Introduction: Adverse childhood experiences (ACE) such as sexual and physical abuse or neglect are frequent in childhood and constitute a massive stressor with long-lasting adverse effects on the brain, mental and physical health.The aim of this qualitative review is to present a concise overview of the present literature on the impact of ACE on neurobiology, mental and somatic health in later adulthood.Methods: The authors reviewed the existing literature on the impact of ACE on neurobiology, mental and somatic health in later adulthood and summarized the results for a concise qualitative overview.Results: In adulthood, the history of ACE can result in complex clinical profiles with several co-occurring mental and somatic disorders such as posttraumatic stress disorder, depression, borderline personality disorder, obesity and diabetes. Although a general stress effect in the development of the disorders and neural alterations can be assumed, the role of type and timing of ACE is of particular interest in terms of prevention and treatment of ACE-related mental and somatic conditions. It has been suggested that during certain vulnerable developmental phases the risk for subsequent ACE-related disorders is increased. Moreover, emerging evidence points to sensitive periods and specificity of ACE-subtypes in the development of neurobiological alterations, e.g., volumetric and functional changes in the amygdala and hippocampus.Conclusion: Longitudinal studies are needed to investigate complex ACE-related characteristics and mechanisms relevant for mental and somatic disorders by integrating state of the art knowledge and methods. By identifying and validating psychosocial and somatic risk factors and diagnostic markers one might improve the development of innovative somatic and psychological treatment options for individuals suffering from ACE-related disorders. Converging evidence from epidemiological and neurobiological studies suggest adverse childhood experiences (ACE) such as sexual and physical abuse and related adverse experiences to be closely related to enduring brain dysfunctions that, in turn, affect physical and mental health throughout the lifespan –3. This At the present time, there is clear evidence that ACE and ACE-related disorders are associated with enduring effects on the structure and function of neural stress-regulatory circuits such as for example the hippocampus, the amygdala or the ACC , 36 and On balance, many studies point at reduced volume of the hippocampus in individuals after ACE compared to non-maltreated individuals , 39–45. Several studies have demonstrated that exposure to specific types of ACE selectively affect sensory systems, which were involved in perceiving the trauma that was experienced , 36. ForIn contrast to the high amount of studies investigating structural brain alterations, only a few have examined functional differences related to ACE. Fortunately, over the last decade, an increasing number of studies have been published using functional neuroimaging (fMRI) techniques to examine possible associations between ACE and alterations in neurocognitive systems.Evidence from animal and human research proposes that the amygdala is critically involved in the detection and processing of salient stimuli, especially in those related to danger . NeuroimACE are supposed to affect psychological and psychosocial mechanisms known to contribute to mental disorders, comprising disturbances in cognitive and affective processing such as a heightened attention toward threatening stimuli , heighteA few studies provide evidence that specific types of maltreatment are associated with greater risk for developing psychopathology than other types. In a meta-analysis, Norman et al.  reportedEmerging evidence suggests that ACE is associated with the development of a wide range of somatic disorders, such as obesity, diabetes, inflammatory bowel diseases (IBD) such as ulcerative colitis (UC) and Crohn Disease (CD) , 105–107Recent studies have suggested a dysregulation of the innate immune system as a possible biological mediator between ACE and adulthood disease. These studies have reported an association between ACE and increased levels of pro-inflammatory markers, most notably of the acute phase protein C-reactive protein (CRP), the cytokines interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) . IBD areACE are complex etiological marker, that appear to vary on impact in terms of type, timing and severity of maltreatment, together with a wide range of vulnerability and resilience cofactors. In the last years, there has been a welcome increase of research on consequences of ACE on neurobiological, psychological and somatic issues. Overall, the available studies indicate an enduring effect of ACE on mental and physical health throughout the lifespan . HoweverJH wrote the article, which CS reviewed and approved 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.


Curved continuous girder bridges (CCGBs) have been widely adopted in the civil engineering field in recent decades for complex interchanges and city viaducts. Unfortunately, compared to straight bridges, this type of bridge with horizontal curvature is relatively vulnerable to earthquakes characterized by large energy and short duration. Seismic damage can degrade the performance of CCGBs, threatening their normal operation and even resulting in collapse. Detection of seismic damage in CCGBs is thus significantly important but is still not well resolved. To this end, a new method based on wavelet packet singular entropy (WPSE) is proposed to identify seismic damage by analyzing the dynamic responses of CCGBs to seismic excitation. This WPSE-based approach features characterizing damage using synergistic advantage of the wavelet packet transform, singular value decomposition, and information entropy. To testify the algorithm, a finite element model of a typical CCGB with two types of seismic damage is built, in which the seismic damage is individually modeled by stiffness reductions at the bottom of piers and at pier-girder connections. The displacement responses of the model to El Centro seismic excitation is used to identify the damage. The results show that damage indices in the WPSE-based approach can correctly locate the seismic damage in CCGBs. Furthermore, the WPSE-based method is competent to identify damage with higher accuracy in comparison with the wavelet packet energy based method, and has a strong immunity to noise revealed by robustness analysis. An array of responses used in this approach paves the way of developing practical technologies for detecting seismic damage using advanced distributed sensing techniques, typically the optical sensors. Curved continuous girder bridges (CCGBs) are key components of urban traffic, extensively used as complex interchanges and city viaducts in civil infrastructure ,2, due tIn recent years, the health assessment of CCGBs after earthquake has attracted much attention in the interest of ensuring safe operation. The visual inspection for local and visible flaw ,9,10 is Representative studies of wavelet transform based damage identification of bridge structures subjected to seismic excitations are described as follows. Wang and Chan  reportedApart from wavelet transform coefficients, various methods based on wavelet entropy, wavelet packet energy entropy, and wavelet singular entropy have been developed to identify damage in bridges in recent years. Ren and Sun  providedMost extant methods have been focused on detecting damage in straight bridges subjected to seismic excitation, with emphasis on revealing abnormality of wavelet coefficients of dynamic responses to characterize damage. However, CCGBs are more complex than straight bridges, because the horizontal curvature makes curved bridges more vulnerable to extensive damage and possible collapse. There are few dedicated methods for detecting seismic damage for CCGBs, especially fairly lacking effective methods that match the advanced distributed optical measurement.To this end, this study develops a method for health assessment and damage identification in CCGBs. A new wavelet packet entropy based method is proposed to identify seismic damage by analyzing the dynamic responses of CCGBs subject to seismic excitation. This method is employed to identify two types of seismic damage in CCGBs, located at the bottom of piers and at pier-girder connections, respectively. Damage identification in numerical CCGB model subjected to El Centro seismic excitation shows the superiority of the proposed method in damage identification accuracy and noise immunity. The proposed method is expected to provide an effective and applicable tool for damage identification using dynamic responses recorded by advanced sensing techniques, especially by distributed optical fiber sensors.The rest of the paper is organized as follows. The fundamentals of wavelet packet, singular value decomposition, and information theory are introduced in This section introduces the fundamentals of the wavelet packet transform (WPT), singular value decomposition (SVD), and information entropy (IE).j, k, and n are the decomposition level, translation factor, and modulation parameter, respectively. The terms The WPT is defined by the following recursive relationships : (1)u2nD=UΛq is number of selected singular value orders.Then, following Equation , the wavWavelet packet singular entropy takes synergistic advantage of WPT, SVD, and IE, with the characteristics manifested in the following aspects: (1) WPSE retains the high time frequency multiresolution characteristics of WPT, permitting highlighting and characterization the intrinsic peculiarity of damage in the full frequency band; (2) WPSE retains the feature space mapping characteristics of SVD, enabling quantificational extraction the linear independent features from the wavelet space; (3) WPSE retains the system complexity metric characteristics of IE, facilitating characterization of the information feature. In summary, WPSE can highlight, extract, and quantificationally characterize the information characteristics, allowing it to be used as a characteristic factor for damage identification.A WPSE-based damage index is defined asquations  and 12)(13)DIWPSn is the total number of sensors distributed in a structure; n damage indices; To demonstrate the damage location more clearly, a threshold value is established with the application of the one-sided confidence limit of urements :(14)DIWPFurther, the damage pre-warning index determine the appropriate effective structural dynamic responses from measuring various types of responses in different directions;(b)select the optimal wavelet parameters used in WPT, including wavelet basic function and decomposition level;(c)choose the dominant order of singular values in calculating WPSE to eliminate the influence of noise.Step 3:Identify damage in CCGBs with the constructed warning curve according to The basic procedures of the WPSE-based methodology for damage identification of CCGBs subjected to seismic excitation are as follows:These procedures are illustrated in The damage model of a real investigated CCGB is elaborated in this section. Two typical damage types are considered, with damage individually located at the bottom of piers and at pier-girder connections. Sixteen damage scenarios are configured. The El Centro seismic excitation is applied to generate the displacement dynamic responses, which are measured by sensors arranged on the curved bridge.The geometry of the CCGB, including the plane, elevation, and cross sections, is illustrated in Two typical damage instances, Damage I at the bottom of a pier and Damage II at a pier-girder connection, are introduced into the FE model to create damage cases. The damage is modeled by reducing the stiffness of relevant elements in damage areas, as shown in The El Centro earthquake record from the El Centro Earthquake webpage  is adoptx- and y-direction displacements of each pier are recorded by specific sensors, marked as n represents the pier ID in The sensor arrangement is illustrated in x- and y-directions, represented by The WPSE-based damage identification method is utilized to identify Damage I and II in the CCGB described in Structural