Tobacco companies have made extensive efforts to build alliances against comprehensive smoke-free legislation. This article analyses the interaction between actors who opposed the development of the European Council Recommendation on smoke-free environments.Drawing on data from 200 policy documents and 32 semistructured interviews and using qualitative textual analysis and organisational network analysis, opponents’ positions on, and responses to, the policy initiative, strategies to oppose the policy, and efforts to build alliances were investigated.The non-binding nature of the policy, scientific evidence and clear political will to adopt EU-wide measures combined to limit the intensity of commercial sector opposition to the comprehensive EU smoke-free policy. Most tobacco companies, led by the Confederation of European Community Cigarette Manufacturers (CECCM), voiced reservations against the proposal, criticised the policy process and fought flanking measures on product regulation. However, some companies focused on instigating harm reduction debates. These divergent approaches and the reluctance of other commercial actors to demonstrate solidarity with the tobacco sector prevented the establishment of a cohesive commercial sector alliance.The comparatively limited opposition to EU smoke-free policy contrasts with previous accounts of tobacco industry resistance to tobacco control. While context-specific factors can partially explain these differences, the paper indicates that the sector's diminished credibility and lack of unity hampered political engagement and alliance building. Industry efforts to emphasise the benefits of smokeless tobacco during smoke-free policy debates highlight the potential of harm reduction as a gateway for tobacco companies to re-enter the political arena. Since Ireland's comprehensive smoke-free policy in 2004, substantial progress has been made across the EU in protecting citizens from secondhand smoke (SHS) in public places. Corresponding national policy debates have confronted industry lobbyists with an unprecedented challenge,7i to the development of the Council Recommendation for smoke-free environments, which recommends that member states “provide effective protection from exposure to tobacco smoke in indoor workplaces, indoor public places, public transport and, as appropriate, other public places”.This article analyses the responses of tobacco companies, tobacco-related and other commercial actorsThe study used documentary and interview data to investigate opposition to comprehensive EU smoke-free policy. Approximately 200 publicly available documents were reviewed, including policy drafts, minutes of meetings, consultation submissions, briefings and reports. Documents offering indepth information about stakeholders’ views were thematically analysed. To investigate interactions between the various actors opposing EU smoke-free policy, all organisational responses submitted to the Commission's public consultation on smoke-free environments (n=176) underwent a structural network analysis, including those from 35 organisations which the Commission categorised as tobacco-related organisations. Each organisation was registered as a node and assigned attributes, including organisation type and position on the proposal. Two organisations were recorded as having a relationship if: (1) A was mentioned as a collaborating partner on the website or consultation submission of B; and/or (2) A cited three or more references in its submission which were also cited by B; and/or (3) plagiarism detection software showed that A's submission was at least 40% identical to B's. Data were analysed using UCINet V.614Qualitative data were gathered through semistructured, narrative interviews with 35 individuals involved in negotiating the Recommendation. Interviewees were purposively sampled from a list of key individuals identified via the documentary review. Forty eight individuals were contacted, of which six declined and five did not respond. The final sample included decision makers , representatives of health advocacy organisations (n=13), professional organisations (n=1), scientific institutions (n=4), social partner organisations , the tobacco sector (n=4), ventilation industry (n=1) and other commercial sector bodies (n=3). Based on documentary review and informal pilot discussions, an interview topic guide was developed, combining a narrative part and open-ended questions. Interviews were conducted by HW between March and July 2011, asking interviewees to recall experiences of engaging in a process undertaken 5–8 years previously. Twenty two interviewees, who wanted to remain anonymous, suggested descriptors for use in publications, with the desire for anonymity reflected in some vague descriptors used below. Interviews were recorded and transcribed verbatim, except interviews with two tobacco industry representatives who preferred that the interviewer took notes. Using QSR NVivo,17With high non-response and rejection rates among tobacco industry representatives , and despite intensive recruitment efforts, only four interviewees could provide internal insights into the industry's responses. This may reflect reluctance to engage with a researcher who had previously examined industry efforts to obstruct policy.Contrasting fierce industry opposition against national smoke-free policies,While various actors publicly voiced reservations about comprehensive smoke-free policy, the data suggest that the strongest counterarguments originated from tobacco manufacturers and tobacco-related actors. They advocated for an “EU-wide smoking ban with exemptions”, designated smoking rooms and exclusion of hospitality venues, private clubs, research laboratories and residential places.Despite efforts to advance a unified response, tobacco companies seemingly struggled to fully align their positions. Contrasting the CECCM-led alliance, Philip Morris’ (PMI) response supported “a total smoking ban…in general public indoor spaces, such as stores, banks, hospitals, public buildings, and public transportation” without suggesting exemptions for specific venues.Everything the tobacco industry has been doing in the last few years is…trying to find as many policy hooks as they can, including [the Recommendation], in order to build up support for snus as a safer alternative or even as a kind of a smoking cessation aid. (Lobbyist)Complementing documentary data, the interviews provided evidence of tobacco industry attempts to instigate, join and frame debates on, and direct attention towards, harm reduction. Such debates were conceived as offering unique public relation opportunities to reposition tobacco companies as reasonable and legitimate stakeholders who could add value to the policy process, lobby for removal of restrictions on smokeless tobacco products, and depict harm reduction policies as necessary adjuncts to prohibiting smoking in public places.While keen to avoid being perceived as strongly opposing a policy aimed at protecting citizens from SHS harm, the CECCM-led tobacco industry alliance worked to hamper the policy process and derail the initiative. CECCM's submission strongly criticised DG SANCO staff for an approach to stakeholder consultations depicted as biased and undemocratic, and instead called for “dialogue and consultation with all interested stakeholders, including the tobacco sector” and for what it framed as a more inclusive approach to policy development.Finally, interviews revealed considerable increases in tobacco companies’ activity in the closing stages of the process. This intensification was triggered by the unexpected inclusion in the policy draft of calls to revise the Tobacco Products Directive and assess future EU policies regarding plain packaging and graphic health-warning labels.In fighting comprehensive smoke-free policy, opponents argued that health risks from SHS were ‘relatively minor’,These factors, however, only partially explain the limited efforts of opponents in countering the development of the Recommendation. Arguably as important was the EU's restricted scope to adopt binding smoke-free legislation due to its lack of legal remit to harmonise national public health legislation, such that opponents did not perceive the initiative as a substantial threat. One tobacco industry representative, for example, reported that “the pressure came off the cooker” and considerable downscaling of lobbying efforts as soon as it was clear that binding legislation would not be supported in the Council of the EU. Similarly, representatives of other industries reported that the Recommendation was perceived as non-threatening and of a low priority, reflecting the implausibility of legal consequences and low probability of translation into national legislation.Notwithstanding their general reticence about increasing EU regulation, membership links and previous collaboration with tobacco companies, the data indicate limited opposition to comprehensive EU policy from social partners and pan-European trade associations, like BusinessEurope (an organisation representing industry associations across Europe), the European Trade Union Confederation (ETUC), the European Federation of Food, Agriculture and Tourism Trade Unions (EFFAT), and the European trade association representing hotels, restaurants and cafés (HOTREC).In addition to the low priority afforded to the initiative by pan-European organisations given the unlikelihood of binding legislation, structural features of EU policymaking seemed to further hamper alliance building. Pan-European commercial sector organisations struggled to reach consensual positions on the policy proposal, with several interviewees reporting cumbersome discussions. A tobacco industry representative, for example, reported that HOTREC had been “between a rock and a hard place” in balancing views of national members opposing the policy  and those willing to support the EU initiative . Reflecting on similar difficulties of aligning diverse members, a representative of another umbrella organisation reported that having “to coordinate things with everyone…[often led to]…very general and…non-committal” positions.Pan-European organisations also seemed to carefully weigh the benefits of opposing a specific EU-level initiative against a more general, long-term aspiration to be perceived as reasonable, progressive and constructive stakeholders, supportive of EU-wide cooperation. Strategic considerations about the impact of their responses on their public image and desires to not appear as “a kind of old-school, old type” but “modern” business  influenced organisational decisions on whether to demonstrate agreement with tobacco-industry organisations. While reporting that they had generally been opposed to comprehensive EU smoke-free policy and met with tobacco company representatives to exchange information and coordinate responses, several social partners and non-tobacco trade representatives recounted a reluctance to position themselves as tobacco industry allies. It is important to note that the guidelines to FCTC Article 5.3 were adopted at the third session of the Conference of the Parties in November 2008, that is, when EU smoke-free policy was negotiated.Previous research extensively documents tobacco industry efforts to combat tobacco control policies, including national smoke-free policies and the regulation of tobacco advertising and tobacco products at EU level.Two context-specific factors are particularly important in interpreting the findings. First, the interview data show that the non-binding nature of the proposed policy considerably shaped opponents’ responses to the proposal. The fact that the EU is not granted competence to harmonise national public health legislationWhile the importance of context-specific factors means that the findings cannot be generalised, our study provides valuable insights into the complexities of tobacco companies’ engagement with each other and other sectors, the dynamics which shape opposition to tobacco control policy at EU level, and the impediments to developing effective EU tobacco control policy. Our indepth analysis of the diverse approaches which opponents pursued in countering comprehensive EU smoke-free policy highlights the contrast between some tobacco companies’ decision to overtly oppose comprehensive smoke-free policy and the seemingly more circumscribed opposition by other actors. The findings demonstrate the difficulties opponents faced when justifying their resistance to EU smoke-free policy and their limited success in reiterating well-known arguments and tactics to counter tobacco control. The CECCM-led alliance subsequently focused its efforts on disputing any reference in the policy document to binding EU level tobacco regulation, arguing for more consultations with tobacco industry representatives, criticising DG SANCO's approach to policy development and instigating division among European Commission DGs. Importantly, the findings point to a considerable reluctance among non-tobacco actors, notably pan-European organisations representing the commercial sector, to openly join such opposition and publicly counter the policy proposal. This contrasts with historical accounts depicting umbrella organisations as serving tobacco industry interestsComplementing previous research, which analyses tobacco companies’ differential political positions and strategies in opposing tobacco taxation in the Czech Republic,60Our study is, to the best of our knowledge, one of threeOpposition to EU smoke-free policy was of limited scope and intensity, reflecting the low priority that opponents afforded to the non-binding policy proposal, reluctance to oppose a policy initiative which was clearly framed around public health and strategic efforts to present themselves as constructive stakeholders.Tobacco companies pursued different lobbying strategies, with a Confederation of European Community Cigarette Manufacturers (CECCM)-led alliance opposing the policy, and Philip Morris’ (PMI) and International Smokeless Tobacco Company Inc (ISTC) using the debates on EU smoke-free policy to initiate harm reduction debates.Hoping to avoid being associated with the tobacco industry and facing difficulties in building consensus among members, pan-European business organisations that had previously allied with the tobacco industry did not publicly oppose EU smoke-free policy.By analysing data from interviews with tobacco industry representatives and tobacco control opponents, this paper provides unique insights into the complex dynamics of tobacco control opposition.The joint depiction of various tobacco-related actors as ‘big tobacco’ risks over-simplifying the complexities of interaction and alliance building among tobacco control opponents.Our findings highlight the potential oversimplification of portraying diverse tobacco companies and related organisations as a monolithic body. The presentation of such actors as ‘big tobacco’


