Neurodegenerative and psychiatric diseases (NPDs) are associated with chronic or flaring brain inflammation and infiltrations of cytotoxic lymphocytes in the inflamed brain parts, causing mild to severe or even lethal brain damage. NPDs thus show features of autoimmune diseases (De Haan P et al., Frontiers in Psychiatry 8: 46, 2017). An autoimmune disease can be effectively treated by tolerization, in which the immune response to the primary self-components that are targeted by the adaptive immune system e.g. the primary self-antigens, pSAgs of the disease is reversed into an immune tolerance response. We at Amarna have developed a replication-defective gene delivery platform named SVec that based on its safety, non-immunogenicity and tolerogenicity properties is highly effective in inducing immune tolerance to pSAgs of autoimmune diseases. For one of the major NPDs, multiple sclerosis (MS) we have recently demonstrated that SVec-mediated expression of myelin oligodendrocyte glycoprotein (MOG), the pSAg of the disease, in liver cells of mice protects the tolerized animals from developing the disease. SVec is therefore excellently suited for developing effective tolerization therapies for patients with autoimmune diseases including NPDs.In order to design effective tolerization treatments for other NPDs, the pSAgs of the disease need to be identified. A pSAg of an autoimmune disease is predominantly or exclusively expressed in the affected tissue and all patients have a cellular immune response to the pSAg. Induction of an immune response to the pSAg in an animal results in the development of the disease. In order to identify pSAgs of autoimmune diseases traditionally, animals were immunized with the pSAg protein or peptides derived thereof in the presence of complete Freund's adjuvant (CFA) to enhance the immune response to the pSAg. This method, however, is highly labour-intensive, time-consuming and costly. We developed a replication-competent Alphaviral replicon vector system named AlphaSelect to reliably and cost-effectively test candidate pSAgs for their capacity to induce autoimmune disease symptoms in animals (De Haan P et al., Human Vaccines & Immunotherapeutics 17: 14-21, 2020). Recombinant AlphaSelect vectors encoding candidate pSAgs of amyotrophic lateral sclerosis (ALS) have been made and tested in animals to induce the characteristic disease symptoms. Here we report on the results of the initial immunization studies and on the feasibility of this approach to reach our long term ambition of developing effective tolerization therapies not only for NPDs, but also for other autoimmune diseases such as obesity, diabetes mellitus, atherosclerotic cardiovascular disease, arthritis, inflammatory bowel diseases and chronic obstructive pulmonary disease.

Viruses have evolved to efficiently express their genes in host cells, which makes them ideally suited as gene delivery vectors for gene and immunotherapies. Replication competent (RC) viral vectors encoding foreign or self-proteins induce strong T-cell responses that can be used for the development of effective cancer treatments. Replication-defective (RD) viral vectors encoding self-proteins are non-immunogenic when introduced in a host naïve for the cognate virus. RD viral vectors can be used to develop gene replacement therapies for genetic disorders and tolerization therapies for autoimmune diseases and allergies. Degenerative/inflammatory diseases are associated with chronic inflammation and immune responses that damage the tissues involved. These diseases therefore strongly resemble autoimmune diseases. This review deals with the use of RC and RD viral vectors for unraveling the pathogenesis of immune-related diseases and their application to the development of the next generation prophylactics and therapeutics for todays’ major diseases.

Viruses efficiently transfer and express their genes in host cells and evolve to evade the host's defense responses. These properties render them highly attractive for use as gene delivery vectors in vaccines, gene, and immunotherapies. Among the viruses used as gene delivery vectors, the macaque polyomavirus Simian Virus 40 (SV40) is unique in its capacity to evade intracellular antiviral defense responses upon cell entry. We here describe the unique way by which SV40 particles deliver their genomes in the nucleus of permissive cells and how they prevent presentation of viral antigens to the host's immune system. The non-immunogenicity in its natural host is not only of benefit to the virus but also to us in developing effective SV40 vector-based treatments for today's major human diseases.

