Alectoris rufa young/adult ratio and summer abundance were related to the intensity of management , harvest intensity or habitat in Central Spain. We hypothesized that partridge abundance would be higher where management practices were applied more intensively. Variation in young/adult ratio among estates was best explained by habitat, year and some management practices. Density of feeders and water points had a positive relationship with this ratio, while the density of partridges released and magpies controlled were negatively related to it. The variables with greatest relative importance were feeders, releases and year. Variations in post-breeding red-legged partridge abundance among estates were best explained by habitat, year, the same management variables that influenced young/adult ratio, and harvest intensity. Harvest intensity was negatively related to partridge abundance. The other management variables had the same type of relationship with abundance as with young/adult ratio, except magpie control. Variables with greatest relative importance were habitat, feeders, water points, releases and harvest intensity. Our study suggests that management had an overall important effect on post-breeding partridge abundance. However, this effect varied among tools, as some had the desired effect (increase in partridge abundance), whereas others did not or even had a negative relationship (such as release of farm-reared birds) and can be thus considered inefficient or even detrimental. We advise reconsidering their use from both ecological and economical points of view.The reduction of game and fish populations has increased investment in management practices. Hunting and fishing managers use several tools to maximize harvest. Managers need to know the impact their management has on wild populations. This issue is especially important to improve management efficacy and biodiversity conservation. We used questionnaires and field bird surveys in 48 hunting estates to assess whether red-legged partridge  The collapse of commercial fisheries and large-scale population declines of other harvested species are particular cases of the current biodiversity crisis that should be dealt within a conceptual framework integrating both ecological and social information Hunting is practiced in many regions throughout the world either for recreation or for subsistence, and it also has an important economic dimension in many areas, sometimes contributing importantly to rural economies Traditional hunting systems have been increasingly altered and sometimes completely replaced by models based on more intensive management. Management focused on increasing or maintaining post-breeding game populations is thus carried out in many areas, particularly when economic interests are strong Some of these hunting management practices are controversial and may lead to conflicts between stakeholder groups. For example, predator control has a strong negative connotation for many non-hunters Several studies have shown that game management is usually related to higher abundances of target game species e.g. . This maAlectoris rufa L. is one of the most important small game species numerically and economically In Spain, the red-legged partridge The intense use and demand of red-legged partridge in Spain We studied the relationship between post-breeding partridge abundance (i.e. the huntable stock) and the intensity of various management tools currently applied in hunting estates , whilst also taking into account two other variables that are known or expected to affect game densities (habitat and harvest levels). We hypothesized that partridge abundance would be higher where management practices were applied more intensively, but we were particularly interested in knowing which contributed more to increase abundance, if applied simultaneously. We discuss our results in terms of their application to improve sustainable exploitation and conservation of a socio-economically important game species with declining wild populations.2 . Land was mostly privately owned and censuses were carried out with the approval of the person responsible of the game activity within the estate.We studied 48 hunting estates located in Central Spain, in latitudes ranging from 37.98N to 40.33N and longitudes from 6.48W to 2.11W, ED 1950 see . Estate Game management in the studied estates was mainly aimed to red-legged partridge hunting. We selected privately managed estates, which are the great majority in the region 87% , either We estimated partridge abundance from field observations in each hunting estate, using a point count method 2 = 76%, n = 33).From this information, we calculated a summer partridge abundance index (hereafter partridge abundance) as the sum of recorded partridges within a radius of 300 m at each observation point, divided by the number of observation points monitored in each estate. We selected this simple abundance index instead of e.g. estimating densities from distance sampling, because we were mainly interested in relative abundance estimates that were robust enough to be comparable among hunting estates. Distance sampling estimates need a minimum of observations to reliably estimate detection functions Additionally, we calculated for each estate the ratio of young to adult partridges observed (hereafter young/adult ratio), when information about partridge age was available . Young/adult ratios can increase with increased young production or with increased adult mortality. Thus, it cannot be considered as directly equivalent to partridge productivity (i.e. the number of young produced by each pair). However, we considered this ratio as indicative of productivity for the goal of this study, as young mortality is much higher than adult mortality during summer months Fieldwork was carried out from mid June to early August, during the red-legged partridge chick rearing period Cistus sp., Halimium sp., Retama sphaerocarpa, Rosmarinus officinalis, with a strong component of Quercus coccifera and holm oak Quercus ilex, the latter sometimes achieving full tree height), woodland , or dehesa (areas of sparse oak woodland which may be cultivated or grazed underneath). We also calculated habitat Shannon diversity index for each estate from the mean percentage for every habitat category in the estate, and Simpson diversity index We recorded habitat data during bird surveys as the percentage of each habitat type within a radius of 100 m at each observation point, and then calculated habitat for each estate as the average for all surveyed points. We considered the following habitat variables, defined with functional and management meaning for red-legged partridge We collected management data through face-to-face questionnaires with game managers that voluntarily participated in the study. For each estate, we obtained data on estate size, number of feeders , number of artificial water points (see below), number of foxes and magpies annually killed , number of farm-bred partridges annually released and red-legged partridge annual harvest (i.e. number bagged). These data usually corresponded to the hunting season previous to the measure of summer abundances. All variables were expressed per estate surface for analyses.2. We found that the number of small water points and the number of feeders were highly correlated in the estates  for four of the estates . Two further estates also lacked information on the number of magpies killed. Given that comparisons of AIC between different models are only possible if the same sample is used in all models Following current legislation and ethical rules for human research in Spain, data from questionnaires used in this work does not need approval from an ethics committee, as the data gathered are not related to biological or medical human data Analyses were carried out with R 2.13 We wanted to evaluate which management variables explain summer red-legged partridge abundance while removing the expected effect of other variables (such as habitat). Therefore, we carried out analyses in two steps. First, we tested certain models including habitat variables, based on prior information about habitat needs for the species An initial exploration of data showed that abundance was positively correlated with young/adult ratio (see results). We evaluated the effect of management variables, the selected habitat variables  and yearFactors affecting variation in response variables (abundance or young/adult ratio) were analysed using GLMs. We fitted response variables to a Gaussian distribution. In all models, we included the variable “number of count points divided by estate area” as a weight, to control for the potential effect of bird census intensity on the abundance estimate. We checked for normality of residuals of the final models. In those, we also calculated the variance inflation factor VIF,  with theSummer red-legged partridge abundance was linearly and positively related to young/adult ratio in our sample of hunting estates (n = 33), the latter explaining 65% of the variance in summer abundance .Of all models tested to explain temporal and habitat variations in partridge abundance, the best model included Mediterranean scrub and arable land. Within delta AICc values <2 were also those models including year, or estimates of landscape diversity. However, the combinations of arable land, Mediterranean scrub, landscape diversity and year were much worse  . Of the Best models explaining variations in partridge young/adult ratio included one habitat variable (Mediterranean scrub), year (with higher young/adult ratio observed the second than the first study year), and several management variables: the density of feeders, big water points, intensity of release of farm-reared birds, harvest intensity and magpie control intensity . DensityBest models explaining variations in summer partridge abundance included the same variables explaining partridge young/adult ratio except magpie control intensity, which was substituted by fox control intensity . FeedersWe did not detect problems of autocorrelation of residuals in any of the final best models selected . The provision of food and water has been frequently suggested for gamebird population improvement Positive effects of water availability on survival or population dynamics have been found in other Mediterranean galliformes A. rufa and A. chukar in wild populations, due to lack of genetic control of released birds Importantly, partridge young/adult ratio (and thus summer abundance) decreased with increasing numbers of farm-reared partridges released. It may be argued that more partridges are released in areas with poor productivities, but in any case, what is clear from our results is that releases are not effectively increasing summer abundances. It may also be argued that the main goal of releases is just to increase partridge bags in the short term and not to increase summer abundance, and thus effectiveness may not be measured in terms of summer abundance. Indeed, harvest intensity had also a negative strong effect on partridge young/adult ratio and summer abundance. At a larger scale, an increase in hunting pressure has also been found to have a significant effect on the population decline observed in Spain since 1970 Tetrax tetrax , which suggests that our measured variable has indeed a biological meaning . Considering the widespread use of predator control in Central Spain . This agrees with the concerns of Decision making in conservation and management is frequently done under a great degree of uncertainty, and with multiple objectives . Managers frequently have to face trade-offs or dilemmas about how and where to invest in management, which tool to promote or in which circumstances. In many cases, these decisions are taken subjectively, or based on general assumptions. In some cases, investment is made in all potential management tools (like is the case of red-legged partridges), without a real evaluation of the relative costs and benefits of each individual tool.Our results provide critical information to help managers in their decision making, and could be used, if coupled with an evaluation of the economic cost of each tool, to build an open decision framework for managers, which could be applied to optimise both economic objectives and wild game sustainability.In the case of red-legged partridges in Central Spain, our results indicate that the best strategy to reinforce wild populations would be to concentrate in improving food and water availability, either directly through providing supplementary food and water (as currently done) or indirectly through improving habitat quality, which could be expected to be a more efficient, stable and profitable long-term strategy More generally, our results indicate that some game management practices are more efficient than others, and that their joint application may not lead to additive results. Studies identifying the relative importance of individual management tools when applied simultaneously may thus help evaluating the relative economic and conservation value of different managerial scenarios.Table S1Spearman correlation coefficients among the management variables considered.(DOC)Click here for additional data file.