The use of genomic technologies for the molecular characterization of tumors has propelled our understanding of cancer biology and is transforming the way patients with cancer are diagnosed and treated. More than any other field of medicine, oncology has benefited from recent revolutionary advances in nucleic acid sequencing technology and genomic analysis. For increasingly lower costs and turnaround times, one can profile the full spectrum of genomic alterations in a tumor sample, including sequence mutations, copy number changes and structural rearrangements . ResearcBRAF) gene from systematic gene sequencing [Research initiatives such as TCGA and ICGC have led to an improved understanding of cancer biology through the genomic analysis of tumors procured from anonymized patients possessing many different types of cancer. A major goal of these and similar efforts is to identify genes and pathways important to cancer progression in order to design better therapies and interventions. As an early example, the identification of recurrent somatic mutations in the serine/threonine-protein kinase B-raf  mutations in lung cancer was prompted by the clinical observation that a subset of patients exhibited a major response when administered tyrosine kinase inhibitors in clinical trials –18. TodaTSC1) and serine/threonine-protein kinase mTOR/mammalian target of rapamycin (MTOR) mutations in bladder cancer patients responding to everolimus and a mutation in serine/threonine-protein kinase A-Raf (ARAF) in a lung cancer patient responding to sorafenib [Recently, the analysis of ‘exceptional responders’ - cancer patients with an unexpected complete and/or durable response to therapy - has proven to be particularly effective. Studying exceptional responders can reveal specific genomic alterations accounting for their exquisite sensitivity to certain drugs. One can hypothesize that other patients whose tumors bear similar alterations will benefit from the same drugs. Recent successful applications of whole-genome or exome sequencing in exceptional responders include the identification of tuberous sclerosis 1 protein/hamartin  to guide treatment. If a sensitizing mutation in EGFR or a rearrangement in ALK is identified, treatment with an inhibitor of EGFR or ALK is recommended. While drugs targeting these alterations are FDA approved, several targeted agents have demonstrated activity against lung cancers harboring other genetic alterations. For instance, cabozantinib has demonstrated activity in lung cancers harboring rearrangements in the gene encoding the tyrosine-protein kinase receptor Ret (RET), and crizotinib has demonstrated activity in lung cancers harboring amplifications in the gene encoding the hepatocyte growth factor receptor (MET) or rearrangements in the ROS1 gene encoding tyrosine-protein kinase ROS [KRAS and NRAS is recommended to assess whether patients might benefit from EGFR-directed monoclonal antibody therapies utilizing the drugs cetuximab or panitumumab. For patients with melanoma harboring V600 mutations in BRAF, treatment with vemurafenib or dabrafenib is recommended.Based in part on the identification of predictive clinical biomarkers from retrospective analyses, there are several tumor types for which prospective mutation profiling as a diagnostic tool is now a standard of care. Patients with metastatic non-small cell lung cancer are routinely tested for nase ROS –34. For Molecular-diagnostics labs have traditionally relied on low-throughput, mutation-specific methods for DNA profiling in patients because there were so few actionable genetic alterations that altered treatment decisions in the clinical care of patients. However, given the growing number of biomarkers and clinical trials studying targeted agents, the approach of testing one mutation at a time is unsustainable. NGS-based assays are replacing these more-focused tests in both academic and commercial settings. The benefits of this are obvious. First, a single NGS test can encompass all ‘actionable’ targets, eliminating the need for multiple parallel tests for different mutations and enabling more-efficient workflows and tissue utilization. Second, the entire coding sequence of target genes can be assayed (rather than only pre-specified sites), facilitating the detection of both common and rare mutations in oncogenes and tumor-suppressor genes. Third, NGS enables the detection of additional classes of genomic alterations, such as copy number changes and structural rearrangements. Finally, subclonal events in heterogeneous tumors can be detected more reliably owing to the high sensitivity afforded by NGS.Some academic centers have implemented pilot programs for the comprehensive genomic characterization of tumors from selected patients by means of whole-genome or exome sequencing (DNA-Seq) and transcriptome sequencing (RNA-Seq) ,36. ThroOwing to practical barriers in the implementation of this comprehensive approach for all patients, namely its high cost and low throughput, large-volume molecular-diagnostics labs have focused instead on targeted sequencing of key cancer-associated genes as a feasible and economical alternative –42. FurtIn order for genomic insights and technologies to be truly transforming in oncology, we must develop and implement high-throughput molecular-diagnostic tests that exhibit both clinical validity and clinical utility. Assays must achieve rapid turnaround times at low cost, encompass all classes of sequence-based mutations and structural alterations, and operate on small biopsies, formalin-fixed paraffin embedded (FFPE) tissue and cytological specimens . FurtherTERT), which rank among the most frequently observed mutations in all cancers [The development of assays compatible with low-quality specimens and low input DNA amounts remains a technical challenge. For these suboptimal samples that are routinely encountered in clinical settings, comprehensive genomic analysis is not always possible. PCR-based capture technologies can produce deep-coverage sequence data with very little DNA, but targets are generally small, and the detection of copy-number alterations and structural rearrangements is compromised. With slightly more DNA, hybridization-based capture assays enable an expanded number of target genes and alteration types but still can exclude important regions of the genome. Even whole-exome sequencing, encompassing all protein-coding genes in the genome, will miss many structural alterations as well as nucleotide substitutions involving regulatory regions such as the promoter of the gene encoding telomerase reverse transcriptase -compliant laboratories, governed by the Centers for Medicare & Medicaid Services (CMS). Accordingly, extensive documentation and technical validation of sensitivity, accuracy and reproducibility are required before any genomic assay can be prospectively administered and/or billed to healthcare payers. While some institutions have initiated programs where large-scale genomic analysis is performed in research labs, followed by confirmatory testing in CLIA-compliant labs, this approach is unsustainable when NGS-related costs cannot be recovered through reimbursement.While use of comprehensive molecular profiling has produced success stories, such as the remarkable responses to erlotinib for patients with EGFR-mutant lung cancers or crizotinib for patients with ALK-positive lung cancers, these findings have also resulted in challenges and questions concerning how to proceed.First, comprehensive molecular profiling using any of the assays or technologies described above will yield information about both well-known molecular drivers in cancer and countless more alterations whose biological and clinical significance is unclear. It is often difficult for a clinician to make the crucially important distinction between key ‘driver alterations’ that should impact treatment and less-relevant ‘passenger mutations’. Some publicly available knowledge banks have been created to help doctors and patients interpret the significance of many commonly seen alterations, such as My Cancer Genome developed at the Vanderbilt-Ingram Cancer Center. While these sites are excellent resources to learn about clinically and pathologically annotated alterations, they are not comprehensive and cannot be expected to include information on all possible alterations that a clinician could encounter. Similarly, there is little guidance for the management of patients with multiple driver alterations in more than one gene. One might rationally assume to administer combinations of targeted therapies; however, without knowing the toxicity profiles or optimal dosing and scheduling of the combination, this strategy is problematic.Also, the use of targeted therapies in the treatment of cancer is rarely straightforward, even in the presence of individual well-characterized driver alterations. NGS testing of heterogeneous tumors can reveal targetable mutations at subclonal allele frequencies, the clinical consequences of which are uncertain. Furthermore, the biological and clinical context can be extremely important. For instance, while patients with melanomas harboring BRAF V600E mutations almost always respond to inhibitors of BRAF, this direct treatment approach has not been replicated for patients with BRAF-mutant colon cancers. Colon cancers harboring BRAF V600E mutations do not respond to single-agent vemurafenib - this is thought to be related to feedback reactivation of alternative or upstream signaling pathways ,49. StudWhich specimens to analyze and when to analyze them are also questions that arise in the clinical care of patients. Given emerging data about tumor heterogeneity in metastatic disease, it is unclear that a single biopsy of a single site of metastatic disease will accurately capture driver alterations that would most impact a patient’s clinical care. Additionally, it is unclear that biopsies obtained at the time of diagnosis remain relevant after patients have developed acquired resistance to targeted therapies or have developed recurrent disease after initial therapies for early-stage cancer. While biopsies taken at the time of acquired resistance are becoming standard in many clinical trials of targeted agents, these biopsies are not routinely performed in the clinical care of patients.Most importantly, targeted therapies are not curing patients of their cancer. While targeted therapies have yielded promising, dramatic and life-changing responses for many patients with cancer, the reality is that these responses are generally short lived. For instance, the average responses to erlotinib among patients with EGFR-mutant lung cancer and vemurafenib for patients with BRAF V600E mutant melanoma are 11 months and 5 months, respectively ,50. All The societal and ethical implications of comprehensive genetic testing must also be considered in this rapidly changing technological landscape. While focused diagnostic tests including targeted ‘hotspot’ panels utilize only tumor-derived DNA, more-comprehensive NGS approaches typically require germline DNA to distinguish between novel somatic mutations and inherited variants. The use of germline DNA poses the risk that incidental findings could be revealed relating to inherited susceptibility to cancer or other diseases. This has led institutions to consider different strategies of informed consent and pre-test genetic counseling. However, given the time pressures of clinical care, the ability to perform thorough genetic counseling in a routine fashion is limited. For those patients in whom an inherited predisposition to cancer is discovered, care must be taken to ensure the patients’ autonomy and privacy, protect them and their families from possible discrimination and also manage the unintended emotional and psychological consequences that such a diagnosis brings.Additionally, it remains unclear whether NGS-based molecular profiling is cost-effective or even clinically effective in the care of patients outside of lung cancer, melanoma and colon cancer. As a result, insurance companies have demonstrated a reluctance to reimburse the cost of comprehensive testing in many tumor types. While large academic cancer centers might be able to offset costs temporarily through grants and philanthropic contributions, broad access to these tests for patients in the community has not been achieved. Demonstrating the general clinical utility of NGS-based molecular profiling is essential in this regard. A related hindrance is that, when actionable mutations are detected in unexpected tumor types, insurance companies are often unwilling to reimburse the off-label administration of therapies approved in other diseases.While genomic knowledge has been successfully applied to direct clinical decisions in several cases, the implementation of clinical genomic workflows for oncology has proven to be complex. Nevertheless, the introduction of NGS technology has the potential to transform clinical oncology, and it is incumbent upon clinicians, scientists, regulators, payers and patients to work collectively to overcome the obstacles that stand in its path.


