sympatric prey, predators or competitors may occur , 114 whiEchinococcus multilocularis in foxes is a prominent example. The adult fox tapeworm is sub-microscopic and infects foxes and, less efficiently, dogs. The larval form infects several wild rodents. In villages and small towns in central Europe, foxes are responsible for environmental E. multilocularis egg contamination in the vicinity of human beings, leading to infection risk if human beings accidentally ingest viable eggs , suggesting that the inherent cost of declaring a brucellosis-infected zone would be far lower than current management to avoid Brucella abortus spillback to cattle  can and have been implemented by countries or states in order to define sub-populations of varying health statuses for disease control. This could become one of the best solutions for disease control at the wildlife-livestock interface in the future . Also, E. multilocularis, and tick-borne diseases. In the future, it is likely that this list will become longer as new scenarios and disease control needs emerge. Future wildlife disease control efforts will probably rely on a better understanding and modeling of wildlife–pathogen interactions (M. bovis vaccines (Most current monitoring and disease control efforts in wildlife are directed toward only a few relevant diseases, including rabies, ASF, CSF, FMD, CWD, brucellosis, TB, ractions , thus imvaccines , 132 andvaccines , 133 supFinally, from a global point of view, disease control schemes should be aimed at the accomplishment of a balance. Most of the above-mentioned examples of shared wildlife diseases are resultant of unbalanced situations in which, for instance, wildlife has increased in numbers, often as the result of anthropogenic factors [such as rural abandonment or land use changes ]. Any prThe 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.


While many major strides have been made in the last three decades concerning the synthesis, characterisation and properties of individual nanoobjects, their technological uptake necessarily requires integration into devices. To achieve this, detailed characterization, design and control of the interface between nanoobjects and their environment is essential. Nanomaterials differ significantly from their bulk counterparts and one important aspect of this is their high surface to volume ratios. For this reason interface properties typically dominate the behavior of nanomaterials. As a consequence, one immediately faces the problem of designing and controlling surfaces and, in case of embedded or layered systems, interfaces on atomic scales.Engineering of electrical contacts is required for nanostructure integration. The performance of electronic devices based on nanostructures is affected by a potential barrier at the metal–nanostructure contact, thus investigation of the metal–nanostructure interaction aiming at understanding the physics and chemistry of this interface is critical for the design of optimal devices –3. PotenNanostructured materials show a great application potential in the areas of nanoelectronics, catalysis, and light harvesting/energy storage, an excellent example being the capability of titanate nanoribbons to simultaneously harvest and store energy in photobatteries . In suchAddressing design, control and characterization of nanoscale interfaces and the development of new tools to facilitate this is a vast area of research, requiring multidisciplinary collaboration between chemists, physicists, biologists and engineers across a wide range of disciplines. It is clearly a very broad topic for a single Thematic Series, and for this reason the current volume has been jointly edited by Carla Bittencourt, Chris Ewels and Arkady Krasheninnikov, with the reviews covering key areas of advanced synthesis methods, controlled functionalization and post-processing routes, developments in nanoscale characterization tools as well as theoretical modeling of structure and dynamics, and electronic properties. Finally some reviews cover key examples of successful nanomaterial integration.2 deposition on carbon nanotubes [Recent drives to improve and refine nanomaterial synthesis routes have adopted different approaches. The increasingly important role played by organic chemists in the field of nanocarbon design has led to new families of atomically designed materials such as cycloparaphenylenes, the ultimate “short nanotubes” . At sliganotubes . Post-granotubes , the filanotubes , or the anotubes .Graphene and related materials such as nanotubes, polycrystalline nanoparticles  and carbRecent developments in X-rays collimation optics triggered the interest in synchrotron radiation-based techniques for the study of nanostructures. In this context, X-ray microscopy has shown to be a powerful tool for chemical analysis of radiation-sensitive nanomaterial . CombiniAnother important characterization tool for nanomaterials has emerged from the introduction of aberration correctors in transmission electron microscopes (TEM), which brings both increased spatial resolution and energy resolution allowing imaging at low voltage, crucial for nanomaterials which are sensitive to electron irradiation. Currently, the atomic structure of different nanomaterials can be revealed with a resolution of better than 0.1 nm, in addition to elemental analysis .The analysis of experimental results can significantly profit from the comparison of the images to the results of first-principles calculations. Similarly, for the precise interpretation of experimental scanning tunneling microscopy (STM) and atomic force microscopy (AFM) images, the microscopy knowledge about the local electronic structure of the system is extremely important. Full understanding of the electronic structure and properties of a large number of solids can be credited to first principles simulations, and more specifically density-functional theory (DFT) approaches. In addition, using DFT-based molecular dynamics, the manipulation of nanostructures by SPM tools and the changes made to the system by the characterization tools, e.g., the production of defects under electron irradiation and their evolution over time, can be simulated.An impressive example of how STM experiments and DFT calculations together can unravel the atomic structure of the material is given in the article by J. A. Lawlor and M. S. Ferreira  focused First-principle calculations have also provided insight into the electronic properties of low-dimensional materials, for example graphene doping by surface adsorbates , and nanAn important remark is related to the background of all contributions of the present volume. They represent a cross-section of research subjects from members of the European COST Action NMP 0901 “NanoTP”, which ran from 2009 to 2014. The project brought together chemists, physicists, biologists and engineers across a wide range of disciplines, to share their common interest and pool their skills in the field of atomic-scale interface design. At its end the project had nearly 300 members drawn from 26 countries COST countries, along with members in Russia, Australia, Armenia, Belarus, Mexico and Canada. The network allowed researchers to mutually visit laboratories, organise training schools and workshops on diverse subjects including new spectroscopy and microscopy tools, latest developments in atomic scale simulations, and nanocarbon design and application. It helped to launch the ongoing “GraphITA” graphene conference series –32.Within its broad scientific and technical objective, secondary objectives were identified including synthesis of novel nanostructures, engineering of metal-nanostructures contacts, development of new tools for characterization and manipulation nanostructures, atomic-scale quantum chemical modelling, and integration in potential devices. All of these are reflected in the 26 reviews presented here. They appropriately demonstrate the richness and diversity within the NanoTP Action, as well the active participation and enthusiasm of the project members which made the project such a success. This collection of articles is a fitting conclusion to the project, and we hope the collection will also serve to stimulate amongst its readers the most important motivation of all: scientific curiosity.Last but not least, we would like to thank the Production Team of the Beilstein-Institut for their enthusiasm, encouragement, patience and engaged editorial support.Carla Bittencourt, Chris Ewels and Arkady V. KrasheninnikovMons, Nantes and Helsinki, June 2015