We present theoretical arguments that suggest plankton species may respond rapidly to selection on mildly beneficial mutations due to exceptionally large population size, and consider the circumstantial evidence that supports our inference that selection may be particularly important for these species. We also review all primary population genetic studies of open ocean zooplankton and show that genetic isolation can be achieved at the scale of gyre systems in open ocean habitats (100s to 1000s of km). Furthermore, population genetic structure often varies across planktonic taxa, and appears to be linked to the particular ecological requirements of the organism. In combination, these characteristics should facilitate adaptive evolution to distinct oceanographic habitats in the plankton. We conclude that marine zooplankton may be capable of rapid evolutionary as well as ecological responses to changing ocean conditions, and discuss the implications of this view. We further suggest two priority areas for future research to test our hypothesis of high evolutionary potential in open ocean zooplankton, which will require (1) assessing how pervasive selection is in driving population divergence and (2) rigorously quantifying the spatial and temporal scales of population differentiation in the open ocean.Open ocean zooplankton often have been viewed as slowly evolving species that have limited capacity to respond adaptively to changing ocean conditions. Hence, attention has focused on the ecological responses of zooplankton to current global change, including range shifts and changing phenology. Here, we argue that zooplankton also are well poised for Recent attention has focused on the ecological responses of open ocean zooplankton to current global change, including range shifts and changing phenology. Here, we argue that marine zooplankton also are well poised for evolutionary responses to global change. The oceans are changing on a global scale and, in some cases, at rates greatly exceeding those observed in the historical and recent geological record  can quickly drive beneficial mutations to fixation at population sizes typical of marine zooplankton , whereas genetic variation in Homo sapiens and Arabidopsis  conformed to a background selection model, with a large number of segregating deleterious polymorphisms . In these cases (and others), barriers to dispersal in the open sea occur at oceanographic regions with sharp transitions in both biotic and abiotic properties , while the most oceanic lineage entirely lacked population genetic structure across the same sampling sites. Similar ecophysiological mechanisms likely control dispersal among populations of open ocean zooplankton, though there are no studies that have examined this for truly oceanic taxa. What constitutes optimal habitat will vary across species, with the nature of dispersal barriers highly dependent on the ecological niche of the organism. Therefore, it is more appropriate to consider population genetic structure a species-specific trait that varies across holoplanktonic taxa, as does any other ecological or life history trait, than to view the zooplankton as monotypically ‘high dispersal’ species.Third and finally, increasingly it appears that population genetic structure is species-specific in marine zooplankton, with closely related species often showing very different patterns of differentiation across the same pelagic habitats Goetze . These rThis emerging view of the complexity of dispersal in marine zooplankton extends a number of observations that have been made in other marine species to the pelagic habitat. Panmixia was once expected for all marine species with large population size and a planktonic phase of the life history , we hypothesize that open ocean zooplankton have large adaptive potential and may be capable of strong evolutionary responses (genetic adaptation) to changing ocean conditions. These responses could be rapid, occurring on ecological time scales  technologies have made possible the rapid generation of large-scale sequence data from nonmodel organisms at reasonable cost. We expect that these technologies . One cost-effective approach to examining adaptive variation in natural populations is to study functionally important traits related to, for instance, morphology or life history. Differentiation at these quantitative traits is typically expressed as In order to rigorously quantify the spatial and temporal scales over which differentiation occurs in the open ocean, we need to substantially increase the types and amounts of data that are collected. First, it is critically important to be able to compare genetic results across marker types in order to obtain rigorous estimates of dispersal through Bayesian, Maximum Likelihood, or coalescent analytical techniques (e.g., Pritchard et al. Finally, a wide range of questions on the fundamental basis of evolutionary change in marine zooplankton remain to be addressed. We hope that the ideas outlined here, in combination with the suite of high-throughput genomics techniques now available for development of genome-wide markers in nonmodel species will catalyze research in this field.


Immunologic changes of pregnancy may increase susceptibility to certain intracellular pathogens. A key component of the response to emerging infections is consideration of special populations, including pregnant women. Successful pregnancy depends on adaptation of the woman's immune system to tolerate a genetically foreign fetus. Although the immune system changes are not well understood, a shift from cell-mediated immunity toward humoral immunity is believed to occur. These immunologic changes may alter susceptibility to and severity of infectious diseases in pregnant women. For example, pregnancy may increase susceptibility to toxoplasmosis and listeriosis and may increase severity of illness and increase mortality rates from influenza and varicella. Compared with information about more conventional disease threats, information about emerging infectious diseases is quite limited. Pregnant women's altered response to infectious diseases should be considered when planning a response to emerging infectious disease threats. As strategies to deal with emerging infectious disease threats are developed, a key component is consideration of special populations, including pregnant women , collectively described as the maternal-fetal interface. On the fetal side of the interface, the placenta and membranes enclose the fetus and are derived entirely from fetal tissue. Forming a specialized epithelial surface within the placenta, fetal syncytiotrophoblast cells directly contact maternal blood for nutrient exchange. On the maternal side of the interface, the uterine tissue in contact with the placenta and fetal membranes, the decidua, is rich in specialized maternal immune cells including lymphocytes and macrophages  lymphocytes, which provide costimulation and induce replication of the B cells. The Th2 response during pregnancy results in vigorous antibody-mediated immunity to pathogens  lymphocytes and the cytokines they release. The most important effectors of the cell-mediated immune response, cytotoxic T lymphocytes, are the main immune cells that recognize foreign antigens on the surface of infected "self" cells. Cells infected with viruses or other intracellular pathogens are cytotoxic T lymphocytes' most common targets. The cell-mediated immune response is critical for controlling such pathogens because their intracellular location shelters them from antibody binding.Emphasis on cell-mediated immunity versus humoral immunity changes according to the type of T-helper lymphocytes responding to an infectious threat. Multiple factors, including the cytokine environment and costimulatory molecules present during activation of the T-helper cell, determine the development of either Th1- or Th2-helper phenotype. One hypothesis is that, in addition to hormonal factors that affect the Th1-Th2 balance, macrophages present at the maternal-fetal interface release predominantly Th2-stimulating cytokines and contribute to the overall dominance of humoral immunity during pregnancy  is more common and severe in pregnant women. Accounts of measles outbreaks before an effective vaccine was available indicate that pregnant women may be more severely affected. For example, the investigation of an outbreak of measles in Greenland in 1951 showed that mortality rates were higher among pregnant women than nonpregnant women. Pregnant women were also more likely to experience heart failure , as well as pathogens that could potentially be used as biologic weapons. Unfortunately, information about how pregnant women are affected by many of these novel and emerging infections is limited.During the worldwide outbreak of SARS in 2003, several countries reported cases in pregnant women. Although these numbers were too small to enable definitive conclusions as to whether SARS was more severe among pregnant than nonpregnant women, some evidence indicates that it may be. The largest case series of pregnant women with SARS was from Hong Kong Special Administrative Region, People's Republic of China, where 12 pregnant women with SARS were admitted to 5 public hospitals; 3 of them died, giving a case-fatality rate of 25%  before vaccination and disease eradication indicates that pregnant women are more susceptible to variola infection and have more severe disease  has long been identified as a cause of pneumonia in immunocompromised persons. Pneumocytis pneumonia was first identified in malnourished children in European orphanages during World War II and was later associated with severe immunosuppression in HIV-infected persons , the causative agent, is transmitted by inhalation of material from infected birds or by exposure to infected amniotic fluid or placentas of sheep or goats. Although each year, ≈75–100 cases of psittacosis occur in the United States, only 14 cases of psittacosis have been reported in pregnant women, including a recent case in a pregnant Montana sheep rancher. Illness during pregnancy can be quite severe, mimicking HELLP  syndrome but without hypertension. Most women rapidly recover after pregnancy (Changes in immune function during pregnancy alter a pregnant woman's susceptibility to and severity of certain infectious diseases. These alterations are particularly problematic because physicians may hesitate to provide prophylaxis or aggressive treatment to pregnant women because of concerns about effects on the fetus. For example, despite the 1997 recommendation that women who would be in their second or third trimester of pregnancy during influenza season receive the inactivated influenza vaccine, among women 18–44 years of age, reports of having received the influenza vaccination during the past 12 months were fewer for pregnant than for nonpregnant women (Compared with what is known about conventional disease threats, knowledge about currently recognized emerging infectious diseases is quite limited. Soon we will likely be faced with novel pathogens about which little or nothing is known. Because the effects of emerging infections in pregnant women might differ from those in the general population, pregnancy must be considered a potential risk factor for disease susceptibility as well as for illness and death. Unfortunately, pregnancy issues are often not well addressed in outbreak investigations, ongoing prospective studies, or emergency preparedness planning. Future scientific inquiry and medical investigations must include pregnancy-related issues as a vital component.