The enteric disease of swine recognized in the early 1970s in Europe was initially described as “epidemic viral diarrhea” and is now termed “porcine epidemic diarrhea (PED)”. The coronavirus referred to as PED virus (PEDV) was determined to be the etiologic agent of this disease in the late 1970s. Since then the disease has been reported in Europe and Asia, but the most severe outbreaks have occurred predominantly in Asian swine-producing countries. Most recently, PED first emerged in early 2013 in the United States that caused high morbidity and mortality associated with PED, remarkably affecting US pig production, and spread further to Canada and Mexico. Soon thereafter, large-scale PED epidemics recurred through the pork industry in South Korea, Japan, and Taiwan. These recent outbreaks and global re-emergence of PED require urgent attention and deeper understanding of PEDV biology and pathogenic mechanisms. This paper highlights the current knowledge of molecular epidemiology, diagnosis, and pathogenesis of PEDV, as well as prevention and control measures against PEDV infection. More information about the virus and the disease is still necessary for the development of effective vaccines and control strategies. It is hoped that this review will stimulate further basic and applied studies and encourage collaboration among producers, researchers, and swine veterinarians to provide answers that improve our understanding of PEDV and PED in an effort to eliminate this economically significant viral disease, which emerged or re-emerged worldwide. In 1971, British veterinary clinicians noted the appearance of a previously unrecognized enteric disease in growing and fattening pigs , 3. In 1Since the 1990s, PED cases have become rare in Europe and PED virus (PEDV)-associated diarrhea has been usually observed in adult pigs, whereas suckling piglets were generally spared or developed only mild symptoms . PED wasThis paper is a brief review focusing on current understanding of the molecular biology, epidemiology, diagnosis, and pathogenesis of PEDV, as well as control strategies to prevent PEDV infection.Alphacoronavirus within the Coronaviridae family placed with the Arteriviridae family in the order Nidovirales on the basis of similarities in genome organization and replication strategy , without typical S protein genetic signatures of the epidemic G2 virus were reported. They phylogenetically clustered closely to novel Chinese strains in the G1b subgroup based on the similarities of the S gene or with the emergent US PEDV strains in the G2 group based on the whole genome characteristics . Novel vThe first PED epizootic in South Korea was confirmed in 1992 . HoweverAfter South Korea experienced severe outbreaks of the foot-and-mouth disease (FMD) in 2010–2011, there was a state of lull during PED emergence. The prevalence of PEDV infections was occasional with only intermittent outbreaks in South Korea from 2011 to early 2013. This epidemic situation likely resulted from the mass culling of more than 3 million pigs (one-third of the entire domestic pig population) in South Korea during the 2010–2011 FMD outbreaks. However, beginning from November 2013, severe PED epidemics increased remarkably and swept through more than 40 % of pig farms across mainland South Korea . Four moIn March of 2014, novel variant G1b PEDV isolates have been found in South Korea, which were similar to the variants reported in China, the United States, and recently in several European countries . They haPEDV infection among pigs occurs principally by a direct or indirect fecal-oral route. Airborne transmission may also play a role in PEDV dissemination under certain conditions . PEDV caAfter an acute (epidemic) outbreak, PEDV may disappear, remain in the farrowing unit because of inadequate hygiene management , or persist in pigs in weaning or growing-finishing units where the virus is circulating, causing mild post-weaning diarrhea with very low mortality rates. In this endemic status, if newly born pigs are unable to obtain sufficient levels of maternal immunity from their dams due to incomplete sow vaccination or defective lactation performance owing to mastitis or agalactia, the virus circulating on the farm will infect susceptible piglets, which serve as the source of recurrence of epidemic outbreaks leading to a high number of pig deaths , 91. SucPEDV can infect pigs of all ages, causing watery diarrhea and vomiting accompanied by anorexia and depression. Morbidity approaches 100 % in piglets, but can vary in sows . The incGross lesions are confined to the gastrointestinal tract and characterized by distended stomach filled with completely undigested milk curd and thin, transparent intestine walls with accumulation of yellowish fluids , 92, 95.PEDV replicates in the cytoplasm of villous epithelial cells throughout the small intestine, destroying target enterocytes as a result of massive necrosis or apoptosis. These processes lead to villous atrophy and vacuolation as well as a marked reduction in the enzymatic activity , 55. Thiin situ hybridization, electron microscopy, virus isolation, enzyme-linked immunosorbent assays (ELISA), and various reverse-transcription polymerase chain reaction (RT-PCR) techniques, have been used. Taking into account their fast turnaround times and sensitivity, conventional and real-time RT-PCR systems available as commercial kits are most widely used for PEDV detection during epidemic or endemic outbreaks, as well as for quarantine or slaughter policies. In addition, nucleotide sequencing of the S gene region may be useful in determining the genotype of PEDV circulating in herds. The combination of RT-PCR and S gene sequencing could well become the optimal tool for monitoring genetic diversity among PEDV isolates.Since signs of the PEDV infection were clinically and pathologically indistinguishable from those caused by TGEV and the recently described porcine deltacoronavirus , 99, 100A number of serological assays have been used for the detection of PEDV antibodies, including indirect fluorescent antibody (IFA) staining, ELISA, and virus neutralization (VN) tests. Due to the special protection strategy (passive immunization) for neonatal piglets against PEDV, determining the presence or absence of anti-PEDV antibodies may be meaningless in sow herds. Instead, measuring quantities (or titers) of neutralizing antibodies against PEDV or, especially, against the S protein in serum and colostrum should be necessary to monitor the immunity level following sow immunization. In this regard, VN test could be essential for estimating levels of protective antibodies, which piglets would receive from sows. However, this method is time-consuming and cannot selectively detect only secretory IgA antibodies representing mucosal immunity. In contrast, IFA and indirect ELISA approaches for antibody detection are equally specific but less time-consuming and easier to perform than VN test. Most assays that are currently in use have been developed on the basis of either whole virus –103 or vOne of the most important measures for prevention and control of acute PED outbreaks is strict biosecurity that blocks in principle the entrance of PEDV into pig farms (fattening and farrowing units) by minimizing introduction of any material or any person, which could be in contact with the virus. To accomplish this, disinfection must be thoroughly applied to all fomites, personnel, and external visitors that could be contaminated with PEDV. Although PEDV is inactivated by most virucidal disinfectants , PEDV RNPEDV is a transboundary virus that seems to spread readily to neighboring or distant countries even across continents. Since PEDV is not a World Organization for Animal Health reportable disease, quarantine inspection might not properly implement potential sources or routes that mediate virus transmission between countries. During large-scale severe PED epidemics in adjacent or trading countries, quarantine  procedures should be adequately reinforced with a particular attention to any risk factors of international disease transmission in order to prevent the entrance of PEDV as well as other emerging or re-emerging pathogens. On the basis of available genetic and phylogenetic data on global PEDV strains, I propose several prospective paths through which PEDV could have spread to Asia and North America Fig. . The claVaccination of sows is a fundamental tool in a strategy to control and eradicate PED during epidemic or endemic outbreaks. Piglets are then protected by a transfer of maternal antibodies via colostrum and milk from immune dams. Although PED first emerged in Europe, the disease has not caused sufficient economic losses to justify the vaccine development. In contrast, PED outbreaks in Asian countries have been serious, and therefore several PEDV vaccines have been developed. In China, CV777-attenuated or -inactivated vaccines have been routinely applied against PED. Attenuation of the virulence of the Japanese PEDV strain 83P-5 was achieved after 100 passages in Vero cells . SubsequThe low to moderate effectiveness of current PEDV vaccines appears to be due to antigenic, genetic (>10 % amino acid variation between respective S proteins) and phylogenetic (G1 vs. G2) differences between vaccine and field epidemic strains , 49, 59.In acute PED outbreaks with rapidly increasing mortality rates, we may consider intentional exposure (feedback) of pregnant sows to the autogenous virus using watery feces or minced intestines from infected neonatal piglets, which will artificially stimulate rapid lactogenic immunity and, hopefully, shorten the outbreak on the farm . HoweverE. coli expressing scFvs were verified to neutralize PEDV in vitro [E. coli harboring scFvs might be an alternative prophylactic measure against PEDV infection. Pharmacological, biological, or natural agents that shorten epithelial cell renewal by stimulating proliferation or reorganization of crypt stem cells could be potential therapeutic targets to reduce PEDV-associated mortality from dehydration following severe villous atrophy [in vitro, are of interest for their potential to treat PED [Artificial passive immunization by oral administration of specific antibodies represents an attractive approach against gastrointestinal pathogens such as PEDV. Chicken egg yolk immunoglobulin against PEDV or the S1 domain has been found to protect neonatal piglets following challenge exposure , 114. Thin vitro . This su atrophy . For ins atrophy . Broad-sreat PED . Chemicareat PED , 56. In For the last two or three decades, PED has continued to plague the pork industry in Europe and Asia. In early 2013, the disease struck North America resulting in great economic losses, especially in the United States. Shortly thereafter, massive nationwide PED outbreaks reoccurred in South Korea, Japan, and Taiwan. PED is now globally recognized as an emerging and re-emerging disease and it has become a major financial issue for the swine industry worldwide. Despite geographically limited PED studies in Europe and Asia, a better knowledge of the virology, pathogenesis, immunology, epidemiology, and vaccinology has been gained. Since the emergence of PED in North America, much has been learned from research in the United States. Nevertheless, further studies are warranted to decipher the virus and the associated disease, and more extensive academic and practical studies are needed for a comprehensive understanding of the molecular and pathogenic biology of PEDV in order to develop effective vaccines and establish control measures including biosecurity in affected areas. Lastly, we should bear in mind that a combined application of vaccines, biosecurity protocols, and husbandry management must be the key step to prevent and control PED. Integrated and coordinated efforts in various disciplines among researchers, swine veterinarians, producers, swine industry specialists, producer associations, and authorities are required to achieve effective implementation of necessary measures.