damage identification is a typical inverse problem, with structural dynamic response as the object of analysis. There are many kinds of measurement data in practical engineering, such as acceleration, velocity and displacement. It is important to select an appropriate response type that is effective and sensitive to damage. In this study, The damage index Equation , is calcThe wavelet basis function and the decomposition scale are the most common wavelet parameters, both having important influences on the wavelet analysis results. The essence of wavelet analysis is to project the signal to a wavelet basis function, and the wavelet coefficients obtained characterize the dynamic features of the original signal. If the wavelet basis function is not properly selected, the unsuitable wavelet coefficients obtained reduce the accuracy of wavelet analysis. Simultaneously, the larger the decomposition scale, the higher the time frequency resolution of the signal, contributing to higher accuracy of the wavelet analysis. Nevertheless, this maneuver also increases the amount of calculation, resulting in information redundancy and a decrease in wavelet analysis efficiency. Therefore, the choice of optimal wavelet parameters is constantly an important topic in the field of wavelet analysis .s. For a given signal, the wavelet basis function corresponding to the lowest cost function value M is regarded as the optimal wavelet base. The decomposition scale can be considered comprehensively based on the cost function value and the calculation efficiency.Currently, no uniform standard exists to determine optimum wavelet parameters. The Shannon entropy based cost function method is an effective method for determining the wavelet basis function. In this calculation, a Shannon entropy based cost function  is definIn this study, twelve candidate wavelet basis functions, identified as db2, db4, db10, db15, sym2, sym4, sym6, sym8, rbio3.5, rbio3.9, rbio4.4, and rbio6.8, and five candidate optimal decomposition scales ranging from 3 to 7 are considered. The displacement response of the CCGB excited by El Centro seismic wave is taken as the analysis object. The values of cost function M are calculated using Equation , with thAs discussed in i represents the position or number of the measuring point. The index To make it applicable for identifying Damage II, the Identification results of Damage II using the It must be noted that, although the On the basis of the Specifically, those thresholds of the For a performance illustration, the proposed ) method  involvinThe identification results for all piers using The effect of seismic excitation is investigated to confirm the adaptability of the proposed method. The strong Whittier Narrows earthquake  is adoptIn practical applications, measured dynamic responses are inevitably polluted by noise. In the assessment of any damage identification method, effective identification of damage under noisy conditions is essential. Here, structural damage identification is undertaken using displacement responses with Gaussian white noise, and the robustness against noise of the WPSE-based damage detection method is discussed.SNR = 60 dB. To label the noise level, a signal-to-noise ratio (SNR) definition is presented asSVD is one of the most effective denoising tools. It can basically eliminate random noise and retain most of the useful information by choosing appropriate orders of singular values. This adaptive anti-noise technique can be used to enhance the robustness of WPSE-based indices because the WPSE inherits the characteristics of SVD, as discussed in SNR = 60 dB are shown in q equals 5 rather than the total number of singular values in Equations Ux with SN is the total number of Monte Carlo simulations; M is the total number of occurrences of warning during N simulations; Monte Carlo simulations are performed at different SNR levels to determine the maximum noise immunity and damage identification accuracy of the proposed method in noisy conditions. Two indices, the missing report rate (In Equations  and 21)21), MRR SNR = 50 dB, implying that the identification of damage below 5% will result in a greater error. In terms of SNR = 50 dB, but the accuracy of early warning of damage decreases quickly to less than 80% for SNR = 40 dB. Therefore, the damage indices SNR≥ 50 dB.In this study, To simplify the application of If we select SNR≥ 50 dB. The benefit of the WPSE-based method was further clarified by comparisons with the WPE-based method. The comparison results showed that the WPSE-based method recognized the location of damage with higher accuracy. Thus, the proposed WPSE-based approach is reliable and applicable to seismic damage identification in CCGBs. This WPSE-based approach holds significant promise as a support to advanced distribution sensing techniques, especially to the distributed optical fiber sensors used in SHM of critical civil infrastructures.In this study, two WPSE-based evaluation indices were proposed to identify seismic damage in CCGBs, by taking synergistic advantage of the wavelet packet transform, singular value decomposition, and information entropy. The effectiveness of the proposed approach was numerically verified by a finite element model of a real CCGB subjected to El Centro seismic excitation. Numerical results showed that the two WPSE-based indices,


Opioids remain among the most effective pain-relieving therapeutics. However, their long-term use is limited due to the development of tolerance and potential for addiction. For many years, researchers have explored the underlying mechanisms that lead to this decreased effectiveness of opioids after repeated use, and numerous theories have been proposed to explain these changes. The most widely studied theories involve alterations in receptor trafficking and intracellular signaling. Other possible mechanisms include the recruitment of new structural neuronal and microglia networks. While many of these theories have been developed using molecular and cellular techniques, more recent behavioral data also supports these findings. In this review, we focus on the mechanisms that underlie tolerance within the descending pain modulatory pathway, including alterations in intracellular signaling, neural-glial interactions, and neurotransmission following opioid exposure. Developing a better understanding of the relationship between these various mechanisms, within different parts of this pathway, is vital for the identification of more efficacious, novel therapeutics to treat chronic pain. The MOPr is a G-protein coupled receptor that couples to inhibitory heterotrimeric G-proteins (Gductance . MOPr exductance .In vitro electrophysiology studies have shown that opioids reduce the frequency of spontaneous mIPSCs in PAG (in vivo studies. Microinjection of bicuculline (GABAA agonist) into the PAG produces antinociception, which suggests that GABA is being tonically released  that hyperpolarize GABAergic neurons in the PAG producing an overall decrease in GABAergic neuron activity Figure . AlternaThe overall effect of MOPr activation in the PAG is an increase in output to the two distinct cell types within the RVM: off-cells and on-cells . The actAt each level of this pathway, a myriad of cellular effects drives the physiological changes mentioned above, and are highly correlated with the development of opioid tolerance. One of the most studied mechanisms involves regulation and signaling at the MOPr. Current research demonstrates that while MOPr is a key player in the development of antinociceptive tolerance, mechanisms beyond simple receptor desensitization, including alterations in neurotransmission and β-arrestin dependent signaling, are also critical. Furthermore, MOPr expression in non-neuronal cells, specifically on microglia and astrocytes within the spinal cord, and more recently within the PAG, greatly contributes to the development of opioid tolerance.Evidence suggests that tolerance is due to changes in the properties of GABAergic neurons in the PAG . First oHowever, the development of tolerance within the PAG produces downstream effects along the descending pain pathway. This is evidenced by the fact that direct injection of morphine into the PAG affects RVM signaling, suggesting that their activity is in fact coupled . While aThe neurophysiological mechanisms of tolerance in the PAG are mediated by MOPr uncoupling from downstream G-protein mediated signaling Figure . One keyGABA release is also regulated by signaling through phospholipase A2-mediated activation of voltage gated potassium channels Figure . This siDirect activation of the MOPr results in the Gα subunit-mediated inhibition of the AC-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway Figure . Howeveri/o-proteins from the receptor and a switch to coupling with Gs-proteins  or intra-PAG injection, during morphine pretreatment has been shown to block the development of morphine tolerance , is particularly enriched in supraspinal midbrain regions known to regulate spinal nociception . There iN-methyl D-aspartate receptors (NMDArs) have been heavily implicated in the development of both spinal-mediated hyperalgesia and opioid tolerance. NMDAr antagonists or the targeted disruption of the NR2 subunits, NR2a and NR2b, attenuates opioid tolerance  is found to be upregulated following repeated intrathecal morphine administration, and this effect is mediated by activation of PI3K/AKT following MOPr activation . AdminisOver the past few decades, researchers have discovered that opioids are potent activators of immune cells within the CNS, and this inflammation is a strong contributor to the development of opioid tolerance . SpecifiOne prominent pro-inflammatory signaling cascade that has been implicated in opioid tolerance involves the immune receptor, toll-like receptor 4  and IL-1β, which are known to increase neuronal AMPA and NMDA receptors, as well as down regulate GABA receptors . Within Another potential point of cross talk is via purinergic receptors, specifically P2X4 and P2X7, which are primarily expressed on microglia. These receptors are also capable of upregulating pro-inflammatory cytokines, and blocking their activity in the spinal cord attenuates morphine tolerance . P2X4 acThe descending pain pathway is a critical modulator of nociception and plays an important role in mediating endogenous and exogenous opioid-induced analgesia. Because of this, it is highly implicated in allostatic cellular and molecular changes following repeated opioid use that lead to the development of tolerance. While this review has touched on a number of those changes at each level of the descending pain pathway, including desensitization of MOPr, altered cellular excitability and signaling, and induction of immune-competent cells, we do not yet have a complete understanding of all the factors that might be contributing to opioid tolerance.Much of the literature on opioid tolerance has focused the effects of morphine on this system. Future research must expand to include other commonly used opioids, especially in light of the increasing use of oxycodone and fentanyl, as each of these has widely different pharmacokinetic and signaling profiles, and may have differential effects on each level of the PAG-RVM-DH pathway. Indeed, studies looking at cross-tolerance between opioid analgesics suggest that differences in the distribution of the drug within the pain pathway may be differentially contributing to the development of tolerance. Furthermore, the cellular signaling pathways initiated within these spinal and supraspinal regions following administration of different opioids are known to vary.Finally, the research design of the studies related to opioids and tolerance has varied widely in terms of not only the drugs used, but also routes of administration, length of exposure, and use of biological systems. Also, the majority of studies on opioid tolerance have focused on males and have largely excluded females. Given that males show greater morphine potency, tolerance, and activation of neurons from PAG to RVM following morphine, as compared to females , it is iLL, AF, and EB contributed to the writing and editing of this 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.