The risk of unexpected uterine leiomyosarcoma (LMS) following surgery for presumed benign leiomyoma is quoted to be between 1 in 498 and 1 in 5000. The objectives of the present study were to determine the prevalence of uterine LMS in a specific patient population and the rate of diagnosis of occult uterine LMS and to evaluate the risk of unintended morcellation of LMS in Saskatchewan.This study was a Canadian Task Force Classification II-2 multicentre retrospective cohort study in academic-affiliated tertiary care centres. All women with the histopathologic diagnosis of uterine LMS in Saskatchewan between January 2000 and December 2014 were included. Women with metastatic LMS at diagnosis or other types of uterine sarcomas were excluded. Data including patients' characteristics, clinical presentation, physical examination findings, imaging, pathology reports, surgical interventions, and survival outcomes were reviewed.A total of 28 patients had a confirmed histopathologic diagnosis of LMS over the 15-year study period. Approximately 26 212 hysterectomies were performed in Saskatchewan over the same time frame. The prevalence of uterine LMS in this patient population over the study time frame is estimated to be one in 853. Mean age at diagnosis was 53.8 ± 10.0. Medical records of 25 patients could be retrieved, and 15 cases (60%) had an occult diagnosis. There were five cases of unintended morcellation (one power, four mechanical). Survival outcomes were comparable in women with unintended morcellation of occult disease and in those without morcellation.This study contributes to the existing body of literature on morcellation of occult LMS, and it ascertains the rate of LMS in a patient population. The results of this study provide valuable information to health care professionals, policy makers, and women in Saskatchewan so that they may make more informed decisions concerning uterine masses.On estime que le risque de léiomyosarcome (LMS) utérin inattendu après une chirurgie pour un léiomyome présumé bénin se situe entre 1 sur 498 et 1 sur 5 000. Cette étude visait à déterminer la prévalence du LMS utérin chez une population particulière de patientes et le taux de diagnostic de LMS utérin occulte ainsi qu’à évaluer le risque de morcellement accidentel d’un LMS en Saskatchewan.Il s’agissait d’une étude de cohorte rétrospective multicentrique de niveau II-2 selon les critères du Groupe d’étude canadien menée dans des centres universitaires de soins tertiaires. Nous avons retenu toutes les femmes ayant reçu un diagnostic histopathologique de LMS utérin entre janvier 2000 et décembre 2014 en Saskatchewan, et avons exclu celles qui avaient des métastases au moment du diagnostic et celles qui étaient atteintes d’autres types de sarcomes utérins. Nous avons examiné des données comme les caractéristiques des patientes, les signes cliniques, les observations à l’examen physique, les rapports d’imagerie et de pathologie, les interventions chirurgicales et la durée de survie.En tout, 28 patientes ont fait l’objet d’un diagnostic histopathologique confirmé de LMS pendant les 15 années de l’étude. Durant cette même période, environ 26 212 hystérectomies ont été pratiquées en Saskatchewan. Nous estimons la prévalence des LMS utérins dans le cadre de l’étude à 1 sur 853. L’âge moyen au diagnostic était de 53,8 ± 10,0 ans. Nous avons pu obtenir les dossiers médicaux de 25 patientes, et 15 LMS (60 %) étaient des cas occultes. Il y a eu cinq cas de morcellement accidentel (un avec un appareil électrique, quatre avec des instruments manuels). La durée de survie des femmes présentant une tumeur occulte était semblable que la tumeur ait été morcelée ou non.Cette étude s’ajoute à la littérature sur le morcellement des LMS occultes et évalue le taux de LMS dans une population de patientes. Les résultats offrent aux professionnels de la santé, aux responsables des politiques et aux Saskatchewanaises des renseignements précieux qui les aideront à prendre des décisions éclairées concernant les masses utérines.

Gene therapy has been shown to be a feasible approach to treat inherited disorders in vivo. Among the currently used viral vector systems, adeno-associated virus (AAV) vectors are the most advanced and have been applied in patients successfully. An important drawback of non-integrating AAV vectors is their loss of expression upon cell division, while repeating systemic administration lacks efficacy due to the induction of neutralizing antibodies. In addition, a significant percentage of the general population is not eligible for AAV-mediated gene therapy due to pre-existing immunity. Development of additional viral vectors may overcome this hurdle. Simian virus 40 (SV40)-derived vectors have been reported to transduce different tissues, including the liver, and prevalence of neutralizing antibodies in the general population is very low. This renders recombinant SV40 (rSV40) vector an interesting candidate for effective (re-)administration. Clinical use of SV40 vectors is in part hampered by less advanced production methods compared to AAVs. To optimize the production of rSV40 and make it better suitable for clinical practice, we developed a production system that relies on Cre recombinase-mediated removal of the bacterial plasmid backbone.