Oxytocin (OT) is a neuropeptide synthesized primarily by neurons of the paraventricular and supraoptic nuclei of the hypothalamus. These neurons have axons that project into the posterior pituitary and release OT into the bloodstream to promote labor and lactation; however, OT neurons also project to other brain areas where it plays a role in numerous brain functions. OT binds to the widely expressed OT receptor (OTR), and, in doing so, it regulates homeostatic processes, social recognition, and fear conditioning. In addition to these functions, OT decreases neuroendocrine stress signaling and anxiety-related and depression-like behaviors. Steroid hormones differentially modulate stress responses and alter OTR expression. In particular, estrogen receptor β activation has been found to both reduce anxiety-related behaviors and increase OT peptide transcription, suggesting a role for OT in this estrogen receptor β-mediated anxiolytic effect. Further research is needed to identify modulators of OT signaling and the pathways utilized and to elucidate molecular mechanisms controlling OT expression to allow better therapeutic manipulations of this system in patient populations. The nonapeptide hormone, oxytocin (OT), has gained widespread attention as a potential therapeutic agent in a myriad of disorders, including autism spectrum disorder, schizophrenia, and addiction . OT is pOtr promoter sequence, facilitated the identification of OTR expressing cells in additional brain regions, e.g., the median raphe nucleus and the lateral hypothalamus. This mouse model has been valuable in the identification of the phenotype of the cells expressing OTRs. For instance, OTRs have been found in serotonergic neurons , which are G protein-coupled receptors that, upon binding to OT, activate the Gq protein subunit and ultimately excite the cell. Autoradiographic studies have identified OTR expression in several regions of the rat brain, including the olfactory system, basal ganglia, hippocampus, central amygdala, and hypothalamus . The gen neurons , implicaCentral OT is important in homeostatic processes, such as thermoregulation , food inAdditional insights into the function of OT and the OTR are gained from the examination of genetically engineered mouse models. Female OT knockout (OTKO) mice show normal parturition and maternal behavior but are unable to nurse their pups demonstrating that in the mouse OT is not necessary for maternal behavior or labor but is essential for milk ejection . CompareUnlike OTKOs, OTRKO males display increased aggression in the resident-intruder task . It is pFurthermore, selective knockout of OTRs in the LS showed that OT plays a bi-directional role in fear regulation dependent on social context. Animals exposed to a non-fearful conspecific  or to sVarious factors influence OT signaling. OTRs are largely expressed centrally but their regional localization varies across species. For instance, mice and rats both express OTRs in the ventromedial nucleus of the hypothalamus VMH , 8], but, but8)],Oxytocin release from neurons of the PVN and the presence of OTRs within the PVN suggests the possibility that OT can directly modulate the stress reactive hypothalamic–pituitary–adrenal (HPA) axis. The HPA axis responds to stressors and activates neurons residing in the PVN causing increased synthesis and secretion of corticotropin-releasing factor (CRF). The release of CRF into the hypophyseal portal system enhances synthesis and release of adrenocorticotropic hormone (ACTH) from the anterior pituitary. In turn, ACTH acts on the adrenal cortex to stimulate release of glucocorticoids [cortisol in humans and corticosterone (CORT) in rats and mice]. Increased levels of circulating glucocorticoids can further inhibit HPA axis activity via glucocorticoid and mineralocorticoid receptors in the brain as well as acting upon specific brain sites to modulate behaviors .Oxytocin can putatively impact several sites within the HPA axis. PVN neurons that project to the median eminence release OT into the hypophyseal portal vasculature to stimulate adrenal glucocorticoid release by potentiating the actions of CRF at the anterior pituitary level in a similar fashion to the closely related neuropeptide vasopressin . By contEndogenous OT levels are also sufficient to alter HPA axis reactivity. ICV injection of OTA showed elevated ACTH and CORT levels prior to behavioral testing suggesting that endogenous OT levels can suppress HPA axis reactivity , 35. AddInterestingly, OT also promotes social buffering in response to stress, similar to the effect seen with fear . Female Oxytocin is strongly implicated in social bond formation and social behavior [for review see Ref. ], but maOxytocin signaling during early development may contribute to later anxiety. Prairie vole pups exposed to a single injection of OT on postnatal day 1 demonstrated an increase in serotonergic axon density in the anterior hypothalamus, cortical amygdala, and VMH but not in the PVN or medial amygdala. Such effects on serotonergic neurons could be a mechanism by which OT affects emotional behaviors, since serotonin is strongly linked to mood, and serotonin dysregulation is seen in depression and anxiety disorders .In adult animals, OT administration reduces anxiety-related behaviors in the elevated plus maze , 45, 46 Oxytocin also reduces measures of depression in the forced swim test (FST) and tail suspension test. In FST, rats treated with an OT analog spent less time immobile and more time swimming and climbing the walls of the chamber than saline-treated animals indicating an antidepressant effect . Similarin vitro studies found that progesterone could inhibit OT binding to the OTR  mice in both males  and steroid receptor coactivator (SRC)-1 were found to occupy the Ot promoter, leading to increased acetylation of histone H4 in the presence of 3β-diol. Taken together, the data suggest that in the presence of 3β-diol, ERβ binds the Ot promoter and recruits ligand-dependent coactivator SRC-1, which binds CBP, and forms a functional complex that acetylates histone H4 to drive Ot gene expression (Ot promoter, the co-activators recruited, and how this interaction modulates HPA axis function in vivo.Recent studies have begun to investigate the complex interaction between ERβ and the a et al.  demonstreceptors . Treatmeccupancy , 74. In Oxytocin has a wide range of roles in the brain and allows interesting and important directions for research. Current data suggest that the OT neurons of the PVN provide the principal OTergic innervation of the forebrain. The function of OT, through OTRs, is regionally specific; however, the localization of OTRs varies across species, age, and sex, so separating the effect of these variables is necessary to determine how animal studies translate to humans. Modulators of the OT system, particularly the steroid hormones, also provide additional regulatory targets since OT modulates HPA axis reactivity and participates in many diverse functions. In particular, ERβ is expressed by many neurons of the PVN, and ERβ activation increases OT synthesis and reduces anxiety and neuroendocrine responses in animals. Hence, such targets may be fruitful directions for future focus.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.