Recently, many studies have been conducted to investigate the effects of exergames on the social well-being of older adults.The aim of this paper is to synthesize existing studies and provide an overall picture on the social effects of exergames on older adults.A comprehensive literature search with inclusive criteria was conducted in major social science bibliographic databases. The characteristics of exergames, participants, methodology, as well as outcome measurements were extracted from the relevant studies included in the review. The bibliometric and altmetric outreach of the included studies were also investigated.A total of 10 studies were included in the review, with 8 studies having used the Nintendo Wii platform. Most of the studies recruited healthy older adults from local communities or senior activity centers. Three groups of social-related outcomes have been identified, including emotion-related, behavior-related, and attitude-related outcomes. A metric analysis has shown that the emotion-related and behavior-related outcomes received high attention from both the academic community and social media platforms.Overall, the majority of exergame studies demonstrated promising results for enhanced social well-being, such as reduction of loneliness, increased social connection, and positive attitudes towards others. The paper also provided implications for health care researchers and exergame designers. Significant population aging has been experienced by countries worldwide. In 2015, a report from the United Nations . Older aWith the advent of digital technology, exergames, which combine digital gaming and physical exercise, are commonly used as daily exercise programs . DespiteThe social effects of exergames have drawn considerable attention from researchers -19. ExerConsidering the major concerns regarding older adults with social disorders, it is important to have an overview on whether exergames may serve as an effective intervention for the social well-being of this group of people. In the literature on exergaming, many systematic reviews examined only on the physical and cognitive benefits of exergaming on older adults -25. SomeThe current review adopted the definition of an exergame from Oh and Yang  who defiA comprehensive literature search was conducted in major social science bibliographic databases, including PsycINFO, PubMed, CINAHL, and ScienceDirect. Potential studies were identified by the combination of exergame terms (exergame OR Wii OR Kinect OR active video game), social terms , and ageing terms (aging OR aged OR elderly OR older OR senior). Reference lists of the included studies and relevant reviews were also inspected for additional studies to be included in the current systematic review. A total of 319 articles published before 22 January 2017 were retrieved for review and analysis. All articles were assessed using either the title, abstract, or full text to determine their eligibility in the systematic review conducted in this paper. The articles without full texts in any of the databases listed above excluded if the full texts could not be retrieved using online search engines or by contacting the authors directly. After identifying the final list of included studies, the characteristics of exergames, participants , methodology (study design and duration), as well as outcome measurements were extracted from the studies. Screening and data extraction was performed mainly by one reviewer, while a second reviewer was assisted by checking and editing the extracted data.To investigate the bibliometric score (in terms of citation count), the altmetric score, and the social media presence of the articles included in the systematic review, we collected citation counts from Scopus , as wellAccording to the inclusive criteria, a total of 10 studies were eligible to be included in the final review process. Cooking Mama and party simulation-styled Wii Party.Of the 10 studies investigated, 8 investigated social effects of exergames using Nintendo Wii, while 2 studies used Microsoft Xbox Kinect. Both platforms are the most popular exergaming platforms in the current market, and both offer console-based devices and games which make exergaming possible in the home setting. In terms of game topics, it is interesting to note that half of the studies (n=5) applied games from Nintendo Wii Sports package -43. The Most of the studies recruited healthy older adults from local communities or senior activity centers. There were 2 studies ,44, howeFive of the 10 studies applied poststudy qualitative methods to assess the social effects of exergames, such as semistructured interview, semistructured group interview, or focus group discussion. Among those applying quantitative methods, 4 studies tested the effects between exergames and other control conditions. For example, Wu et al , Jung etThe findings of the included studies have identified several social-related outcomes. Based on the different natures of the outcomes, they were categorized into three groups: emotion-related, behavior-related, and attitude-related.Loneliness was identified to be the main emotion-related social outcome affected by exergames. Jung et al conducted a study to assess the potential of Nintendo Wii in improving the quality of life among older adults in a long-term care facility . Their rAdditionally, Xu et al found a significant decrease in loneliness among older adults after exergaming, although little differences were found across different play types or age groups (young-old vs old-old) . In the Wu et al  presenteCurrently, the study from Agmon et al  has receAgmon et al had a high count of 180 Mendeley readers, but since Altmetric.com does not include Mendeley readers in its score, this is not reflected as this study had a score of 3 . Again, While previous reviews have synthesized the psychosocial effects of exergames ,28,29, tAlthough the social benefits of exergames are often discussed in the literature on exergames, there has been no particular review found on this topic. By summarizing the existing original studies, the findings from the current review showed that exergames could be an effective intervention for social improvements among older adults. The review supported the finding that exergames were able to reduce the loneliness level among older adults. The decrease in loneliness was perhaps not due to playing the exergame itself, but rather due to the increased interactions between the participants and other players. In a large sample survey study, Lee and Ishii-Kuntz  indicateAttitude-related social outcomes are a new finding which have not been reported in previously published exergame reviews (eg, those published by Chao et al  or MatalThis review showed that that Nintendo Wii was the most frequently used exergaming platform in the included studies. This finding is supported by another review , which rAlthough most of studies targeted healthy older adults, two studies examined the social effects on older adults with physical disabilities ,44. PhysThe study designs included in this study varied in rigor, with 6 studies applying a within-group design, and 4 studies applying between-group design by comparing exergames with a control condition. Although the majority of studies showed promising results for the use of exergames for social enhancement, the conclusions need to be interpreted with caution due to the limited number of randomized controlled trials. The included studies were either predominantly small pilot trials or feasibility studies; they lacked the adequate sample sizes needed for a powered efficacy trial. Meanwhile, half of the studies used qualitative methods for data collection. Without validated quantitative instruments, their findings do not have the capacity to detect significant changes in social outcomes.There are some limitations in the review. Due to the limited number of identified studies, the systematic review included articles with both qualitative and quantitative analyses. The quality assessment of the included studies was difficult to conduct, and it was not possible to produce mean effect sizes via a meta-analysis. However, the review was broad in scope and included a diversity of study conditions and social outcome measures. Another limitation is that the key conclusions should be interpreted with caution due to the small number of included studies. Furthermore, relevant studies may have been unintentionally excluded because of the specific keywords used and the databases selected. Lastly, a publication bias, particularly language bias, might have occurred because we restricted the search to English language publications.