Type 1 diabetes mellitus (T1DM) is due to the selective destruction of islet beta cells by immune cells. Current therapies focused on repressing the immune attack or stimulating beta cell regeneration still have limited clinical efficacy. Therefore, it is timely to identify innovative targets to dampen the immune process, while promoting beta cell survival and function. Liver receptor homologue-1 (LRH-1) is a nuclear receptor that represses inflammation in digestive organs, and protects pancreatic islets against apoptosis. Here, we show that BL001, a small LRH-1 agonist, impedes hyperglycemia progression and the immune-dependent inflammation of pancreas in murine models of T1DM, and beta cell apoptosis in islets of type 2 diabetic patients, while increasing beta cell mass and insulin secretion. Thus, we suggest that LRH-1 agonism favors a dialogue between immune and islet cells, which could be druggable to protect against diabetes mellitus.

Neurodegenerative and psychiatric diseases (NPDs) are today's most important group of diseases, surpassing both atherosclerotic cardiovascular disease and cancer in morbidity incidence. Although NPDs have a dramatic impact on our society because of their high incidence, mortality, and severe debilitating character, remarkably few effective interventions have become available. The current treatments, if available, comprise the lifelong intake of general immunosuppressants to delay disease progression or neurotransmitter antagonists/agonists to dampen undesired behaviors. The long-term usage of such medication, however, coincides with often severe adverse side effects. There is, therefore, an urgent need for safe and effective treatments for these diseases. Here, we discuss that many NPDs coincide with subtle chronic or flaring brain inflammation sometimes escalating with infiltrations of lymphocytes in the inflamed brain parts causing mild to severe or even lethal brain damage. Thus, NPDs show all features of autoimmune diseases. In this review, we postulate that NPDs resemble autoimmune-driven inflammatory diseases in many aspects and may belong to the same disease spectrum. Just like in autoimmune diseases, NPD symptoms basically are manifestations of a chronic self-sustaining inflammatory process with detrimental consequences for the patient. Specific inhibition of the destructive immune responses in the brain, leaving the patient's immune system intact, would be the ultimate solution to cure patients from the disease. To reach this goal, the primary targets, e.g., the primary self-antigens (pSAgs) of the patient's chronic (auto)immune response, need to be identified. For a few major NPDs, immunological studies led to the identification of the pSAgs involved in the autoimmune damage of specific brain parts. However, further research is needed to complete the list of pSAgs for all NPDs. Such immunological studies will not only provide crucial insights into NPD pathogenesis but also ultimately enable the development of a new generation of safe and effective immunotherapies for NPDs. Interventions that will dramatically improve the life expectancy and quality of life of individual patients and, moreover, will significantly reduce the health-care costs of the society in general.

Replication-defective (RD) recombinant simian virus 40 (SV40)-based gene delivery vectors hold a great potential for clinical applications because of their presumed non-immunogenicity and capacity to induce immune tolerance to the transgene products in humans. However, the clinical use of SV40 vectors has been hampered by the lack of a packaging cell line that produces replication-competent (RC) free SV40 particles in the vector production process. To solve this problem, we have adapted the current SV40 vector genome used for the production of vector particles and generated a novel Vero-based packaging cell line named SuperVero that exclusively expresses the SV40 large T antigen. SuperVero cells produce similar numbers of SV40 vector particles compared to the currently used packaging cell lines, albeit in the absence of contaminating RC SV40 particles. Our unique SV40 vector platform named SVac paves the way to clinically test a whole new generation of SV40-based therapeutics for a broad range of important diseases.

It has long been recognized that sustained or repeated child maltreatment has lasting psychological and emotional effects on the victims. This has helped to inform the criminal and civil justice systems how best to deal with perpetrators of abuse, as well social and health services when treating the victims. However, what is generally less well recognized is that physical and emotional abuse has a lasting and potentially non-reversible effect on brain function.We conducted a literature review on the forensic, mental, psychological, and pathophysiological impact of child maltreatment and discuss the implications of child maltreatment as a potential mitigating factor in criminal court in cases where victims of abuse become perpetrators themselves.Repeated exposure to traumatic experiences changes the responsiveness in the hypothalamus-pituitary-adrenal axis with lasting consequences in the developing brain for structures, such as the hippocampus and amygdala. These physiological changes are thought to cause a range of mental disorders, which are associated with poor affect regulation, anxiety, depression, and substance abuse.The importance of developing our understanding of the long-term effects of child abuse and neglect cannot be overestimated as the result of child maltreatment will perpetrate criminal acts since offenders have higher rates of mental illness than the general community.