Stroke often aggravated age-related cognitive impairments that strongly affect several aspects of quality of life. However, few studies are, to date, focused on rehabilitation strategies that could improve cognition. Among possible interventions, aerobic training is well known to enhance cardiovascular and motor functions but may also induce beneficial effects on cognitive functions. To assess the effectiveness of aerobic training on cognition, it seems necessary to know whether training promotes the neuroplasticity in brain areas involved in cognitive functions. In the present review, we first explore in both human and animal how aerobic training could improve cognition after stroke by highlighting the neuroplasticity mechanisms. Then, we address the potential effect of combinations between aerobic training with other interventions, including resistance exercises and pharmacological treatments. In addition, we postulate that classic recommendations for aerobic training need to be reconsidered to target both cognition and motor recovery because the current guidelines are only focused on cardiovascular and motor recovery. Finally, methodological limitations of training programs and cognitive function assessment are also developed in this review to clarify their effectiveness in stroke patients. Sedentary older adults are prone to cardiovascular diseases, such as stroke  and types of exercise as well as timing of training initiation after stroke onset. Indeed, they might differ in their effectiveness to improve cognition and motor functions. Therefore, classic recommendations for aerobic exercise are reconsidered to target cognition as well as motor and cardiovascular functions. Finally, we discuss about the methodological limitations that may hamper the understanding of aerobic training effects on cognition both in human and animal studies.Cognitive disorders , motor dysfunctions  and cardiorespiratory fitness weakness are frequently observed in both elderly and stroke people. However, the severity of these impairments is exacerbated due to ischemia in older stroke patients , insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor  might also explain the influence of aerobic training on neuroplasticity and cognition in Human but this point remains controversial  the VEGF, which is a protein whose main role is to stimulate angiogenesis; (ii) the BDNF, which is a critical protein involved in brain plasticity. Indeed, this neurotrophin mediates numerous proteins expressions and molecular pathways, i.e., synapsin-I and synpatophysin, both involved in synaptic transmission and neurotransmitter release, C-AMP response element-binding protein (CREB) and beta calcium/calmodulinedependent kinase II (β-CaMKII) both contributing in long-term potentiation (LTP). BDNF thus promotes synaptic plasticity, neurogenesis and neuronal survival within the hippocampus  pathway , also involved in LTP within hippocampus  and by increase of both CREB phosphorylation and newborn cell survival  recombinant tissue plasminogen activator (rt-PA) was approval by the USA Food and Drug Administration for the treatment of acute ischemic stroke in human  on cognitive recovery remains to be further investigated .Low-intensity aerobic training seems to be more effective than high-intensity training to promote cognitive health. Indeed, animal studies have shown that low-intensity training induced superior positive effects on spatial learning task in a water maze test and on both object recognition and location tasks than intense training  to improve cognitive functions. Indeed, spatial learning and retention were better improved with the late training and was associated with the upregulation of BDNF , forced (treadmill) and voluntary (running wheel) training have similar beneficial effects on cognitive function after cerebral ischemia as indicated by improvement of both object recognition and location tests  and 6-min walk performances, that remained higher 1 year after the end of training compared with baseline value . However, it was demonstrated that high-intensity exercise could improve aerobic fitness by increasing the peak oxygen uptake  and presented a low relative risk for adverse effects , might find a compromise in rats by proving that training with gradually increased intensity on treadmill (from 5 to 26 m/min) could better improve motor function and produce higher hippocampal BDNF with lower stress compared to both stably low and high intensity training  were the most frequently used parameters , which is related to memory and learning  in both elderly and stroke people that mediate beneficial neuroplasticity in brain areas involved in cognitive functions. We also identify some methodological limitations in both human and animal studies such as the standardization procedure of aerobic exercise intensity and the chosen cognitive tests, depending on the target population, that remain one of the most important concerns. Moreover, our review article suggests that the combination of aerobic training with other exercises/therapies or treatments represent a promising strategy with strong clinical perspectives. Importantly, this review highlights the lack of a firm guideline for exercise recommendations targeting recovery of cognition in stroke patient. Therefore, no standard aerobic protocol has yet been established as a commonly accepted reference regarding intensity, timing of training initiation and exercise type. It also appears that investigating changes in cognitive-motor interplay are critical to develop appropriate rehabilitation to improve both cognition and motor control after stroke.All authors listed, have made substantial, direct, and intellectual contribution to the work, and approved its final version for publication. JL and J-JT: conceived the review focus, conducted literature review, summarized, and finalized the manuscript. PD: summarized, and finalized the manuscript. AC, CP-B and JL: reviewed literature, wrote first draft, and finalized the manuscript. All authors approved final version of manuscript.This work was supported by Aix-Marseille Université (AMU) and Centre National de la Recherche Scientifique (CNRS).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.


Parasitic helminths reside in immunologically-exposed extracellular locations within their hosts, yet they are capable of surviving for extended periods. To enable this survival, these parasites have developed complex and multifaceted mechanisms to subvert or suppress host immunity. This review summarises current knowledge of immune modulation by helminth parasites of ruminants and the parasite-derived molecules involved in driving this modulation. Such immunomodulatory molecules have considerable promise as vaccine targets, as neutralisation of their function is predicted to enhance anti-parasite immunity and, as such, current knowledge in this area is presented herein. Furthermore, we summarise current evidence that, as well as affecting parasite-specific immunity, immune modulation by these parasites may also affect the ability of ruminant hosts to control concurrent diseases or mount effective responses to vaccination.


Optical coherence tomography (OCT) is a rapid noncontact method that allows in vivo imaging of the retina and it has become an important component in clinical practice. OCT is a useful ancillary tool for assessing retinal diseases because of its ability to provide cross-sectional retinal images and quantitatively analyze retinal morphology. The introduction of spectral-domain OCT provided major improvements in image acquisition speed and image resolution. Future studies will address how these major technologic advances will impact the use of OCT in research and clinical practice. Optical coherence tomography (OCT) was introduced in 1991 as a noninvasive in vivo ophthalmic imaging technique for facilitating retinal thickness measurement . This hiOCT is an interferometric imaging technique that generates cross-sectional images by mapping the depth-wise reflections of low-coherence laser light from different kinds of tissue. Spectral-domain OCT (SD-OCT) or Fourier-domain OCT refers to Fourier transformation of the optical spectrum of the low-coherence interferometer. The optical spectrum output of an interferometer exhibits peaks and troughs, and the period of such modulation is proportional to the optical path differences in the interferometer. Imaging of multilayer objects, such as the retina, results in various modulation periodicities representing the depth of each layer. SD-OCT identifies the retinal thickness from the RPE to the inner limiting membrane .The most important advantage of SD-OCT compared with conventional time-domain OCT (TD-OCT) technique is the increased scanning speed , 4, 5. WThis report is an evidence-based review of the increasing role of OCT in the diagnosis and management of ocular disorders, particularly in age-related macular degeneration (AMD), diabetic macular edema (DME), and retinal vein occlusion (RVO).Macular edema, whether associated with diabetic retinopathy, uveitis, retinal vascular diseases ), or postcataract (Irvine-Gass) macular edema, or in idiopathic cases, can lead to severe visual loss if undetected and untreated. Current diagnostic techniques for assessing macular edema, including biomicroscopy, fundus photography, and fluorescein angiography (FA), are widely used, but interpretation of the results can be subjective, and subtle changes in retinal thickness in early stage macular edema may not be evident using these techniques.OCT is considered the best reference standard for detecting and quantifying macular edema compared to ultrasound, retinal thickness analysis, and scanning laser ophthalmoscopy , 7. CompCystoid macular edema  can be s DME is the most common cause of moderate visual loss in patients with diabetes . In 1998The prevalence of DME increases from 0% to 3% in individuals with recent diagnoses of diabetes to 28% to 29% in those with diabetes for longer than 20 years , making DME had been characterized as focal or diffuse based on clinical examination and FA findings . OCT allThe OCT findings were well correlated with other evaluation techniques for DME. Although there is a moderate correlation between the retinal thickness measured by OCT and visual acuity (VA), OCT cannot replace the VA because there is a high degree of variability . MacularTo investigate the relationship between VA and the CRT measured by OCT, 251 eyes of 210 patients with DME were enrolled in a cross-sectional and longitudinal randomized clinical trial [OCT also allows analysis of the integrity of the outer retinal layers in DME. Various studies have reported that the integrity of the outer retinal layer is linked directly to the visual prognosis –29. DisrMany treatment options exist for treating DME, such as focal therapies laser , pharmacIn 1985, the Early Treatment Diabetic Retinopathy Study (ETDRS) defined clinically relevant macular edema. The study further reported that focal or grid photocoagulation of eyes with clinically relevant macular edema reduced the 3-year risk of losing three or more lines of VA by 50%, from 30% in the control group to 15% in the laser group .Treating DME with peribulbar triamcinolone acetonide did not significantly improve the central subfield macular thickness measured by OCT or VA . IntraviThe newest frontier in treating DME involves the use of anti-VEGF agents. Ranibizumab , a monoclonal antibody fragment with binding affinity for VEGF-A, has been evaluated for treating DME . Pilot sSD-OCT is becoming an integral component in the diagnosis and management of AMD. OCT testing provides qualitative and quantitative assessment