Leishmania (L.) major, Leishmania tropica, Leishmania braziliensis, L. mexicana, and L. amazonensis. The burden of CL in terms of morbidity and social stigmas are concentrated on certain developing countries in Asia, Africa, and South America. People with asymptomatic CL represent a large proportion of the infected individuals in the endemic areas who exhibit no lesion and can control the infection by as yet not fully understood mechanisms. Currently, there is no approved prophylactic control measure for CL. Discovery of biomarkers of CL infection and immunity can inform the development of more precise diagnostics tools as well as curative or preventive strategies to control CL. Herein, we provide a brief overview of the state-of-the-art for the biomarkers of CL with a special emphasis on the asymptomatic CL biomarkers. Among the identified CL biomarkers so far, direct biomarkers which indicate the actual presence of the infection as well as indirect biomarkers which reflect the host's reaction to the infection, such as alterations in delayed type hypersensitivity, T-cell subpopulations and cytokines, adenosine deaminase, and antibodies against the sand fly saliva proteins are discussed in detail. The future avenues such as the use of systems analysis to identify and characterize novel CL biomarkers are also discussed.Cutaneous leishmaniasis (CL) is an immune-mediated skin pathology caused mainly by  Leishmania genus which pose serious public health problems in the endemic regions across 98 countries  or Lutzomyia  genera or an amastigote form inside parasitophorous vacuoles within the phagocytic cells (mostly macrophages) of its vertebrate host  phagocytosis and killing of the parasites (ii) formation of neutrophil extracellular traps (NETs) and releasing different microbicidal agents associated with DNA backbone . LST remains a powerful biomarker to distinguish the subpopulations with respect to asymptomatic CL. Moreover, LST results can be interpreted as an indicative of the developed immunological memory  in the outcome of CL  and Th17 cells have also been identified to regulate immunity to CL. Treg are known to function as the sustainers of tolerance and preventers of excessive damages during the inflammatory responses  found in macrophages mediates deamination of the anti-inflammatory nucleoside adenosine to inosine. The level of ADA has been shown to be significantly increased in sera and lymphocytes of CL patients  were higher than those with chronic CL (duration ≥2 years) and also in the skin of the uninfected people . In CL, arginine acts as a double-edged sword so that while it is needed for nitric oxide (NO)-mediated parasite killing, it can promote polyamine-mediated parasite replication  has been recently tested as a putative biomarker of exposure to sand fly in Tunisia  have been shown to be comparable in the peripheral blood of the healed and the asymptomatic individuals in an endemic area of L. major infection in Iran. However, mRNA level of the Treg transcription factor FOXP3 was shown to be higher in the asymptomatic group compared to the symptomatic patients. The mRNA level of IL-17 transcription factor RORC, on the other hand, was not significantly different between the groups , were substantiated. Similarly, the levels of IL-2, IL-12p70, IL-13, and IL-18 were shown to be significantly higher in all LST+ individuals but not in LST- cohort. Once validated, such biomarkers could be potentially used to stratify patients for different clinical/sub-clinical manifestations of the disease.Recently, a cross-sectional immune profiling study on PBMC of individuals living in endemic areas of Ph. papatasi could act as a serum biomarker of exposure , or proteins thereof may be used to determine the risk of contact, vector surveillance and as such could inform the disease management programs  species has also been reported  parasites in tissues and its availability in tissues accessible to sand flies were demonstrated in that study. By considering two body sites consisting of the border of an active lesion and a normal skin site, viable parasites were detected in the unaffected skin of these individuals (Rosales-Chilama et al., Leishmania species persist inside the skin of the asymptomatic individuals. To substantiate this, efforts should be made to analyze the skin and blood of the asymptomatic subjects with confirmed DTH, in the highly endemic areas.There is a dearth of documents regarding the parasite detection in either skin or blood of the asymptomatic patients of the Old World CL. Of note, the detection of L. tropica as compared with the healthy normal skin (Masoudzadeh et al., L. tropica infection during the acute phase of the disease. Another recent transcriptomics study of lesion biopsies from L. braziliensis patients has identified B cell activation and immunoglobulin transcript signatures in the lesions, depending on the presence or absence of the parasite transcripts (Christensen et al., Leishmania species will be identified (Patino and Ramírez, While few indicative biomarkers of CL infection and immunity have been proposed using conventional approaches, systems-level data integrating different layers of information on host response to CL infection are scarce. Such information can provide a comprehensive understanding of the disease and may presumably identify key biomarkers of CL disease and immunity. The recent technological advances in high-throughput omics technology combined with systems biology approaches have begun to provide new insights into pathogenesis of different diseases, and to unravel the molecular signatures of vaccine-induced responses in humans. Recently, whole genome transcriptomics analysis has been applied to CL. We have reported the transcript changes in the lesion of patients infected with Once available, such key information on biomarkers could inform the design of rational diagnostics and intervention strategies to control CL. In particular, this can help to screen the populations living in the endemic areas for asymptomatic individuals, which can in turn provide invaluable information on how gender, ethnic or geographical factors would affect the asymptomaticity. To achieve this, accurate, non-invasive and rapid methods with the ability to distinguish latent CL from the asymptomatic CL are needed. It should be noted that more awareness at global and national levels is also required to pave the way for developing cost-effective diagnostics and intervention strategies for the expanding inflicted populations.FB, AH, and SR drafted the manuscript. All the authors provided critical feedback on the manuscript prior to publication and have agreed to the final content.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.


Lactobacillus and Bifidobacterium, many of them with probiotic characteristics, in the intestinal environment. Current knowledge on genetics and biosynthesis pathways of EPS in lactic acid bacteria and bifidobacteria, as well as the development of genetic tools, has created possibilities to elucidate the interplay between EPS and host intestinal mucosa. These include the microbiota that inhabits this ecological niche and the host cells. Several carbohydrate recognition receptors located in the intestinal epithelium could be involved in the interaction with bacterial EPS and modulation of immune response; however, little is known about the receptors recognizing EPS from lactobacilli or bifidobacteria and the triggered response. On the contrary, it has been clearly demonstrated that EPS play a relevant role in the persistence of the producing bacteria in the intestinal tract. Indeed, some authors postulate that some of the beneficial actions of EPS-producing probiotics could be related to the formation of a biofilm layer protecting the host against injury, for example by pathogens or their toxins. Nevertheless, the in vivo formation of biofilms by probiotics has not been proved to date. Finally, EPS produced by probiotic strains are also able to interact with the intestinal microbiota that populates the gut. In fact, some of these polymers can be used as carbohydrate fermentable source by some gut commensals thus being putatively involved in the release of bacterial metabolites that exert positive benefits for the host. In spite of the increasing knowledge about the role that these surface molecules play in the interaction of probiotic bacteria with the gut mucosal actors, both intestinal receptors and microbiota, the challenging issue is to demonstrate the functionality of EPS in vivo, which will open an avenue of opportunities for the application of EPS-producing probiotics to improve health.Exopolysaccharides (EPS) are surface carbohydrate polymers present in most bacteria acting as a protective surface layer but also interacting with the surrounding environment. This review discusses the roles of EPS synthesized by strains of  Bacteroides fragilis  located into the cytoplasm or structural polymers (glycans) which form part of the microbial envelope. Glycan polymers can be tightly linked to the cell surface forming a capsular polysaccharide, (CPS), loosely attached to the extracellular surface, or secreted to the environment as exopolysaccharides (EPS). In bacteria these polymers have received special attention because they play a relevant role in the interaction between pathogens and the host, being in some cases virulent factors responsible of pathogenesis , together with some from genus Bifidobacterium (phylum Actinobacteria) are used in the formulation of foods that promote health, which are known as functional foods  are a particular group of Gram-positives that are involved in many natural food fermentations but there are also widely used in food manufacture industry. In addition, a few species of this group mainly those belonging to genus EPS-producing LAB have a long tradition of use in dairy fermentations because the polymers help to improve the texture, mouthfeel, and viscosity of fermented milks, being also effective fat-replacers, avoiding syneresis, or whey separation  and heteropolysaccharides (HePS), are synthesized by LAB or bifidobacteria  and N-acetyl-galactosamine , fucose, glucuronic acid, glycerol, or mannose can also be present , involved in the biosynthesis of the repeating