People with major depressive disorder (MDD) are more prone to experiencing moral emotions related to self-blame, such as guilt and shame. DSM-IV-TR recognizes excessive or inappropriate guilt as one of the core symptoms of current MDD, whereas excessive shame is not part of the criteria for MDD. However, previous studies specifically assessing shame suggested its involvement in MDD. In the first part of this review, we will consider literature discussing the role of self-blaming moral emotions in MDD. These self-blaming moral emotions have been purported to influence people when they make social and financial decisions in cognitive studies, particularly those using neuroeconomical paradigms. Such paradigms aim to predict social behavior in activities of daily living, by using important resource tangibles  in laboratory conditions. Previous literature suggests that guilt promotes altruistic behavior via acting out reparative tendencies, whereas shame reduces altruism by means of increasing social and interpersonal distance. In the second part of this review, we will discuss the potential influence of self-blaming moral emotions on overt behavior in MDD, reviewing clinical and experimental studies in social and financial decision-making, in which guilt, and shame were manipulated. This is not a well-established area in the depression literature, however in this opinion paper we will argue that studies of moral emotions and their impact on behavioral decision-making are of potential importance in the clinical field, by linking specific symptoms of a disorder to a behavioral outcome which may lead to stratification of clinical diagnoses in the future. But they whose guilt within their bosoms lie, Imagine every eye beholds their blame, For Lucrece thought he blush’d to see her shame.The Rape of Lucrece, William ShakespeareMoral emotions  are discussed as of a critical evolutionary importance  guilt has been recognized as one of the distinctive clinical symptoms of MDD, especially of the melancholic subtype  were significantly higher in a population of people with MDD fully remitted from symptoms compared with healthy subjects . Tangney and colleagues have investigated the role of shame in predicting depressive symptoms in healthy people using the Test of Self-Conscious and Affect (TOSCA) scale, which assesses indirect manifestations of affect on behavior. Their research group conceptualized shame with attempts to deny, hide or escape the shame-eliciting situation; as a result leading to interpersonal separation and increased social distance . Attempts to differentiate between these moral emotions aimed at a distinction based on: (a) types of eliciting events, (b) social audience, and (c) the extent to which a person attributes the failure, within the context of the eliciting event, to the self, or the behavior Tangney, . Guilt iOn the other hand, recent work by O’Connor and colleagues provides evidence against the general association of guilt-proneness with behavioral self-blame by identifying characterological forms of empathy-based guilt . This result was obtained whilst using various control conditions [other-critical emotions such as indignation  confirmed the fMRI evidence for involvement of the frontopolar and septal region in prosocial sentiments  analysis and reported selective decoupling between subgenual anterior cingulate cortex (sgACC) and right superior anterior temporal lobe for guilt compared with indignation in people with MDD, fully remitted from symptoms  and overall pathophysiology. Studies investigating the functional neuroanatomy of guilt and/or shame in patients with MDD are very limited. Current findings suggest that overgeneralization of guilt in MDD is associated with functional disconnection of the anterior temporal cortex and the sgACC, frontopolar, hippocampal, and hypothalamic regions showing guilt-selective disconnection. Previous behavioral studies suggest that there is a need to investigate the functional neuroanatomy of shame in patients with MDD. We provide a summary table of the cited literature for this section Table .In the previous section, we showed that it was difficult to dissociate the roles of guilt and shame in MDD on the basis of the current behavioral or functional neuroimaging evidence. In social psychology, both guilt and shame are conceptualized to be within the same category of moral emotions which help reduce socially undesirable behavior Tangney, . HoweverUnderstanding the relationship between MDD, guilt, shame, and altruism may be more complicated than previously proposed. Both Tangney and O’Connor’s hypotheses have certain limitations. Tangney’s hypotheses are derived mainly from undergraduate student populations, which limits their validity for patients with MDD, whereas O’Connor’s hypotheses may underestimate the role of shame in interpersonal decision-making in MDD. Both of these assumptions rely on self-reported hypothetical behavior and experience, but not actual decision-making. Although one may argue for a direct relationship between choice preferences and actual behavior, the results of choice behavior paradigms are usually confounded by factors such as social desirability. Therefore, computerized behavioral paradigms may have more ecological validity in terms of modeling how people interact with their social environment. Here, we propose a neuro-behavioral economical approach to investigate the extent to which guilt and shame influence social and financial decisions in patients with MDD.Neuroeconomical paradigms make use of quantifiable resource tangibles such as time, money, or amount of water to survive, so that they activate daily life decision-making mechanisms. These experiments are referred to as “games.” Neuroeconomical game is defined as; “a decision problem with structure so that one’s payoffs can depend on one’s own choices and some other input”  consistently made more ethical choices across different domains .The so-called “Prisoner’s Dilemma” (PD) is the most frequently used neuroeconomical paradigm to investigate interpersonal cooperation in uncertain environments. Most commonly in PD, participants interact with a single partner over a one-shot or an iterated decision between cooperation and defection, in which the payoffs for any combinations of these decisions are determined on a pre-defined matrix  in two respects. Firstly, the financial impact of charitable donation behavior is far reaching, relative to the other paradigms which are based on the interaction between two individuals. Secondly, it measures individuals’ responsiveness to concrete societal causes rather than a universally accepted abstract norm such as “fairness” on which the altruistic punishment in the UG is based. In the case of charitable donations, survivor guilt may motivate individuals to restore a sense of fairness with respect to a specific societal issue, by making donations to those who are in need . A recent study showed that guilt induction, irrespective of health concerns, significantly increased the amount of money people were willing to donate to cancer research  in regions of the brain associated with behavioral planning  on interpersonal cooperation in the PD as well as altruistic punishment in the UG. It was shown that acute tryptophan depletion significantly impaired interpersonal cooperation in healthy subjects. Participants in a PD game showed reduced cooperation with playing partners who they were encountering for the first time, but not during subsequent encounters (Wood et al., In their seminal study, Greene et al.  showed tThe role of the dorsal ACC in evaluation of players in interpersonal economic exchanges has been a topic of interest as such evaluations are important in guiding decisions whether or not to punish unfair players. During their evaluation of the active participants, observers activated anterior insula and ACC when fair participants received unfair electric shocks, whereas activations related to empathic concern were significantly reduced in male participants when they observed unfair players receiving electric shocks (Singer et al., The lesion literature suggests that ventral parts of the prefrontal cortex are involved in neuroeconomic decisions. Koenigs and colleagues showed that vmPFC is an important region guiding altruistic punishment decisions in the UG (Koenigs et al., Charitable donation paradigms are also important for neuroeconomical studies. Moll et al.  not onlyIn this section, we have reviewed literature (summarized in Table In the present opinion paper, we have argued that MDD is associated with elevated proneness to guilt and shame. We highlighted the possible overlap in the functional neuroanatomy of guilt and shame with regions known to be functionally abnormal in MDD, and emphasized a need for more neuroimaging studies to dissociate the functional neuroanatomy of guilt and shame in patients with MDD. Following the suggestions of previous authors, we considered the idea that guilt and shame can be differentiated based on their influence on social-economical decision making. We showed that there is converging literature supporting a positive impact of guilt on altruistic decisions; however literature on the impact of shame is inconclusive. Studies of social-economical decision making to date have focused on healthy populations. Considering coexisting emotional processing abnormalities in MDD, it is more challenging to dissociate the impact of moral emotions on social-economical decision making in clinical populations. In order to address this issue, we have argued in this opinion paper that using functional imaging with neuroeconomical paradigms could be an important direction for future psychiatric research.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.