of various AMD presentations and detects early nonexudative AMD changes, such as drusen and RPE atrophy, and exudative AMD findings, such as intraretinal fluid, RPE detachment, retinal angiomatous proliferation, and choroidal neovascular membranes . OCT mayThe ability to detect intraretinal fluid on OCT images is an effective way to guide treatment and retreatment, because intraretinal fluid is associated with active neovascular membranes. OCT imaging features that may be associated with choroidal neovascularization include thickening or fragmentation of the RPE, choriocapillaris, intraretinal and subretinal fluid accumulation related to neovascularization exudation, and pigment epithelial detachment  Figure . Serial RVO is a common, sight-threatening retinal vascular disorder and the second most common retinal vascular disease after diabetic retinopathy. The clinical characteristics, prognosis, and response to treatment are affected by the location of the occlusion in the retinal venous vasculature, the presence of macular edema, and the extent of the retinal nonperfusion. The location of the occlusion may affect the severity of the clinical manifestations.Macular edema, a major cause of visual loss in patients with RVO , involveSome parameters seen on SD-OCT seem to be correlated negatively with visual recovery after RVO, such as foveal thickness, serous retinal detachment, central cystoid spaces, and pigment epithelial changes. When a very thick fovea (more than 700 microns) is seen on SD-OCT, an ischemic form of CRVO should be suspected . Loss ofThe current management of macular edema secondary to CRVO relies on two different approaches involving intravitreal therapy: anti-VEGF therapy , 50 or sSD-OCT technology has revolutionized the evaluation and treatment of macular edema. The technology facilitates quantitative assessment of the degree of retinal thickness, which has proved to be useful in the diagnosis, management, and follow-up of patients with macular edema, including assessment of foveal microstructural changes. OCT is a highly accurate and reproducible method for diagnosing macular edema that compares well to the standard clinical examination, ETDRS photos, and FA. Although it is useful for anatomic study, OCT is not an effective surrogate for functional tests such as VA measurement in the assessment of macular edema . Future


Many stillbirths result from pregnancy complications, whose root cause is abnormal development and function of the placenta . In ordeWe have developed a method for step-wise differentiation of such hPSCs, first into trophoblast precursor cells and then into terminally differentiated, functional trophoblast, including multinucleated syncytiotrophoblast (STB) and invasive extravillous trophoblast (EVT). These two cell types are the functional units of the placenta: STB carry out nutrient and gas exchange, while the EVT invade the maternal uterus and establish blood flow to the feto-placental unit. Our differentiation method is both reproducible and highly efficient, with >95% of cells becoming trophoblast in the culture dish, based both on expression of specific genes and on functional assays such as secretion of the pregnancy hormone, hCG. We recently applied this method to hPSCs carrying a chromosomal aneuploidy, Trisomy 21 (T21). It is known that trophoblast isolated from T21 placentas have a defect in differentiation into multinucleated, hCG-secreting STB . We aske


For thousands of years, humans have safely consumed microorganisms through fermented foods. Many of these bacteria are considered probiotics, which act through diverse mechanisms to confer a health benefit to the host. However, it was not until the availability of whole-genome sequencing and the era of genomics that mechanisms of probiotic efficacy could be discovered. In this review, we explore the history of the probiotic concept and the current standard of integrated genomic techniques to discern the complex, beneficial relationships between probiotic microbes and their hosts. A multitude of autochthonous  commensal bacterial species inhabit the mucosal surfaces of the gastro-intestinal tract (GIT), as well as those of the nose, mouth and vagina. It has long been held that the consumption of allochthonous (transient) beneficial bacteria, either through food products or supplements, has a positive influence on general health and well-being of the host via commensal interactions with the GIT immune system and resident microbiota. These beneficial microorganisms, known as probiotics, are defined by the World Health Organization as “live microorganisms, which when administered in adequate amounts, confer a health benefit upon the host” FAO/WHO . Over thFor millennia, humans have consumed microorganisms via fermented foods, which served to prevent putrefaction as well as increase sensory aspects in the food. Some of the first fermentations were likely the result of serendipitous contaminations in favourable environments resulting in soured milk products such as kefir, leben, koumiss, yogurt and sour cream—products that are still consumed worldwide Hosono . FurtherThe Prolongation of Life: Optimistic Studies, Metchnikoff (Lactobacillus delbrueckii subsp. bulgaricus) was recoverable from faeces; reduced the prevalence of putrefactive toxins; and aided in the treatment of colitis following transplantation to the large intestine  from human faeces that could survive passage through the GIT  belonging to the phylum Firmicutes , an exponential advancement in sequencing processing, genome assembly and annotation technologies, at increasingly economical pricing, has yielded well over a thousand publicly available genomes  survival through gastrointestinal transit and adhesion to intestinal epithelia; (ii) competitive exclusion and antimicrobial activity; and (iii) modulation of the host GIT immune system Fig. .Fig. 3aLactobacillus casei demonstrated a marked increase of mono-unsaturated fatty acids in response to acidification ] source significantly increased acid survival by incorporating the oleic acid into the membrane, which is reduced to stearic acid (C18:0) in the acidified environment  encoding a functional HPK was knocked out, resulting in a mutant with increased sensitivity to acid stress compared to the parent strain. Furthermore, transcriptomic analysis via DNA microarray comparing the lba1524 mutant to wild-type demonstrated an impact on 80 genes  transporter efflux pumps , was recently identified through in silico genome analysis and functionally characterized , lactulose, fructo-oligosaccharide and inulin, are resistant to gastric acidity, hydrolysis and gastrointestinal absorption  were found to be induced in the murine GIT  resulted in a 100- to 1,000-fold decrease in persistence capacity of the L. plantarum lp_2940 knockout mutant in a mouse model. In L. acidophilus NCFM, in silico genome screening lead to the selection of five putative adhesion cell surface proteins, including a fibronectin binding protein (FbpA), S-layer protein (SlpA), mucin-binding protein (Mub) and two R28 homologues involved in streptococcal adhesion  contains domains that bind to laminin and collagens  pilus-encoding gene cluster has been identified in Bifidobacterium breve UCC2003  which produced modified teichoic acids with less d-alanine than the parent strain  in situ , revealed an immunomodulatory phenotype using in vitro bacterial-DC co-incubation assays, suggesting the potential of multiple unknown MAMPs associated with the S-layer of L. acidophilus NCFM. Researchers are also trying to understand the complex dynamic of host-microbe crosstalk by using whole transcriptome profiling of human intestinal epithelia upon exposure to probiotics. In one study, transcriptomes were obtained from the mucosa of the proximal small intestines of healthy volunteers exposed to probiotic L. acidophilus, L. casei, and L. rhamnosus (van Baarlen et al. L. acidophilus NCFM corroborated these data (O’Flaherty and Klaenhammer Bifidobacterium bifidum PRL2010 transcriptome analyses with both in vitro human cell lines and in vivo murine models demonstrated the capacity for strain PRL2010 to modulate host innate immunity (Turroni et al. Like many probiotic effectors, most MAMPs are found on the cell surface of Gram-positive microbes. Recently, the genomes and proteomes of several lactobacilli were bioinformatically screened to create a secretome database cataloging the various extracellular proteins in LAB (Kleerebezem et al. While the paradigm of discovery based genomics in probiotic LAB has uncovered vital aspects of probiotic mechanisms, it has also revealed the complexity of these interactions with the resident microbiota and the mucosal immune system. But with this challenge has come great opportunity. For example, probiotic bacteria are now being explored as suitable models for vaccine/drug delivery, due to their close association with host immunity and immunomodulatory action (Kajikawa et al.


Low back pain with resultant loss of function, decreased productivity, and high economic costs is burdensome for both the individual and the society. Evidence suggests that intervertebral disc pathology is a major contributor to spine-related pain and degeneration. When commonly used conservative therapies fail, traditional percutaneous or surgical options may be beneficial for pain relief but are suboptimal because of their inability to alter disc microenvironment catabolism, restore disc tissue, and/or preserve native spine biomechanics. Percutaneously injected Multipotent Mesenchymal Stem Cell (MSC) therapy has recently gained clinical interest for its potential to revolutionarily treat disc-generated (discogenic) pain and associated disc degeneration. Unlike previous therapies to date, MSCs may uniquely offer the ability to improve discogenic pain and provide more sustained improvement by reducing disc microenvironment catabolism and regenerating disc tissue. Consistent treatment success has the potential to create a paradigm shift with regards to the treatment of discogenic pain and disc degeneration. It is well documented that low back pain is a common and debilitating condition. Costs related to its medical expenses and lost wages exceed $100–200 billion annually in the United States alone. The intervertebral disc has been identified as the single most common pain generator for low back pain . There iWhen defining intervertebral disc (IVD) pathology, it is prudent to first briefly review normal disc anatomy and physiology. In simple terms, a disc is composed of the nucleus pulposus (NP), annulus fibrosis (AF), and the vertebral endplate. A remnant of the notochord, the NP contains randomly organized collagen and elastin fibers embedded in a highly hydrated gel-like matrix rich in proteoglycan (PG), which is synthesized by chondrocyte-like intervertebral disc (IVD) cells. The nuclear matrix is about 70–90% water, which is contained within the domains of proteoglycan. The NP plays an important role in spine flexibility and axial load distribution. The surrounding annulus (AF) is composed of parallel collagen fibers arranged in concentric lamellae (10–20 sheets). The cells of the AF are morphologically and phenotypically similar to fibroblasts. The annulus provides tensile strength and works in conjunction with the nucleus to absorb shock. The third component, the cartilaginous vertebral endplate, is composed of both hyaline and fibrocartilage and is intimately involved in connecting and anchoring the disc to the vertebral body –6. InnerThe IVD is a dynamic structure. The disc microenvironment appears to rely on a harmonious balance between anabolic and catabolic factors important for normal