Consumers' adoption of fuel-efficient vehicles is crucial to saving energy in road transport. Investigating reasons for the gap between intention and revealed behavior can contribute to more effective, faster, and less costly market penetration of efficient vehicles. We compare psychological determinants of the stated importance of fuel consumption versus determinants of actual behavior. Swiss survey data of potential new car buyers and of owners of recently purchased new vehicles were used. Car-purchase behavior is represented by four different proxies. Each differently accounts for needs and resources indicated by household type and socioeconomic status. The results indicated that intention (the stated importance of fuel consumption) is explained to a lower degree than behavior; it is mainly expressed according to an inner feeling of obligation (personal norm) and influenced by symbolic motives (to express one's self and social position through the car). For revealed purchase behavior, the evaluation of less vehicle power and smaller size and perceived behavioral control added considerable explanatory power. As the intention for the next car purchase will mostly still be rather vague, it might not be influenced yet by variables that are very close to behavior. With regard to the promotion of fuel-efficient vehicles, policy measures to influence both intention and behavior are outlined.

SummaryHouseholds exert an important influence on total greenhouse gas (GHG) emissions. Therefore, their consumption behavior is of interest in evaluations of climate policy options and projections of future emission paths. While most evaluations of household consumption and its emissions are based on expenditure only, we use a household consumption model based on functional units (e.g., kg food, person kilometers, living square meters). The goal of this article is to assess changes in consumption with increasing affluence level of households and to compare the allocation of GHG emissions to monetary versus functional units. We find that (1) the model based on functional units provides good bottom‐up estimates for greenhouse emissions of Swiss households; (2) quality (price per functional unit) increases with income for many consumption categories, and therefore using functional instead of monetary units leads to a lower increase of greenhouse gas emissions with income; (3) the relevance of GHG emissions from goods and mobility will increase. We conclude that using household models based on monetary units only overestimates the impact of marginal consumption and neglects the potential of decoupling income and environmental impact by consuming better instead of more. For sustainable consumption, research and policy should aim at preventing goods of higher quality from having higher environmental impact in order to benefit from the increasing quality orientation with rising income.

RNA-mediated gene silencing or RNA silencing is a gene regulation mechanism in eukaryotes involved in RNA-mediated sequence-specific RNA degradation in the cytoplasm followed by chromatin remodelling in the nucleus that plays a crucial role in differentiation and developmental processes. Besides, the RNA silencing machinery serves as an innate defense response against viruses and transposons and is responsible for down regulating transgene expression. Therefore, the induction of RNA silencing in producer cells limits the production of recombinant proteins and of (recombinant) virus particles for use as vaccines or in gene therapies. Viruses counteract the RNA silencing response by expressing RNA silencing suppressors (RSSs) in infected cells, thereby allowing the virus to reach higher titers. We employed viral RSSs for improving the productivity of eukaryotic production systems. We here describe the application of viral RSSs in enhancing the production of pharmaceutical proteins including monoclonal antibodies and virus particles in mammalian cell lines.

Life cycle assessment (LCA) according to ISO 14040 standard (ISO-LCA) is applied to assess the environmental impact per functional unit of new or modified products. However, new or modified products can also induce demand changes-so-called rebound effects. If overall environmental impact is of interest, there is a need to assess the potential magnitude of such rebound effects and to allow recommendations on how to mitigate these effects. To do so, this study proposes to complement the constant demand assumption (implicitly assumed by the ISO-LCA), commonly known as the ceteris paribus assumption, with a consumption-as-usual assumption allowing a systematic stepwise inclusion of rebound effects.We base our results on a formal description of household consumption. To indicate the relevance of the proposed integration of rebound effects, different comparative LCAs are reviewed and the concept is applied to mobility as illustrative examples.Based on a description of household demand and consumption feedback loops, we propose the consumption-as-usual concept, which in contrast to the constant demand assumption assumes that (1) the use of household resources for consumption does not change and (2) preferences remain the same. Household resources for example are purchasing power (we assume that households do not work less), time, and living space. We outline how this concept allows integrating potential rebound effects into ISO-LCA by considering three different cases of reallocating freed household resources. To illustrate the use of the consumption-as-usual concept, we draw implications for different comparative LCAs from the literature and illustrate cases with income and time rebound for different personal travel modes.The consumption-as-usual concept is applicable to a broad range of product modifications and allows an important complementation of the LCA regarding rebound effects. For products with various changes in the need for household resources, the assessment becomes however a challenging task. The limits of the consumption-as-usual concept are mainly given by its two underlying assumptions. Therefore, new or modified products with the potential to change consumer preferences or even the amount of household resources used for consumption go beyond this concept.The integration of rebound effects is feasible for many comparative LCAs. It helps in increasing the reliability of the assessment of overall environmental impact reduction through new or modified products. In addition, a basis is provided with which to mitigate rebound effects and give appropriate recommendations to product users.Potential rebound effects should be included in LCA in order to guide consumers and policy towards sustainable consumption. We recommend the consumption-as-usual concept for this purpose. To predict rebound effects under consumption as usual instead of outlining potential amplitudes, further research on household preferences is needed and an optimisation model should be applied for household consumption. However, even if data are available for such a prediction, the assessment of potential rebound effects is still recommended in order to recognise dangers and opportunities in consumption changes.