Exciting challenges await the field, including determining the most relevant miRNA targets and parlaying our current understanding of viral miRNAs into new therapeutic strategies. To accomplish these goals and to better grasp miRNA function, new in vivo models that recapitulate persistent infections associated with viral pathogens are required.MicroRNAs (miRNAs) are small RNAs that play important roles in the regulation of gene expression. First described as posttranscriptional gene regulators in eukaryotic hosts, virus-encoded miRNAs were later uncovered. It is now apparent that diverse virus families, most with DNA genomes, but at least some with RNA genomes, encode miRNAs. While deciphering the functions of viral miRNAs has lagged behind their discovery, recent functional studies are bringing into focus these roles. Some of the best characterized viral miRNA functions include subtle roles in prolonging the longevity of infected cells, evading the immune response, and regulating the switch to lytic infection. Notably, all of these functions are particularly important during persistent infections. Furthermore, an emerging view of viral miRNAs suggests two distinct groups exist. In the first group, viral miRNAs mimic host miRNAs and take advantage of conserved networks of host miRNA target sites. In the larger second group, viral miRNAs do not share common target sites conserved for host miRNAs, and it remains unclear what fraction of these targeted transcripts are beneficial to the virus. Recent insights from multiple virus families have revealed new ways of interacting with the host miRNA machinery including noncanonical miRNA biogenesis and new mechanisms of posttranscriptional  In recent years, non-protein-coding regulatory RNAs have been the subject of increasing interest in both prokaryotic and eukaryotic fields. A new understanding of the mammalian genome is emerging where a majority (50%–85%) of the genome is transcribed with at least some noncoding RNA (ncRNA) transcripts being functionally relevant C. elegansDrosophila research communities, RNAi commonly functions to defend hosts against harmful nucleic acids such as endogenous transposons or exogenous viruses (reviewed in RNA interference (RNAi), the process whereby small ncRNAs (<30 nts) serve to direct gene silencing via specific protein machinery, is evolutionarily conserved throughout most eukaryotes. Discovered in studies of the nematode Saccharomyces cerevisiae, and some loss-of-functions in either RNAi or CRISPRs have been associated with the gain of beneficial foreign genetic elements Prokaryotes also possess a nucleic acid-based defense called Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs). Like RNAi, CRISPRs can be thought of as a nucleic acid-based adaptive immune response providing protection against plasmids, transposons, or phage. Similar to RNAi, some bacterial CRISPR systems use double stranded RNA (dsRNA) and RNAse III enzymes in the process of generating effectors that silence gene expression, typically through cleavage of targeted DNA C. elegans via forward genetic screens designed to identify genes involved in larval stage development Drosophila), comprise the Microprocessor complex that binds to the pri-miRNA, where multiple structural cues position cleavage towards the base of the hairpin stem. The newly liberated ∼60 nt hairpin pre-miRNA is then exported from the nucleus to the cytoplasm via the RAN-GTPase Exportin 5. Once in the cytosol, the pre-miRNAs are cleaved by the RNAse III-like endonuclease Dicer. Dicer-mediated cleavage produces a transient ∼22 nt duplex RNA, of which one strand (the miRNA or “guide” strand) is stably incorporated into the RNA-induced silencing complex (RISC). The other strand, called the “star” (*) or “passenger” strand, is less likely to associate with RISC and consequently is typically found at several-fold lower steady state levels. RISC is a multiprotein complex of which a key component is an Argonaute (Ago) protein. Ago-loaded miRNAs (miRISC) typically bind to target transcripts and repress gene expression. However, notable exceptions including translational activation under stress conditions and modulation of hepatitis C virus (HCV) replication have been reported miRNAs are small, approximately 22 nt RNAs that typically silence gene expression by directing repressive protein complexes to the 3′ untranslated region (UTR) of target messenger RNA (mRNA) transcripts. The first miRNAs were discovered in Mounting evidence suggests that miRISC functions by inducing an initial blockade to translation followed by enhanced turnover of repressed target transcripts RNAi likely arose as a primary defense against harmful genetic elements such as viruses, yet in an interesting evolutionary twist, divergent viruses co-opted miRNA expression for pro-viral purposes. DNA viruses account for the majority of known virus-encoded miRNAs with the herpesvirus family encoding most known viral miRNAs. Herpesviruses dominate both in terms of absolute number of known virus-encoded miRNAs and in the average number of miRNAs encoded per virus . Herpesviruses comprise an extended family of large genome DNA viruses whose defining trait is the ability to undergo long-term, and often life-long, latent infections. Latency is a specialized type of persistent infection where only a few viral gene products are expressed allowing for efficient evasion of the immune response. Latency is fully reversible, and with appropriate cues, the virus initiates the lytic mode of infection comprising full viral gene expression and culminating in the production of infectious virus and lysis of the host cell Several aspects of the herpesvirus life cycle are instructive for understanding the other virus families that encode miRNAs. miRNAs are likely invisible to the adaptive immune response—a valuable trait for viruses that undergo persistent infection cis cleavage of the genome, antigenome, or mRNAs mediated by the miRNA processing machinery Naturally occurring positive or negative sense RNA or dsRNA genome viruses that express miRNAs are not widely accepted. In fact, until recently, it had been speculated that viruses with RNA genomes would not encode miRNAs due to negative effects on fitness that would be incurred with As mentioned above, the viruses most likely to encode miRNAs will have nuclear and DNA components to their lifecycle and have the ability to establish persistent infections. However, it's clear that not all viruses that meet these criteria encode miRNAs. As least one type of human papillomavirus  does not encode miRNAs Virus-encoded miRNAs can be grouped into two classes: those that are analogs of host miRNAs and those that are viral specific. Similar to some virus-encoded regulatory proteins, a subset of viral miRNAs have evolved to mimic host effectors. Viral miRNAs that mimic host effectors are referred to as “analogs.” The 5′ end of a miRNA (∼nucleotides 2–8), called the “seed” region, plays an especially important role in directing RISC to mRNA targets. It is estimated that ∼60% of regulation by a particular miRNA is due to binding with perfect seed complementary to the target transcript Although more than 250 virus-encoded miRNAs are known Preventing cell death seems like an obvious advantage to viruses that take up persistent or latent infections in long-lived cells. Host miRNAs play a major role in maintaining cellular homeostasis, and not surprisingly, numerous host miRNAs are implicated in the regulation of cell death , are associated with tumorigenesis. Inducing tumors is likely not a primary advantage for these viruses but rather an accidental off consequence of the need to alter the cell cycle, prevent cell death, and avoid the immune response We have shown that one of the BLV miRNAs functions as an analog of the host miRNA miR-29. miR-29 has been shown to function as either an oncogene or a tumor suppressor depending on the