units, proteins related to polymerization and export of these units, as well as other genes with unknown functions including mobile elements. In addition, genes involved in the regulation of HePS synthesis are detected in lactobacilli but not in bifidobacterial eps clusters , nucleotide oligomerization domain-like receptors (NODs), RIG-1-like receptors (RLRs) and lectin receptors recognizing glycan-containing molecular structures. There are three classes of innate lectin receptors; the galectins which bind to beta-galactosidase sugars, the siglecs which bind to sialic acid containing glycans, and the C-type lectin receptors (CLRs) which bind to various glycans. The CLRs belong to the superfamily of C-type lectins comprising more than 1,000 proteins with a common protein fold known as the carbohydrate recognition domain (CRD) and have diverse functions in homing and trafficking of immune cells, T cell activation, innate recognition of microbes and regulation of immune responses.The secreted soluble CLRs involved in innate immunity include SP-A, SP-D, mannose binding lectins (MBLs) also termed mannose-binding protein (MBP), that can bind to microbes and promote direct or indirect killing, phagocytic uptake, or activation of the lectin pathway of complement activation  EPS  the metabolic labeling of the bacterium using 3H nucleotides  using the endogenous biosynthetic machinery of the bacterium. The azide group permitted the incorporation of a fluorescent label, which could then be used to detected and quantify the fluorescent bacteria in vivo , was constructed. TEM images showed the wild-type strain was surrounded by a dense capsular layer, which was absent in the ΔcpsD mutant, revealing the presence of multiple fimbriae  in which we constructed an isogenic mutant strain (S89L) which differs to wild-type in a single nucleotide in the gene involved in the chain-length determination. The mutant S89L has a “ropy” phenotype due to the higher production of a high molecular weight (HMW), rhamnose-rich EPS fraction  in comparison with the parental strain . Bacteria, archaea, fungi, protozoa and viruses are the main inhabitants of our intestine, in which they stablish commensal, mutualistic or parasitic relationships with the host that shape our physiology. Among them, bacteria are the most abundant members, being Firmicutes and Bacteroidetes the main bacterial phyla, although other less abundant, such as Actinobacteria or Proteobacteria, are also present. All together, these microbes provide us with a significant metabolic potential encoded in the metagenome; thus the intestinal microbiota has been postulated as another organ of the human body  prebiotics. In this case their large molecular weight could difficult the enzymatic hydrolysis, although the lack of human enzymatic activities specific for these polymers could also be a plausible explanation. Additionally, several studies have demonstrated that the simple primary structure allows them to be fermented by fecal microbiota. As an example, is the case of the HoPS isolated from in vivo. Wistar rats fed with EPS-producing B. animalis subsp. lactis or B. longum showed an increase in the levels of intestinal bifidobacteria analyzed in caecum content and feces (Salazar et al., L. rhmanosus GG in the gut was higher, than its mutant strain lacking the polymer, due to the protective role of the HePS against immune innate factors (Lebeer et al., In vitro models also showed that the susceptibility of HePS to biodegradation in the gut depends on the physical-chemical composition of the polymer. In general, simplest structures of the repeating units building these polymers are easily hydrolysed by enzymes. As example, the HePS from Streptococcus thermophilus Sfi39 and Sfi12 were degraded in higher extent than those of Lactobacillus sakei 0-1 and Lactobacillus helveticus Lh59, which had more complex structures (Ruijssenaars et al., in vitro fermentations (Salazar et al., In the case of HePS, their more complex structure could lead to a high resistance to gastrointestinal digestion but this has not been demonstrated Lactobacillus and Bifidobacterium seem to be resistant to gastrointestinal digestion, but they are susceptible of biodegradation by the intestinal microbiota, this feature being dependent on the physicochemical characteristics of the EPS as well as on the pool of glycolytic enzymes harbored by gut microorganisms. Besides, monosaccharides released from biodegradable EPS could also have influence on microbiota composition through cross-feeding mechanisms (Rios-Covián et al., In summary, HoPS and HePS synthesized by in vitro and in vivo models. One of the traditional phenotypic features tested during in vitro probiotic characterization is the ability to co-aggregate with pathogens, since it is believed that co-aggregation decreases the accessibility of the harmful microorganisms to the intestinal epithelium. In this regard, high EPS producing strains of Lactobacillus delbrueckii subsp. bulgaricus were shown to be associated with a high co-aggregation ability with Escherichia coli (Aslim et al., E. coli-induced inflammatory response by the EPS of L. delbrueckii TUA4408L on porcine intestinal enterocytes (Wachi et al., Bacillus cereus toxins on the monolayer integrity of Caco-2 human colonocytes (Ruas-Madiedo et al., Candida of L. rhamnosus GG, its EPS deficient mutant, and the purified EPS was analyzed showing that the EPS layer might be involved in the reduction of hyphal formation and in the competition with Candida during adhesion to vaginal epithelial cells (Allonsius et al., Lactobacillus paraplantarum BGCG11, and its non-EPS-producing isogenic strain NB1, against seven opportunistic pathogens was determined on HT29-MTX cell monolayers, pointing out that the EPS of the parental BGCG11 strain could be involved in the reinforcement of the innate mucosal barrier by hindering the contact of pathogens, such as E. coli or Listeria monocytogenes, with the epithelial cells (Živković et al., Lactobacillus paracasei subsp. paracasei BGSJ2-8 EPS-deficient mutant, producing an EPS with a different composition than the wild type, indicated that the wild type EPS is essential to reduce E. coli association with Caco-2 cells, while the mutant strain was not able to counteract this interaction (Živković et al., The antagonistic effect of EPS-producing lactobacilli and bifidobacteria, and in particular cases their isolated EPS, against microbial pathogens has been studied using in vivo experiments, different animal models have been used to study how EPS-producing strains interfere with pathogen activity. For instance, L. johnsonii FI9785, a poultry-derived isolate producing two different exopolysaccharides, was effective to supress colonization and persistence of Clostridium perfringens in poultry and to reduce colonization by E. coli of the small intestine (La Ragione et al., B. longum subsp. infantis 35624, which synthesizes an EPS containing galactose, glucose, galacturonic acid, and 6-deoxy-L-talose, decreases the effect of Salmonella on brush border enzyme activity and weight loss in mice (Symonds et al., B. breve UCC2003, and their isogenic non-EPS producing isogenic mutant, are those providing more solid evidence of the role of EPS in pathogen antagonism (Fanning et al., C. rodentium, suggesting a role of EPS in conferring the health benefits normally associated with probiotic strains.Regarding Lactobacillus and Bifidobacterium might act as effector surface macromolecules that are directly involved in the interaction of these bacteria with different host cells as well as with the microbial community that occupy the intestinal lumen. In order to understand these complex interactions is pivotal to gain more insight into the biosynthetic machinery of these polymers, as well as to determine which are the key properties of the EPS that are important for their biological functions. Thus, combination of high-throughput techniques allowing to take advantage of the information codified in the genomes of the producer strains, in combination with powerful analytical techniques and in vitro models better reflecting the in vivo physiology of the host, will be valuable tools for the establishment of this correlation genes-structure-function. This knowledge would facilitate use of synthetic biology to obtain tailor-made polymers, opening up possibilities to enhance attributes considered positive for health. It would allow to improve the production of polymers with known benefits given that, due to their low production yield, the application of purified polymers is not affordable to date. Furthermore, the biodiversity of EPS synthesized by bacteria from nature can be further explored to find novel biotechnological applications. Thus, research on bacterial EPS is a relevant topic with considerable prospects for health and biotechnological applications.EPS produced by Box 1Well-known statementsThe genetic determinants for EPS synthesis have been identifiedThe physical-chemical properties of EPS determine their biological propertiesEPS help the survival of the producing bacteria under the gastrointestinal challengesThe EPS surrounding the bacterial surface reduces adherence to intestinal cells due to shielding of surface macromolecules acting as adhesinsIsogenic EPS-producing and non-EPS-producing strains induce different immune responsesEPS are resistant to gastrointestinal digestion and they are susceptible of biodegradation by the intestinal microbiotaSpecific EPS-producing strains act as antagonists of pathogens in the gutFuture perspectivesLactobacillus and BifidobacteriumTo gain insight in the biosynthetic route of EPS formation in To know whether, or not, bacterial EPS activate or inhibit innate immunity via specific pattern-recognition receptorsTo decipher the mechanisms behind the beneficial properties of EPSin vivo demonstrate the EPS production by intestinal lactobacilli and bifidobacteria.To JMW, AM, and PR-M contributed with the organization and structure of the review and NC-B, JMW, AM, and PR-M contributed to the writing. PR-M was in charge of organizing the drafted manuscript and all authors performed a critical revision of the manuscript and approved the final version.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.