An effective consumer-oriented climate policy requires knowing the GHG reduction potential of sustainable consumption. The aim of this study is to draw lessons from differences in consumption between households with high and low GHG emissions. We evaluate a survey of 14,500 households and use a method that allows measuring changes in price level of consumption. Comparing the 10% of households with the highest GHG emissions per capita with the lowest 10% – controlling for differences in expenditure level and household structure – we find a range 5–17 tons of CO2-equivalent per capita and year. The observed differences stem mainly from heating, electricity use, car use, and travel by aircraft. Consumption patterns with low GHG emissions are characterized by less spending on mobility, but more on leisure and quality oriented consumption (leading to higher prices per unit). Further characteristics are: a higher share of organic food, low meat consumption and fewer detached single family houses. Our findings imply that a significant reduction in GHG emissions would be possible by adopting real-world consumption patterns observable in society. The twin challenge is to shift consumption towards more climate friendly patterns, and to prevent any trend towards high emitting consumption patterns.

In this paper, we simulate the car market in order to forecast the effects of feebate systems based on an energy-labeling scheme using categories A to G. Very fuel-efficient (A) cars receive a cash incentive, highly inefficient (G) cars pay additional fees. Consumers have different price elasticities and behavioral options to react to feebates. They can switch to a smaller sized car, but as energy-efficiency varies widely within size segments, they can also stick to the preferred size class and choose a more efficient (smaller) engine. In addition, previously owned cars influence the next car to be chosen. We use an agent-based microsimulation approach particularly suited to predict environmental and market effects of feebates. Heteorogenous agents choose from a choice set drawn from a detailed fleet of new cars. Incentives of €2000 for A-labeled cars induce an additional rated CO2 emission decrease of new car registrations between 3.4% and 4.3%, with CO2 abatement costs between €6 and €13 per ton, and otherwise little undesired market disturbance. The risk of rebound effects is estimated to be low. After adopting the frequencies of consumer segments to a given country, the model presented is applicable to all European car markets.

The NS1 gene of influenza A virus encodes a multi-functional protein that plays an important role in counteracting cellular antiviral mechanisms such as the interferon (IFN), protein kinase R and retinoic acid-inducible gene product I pathways. In addition, NS1 has recently been shown to have RNA interference (RNAi) or RNA silencing suppression (RSS) activity. This study analysed the IFN antagonistic activity of NS1 and the RSS activity for several influenza subtypes: H1N1, H3N2, H5N1 and H7N7. It was shown that the various NS1 proteins were capable of inhibiting the activation of an IFN-responsive promoter. However, differential RSS activity was measured among the NS1 variants. The NS1 protein of strain A/WSN/33 (H1N1) was most potent in suppressing short hairpin RNA-mediated gene silencing. In contrast, NS1 proteins of the highly pathogenic H5N1 strains A/VN/1194/04 and A/HK/156/97 were most potent in complementing the RSS function of the human immunodeficiency virus type 1 Tat protein. These results show that the ability of NS1 to suppress RNAi varies among influenza strains and is likely to contribute to differences in viral replication capacity and pathogenicity.

According to some recent studies, noise from road transport is estimated to cause human health effects of the same order of magnitude as the sum of all other emissions from the transport life cycle. Thus, ISO 14′040 implies that traffic noise effects should be considered in life cycle assessment (LCA) studies where transports might play an important role. So far, five methods for the inclusion of noise in LCA have been proposed. However, at present, none of them is implemented in any of the major life cycle inventory (LCI) databases and commonly used in LCA studies. The goal of the present paper is to define a requirement profile for a method to include traffic noise in LCA and to assess the compliance of the five existing methods with this profile. It concludes by identifying necessary cornerstones for a model for noise effects of generic road transports that meets all requirements.