context + T-cells. Taken together, these observations support the hypothesis that some virus-encoded miRNAs serve a subtle role in evading the immune response.Akin to nonstructural viral proteins that often function to evade the immune response, it seems likely that some viral miRNAs perform a similar role. Theoretically, miRNAs can contribute to immune evasion indirectly by lowering viral protein levels and consequent antigenicity, or directly by suppressing components of the host immune response The restricted gene expression of latency and some other forms of persistent infection represents a successful immune evasion strategy. In addition to encoding direct modulators of the immune response, latently infected cells evade the immune response by expressing a limited number of proteins, providing for reduced antigenicity. Several different herpesviruses encode miRNAs that have been implicated in maintaining latent infection and altering the balance between latent/lytic infection Studies of the KSHV latency system provide some of the most well-documented examples of viral miRNAs regulating the latent/lytic switch  3′UTR are cleaved by the Microprocessor complex resulting in decreased KapB protein expression in latency cis pre-miRNA elements, but in this situation, docking of the Microprocessor complex combined with an alternative transcription initiation site is hypothesized to promote an altered splicing pattern of this mRNA to a form that is subject to reduced translational efficiency Finally, for virus-encoded miRNAs, especially those that are not analogs of host miRNAs, all possible functions must be considered. Several viral miRNAs are found clustered in genomic regions near origins of replication or antisense to ncRNAs. It is possible that completely novel functions in genome replication or regulation of ncRNAs await discovery for viral and host miRNAs. In addition to canonical miRNA repressive activities on nscripts . Pre-miRCurrently, specific challenges and goals of the virus-encoded miRNA field overlap extensively with the broader parent fields of both virology and RNAi. Similar to host miRNAs, it will be imperative to determine which of the reported viral miRNAs possess biologically relevant activities. Several reported viral miRNAs are expressed at low levels compared to the other host and viral miRNAs. Are these miRNAs expressed at higher levels in other contexts, or do they possibly function at lower levels by a currently unknown mechanism? With notable exceptions, there is a striking lack of evolutionary conservation of most viral miRNAs. This could imply that viral miRNAs are a site of rapid evolution, perhaps even a driver of speciation. To help better understand which viral miRNAs are most relevant, it would be useful to have a deeper survey of the viruses that encode miRNAs. Perhaps even more informative will be to understand why some members of the same virus subfamilies do and do not encode miRNAs  and those that are viral specific . MimickiAnother mystery that applies to animal host and viral miRNAs is the observation that most lack perfect complementarity to their target transcripts. It has been hypothesized that miRNAs evolved independently in plants and animals Finally, a major reason for studying virus-encoded miRNAs is to be able to better develop therapeutic interventions. One of the most exciting stories in the world of viruses and miRNAs comes not from a virus-encoded miRNA but rather a host miRNA, miR-122 cis-regulation mediated by components of the miRNA pathway.While this manuscript was in revision, three additional papers documenting functions of KSHV miRNAs were published


EHP provide evidence that exposure to certain POPs in the womb may be associated with an increased risk of developing asthma that persists into young adulthood.Although previous research has suggested that prenatal exposures to persistent organic pollutants (POPs) may be harmful to a child’s developing immune system,“The focus in immunotoxicity studies has often been on immunologic intermediates, such as immune cell counts. This study is unique in that it looks at a long-term clinically relevant outcome,” says Todd Jusko, an environmental epidemiologist at the University of Rochester, who was not involved in the study.Researchers in Denmark assessed the relationship between asthma and prenatal exposure to polychlorinated biphenyls (PCBs), the pesticide hexachlorobenzene (HCB), and dichlorodiphenyldichloroethylene (DDE), a daughter compound of the pesticide DDT. PCBs, HCB, and DDT have been banned or restricted for many years due to concerns over human health effects, but they persist in the environment and the human body. Although levels of these chemicals have been dropping in the human population, diet—especially seafood consumption—remains an important route of exposure.Past studies have reported an association between prenatal exposure to DDE and asthma in children at ages 4Divergent findings may be due in part to the fact that previous studies have not followed up past childhood, say the authors of the new study. “A twenty-year follow-up allows us to distinguish between [true] asthma and asthma-like symptoms, such as wheeze, which may resolve in the first six to seven years of life,” says lead author Susanne Hansen, a doctoral candidate in epidemiology at the Statens Serum Institut in Copenhagen.In the current study, the researchers assessed POP levels in blood samples from 872 pregnant Danish women. All samples were collected between 1988 and 1989, during the 30th week of pregnancy. Although blood levels of POPs in the cohort were higher than concentrations found in the general population today, they were average for women of childbearing age in Denmark in the 1980s, according to Hansen. The researchers used registry data and self-reports on asthma diagnosis and medication use since age 6 to identify asthma cases.At age 20, children of mothers with the highest blood HCB concentrations during pregnancy were nearly twice as likely as children of mothers with the lowest concentrations to have been prescribed asthma medication. This is the first study to report an association between developmental HCB exposure and asthma in offspring, according to the researchers.Similarly, children of mothers with the highest blood levels of one particular PCB congener, dioxin-like PCB-118, were nearly twice as likely as children of mothers with the lowest levels to have used asthma medications. The researchers also observed positive associations of HCB and PCBs with self-reported lifetime diagnosis of asthma, and between HCB and clinical diagnosis of asthma, although these associations were not statistically significant. None of the outcomes were associated with maternal DDE.The mechanisms by which HCB and dioxin-like PCBs might influence asthma remain unclear. However, some researchers suspect PCBs and HCB interact with the aryl hydrocarbon receptor—a protein that, when activated in immune cells, may lead to a suppressed immune response.“The association between dioxin-like PCB 118 and asthma is an interesting one but not sufficient to suggest a general link between all dioxin-like chemicals and asthma,” says Michele La Merrill, a developmental toxicologist at the University of California, Davis. She says the possibility of such a general link is “a provocative question that would be worth continuing to ask in future studies.”PCBs, HCB, and DDT have been shown to readily cross the placenta, making maternal blood levels during pregnancy a good proxy for fetal exposure. However, the current results are not sufficient for the researchers to make any claims about the relative importance of prenatal versus postnatal exposures. POPs can be transferred via breast milk, and the researchers had no data on breastfeeding habits for the mothers in their study. “Maternal serum levels could also be correlated with postnatal exposure,” Hansen says.