disc cell turnover. Growth factors including TGF-b, BMP, GDF-5, and IL-GF are examples of important anabolic factors. Conversely, catabolic metabolism is achieved via catabolic enzymes, inflammatory cytokines, proteinases, and aggrecanases. Examples include IL-1, TNF-alpha, ADAMTS, and MMPs. The disc is relatively avascular, consequently creating a harsh microenvironment  for IVD cells. Similar to most cartilaginous structures, this limited vascular support is a significant contributor to the poor natural regenerative capacity of the IVD when metabolic homeostasis is disrupted , 6.Clinical history and physical exam can be suggestive, but not diagnostic, of intervertebral disc pathology. Advanced imaging and diagnostics are needed for more accurate confirmation , 18. MRILiterature review reveals that these three pathologic disc states found in Along with a proper understanding of disc anatomy and pathophysiology, knowledge of currently available treatment modalities for discogenic LBP is important for enhancing patient outcomes and mitigating risks. If conservative treatment modalities fail, trained clinicians have traditionally used interventional techniques to aid in providing pain relief. However, we propose that interventional therapies should optimally strive to fulfill the following trio of treatment objectives: decline and/or resolution of primary nociceptive disc pain , slowing and/or reversal of the catabolic metabolism (and associated degradation) within the disc microenvironment, and partial or complete restoration of disc tissue  12, 28––28–30. RMany of these therapies have shown some benefit ; however, with the exception of a few listed emerging therapies, each are considered suboptimal in the treatment of discogenic pain because they fail to accomplish the entirety of the three previously stated objectives for disc pathology treatment. Most demonstrate evidence of short-term pain improvements, but many lack evidence of sustained benefit and/or complete relief. Though difficult to study, nearly all (with the exception of biologics) lack cellular support and consequently evidence demonstrating an ability to slow or reverse microenvironment catabolism, regenerate disc tissue, or restore native spine biomechanics. To achieve disc access, many of these interventions may promote a predisposition to further disc pathology with no ability to reverse this negative cascade , 31. FurTraditional conservative, interventional, and surgical treatment modalities used in isolation or in combination have reported successes but are insufficient to accomplish the entirety of the three objectives outlined for the treatment of disc pathology. Percutaneously delivered Multipotent Mesenchymal Stem Cell (MSC) treatment has recently gained attention for its potential to revolutionize the treatment of discogenic LBP and associated disc degeneration. Contrary to traditional interventions, MSC therapy provides the necessary cellular support for regeneration and current research suggests that these treatments may have the ability to uniquely accomplish the three stated objectives for treating disc pathology .It is important to first highlight key fundamental concepts of MSCs prior to discussing their clinical applicability in MSC-based intradiscal treatments. According to the International Society of Cellular Therapy, MSCs are defined by their ability to adhere to plastic under standard tissue culture conditions, express certain cell surface markers and lack expression of other markers, and differentiate into osteoblasts, adipocytes, and chondroblasts under in vitro conditions . InquiryCurrently, tissue engineers and clinicians appear most interested in the application of MSCs derived from bone marrow, adipose, and umbilical cord tissue. Synovium, skeletal muscle, and periosteum have also been rarely sourced . InteresHuman umbilical cord tissue-derived Mesenchymal Stem Cells (HUC-MSCs) may be easily collected from cord tissue and have potential allogeneic application. Perhaps the best source of cells for expansion, HUC-MSCs, could be well suited for allogeneic use because they appear to demonstrate low-immunogenicity . There iResearchers have identified other cell types worth mentioning that may work in concert with MSCs. There has been some interest in a niche of cells located within the disc referred to informally as mesenchymal progenitor cells. Recognition of these cells may be prove beneficial if they can be stimulated by exogenously administered MSCs or other biologics to aid in the regenerative process , 92–94. Once harvested and isolated via centrifugation, MSCs  follow one of a few basic treatment modes/models prior to percutaneous cell implant . Cells cIn vivo models on the other hand are helpful for the study of safety, feasibility, and efficacy of MSC transfer. Several studies using an animal in vivo model have yielded valuable information about how injected cells perform within the disc environment . The pheA third basic cell transplant model exists in which cells may be isolated from a donor and transplanted into a separate recipient . Cells in this model may undergo in vitro culturing and/or may be simply stored without manipulation prior to future transplantation . Unlike Bench-work research has provided positive evidence for the potential benefits of MSC transplant into pathologic discs. Disc microenvironment is important for growth and viability of native IVD cells as well as injected MSCs. In vitro studies combining MSCs and IVD cells have demonstrated bidirectional synergy promoting an anabolic environment. An in vitro study of combined rat IVD-NP cells and human synovial MSCs revealed suppression of genes related to matrix degradative enzymes and inflammatory cytokines . Lui et The application of MSCs in various in vivo animal models has demonstrated similar anticatabolic and disc-tissue regenerative effects. In postnucleotomy rabbit models, injection of BM-MSCs has demonstrated suppression of nuclear collagen type 1 formation (causing fibrosis) as well as restoration of disc height values and MRI signal intensities approaching 81% and 91%, as compared to sham-treated discs at 67% and 60%, respectively , 109.Using a canine in vivo model, Hiyama et al. similarly demonstrated that an injection of BM-MSCs into the nucleus of enzymatically degenerated (postnucleotomy) discs showed increased proteoglycan (PG) content consistent with normal nondegenerated discs at 12 weeks posttreatment; control postnucleotomy discs showed marked decrease in PG , 109. A Since there are clear mechanical differences on the IVD with a quadrupedal animal, human in vivo studies are needed to confirm reduction or elimination of discogenic pain. A pilot investigation by Orozco et al. using in vitro expanded autologous BM-MSCs has demonstrated early safety and feasibility in a percutaneous approach. Ten patients with persistent discogenic LBP, DDD (>50% disc height loss), and an intact outer annulus (per discography) underwent a single injection after failing 6 months of conservative treatment. The authors reported statistically significant improvements in both pain and disability after 3 months, with results persisting at one year. Although there was no recovery of disc height on MRI, water content of the discs significantly improved at one year . In a reThese cited studies have added to the growing body of evidence suggesting that percutaneously injected MSCs may uniquely provide clinical improvements in discogenic pain, reduced microenvironment catabolism, and reduced disc tissue loss. In vitro studies show evidence that MSCs promote anti-inflammatory and anticatabolic effects within the disc microenvironment. Furthermore, in vitro and in vivo studies  have demonstrated MSC and IVD cell proliferation, nuclear matrix production, improved annular integrity, and recovered disc height. In vitro culturing of human MSCs has shown many of these same effects. Though limited clinical studies have been conducted to date, improvements in pain and function following intradiscal MSC injection have been demonstrated. Evidence supporting disc tissue regeneration exists and there is evidence now demonstrating at least partial recovery of disc tissue in both animals and humans, , 112. ThThe efficacy of intradiscally injected MSC based treatments may be potentially enhanced by combination therapies. The concept of an added cell matrix or scaffold has been introduced and has been shown to aid in cell delivery, orientation, differentiation, growth, replication, cellular metabolism, and sustained viability. Naturally occurring biopolymers can be used alone or in combination as hydrogel scaffolds for in vitro and in vivo MSC preparation and treatment. Examples include collagen-type I and solubilized atelocollagen, calcium alginate, chitosan, KLD-12 peptide, carboxymethyl cellulose, and collagen-type II. Hyaluronan-based hydrogels have been frequently used as a scaffold for MSCs with noted success in rat, sheep, and more recently human models , 111. AnThe clinical application of intradiscally injected MSCs has raised potential concerns. Chief among these is the potential for MSC-promoted carcinogenesis. It is well documented that embryonic stem cells (found earlier in cell lineage) can promote tumor formation . HoweverPercutaneous injection of MSCs into the intervertebral disc may uniquely fulfill the objectives of treating disc pathology with clinical improvements in pain and decreased microenvironment catabolism. Some evidence suggests that MSCs may fulfill the final major objective of reversing disc tissue loss. At this time, MSC-based therapies appear most suited for the treatment of discogenic pain secondary to IDD and DDD, serving as an obviation of more invasive surgical interventions. If successfully accomplished early in the disease process, this proactive approach may aid in preserving native spine biomechanics, ideally resulting in less degeneration of the IVD and surrounding tissues structures and again serving as an obviation to future surgical interventions. As the therapy advances, there may be some role for these therapies in improving the disc microenvironment and structural integrity in cases of small disc herniations; some have suggested a similar role in postsurgical discs  , 96, 123A continued collaboration between researchers and physicians will be important in optimizing the therapeutic potential of MSC-based therapies as well as the refinement of optimal MSC cell type, concentration, and supportive componentry. As the field advances, factors unique to both autologous and allogeneic cell preparations should be further studied and considered with each having advantages and disadvantages. Ideally, in vitro cell culturing should be compared to same-day therapies in terms of safety and efficacy. Given the harsh environment of the disc  development of future therapies may include preconditioning cells prior to transplant  to enhance survivability. Additionally, studies are needed to determine an optimal time course for applying MSC treatments. Traditionally, symptomatic patients have been encouraged to pursue conservative therapies, reserving interventional injections for nonresponders and spine surgery for severe, refractory axial pain. This dogma has existed in large part because previous interventional and surgical options have inconsistently provided pain relief and have not offered the same restorative potential that may be present with MSC-based treatments. If demonstrated consistently safe and effective, MSC-based treatment may lead to a paradigm shift towards more aggressive, nonsurgical care for patients with discogenic LBP (IDD/DDD) and other forms of compromised disc integrity.