One approach for reducing alcohol and other drug problems is community-based prevention programs. These programs focus on changing the environment in which a person consumes alcohol rather than the behavior of the individual drinker. Several international and U.S. programs have assessed the effectiveness of such approaches in reducing alcohol-related problems. Some of those analyses have had inconclusive results. Others, however, found reductions in alcohol-related problems such as drunk driving, alcohol-related car crashes and their consequences, the sale of alcohol to underage drinkers, and assault injuries. Nevertheless, several aspects of community-based prevention programs require further study. For some chronic health problems, such as cardiovascular disease (CVD), community-based prevention programs have been effective in reducing those problems . AccordiComparing programs to reduce AOD and CVD as an example, the following four differences arise. First, interventions for high-risk medical conditions, such as changing dietary habits for CVD, operate under the assumption that people suffering from this condition have the power to control their behavior rationally. Conversely, efforts to reduce alcohol-related problems do not necessarily operate under this assumption.Second, both greater needs and greater opportunities exist for regulating behaviors associated with alcohol-related problems than for regulating behaviors associated with CVD. For example, unlike poor dietary habits and smoking, which primarily affect the person exhibiting these behaviors (with the notable exception of secondhand smoke), alcohol consumption impacts the broader community system through traffic and other injuries. At the same time, alcohol consumption occurs within a highly regulatable distribution system of bars, restaurants, and other establishments.Third, the consumption of alcohol often is more closely linked in time and space to the resulting alcohol-related problems  than are poor dietary habits  and the resulting CVD, which may develop only after years of unhealthy eating. And fourth, societal norms associated with drinking differ dramatically from those associated with problematic dietary patterns. For example, because the decisions made by drinking drivers have consequences beyond the drinker him- or herself, the drinker’s behaviors are perceived as legitimate targets for social control and regulation.Thus, although much may be learned from the experiences of CVD prevention programs, the specific methods that effectively reduce chronic health problems may be somewhat less applicable to the reduction of alcohol-related problems. The circumstances that surround alcohol consumption and the generation of alcohol-related problems may present unique challenges and strategic opportunities for the development of community prevention programs. In fact, with these differences in mind, researchers, community stakeholders, and funding agencies have increasingly turned to so-called environmental approaches for reducing alcohol-related problems. This article reviews current research on the effectiveness of such approaches. After briefly describing the theoretical framework of community-based environmental approaches, the article presents programs that have been implemented in the United States and in other countries and discusses future directions in the study of environmental approaches. The prevention projects included in this review were selected using criteria that, in the absence of random assignment to treatment and control conditions,The studies include a careful collection of baseline data during the period preceding the intervention.They target well-defined community-level alcohol-related problems .They have a long-term implementation and monitoring period.They are followed by a final evaluation of changes in target problems.They involve empirically documented, successful or at least promising results in the target problem that are attributable to the intervention.In general, community-based environmental prevention programs as described here  focus not so much on changing the behavior of the individual drinker but on changing the environment in which a person consumes alcohol. The difference between environmental approaches and other approaches can best be clarified by considering the difference between “problem drinking” and “drinking-related problems.” The term “problem drinking” describes the behavior of an individual, and the treatment and prevention of problem drinking address the personal costs of drinking to the individual drinker, such as jeopardizing health, work, and family life. Conversely, “drinking-related problems” refers to consequences of alcohol consumption that affect many other people besides the individual drinker, including family, colleagues, neighbors, and other members of the community. These consequences include the health, social, and economic costs of drinking to the larger community, such as alcohol-related traffic crashes, alcohol-involved violence , and school dropout rates.On the individual level, problem drinkers disproportionately contribute to drinking problems . Nevertheless, because most people who consume alcohol cannot be classified as problem drinkers, the majority of drinking-related problems arise from nonproblem drinkers. As a result, community-based environmental prevention efforts seek to address these wider-scale problems caused by nonproblem drinkers rather than those caused by problem drinkers.Because of this broader scope, environmental approaches to the prevention of alcohol-related problems operate differently than do individual treatment and prevention approaches. For example, environmental approaches focus on the community as a system involving numerous components, including the following:Individual drinkersVendors of alcohol, including both places where alcohol is consumed  and places where alcohol is sold Social events where alcoholic beverages are sold and consumedLocal laws, regulations, and enforcement agenciesLocal medical clinics and treatment facilitiesSocial organizations that may support and promote public health campaigns, including schools and PTAs, churches, business organizations, and social clubs.Other, less obvious components of the community system affecting and affected by alcohol problems include the social networks and family connections through which people “learn” drinking attitudes and behaviors. These social networks may play a particularly important role with underage drinkers, who most likely obtain alcohol through friends and family networks .Although various applications of such a comprehensive environmental approach to the reduction of alcohol problems differ in terms of specific targets and tactics, they are all characterized by several strategies that distinguish them from more traditional information-based approaches that aim to change the alcohol use practices of individuals. First, environmental approaches use the media to target policymakers, as opposed to the general public, in support of program goals. Second, environmental approaches focus on mobilizing the community to make structural and system changes rather than on persuading individuals to change their behaviors. Third, environmental approaches generally target the supply of alcohol through social and market systems as opposed to the demand for alcohol by individuals . In sum,Several interventions conducted outside the United States have provided useful guidelines for environmental prevention efforts. Together with efforts to evaluate so-called natural experiments,The Community Action Project, conducted during the early 1980s in six New Zealand communities, aimed to increase public support for alcohol regulation policies as well as induce changes in attitudes and behavior. The project involved a mass media campaign conducted in two communities, an intensive intervention media campaign and a community organizer in two other communities, and no intervention in two comparison communities  project, conducted in the Australian city of Geraldton between 1992 and 1995, was designed to reduce alcohol-related injuries by focusing not on heavy drinkers or alcoholics but on the general context of use in the community , and the Community Trials project. To illustrate the unique form and direction of environmentally based community prevention efforts, this section discusses these three projects in some detail.The Saving Lives Project, conducted in six Massachusetts communities with the rest of Massachusetts serving as a statistical control, aimed to reduce alcohol-impaired driving and related problems . The speThis project, which included 15 communities in Minnesota and western Wisconsin, aimed to reduce access to alcohol among underage youth a,b. The Analyses of the results of the project showed that merchants increased checks for age identification, reduced sales to minors, and reported more care in controlling sales to youth in the experimental communities b. AttempThe Community Trials Project  was a fiA media and mobilization component to develop community organization and support for the goals and strategies of the project and to increase local news to raise public support for the goals and strategies of the projectA responsible beverage service component to reduce service to intoxicated patrons at bars and restaurantsA sales to youth component to reduce underage access to alcoholA drinking and driving component to increase local driving-while-intoxicated enforcement activityAn access component to reduce the availability of alcohol.Postintervention analysis demonstrated several important improvements in the experimental communities versus the comparison communities. For example, nighttime crashes resulting in injuries were 10 percent lower and crashes in which the driver was found by police to have been drinking were 6 percent lower in the experimental communities than in the comparison communities. Moreover, assault injuries observed in emergency departments declined by 43 percent, and assault injuries requiring hospitalization declined by 2 percent in the intervention communities versus the comparison communities. In addition, as indicated in a random-digit dialed telephone survey of community residents, self-reports of driving after having had “too much to drink” declined by 49 percent and self-reports of driving when “over the legal limit” declined by 51 percent in the experimental communities compared with the comparison communities. And although the drinking population increased slightly in the experimental communities over the course of the study, problematic alcohol use was significantly reduced in those communities . FinallyAlthough the studies reviewed above support the efficacy of community-based environmental prevention approaches, further research is needed to determine which aspects of this approach are the most promising or effective. Four key areas of such research are described below.First, the relative efficacy of different preventive interventions must be assessed so that communities with limited resources can choose between alternative intervention strategies and select the most effective programs for their specific needs. To date, however, the efficacy of individual strategies must be inferred primarily from the programs’ effects on intermediary variables  because any distal effects  reflect the contribution of all interventions working in concert. For example, the combination of increased enforcement of laws