Autism spectrum disorder (ASD) is characterized by deficits in communication and social ability, as well as restricted interests and repetitive behavior. Anxiety is a persistent anticipation or apprehension about one or more situations to which a person is exposed, and affects many people, including children with ASD. Stress, by contrast, is a response to situations that are threatening, uncontrollable, or unexpected. Indices of anxiety are often measured through informants, with parents and teachers serving as the primary sources of reported anxiety in children. However, self-report measures exist, allowing current (state) and persistent (trait) anxiety to be assessed. The current study was designed to evaluate whether children with autism could identify their own levels of anxiety and the degree to which these levels were associated with symptom profile and physiological arousal.Self-reported state and trait anxiety were collected during exposure to different stress paradigms for 40 children  and compared to parent reported social ability  and stress responsivity (cortisol).Significant differences were found between typically developing and children with autism for both state and trait anxiety across all conditions. Associations were identified between severity of parent-reported social impairment and both types of self-report anxiety. No relationship was found between stress  and anxiety in children with autism.Children with autism are able to consistently report their persistent level of anxiety symptoms in stressful situations of benign character. Therefore, the inclusion of such measures may be useful in identifying and tracking symptoms in children with autism under appropriate circumstances. In addition to the primary impairments in reciprocal social communication and repetitive, restricted behaviors in autism spectrum disorder , there aLongitudinal and developmental neuroscience studies reveal that alongside cognitive and neural development, anxiety often follows a developmental trajectory starting early in life and proceeding into adolescence and adulthood ,17. For Cognitive ability and impairment in autism are distinct; thus, severity of impairment in other core domains, such as social difficulty, can be major contributors to anxiety. Additionally, while autism symptom expression and social anxiety symptoms have been associated in early life, including non-autistic children with clinical anxiety disorders displaying autism-like symptoms and developmental traits , the relAlthough a certain amount of anticipatory anxiety is considered adaptive, it can become pathological when the worry is excessive or uncontrollable and begins to exert strong influences on behavior and cognition . IncreasOne of the primary stress systems, the hypothalamic-pituitary-adrenocortical axis (HPA) responds consistently to perceived novel or unfamiliar situations. Cortisol is a glucocorticoid associated with activation of the HPA axis and can serve as an important biomarker of stress to a variety of different stimuli ,28. PrevIt should be noted that, while self-reports have been questioned, parent reports are also inconclusive. Recent studies in adolescents with autism suggest discrepancies across informants (parents versus self-reports) for social functioning  and qualAs part of comprehensive studies examining biobehavioral profiles of children with autism parent report, self-report, and physiological stress data were collected during play with peers , exposurParticipants were recruited through distributed flyers and direct phone contact from a subject tracking system for a playground study examining HPA reactivity to social stressors in children with autism. After completion of the initial playground paradigm participants were recruited by phone for up to two additional studies investigating reactivity to both social and non-social stressors. Only a portion of the participants chose to return for each of the additional studies. The two additional studies had an average latency of 6 months from the initial playground study, with a range of 4 to 9 months. Participants provided saliva samples at 20-minute intervals for later analysis examining salivary cortisol levels at each of the three different stress paradigms described below. Additionally, two separate self-report measures of anxiety were completed at each study session by the participants, while a parent-report measure of social ability was collected at the initial playground session.Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) criteria [The playground study consisted of 40 healthy, age matched, unmedicated, prepubertal, male children 8 to 12 years of age. A total of 19 participants were diagnosed as having autistic disorder, and 21 were typically developing. Diagnosis was based on the criteria  and estacriteria , adminisA portion of the participants from the playground sample, consisting of 15 (71%) of the typically developing children and 11 (58%) of the children with autism, participated in an additional study using a mock MRI as a non-social stressor. A second subset of the playground sample, including 16 (76%) of the typically developing children and 13 (68%) of the children with autism, participated in the TSST-C, a social stress paradigm. Ten (48%) of the typically developing children and five (26%) of the children with autism from the original playground sample participated in both the mock MRI and TSST-C. Study visits were ordered as follows: playground first, followed by MRI, and finally TSST-C. Additional demographic information is included in Table Informed written consent was obtained from parents and verbal assent was obtained from research participants prior to inclusion in the study. Participants were told they would be playing on a playground with peers for the playground paradigm or that they would be participating in a school-like performance task in the TSST-C paradigm . ParticiThe Autism Diagnostic Observation Schedule  is a seThe Wechsler Abbreviated Scale of Intelligence  is a meThe Social Responsiveness Scale  is a 65The State Trait Anxiety Inventory for Children  is a 40The Multidimensional Anxiety Scale for Children  is a 39The playground social stress protocol was the positive social stressor in this study. Participants were asked to play on an enclosed 18.2 × 18.2 m playground with two other age and gender matched children. The 20-minute play period was subdivided into periods of prescribed free and cooperative play was facilitated by a confederate child on the playground communicating with the research staff via concealed audio technology . A totalThe mock MRI is a non-social neutral stressor designed to evaluate reactivity to a situation with significant sensory components, including simulated MRI machine noises and relative confinement inside the scanning bore . ParticiThe TSST-C is a well validated negative social stressor in which participants are asked to make a public presentation to neutral raters. The Trier Social Stress Test  was origAll three studies included a saliva sample taken after exposure to the stressor. The STAIC and MASC were administered to participants during the rest period immediately following this sample. Parents were present during administration of the STAIC and MASC, but were not permitted to assist their child. Research personnel carefully avoided referencing other terms used on the measures when answering participant questions. For example, the word nervous was avoided when describing what jittery means, as nervous appears later in the measure. The use of multiple measures with each participant allowed for better characterization of anxiety along multiple axes; the MASC focusing on anxiety subtypes, while the STAIC explored temporal characteristics.Established salivary collection protocols were carefully followed, including using consistent collection materials and methods, controlling the intake and time of drinks, food, and prescription medications, as well as using standardized procedures ,66. The Between-group analyses for the autism and typically developing groups were performed across demographic (age and IQ) and dependent variables of interest (STAIC and MASC anxiety) using independent two-sample t tests if the assumption of normality held true; otherwise, the equivalent non-parametric test was used. Pearson product correlations were performed to assess the association between self-reported anxiety and parent-reported social functioning (SRS) and physiological levels , respectively. Statistics were analyzed using SPSS version 21 .P = 0.020) and trait  anxiety . Typically developing children showed very strong consistency between the playground and MRI for both state  and trait  anxiety. Trait anxiety between playground and the TSST-C was also highly correlated . Analyses between MRI and TSST-C reported anxiety were not made due to limited sample size of participants shared between the two conditions. Thus, the STAIC appears to be a stable measure for use in children with autism in some stress conditions  the longitudinal consistency of self-reported state and trait anxiety measures used with children with autism; 2) the relationship between both types of self-reported anxiety to parent-reported functioning; and (3) the association between reported anxiety and physiological stress responses. Self-reported anxiety was investigated across the two groups revealing significant differences between the typically developing and autism groups in terms of trait anxiety measured by the STAIC (see Table  the relaP = 0.026), while trait anxiety was strongly related to the same subscale . There were also relationships between the coping subscale and both state  and trait  anxiety in the playground paradigm. Additionally, strong correlations were observed between the MASC social anxiety subscale and trait anxiety at both the MRI  and the TSST-C .Self-rated anxiety on the MASC was also found to be significantly different between typically developing children and those with autism (See Table P = 0.004) as well as trait  anxiety. These results suggest that children with lower levels of overall social ability experience heightened anxiety in various situations when compared to their higher functioning peers. Similarly, the SRS Total Score was also correlated with MRI trait anxiety  and TSST-C trait anxiety . This demonstrates a relationship between parent-reported social functioning and child self-reported anxiety across conditions.An additional relationship was found between anxiety in the playground condition and overall social functioning level measured by total score on the parent SRS measure for both state  when compared to typically developing children. In the TSST-C, however, children with autism showed a lack of responsivity compared to their typical peers, as previously reported [P >0.05). Within group comparisons between the equivalent saliva samples and self-report anxiety showed that in children with autism there was no correlation between cortisol and anxiety on any of the stressors. Typically developing children, however, did show some relationship between cortisol and anxiety at certain time points. For example, salivary cortisol at arrival in children participating in the MRI condition correlated with trait anxiety , whereas in the TSST-C condition arrival cortisol correlated with state anxiety .In regards to salivary cortisol levels, children with autism showed significantly higher activation in response to the playground social stressor (previously reported in ), and toreported . CorrelaThe current investigation explored the long-term stability of self-reported anxiety in autism as well as associations to parent-reported functioning and physiological stress responses. The findings reveal large differences between the typically developing and autism groups in trait anxiety and MASC subscales, providing further support that anxiety is a significant and common condition in children with autism . TheThe lack of correspondence with the TSST-C reported anxiety may be due, in part, to differences in stress response, as the TSST-C failed to evoke a physiological response in children with autism ,67. As mMoreover, the physiological stress under which a child with autism is requested to rate their general anxiety may influence their response. In other words, under conditions in which they exhibit physiological arousal as demonstrated by cortisol elevation they may be better able to rate their trait anxiety as being elevated, while under conditions in which they do not demonstrate physiological arousal such as the TSST-C, children with autism may appraise their overall anxiety as being lower. Interestingly, the children with autism did not consistently endorse state or current anxiety during the time of the stressor. It may be that