Despite numerous promising discoveries in contemporary cancer research, and the emerging innovative cancer treatment strategies, the global burden of malignant diseases is expected to rise, in part due to the rapidly aging human populations. Consequently, there is an urgent need to design and validate new treatments in oncology, especially for cancer types with presently dismal prognosis due to limited treatment options, such as for patients suffering from glioblastoma multiforme. The efforts to address such need rely in large part on the recently introduced powerful technologies including high-throughput screens to uncover genetic and functional vulnerabilities of specific types of human tumors. Such effort also raises hopes to identify molecular targets so far associated with pathologies other than malignancies, and thereby bring about opportunities to repurpose already existing, approved drugs that target such mechanisms. Apart from lower cost of introducing such treatments for oncological diseases, drug repurposing is also good news due to much shorter time needed from target discovery to clinical applications, an aspect particularly valuable in disease contexts such as glioblastoma.in vitro. The screen design differed from previous genome-wide shRNA screens in its subtractive nature, where gene silencings that caused lethality in lung cancer cell lines were “subtracted” from those causing lethality in glioblastoma lines. In this way, the authors identified genes specifically required for glioblastoma growth. Analysis of the gene list revealed a rather unexpected discovery – the top five pathways represented in this gene list involved neuro-transmitter receptor signaling. The authors subsequently demonstrated that one of these pathways, mediated by the dopamine receptor subtype 2 (DRD2), plays a critical role in glioblastoma mitogenic signaling.A successful example of the latter approach has now been reported in a new exciting study published in Oncotarget. Through a series of genome-wide shRNA screens, Li et al.  globallyUpon activation, DRD2 activates the trimeric G-protein complex through a canonical interaction. The G subunit of this complex, in turn recruits a GTPase that hydrolyzes RAP1-GTP, a small G-protein in the Ras superfamily [Importantly, the authors demonstrated that DRD2 antagonists, clinically used as anti-psychotic drugs, harbor anti-glioblastoma activities. Moreover, these activities are synergistic when combined with EGFR inhibition. As discussed in the article, the work offers significant translational implications and suggests that FDA approved anti-psychotic agents may be repurposed as glioblastoma therapeutics. The notion is particularly attractive since these agents are known to cross the blood-brain barrier.Beyond the obvious translational implications of dopamine antagonists as glioblastoma therapeutics, the findings by Li et al. raise additional biological implications. First, it is well-appreciated that glioblastoma is an aggressively infiltrative disease. Yet, it rarely metastasizes beyond the central nervous system (CNS) . The obsAnother puzzle in glioblastoma therapeutic development involved the observation that while Epidermal Growth Factor Receptor (EGFR) dysregulation is critically important in the pathogenesis of glioblastomas , EGFR inTo the extent that dopamine influences reward systems in the brain, emotion, and personality traits, the work further raises the possibility that personality and emotion may impact the risk of cancer development or growth through modulation of dopamine release. Interestingly, studies have related cancer risks to personality types .There is an emerging recognition of the role of neurotransmitters in regulating cancer phenotypes. For instance, Magnon et al. published in a recent Science article that prostate cancers are infiltrated with parasympathetic cholinergic fibers. Moreover, the cholinergic neurotransmitters released by these fibers promoted tumor dissemination . The wor


Consistent with expectations, cocaine users showed abnormal neural activity in several frontostriatial regions during implicit moral picture processing compared to their non-cocaine using peers. This included reduced moral/non-moral picture discrimination in the vACC, vmPFC, lOFC, and left vSTR. Additionally, psychopathy was negatively correlated with activity in an overlapping region of the ACC and right lateralized vSTR. These results suggest that regular cocaine abuse may be associated with affective deficits which can impact relatively high-level processes like moral cognition.Investigations into the neurobiology of moral cognition are often done by examining clinical populations characterized by diminished moral emotions and a proclivity toward immoral behavior. Psychopathy is the most common disorder studied for this purpose. Although cocaine abuse is highly co-morbid with psychopathy and cocaine-dependent individuals exhibit many of the same abnormalities in socio-affective processing as psychopaths, this population has received relatively little attention in moral psychology. To address this issue, the authors used functional magnetic resonance imaging (fMRI) to record hemodynamic activity in 306 incarcerated male adults, stratified into regular cocaine users ( Academic interest in this aspect of our social world has been longstanding, but recent decades have seen a rapid expansion of moral psychological research into a myriad of scientific disciplines. Cognitive neuroscience is one of the more recent members— employing newly emerging neuroimaging techniques to provide novel insights into the neural circuitry engaged by moral cognition.Many differing theoretical perspectives on moral cognition have followed, but one aspect which is widely shared amongst them is the bifurcation of moral decision-making into two relatively distinct, though highly interdependent, processes. The first, which we will refer to as “moral intuition,” consists of relatively automatic and reflexive operations through which an individual attends and responds to the morally-salient aspects of a given stimulus. “Moral deliberation,” by contrast, denotes more deliberative and cognitively taxing processes , dorsolateral prefrontal cortex (dlPFC), temporo-parietal junction (TPJ), cingulate cortices (ACC and PCC), and amygdala  and a proclivity toward immoral behavior. Psychopathy is the most common disorder studied for this purpose—a personality disorder typified by a suite of interpersonal, affective, and behavioral characteristics including grandiosity, callousness, and lack of empathy . Moreover, both current and former cocaine-dependent individuals exhibit many of the same abnormalities in socio-affective processing as psychopaths. Psychopaths exhibit significant deficits in affective empathy and facial affect recognition  and a muted response to all non-drug affective stimuli  to evaluate brain activity during the performance of a moral intuitive processing task by individuals with varying levels of psychopathy and (past) cocaine use. Using the Mind Research Network's mobile MRI system, incarcerated participants were scanned while viewing pictures that did or did not contain moral violations , as well as affectively neutral pictures , and judging whether each picture depicted a scene occurring indoors or outdoors. Participants were therefore exposed to morally salient stimuli, but were not made aware of the moral/non-moral picture distinction . The remaining incarcerated participants were assigned to the Non-Cocaine Using (NCU) group (n = 219). Additionally, a subset of the NCU group was matched with the CU group on age, IQ, psychopathy and severity of use for substances other than cocaine. This subset will be referred to as the Matched Non-Cocaine Users (mNCU). Demographic information on each group is provided in Table This study included 316 incarcerated male volunteers recruited from correctional facilities in New Mexico and Wisconsin. Inclusion criteria were: age between 18 and 50, reading level above 4th grade, IQ above 75, no history of seizures, no current Diagnostic and Statistical Manual of Mental Disorders  was administered in order to gauge the accuracy of other self-report measures. The BIDR Paulhus,  is a 40 IQ was estimated using the Vocabulary and Matrix Reasoning subtests of the Wechsler Adult Intelligence Scale  were selected, largely from the International Affective Picture System .http://www.neurobs.com/).Moral, non-moral, and neutral picture trials were presented in a random order and interspersed with 25 null trials in which a fixation cross was presented for the entire trial. The 100 total trials were presented across two separate runs of the task. Images were rear-projected into the scanner using an LCD projector, controlled by a PC computer. Tasks were designed and presented and responses were recorded using Presentation software  equipped with a 12-element head coil. The EPI gradient-echo pulse sequence  effectively covered the entire brain (150 mm) in 2.0 s. Head motion was minimized using padding and restraint.To correct residual head motion, “bad” images (confounded by motion or radio-frequency spikes) were estimated and removed using ART-Repair . Functional images were spatially normalized to the MNI template and smoothed (8 mm FWHM). Picture presentations  and the rating period for all pictures were modeled as four separate events. Each event was modeled with a six (picture) or four (indoor/outdoor judgment) second hemodynamic response function. Functional images were computed for each participant that represented brain activation associated with viewing moral, non-moral, or neutral pictures. The moral > non-moral picture comparison evaluated brain activation to morally-salient pictures while controlling for general emotional and social content.Imaging data were analyzed using SPM5 . To correct for skew within the CU sample, severity of cocaine use was calculated by taking the square root of the duration of regular use divided by the participant's age , although dimensional models have been proposed more recently . Age, IQ, psychopathy, severity of use for cocaine, alcohol, cannabis, heroin, other opiates, other amphetamines and nicotine, and duration of abstinence for alcohol, cannabis, heroin, other opiates, other amphetamines and nicotine were also entered in as covariates into this regression . There were no significant group differences between CUs, mNCUs, or NCUs in the number of abstinent participants or the duration of abstinence for any substances . Peak coordinates for these regions were drawn from the moral > non-moral functional maps of a separate sample of 36 non-incarcerated participants . For the group comparison of CUs and mNCUs, the cluster threshold was set at ≥ 1026 mm3 (38 contiguous voxels). For the regression analysis within the entire incarcerated sample, the cluster threshold was set more stringently at ≥ 1134 mm3 (42 contiguous voxels).Whole-brain analyses were thresholded at We first examined the main effects of viewing moral > non-moral pictures across all non-incarcerated participants Figure . ConsistRelative to matched non-cocaine users, cocaine users showed significantly reduced hemodynamic activity in the moral > non-moral contrast in several cortical and subcortical regions. Whole-brain analysis revealed reduced activity in both the left subgenual anterior cingulate cortex (sgACC) and right superior frontal gyrus (SFG). Small-volume correction revealed additional areas of hypoactivation. In the left hemisphere, these included the ventral and dorsal striatum (vSTR and dSTR), ventral anterior cingulate cortex (vACC), ventromedial prefrontal cortex (vmPFC), and middle temporal gyrus (MTG). On the right, hypoactivity was observed in the dorsomedial prefrontal cortex (dmPFC) extending into the rostral anterior cingulate cortex (rACC), and ventrolateral prefrontal cortex (vlPFC) extending into the anterior insula (aINS). Hypoactivation was observed bilaterally in the parahippocampal gyrus (PHG) and anterior thalamus (THAL). Cocaine users did not show greater hemodynamic activity in the moral > non-moral contrast, compared to the mNCUs, in any regions of interest (see Table n = 61), duration of abstinence from cocaine was negatively correlated with hemodynamic activity in the right posterior cingulate cortex (PCC) for the moral > non-moral contrast , subgenual cingulate cortex (sgACC), and ventromedial prefrontal cortex (vmPFC). On the right, hypoactivity was observed in the dorsomedial prefrontal cortex (dmPFC) extending into the pregenual anterior cingulate cortex (pgACC), and ventrolateral prefrontal cortex (vlPFC) extending into the anterior insula (aINS). Additionally, cocaine users higher in psychopathic traits exhibited additional neural abnormalities in the right vSTR and more severe aberration in the right pgACC.CUs exhibited reduced activity in the left vSTR during moral vs. non-moral picture processing, when compared to mNCUs. Moreover, within the CU group, hemodynamic activity in the right vSTR was negatively correlated with Factor 1 scores. A plethora of research has established that this reward pathway is involved in the perception of social stimuli and social interaction  levels by blocking presynaptic dopamine transporter  and the other adjacent to the vmPFC (vACC). In both these regions, mNCUs exhibited greater hemodynamic activity for moral vs. non-moral stimuli, while CUs did not. The pgACC is selectively engaged when task-irrelevant information interferes with performance due to its emotional content  of the participant during regular use, the average quantity of cocaine used during periods of regular use and the duration of abstinence from cocaine use prior to scanning. Moreover, devising a substance use measure which applies to individuals who used a drug with some frequency, but fewer than three times a week, would help to normalize the distribution of that measure within the population.Consistent with our second hypothesis regarding the interaction between psychopathy and cocaine abuse, within the cocaine users, higher Factor 1 scores were associated with reduced activity in the right vSTR and rACC. These regions have been implicated separately in both substance abuse and psychopathy researchSeveral studies have identified functional .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.