against drinking and driving and increased coverage of that enforcement has been linked to increases in perceived risk of arrest for drinking and driving . This increased perception of risk has, in turn, been linked to decreases in drinking and driving and subsequent automobile crashes  .Similarly, merchant training, enforcement, and media advocacy, when used in combination, can be effective in reducing underage purchases of alcohol ; traininSecond, little information exists regarding the effectiveness of environmental programs from a cost-benefit perspective. For example, investigators still must determine the return on funds spent on such programs in terms of costs saved for treatment of alcohol-related injuries or lives saved. Communities facing the allocation of scarce resources will be very interested in the answers to such questions.Third, the appropriate geographic focus of specific interventions remains unclear. Thus, future analyses must clarify whether interventions are best implemented at the community level or more locally, at the neighborhood level. One can argue that certain problems  are of a community-wide nature whereas others  are of a more local nature. The questions of who “owns” such problems or who is best equipped to address them are far from resolved.Fourth, more research must be directed toward establishing the efficacy of local prevention programs in racial/ethnic minority neighborhoods, which typically experience different alcohol problems from those of non-minority communities. Toward this end, the National Institute on Alcohol Abuse and Alcoholism is currently sponsoring a project conducted by the Prevention Research Center, based in Berkeley, California, in two largely Hispanic, low-income neighborhoods in northern California to reduce under-age alcohol access, youth drinking, and related problems. This action project, to be conducted over a 5-year period, will include the following components:A community awareness component disseminating information about youth and young adult alcohol access and useA responsible beverage service program focusing on service to minors and intoxicated patrons at on- and off-sale establishmentsAn underage access component providing support for increased police enforcement of underage sales laws and laws regarding provision of alcohol to minors by social hostsAn enforcement component focusing on laws regulating sales to intoxicated people both in establishments and at special events where alcohol is servedA community mobilization effort providing neighborhood support for the other components.This project, though similar to the Community Trials project in terms of its environmental approach, differs from that project in three important regards. First, project interventions are to be implemented at the neighborhood level as opposed to the community level. Second, project interventions have been tailored to address the unique drinking problems and patterns characteristic of neighborhoods with large proportions of low-income ethnic minorities, such as high density of alcohol outlets and associated high levels of youth violence  and inte


Since first isolation in 1947 from the Zika forest in Uganda, Zika virus (ZIKV) has been principally known as a benign agent associated with sporadic human infections in a restricted number of African countries. However, during 2015–2016, an Asian lineage of ZIKV caused an unprecedentedly large outbreak in the Americas and sizeable numbers of exported cases across the globe. In this review, we critically appraise the recent advances in molecular epidemiological studies of ZIKV performed to date, and we highlight the pivotal role played by genomic surveillance in elucidating the origins, dissemination and evolution of the Asian lineage of ZIKV in Asia and in the Americas. The positive sense, single-stranded RNA genomic structure of ZIKV and processing of the single encoded large polyprotein is characteristic of other flaviviruses, such as yellow fever virus and dengue virus. The majority of ZIKV infections are asymptomatic; however, in some cases can have a febrile presentation with mild clinical symptoms, such as, headache, skin rashes and conjunctivitis, which are typically self-limiting, resolve without consequence and do not require anti-viral treatment , which also segregated within the American ZIKV cases  of ZIKV over time using a birth–death skyline approach  and the recent Chinese imported case from the Maldives, a comprehensive survey of ZIKV and other flaviviruses in competent vectors in the Eastern Pacific region is urgently required. This would allow a finer scale delineation of the extant genetic diversity and detailed pathogenesis studies and, critically, to facilitate a rational, evidence-based development of vaccines to stable epitope targets to mitigate further disease outbreaks.In summary, although the ZIKV outbreak in the Americas ended suddenly (Netto


Plant proteases play a crucial role in many different biological processes along the plant life cycle. One of the most determinant stages in which proteases are key protagonists is the plant germination through the hydrolysis and mobilization of other proteins accumulated in seeds and cereal grains. The most represented proteases in charge of this are the cysteine proteases group, including the C1A family known as papain-like and the C13 family also called legumains. In cereal species such as wheat, oat or rye, gluten is a very complex mixture of grain storage proteins, which may affect the health of sensitive consumers like celiac patients. Since gluten proteins are suitable targets for plant proteases, the knowledge of the proteases involved in storage protein mobilization could be employed to manipulate the amount of gluten in the grain. Some proteases have been previously found to exhibit promising properties for their application in the degradation of known toxic peptides from gluten. To explore the variability in gluten-degrading capacities, we have now analyzed the degradation of gluten from different wheat cultivars using several cysteine proteases from barley. The wide variability showed highlights the possibility to select the protease with the highest potential to alter grain composition reducing the gluten content. Consequently, new avenues could be explored combining genetic manipulation of proteolytic processes with silencing techniques to be used as biotechnological tools against gluten-related disorders. Plant proteases have been described to accomplish multiple roles in different physiological processes along the plant life cycle, such as programmed cell death, senescence, abscission, fruit ripening, plant growth, and N homeostasis . In respCereals grains include proteins with many different functions. Around 80% of these proteins are storage proteins, packed in the endosperm together with starch and lipids . These pPharmaceutical, food and beverage, detergent, and biofuel industries have for long time exploited enzyme catalysis in commercial-scale applications, being the use of papain an example of success in food industry over the last four decades . In receIn cereal grains, different proteases have been involved in the germination process . In maizBarley is one of the grain cereals further studied. Proteases, amylases, dextrinases and other hydrolases are crucial for the survival of the seedling until the photosynthesis is finally established . In the HvPap-1 gene in barley transgenic plants alters the metabolite composition of the grain, and modifies the germination process by delaying or accelerating it  -DQ2 or -DQ8 . A lifelDifferent therapeutic alternatives are being developed, as the inhibition of transglutaminase, the antagonism of peptide binding to HLA-DQ2 or HLA-DQ8, the enzymatic detoxification of gluten, or the introduction of natural amino acid substitution to eliminate toxicity . Some otAspergillus niger (AN-PEP) and Sphingomonas capsulate (SC-PEP), with also gliadin degradative capacity, have been characterized   by four hia coli . With thHydrolysis of the different wheat gliadins by purified barley CysProt was observed by western blot analysis. When gliadins from the six wheat cultivars were treated with HvPap-1, -4, -6, and -16 for 1 and 12 h, different degradation patterns by each CysProt were observed . DegradaHvPap-6 gene from barley would have advantages in comparison of oral enzyme therapy since consumers will not be exposed to harmful gluten. Unwanted reactions in celiac patients probably would decrease by consuming lower or even null levels of toxic gluten from transgenic wheat lines. Administration of hydrolytic glutenases as food supplement is an alternative to deliver the therapeutic agents directly to the small intestine, for further degradation of wheat gluten. One step beyond would be to produce transgenic wheat lines combining the overexpression of a CysProt with high gliadin-degrading efficiency and the downregulation of the expression of gliadins using genome editing techniques. The next step should be to evaluate the in vivo efficiency of selected proteases for degrading wheat gliadins in the endosperm. For that, overexpression of these proteases under the control of endosperm specific promoters, such as those from HMW, gamma or alpha-gliadins would be necessary. A recent publication by 1Dy high-molecular wheat glutenin promoter (pHMWg). A significant reduction (up to 67%) of the amount of the indigestible gluten peptides of all prolamin families was achieved. This approach will contribute, in combination with other strategies like CRISPR/Cas9, to provide wheat varieties suitable for celiac and other gluten intolerance patients. Overall, with new goals, the potential of exploring the properties of plant proteases acting along seed germination will open a great field of research, as a biotechnological alternative to provide new prevention strategies in protein-caused disorders. In conclusion, this is an alternative approach for enzymatic gluten degradation to generate gluten-free wheat to manufacture gluten free products.The potential of plant proteases to alter grain content and the mobilization of stored proteins greatly supports their use as biotechnological tools. As an example, gluten proteins responsible of celiac disease and other gluten-related pathologies are suitable targets for these enzymes. For gluten intolerant patients, the only available treatment is a gluten-free diet, which is extremely difficult to maintain due to gluten ubiquity in human diets. Given the negative impact of quality of life of celiac patients, there is an urgent need to develop food suitable for them. In our research, we have provided new information on the potential of plant CysProt to reduce gliadin content. HvPap-6 CysProt ranks as the best candidate given the encouraging results of this study. The final aim would be the development of transgenic lines of wheat overexpressing this CysProt under endosperm specific promoters to reduce gluten toxicity. This cereal would be used afterward to obtain flour with which to elaborate products suitable for humans with this autoimmune enteropathy. Transgenic wheat lines overexpressing the All datasets generated for this study are included in the manuscript and/or the ID, FB, MM, and MD-M conceived the research. SG-C, MG, and MD-M performed the experimental research. All authors contributed to the final version of the manuscript, and read and approved the final 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.