children with autism are unable to discern current states of functioning yet are able to endorse enduring states of being. Sensory sensitivity frequently found in individuals with autism may be interfering with their ability to assess themselves cognitively during an experience. In other words, their sensory reactivity may be paramount and prevent them from being able to adequately identify internal states. Since this is merely conjecture, additional research is needed in this area. Our hypothesis that self-reported anxiety would be consistent across the different study paradigms was therefore only partially correct as consistency was only displayed for trait anxiety and only when the paradigm provoked a stress response.External social cues related to autism symptoms, as well as direct cognitive awareness of one’s own impairments are both potential sources of anxiety for individuals with autism. Specific autism traits such as insistence on sameness  and restOur final hypothesis, that physiological stress measures would be associated with self-report anxiety, proved incorrect, as the data showed no such relationship. Anxiety and physiological stress, while often deeply intertwined, are distinct constructs that may be individually provoked under specific circumstances. The lack of correlation between salivary cortisol and self-report anxiety across all three stressors provides support for the distinctiveness of these constructs in children with autism . In typiThese findings demonstrate that children with average cognitive ability in addition to autism may be able to accurately assess their personal anxiety levels in contrast to some reports that question the usefulness of such measures ,42, or sClinical implications of this study include providing evidence of long-term stability of the trait portion of the STAIC in this population across consistent conditions. Support is also present for using parent-reported functioning measures, such as the SRS, as an alternate method of assessing anxiety risk. As the STAIC and MASC are normalized for typically developing individuals, closer inspections through reliability and validity studies, which have been conducted in typically developing youth , are neeDue to the frequency of occurrence and deleterious effects on quality of life, addressing anxiety in children with autism is highly important. Interventions to reduce anxiety are often warranted and may be especially prudent in populations with lower social functioning, particularly those with self-assessed anxiety relating to social interaction or coping ability. Cognitive behavioral therapy (CBT) has been shown to be effective in children with autism to ameliorate anxiety symptoms ,74,75, aAdditionally, the current results demonstrate the lack of an association between stress, as measured by salivary cortisol, and self-reported anxiety in children with autism. While this has previously been established in regards to the TSST-C, this study extends the finding to stressors that induce physiological reactivity in children with autism. This is an important consideration for research involving biological stress measurements as it strongly indicates that anxiety cannot be used as a predictor of biological reactivity to the environment in autism. Some evidence exists of a relationship in typically developing children between salivary cortisol at arrival and self-report anxiety, and future research would be well guided to examine this further.Despite these interesting findings, there are some limitations to acknowledge. The sample size was somewhat moderate for a few of the experimental conditions. Furthermore, the age range was narrow due to the developmental play paradigm and exclusion criteria for puberty. While diagnostic and study inclusion criteria in the field have changed , the chiIn summary, many children with autism exhibit persistent anxiety that can be captured via self-report measures. Furthermore, children’s anxiety is directly related to their level of social impairment. While no relationship was found between salivary cortisol reactivity and self-report anxiety, it appears that reported anxiety remains consistent between conditions that evoke similar physiological responses. Future research designed to identify children with enduring and impairing anxiety remains an important pursuit. Associations with social, perceptual, and cognitive functioning may identify subgroups at risk and in need of targeted treatment for anxiety in autism.The authors declare that they have no competing interests.DMS collected and interpreted participant data for both self-report and parent-report forms, as well as physiological measures, and also wrote a major portion of the finished manuscript. BAC analyzed and interpreted participant data for both self-report and parent-report forms as well as the physiological data, and also made major contributions and edited the finished manuscript. Both authors read and approved the final manuscript.


Glaucoma is a neurodegenerative disorder in which a variety of complicated mechanisms lead to retinal ganglion cell (RGC) apoptosis and death. According to various theories, factors including elevated intraocular pressure (IOP) and vascular dysregulation primarily contribute to the initial insult during glaucomatous atrophy in the form of obstruction of axoplasmic flow within RGC axons, alteration in optic nerve microcirculation at the level of lamina and changes in the laminar glial and connective tissues. Deprivation of neurotrophic factors, such as the brain-derived neurotrophic factor, and release of a large number of neurotoxic agents within the retina, including glutamate, nitric oxide and free radicals, are the consequences of the above-mentioned events.Currently, IOP reduction is the only proven clinical therapy available for treatment of glaucoma. Nevertheless, signs of progression can be seen in many patients despite well-controlled IOP. In addition, glaucomatous optic neuropathy (GON) has even been observed in patients with “normal” IOP. Therefore, the pathophysiology of glaucoma and particularly normal tension glaucoma is probably beyond IOP alone. Studies on low pressure glaucoma have demonstrated that in most patients higher intraocular pressure is unrelated to poor visual fields.During the past decade, most studies more focused on discovering neuroprotective treatments along with IOP lowering medications in order to prevent ganglion cell death or even reverse the process of cell death. Neuroprotection aims at blocking primary destructive events affecting RGCs or optic nerve fibers, enhancing RGC or optic nerve fiber survival mechanisms and finally repairing damage occurring during the progressive, secondary stage of the injury.The idea of using neuroprotective treatment for management of glaucoma arose from the successful results of employing neuroprotective drugs for treatment of certain neurodegenerative disorders of the central nervous system (CNS) such as Alzheimer’s and Parkinson’s disease. Recent studies have elucidated strong links between mechanisms of cell death in Alzheimer’s disease and glaucoma.Glaucoma is a chronic and slowly progressive disease, therefore assessment of the efficacy of these drugs is a difficult task and requires a long time to be proven. Certain IOP lowering medications, such as brimonidine and betaxolol have been suggested to possess neuroprotective effects. Recently, the low pressure glaucoma study group released the results of comparing brimonidine with timolol with average follow-up of 30 months. This study showed that patients treated with brimonidine were less likely (9%) to experience visual field progression as compared to subjects treated with timolol (39%). IOP reduction was not significantly different between patients assigned to brimonidine or timolol. Although in this study brimonidine, at least indirectly, demonstrated neuroprotective properties, its mechanism of action is not yet fully understood.There are other promising medical approaches for glaucoma neuroprotection which are briefly discussed below:Glutamate antagonists: Glutamate is one of the major neurotransmitters in the CNS and retina and elevated levels have been reported in glaucoma. This transmitter may facilitate RGC apoptosis by activation of N-methyl D-aspartate receptors (NMDA), resulting in calcium influx into RGCs and disruption of the cell wall. Memantine is an anatagonist of NMDA which had earlier been proven to be effective in Alzheimer’s and Parkinson’s diseases. Although animal studies confirmed that this drug protects injured neural cells from death, phase III clinical trials on glaucoma in humans failed to demonstrate any clinical benefit. The study showed that glaucoma progression was significantly slower in patients receiving higher doses of memantine as compared to patients receiving lower doses of the medication, but there was no significant benefit when compared to patients receiving placebo.Neurotrophins: The brain-derived neurotrophic factor (BDNF) and ciliary derived neurotrophic factor (CNTF) have been found to be decreased in glaucoma. These factors suppress the intrinsic apoptotic process and activate survival signals. Several investigators are now exploring the use of gene transfer methods to deliver neurotrophic factors. Encapsulated cell technology helps to implant cells that have been genetically modified to produce neurotrophic factors.Anti inflammatory agents: Recently researchers have investigated the immune system as a means of providing neuroprotection against neuronal damage. Cop-1, otherwise known as glatiramer acetate, copolymer-1 and copax-1, is a synthetic, random oligopeptide composed of amino acids tyrosine, glutamate, lysine, and alanine. Cop-1 is a Food and Drug Administration (FDA) approved drug for treatment of multiple sclerosis. It is a low affinity antigen and can evoke both active and passive T-cell mediated immune responses at various sites of injury. It has been demonstrated that Cop-1 reduces damage caused by mechanical injury to the optic nerve or intravitreally administered glutamate. Another study reported that vaccination with Cop-1 leads to significant reduction in RGC death in rat models of ocular hypertension.Antioxidants: It is hypothesized that oxidative stress can cause RGC death by damaging the trabecular meshwork, the optic nerve head and the retina. Targets of oxidative stress relevant to the development of GON are most probably the mitochondria. Reduced levels of the antioxidant agent glutathione and increased serum lipid peroxidation products have been identified in patients with primary open angle glaucoma. Antioxidants such as vitamin C, vitamin E and Ginkgo biloba extract have been shown to improve visual fields in patients with glaucoma but there is still no strong evidence or long term studies to confirm this idea. Melatonin (N-acetyl-5-methoxytryptamine) is an indoleamine, secreted by the pineal gland, which exerts antioxidant properties. Melatonin has been shown to neutralize free radicals and activate some antioxidative enzymes; it can therefore be categorized under neuroprotective agents.Mitochondrial augmentation: Mitochondrial dysfunction has been implicated in neuronal apoptosis and shown to play a role in experimental glaucoma. One in vitro study provided evidence that mitochondrial dysfunction accompanying RGC death may be induced by glaucoma-related stimuli such as tumor necrosis factor (TNF)-α and hypoxia. The dopaminergic agent, GLC756, has recently been shown to inhibit TNF-α release from activated rat mast cells, suggesting a potential role for glaucoma neuroprotection. Coenzyme Q10 (CoQ10) is a cofactor which plays a crucial role in energy production via the mitochondrial electron transport chain and also has strong antioxidant properties.Stem cells: Several potential applications of stem cells for neuroprotection have been explored. Transplanted mesenchymal stem/stromal (MSC) cells have demonstrated significant neuroprotection in a rat model of glaucoma. These cells can be isolated from a variety of tissues including the adult bone marrow and may have the potential to trans-differentiate into neural cells. However, in order for stem cells to be successful, their use must be coupled with mechanisms by which newly transplanted RGCs can extend their axons to appropriate structures within the brain.Neuroprotective interventions are promising in the treatment of glaucoma as adjunctive therapy to conventional IOP lowering medications. There are animal studies with promising results in protecting neuronal cells but no longitudinal study in human subjects has yet proven these observations. Better understanding of the pathophysiology and mechanisms of glaucoma can help investigators find new modalities of treatments in the future.