This research will be a useful reference for exploring the multi-scale balancing of conservation and development for Winter Olympic regions, and in turn, for concentrated global constructions.Optimizing spatial patterns of land development and minimizing the ecological impact of concentrated construction is the key to realizing regionally sustainable development. The reasonable assessment of the ecological effects of the Winter Olympic construction on areas where the mountainous ecosystem is ecologically sensitive and vulnerable is urgent for responsible urban and regional development. Here, we assess the multi-scale suitability of ecologically compatible development in Winter Olympic regions using the ecological suitability assessment method based on GIS spatial analysis. We found the Chongli District had relatively high ecological structure and function resistances at a basin scale and that the towns where Olympic facilities located also had larger ecological resistance. The integrated suitability assessment showed the prior and moderate zones for suitable large-scale development and utilization in Chongli were smaller than those in other counties. The total loss area of natural ecological systems  for a new ski resort is 117.27 hm The regional development accompanied by land use structure changes is dependent on both territorial characteristics and land use policies8. The analysis of ecological suitability driven by historical, ecological and socioeconomic analysis may contribute to sustainable land management principles10 and the sustainable development of urban and rural areas6. Beijing and the neighboring city Zhangjiakou in Hebei province have announced a joint bid to host the 2022 Winter Olympics, in which events staged on ice will be in Beijing, and the snow events will be in Zhangjiakou. Concerns have been raised regarding the potential ecological impact of the 2022 Winter Olympics on surrounding regions12. The Winter Olympics could not only affect the current landscape pattern but also drive the distribution of regional investment, population layout and redistributed land use patterns in the future. Each part of planning the construction for these events or future land use management represents a significant source of anthropogenic disturbance, while also providing recreation and revenue13. Among the available landscape modifications, some are particularly high-risk and are likely to exert a negative influence on the ecosystem structure and function14. Generally, high-risk situations include a vulnerable natural ecosystem, the presence of sensitive species, proximity to wetlands and water ecosystems, the presence of cultural or historical value requiring strict protection or sites that have been the subject of public protest leading up to the Games. Regional land use management including the current ski areas is a conservation concern because they significantly alter fragile high-elevation ecosystems16. Destruction to natural ecosystems and metabolic imbalances greatly threatens regional ecological security19. Therefore, to ensure effective ecological constraints on development and construction, it is essential to conduct an ecological suitability assessment.The relationship between regional development and ecosystem conservation has been the subject of decades of intensive research21. This method is able to identify the location and ecological boundaries of suitable areas for construction based on grading index factors through GIS-based rule settings such as cellular automata22, fuzzy measures24, analytical network process25, urban growth model26 and spatial clustering10. Overall, the basic principles of existing studies are similar to those utilized in this study: optimizing the adaptive hierarchy through remote sensing and GIS spatial analysis techniques28. This approach has also been extensively applied in the assessment of different kinds of land use types, such as agricultural land29, land habitats for animal and plant species30, landscape evaluation and planning31, ecosystem services sustainability33, and environmental impact assessments34. For example, the regression neural network method and ecological suitability conceptual model were used to evaluate the land suitability35. Additionally, the ecological suitability theories have been expanded by studies examining landscape ecological security patterns and ecosystem services37 and dynamic ecological processing frameworks38. However, most of these ecological constraints add ecological factors to the model10 rather than investigate the relative supporting roles, essentially neglecting the effects of simulation in different scales and the evaluation of ecological loss of construction through ecological field survey.Currently, the ecological suitability evaluation for land construction focuses on the paradox between ecological protection and construction on land for rapid expansionThis paper aims to assess the suitability of ecologically compatible development in Winter Olympic regions by using multi-scale GIS spatial analysis methods. The spatial scales of the study area includes the Yanghe watershed, Chongli District, and the Olympic ski area and ski piste. More particularly, we give priority to the future of independent development in the Yanghe basin, while focusing on risk management and trade-off analysis in the Chongli District. Meanwhile, through evaluating the loss of ecological benefits, we stress the need for better ecological protection and compensation in ski resorts and their areas. To reduce the negative impacts of human activities, to decrease ecological risks, and to add to the quality and fun of the Games in construction areas, this paper examines the spatial dynamic characteristics of a natural ecosystem’s self-organization and capacity to self-update, reflected in terms of the three aspects of ecological elements, ecological importance, and ecological resilience by using the integrated ecological resistance (IER) conceptual model based on ecological circulation theory.Spatial distribution of ecological structure resistance Fig.  determin39. To protect this water source, which is an important supplier of water, and to support the regional ecosystem function, ecological function resistance of land-cover data containing water bodies is directly defined at the highest level. The proportions of the total area for the highest resistance category and lowest resistance category are very small, 5.09% and 3.15%, respectively, while the moderate level and fourth level together account for 64.86% of the total area  in total. The large amount of deforestation required to build multiple ski runs increases soil erosion, destroys plant life, reduces plant species diversity, and causes permanent damage to the ecological environment of the surface soil. Despite having more abundant herbaceous species on primary ski pistes than on others  available for plants from primary to advanced ski pistes. In terms of coverage and growth status, more biomass accumulation and soil reinforcement were found in advanced ski pistes. Additionally, the variations of soil physical and chemical indicators  may directly relate to the changes of multiple ecosystem functions. The results also indicated that the soil moisture content increased and that the soil bulk density decreased with increasing distance from the construction areas.From an ecological perspective, the Field of Play for mountainous areas, which enables freestyle skiing and snowboarding events at the 2022 Winter Olympics, also plays a vital role in water retention, soil conservation and protecting biological diversity. To study the ecosystem function loss in different ski runs, we investigated vegetation coverage Fig. . General40. Land and economic development are accompanied by landscape changes and ecological destruction in the cities that hold the Olympics16, and the 2022 Winter Olympic Games could also bring large-scale regional development and population aggregation in the study area. However, at a regional scale, the spatial ecological suitability results show that the areas with lower integrated ecological resistance are mainly located in plains along the Yanghe River, including Shangyi, Wanquan, Zhangjiakou and Xuanhua counties, where construction will be less of a disturbance to the eco-environment , functions (wf), dynamics(wd), water retention, soil conservation, earthquake, and DEM were the most sensitive parameters (Table in-situ monitoring networks should be strengthened. (5) The accommodation, amusement and other supporting facilities of the Winter Olympics are recommended to move to Chongli and Zhangjiakou urban areas or regions with prior development.For the Winter Olympic venues, the ecological indicators that have relatively low ecological resistance include the slopes, DEM, general geological hazards, earthquake, and vegetation stability. However, the indicators, such as biodiversity protection, water retention, and soil conservation, have high ecological resistance to the ski resort we studied. Hence, a sensitivity analysis of all the parameters controlling suitability results was conducted, with one sub-region  chosen as an illustrative example. The results of sensitivity analysis indicated that the weightings of ecological structures .An ecological suitability evaluation for Winter Olympic construction in mountainous areas involves datasets on topography (DEM), land use, geological disasters , vegetation (NDVI), soil, precipitation, population density, GDP per area, and other factors Fig. . Most ofES), ecological functions (EF), and ecological dynamics (ED). Peng’s conceptual model was described as follows10:ws, wf, and wd are the weights of the indies. According to the literature10, given that structures, functions, and dynamics are interdependent for the entire ecological environment, these three weights were given equal values. Based on regional characteristics, we defined representative indicators and calculation methods. Integrated ecological resistance was evaluated as indicated in Table 46. The weight of each structural index was calculated by the method of Analytical Hierarchy Process (AHP). The ecological function resistance was evaluated by biodiversity protection, water and soil conservation. The classification and weightings of biodiversity protection and water retention refer to the methods in the literature47. Soil conservation was evaluated by the revised universal soil loss equation (RUSLE). Ecological dynamics assessment was reflected by vegetation stability , ecological sensitivity48 (expressed by the distance to sensitive areas), and human disturbance . Generally, the more stable the geology and the flatter the terrain, the lower the ecological resistance to Winter Olympic development by assessing ecological structure resistance. The higher the level of ecological importance, the greater the ecological risk of mountainous area development by determining ecological functional resistance. If the negative relationship between construction sites and ecological function indicators was found, which would lead to the decline of ecosystem service functions. The construction of venues has increased the intensity of disturbances in regional human activities, resulting in a decrease in vegetation stability, leading to ecological fragility and sensitivity. Meanwhile, higher ecological resilience from the ecological dynamic assessment indicates a greater capacity for self-recovery and self-updating to counteract disturbance or damage, as well as less ecological risk and lower ecological resistance to construction.Winter Olympic construction is a process of human activity to overcome the resistance of natural ecosystems for sustainable development. Then, the integrated ecological resistance (IER) has a negative correlation with the level of ecological suitability, which means that the higher the former, the lower the latter. There is a high correlation between construction security risks and ecological risks, which have been calculated and graded into several categories separately to characterize the resistance of ecological structures (The sensitivity analysis of the main indexes and weightings was conducted. For each evaluation, one parameter was selected and increased by 1%. The sensitivity coefficient S of this parameter was defined using formula :2\documeSupporting materials