Aromatase inhibitors have been introduced as a new treatment modality that could challenge clomiphene citrate as an ovulation induction regiment in patients with PCOS. Although several randomized trials have been conducted regarding their use as ovulation induction agents, only few trials are available regarding their efficacy in IVF stimulated cycles. Current available evidence support that letrozole may have a promising role in stimulated IVF cycles, either when administered during the follicular phase for ovarian stimulation. Especially for women with poor ovarian response, letrozole appears to have the potential to increase clinical pregnancy rates when combined with gonadotropins, whereas at the same time reduces the total gonadotropin dose required for ovarian stimulation. However, given that in all of the trials letrozole has been administered in GnRH antagonist cycles, it is intriguing to test in the future how it may perform when used in GnRH agonist cycles. Finally administration of letrozole during luteal phase in IVF cycles offers another treatment modality for patients at high risk for OHSS taking into account that it drastically reduces estradiol levels Aromatase inhibitors are drugs traditionally used for the treatment of hormone responsive advanced breast cancer . The lasSeveral trials have tested the effect of AIs (letrozole or anastrozole) in women with anovulatory  or unexpThis lack of strong evidence is even greater regarding the use of AIs in IVF/ICSI cycles. Only few randomized trials, with limited series of patients, have been conducted up to date and the main research interest has been accumulated in the effect of letrozole in the treatment of poor responders.Only one randomized trial has been conducted up-to-date that evaluated the addition of letrozole in patients with normal ovarian response undergoing IVF or ICSI . DespiteOnly four randomized trials have been published through 2010 with a total of 235 patients with poor ovarian response randomized to receive letrozole combined with gonadotropins or gonadotropins alone as ovarian stimulation protocols in IVF/ICSI cycles (Table The first small randomized trial published in 2004 examined the use of letrozole as part of a low-cost IVF protocol for poor responders. According to this study, letrozole+ rFSH resulted in comparable pregnancy rates with patients treated with a GnRH agonist and rFSH alone . In addiThe first randomized pilot study assessing the effect of administration of letrozole during the luteal phase of stimulated IVF cycles in oocyte donors was published in 2008 . DespiteAnother randomized placebo controlled trial has tested the same hypothesis in 30 egg donors after oocyte retrieval Table . AdminisBoth randomized trials provided evidence that letrozole drastically reduces E2 levels whereas the trial Garcia-Velasco et al., demonstrated that letrozole restores LH secretion sooner in egg donors. In this regard AIs may be of interest not only for egg donors but also in patients at high risk of OHSS who freeze their oocytes/embryos or cancel hCG administration in order to reduce the risk linked to elevated estadiol levels.The last five years letrozole has been introduced as an agent that could be used in cancer patients undergoing ovarian stimulation in order to preserve fertility prior chemotherapy, through embryo or oocyte cryopreservation . The benAt the moment existing evidence for the use of aromatase inhibitors as part of an IVF/ICSI protocol is fragmented and weak. Despite the timeframe of ten years since introduction of AIs as ovulation induction agents, few randomized trials with limited number of patients have been conducted up to date. The lack of solid evidence in this field has been previously underlined , and defBased on the available data it appears that scant scientific evidence exists regarding the role of letrozole during the follicular phase women with normal ovarian response. However, letrozole may serve as an alternative for patients with poor ovarian response. A very important observation from the available data, is that a treatment protocol with GnRH antagonists, letrozole and gonadotrophins, hints a danger for compromising clinical pregnancy rates compared to treatment with gonadotropins in a microdose GnRH agonist protocol. Nonetheless, these negative results should not necessarily be attributed to letrozole, but may indeed be related to the type of analogue used. Microdose GnRH agonist flare protocols have been shown to be an effective treatment for poor responders, with randomized trials supporting significantly higher implantation rates compared to GnRH antagonist protocols ,22. ConsFurthemore, the administration of letrozole during the luteal phase offers a new insight for patients at high risk for OHSS. Despite the fact that the results are based solely on one randomized trial, if they are confirmed by future studies letrozole may have a role as a preventive measure for OHSS.Although a trial published by Tulandi et al. in 2006  supporteCurrent evidence supports aromatase inhibitors can be a safe solution for fertility preservation in cancer patients prior chemotherapy. Although letrozole may have a role in the treatment of poor responder patients, results are based on few evidence and need to be replicated in the future, whereas its use in normoresponders requires even more intensive research. Their use should also include the long agonist protocol for IVF, as this protocol is widely used. Off label inscription shall be re-evaluated from pharmaceutical industry and reproductive associations in order more appropriate randomized studies to be performed.AIs: aromatase inhibitors; IVF: in vitro fertilization; ICSI: intracytoplasmic sperm injection; OHSS: ovarian hyperstimulation syndrome; HMG: human menopausal gonadotropin; FSH: follicular stimulating hormone; GnRH: gnonadotropin releasing hormone; LH: luteinising hormone; hCG: human chorionic gonadotropin.The authors declare that they have no competing interests.EGP wrote the manuscript, NPP wrote the manuscript, PH wrote the manuscript, GP, revised the manuscript, EB revised the manuscript, HT revised the manuscript, BT revised the manuscript. All authors read and approved the final manuscript.


This ironment ,4. The aThe brain is able to maintain a stable perception although the visual stimuli vary substantially on the retina due to geometric transformations and lightning variations in the environment. These transformations comprise (i) local scaling transformations caused by objects of different size and at different distances to the observer, (ii) locally linearized image deformations caused by variations in the viewing direction in relation to the object, (iii) locally linearized relative motions between the object and the observer and (iv) local multiplicative intensity transformations caused by illumination variations. Let us assume that receptive fields should be constructed by linear operations that are shift-invariant over space and/or space-time, with an additional requirement that receptive fields must not create new image structures at coarser scales that do not correspond to simplifications of corresponding structures at finer scales.by necessity, based on Gaussian kernels, Gaussian derivatives or closely related operators [Given the above structural conditions, we derive idealized families of spatial and spatio-temporal receptive fields that satisfy these structural requirements perators ,4. We shperators ,4. Indeeperators  in theirtrue invariance of receptive field responses can be obtained under scaling transformations, affine transformations and Galilean transformations [By complementing such receptive field measurements with selection mechanisms over the parameters in the receptive field families, we show how rmations . Thereby