Reimbursement drug policy in the Netherlands and Bulgaria - comparative analysis

The method of reimbursement provides a fair and affordable high-quality health care. The aim of this review is to compare the reimbursement policies in the Netherlands and Bulgaria. The topics discussed are the authorities and considerations in decision-making for the inclusion of medicines in the positive reimbursement list, the pricing of these drugs, as well as the transparency in decision-making for reimbursement

The Riemannian curvature identities for the torsion connection on Spin(7)Spin(7)-manifold

Curvature properties of the torsion connection preserving a given $Spin(7)$ structure in dimension eight are investigated. It is shown that on compact $Spin(7)$ manifold with exterior derivative of the Lee form lying in the Lie algebra $spin(7)$ the curvature of the torsion connection $R\in S^2\Lambda^2$ with vanishing Ricci tensor if and only if the three-form torsion is parallel with respect to the Levi-Civita connection. In particular the 3-form torsion is harmonic. It is also proved that, in addition, the curvature of the torsion connection satisfies the Riemannian first Bianchi identity if and only if the three-form torsion is parallel with respect to the Levi-Civita and to the characteristic connection simultaneously. In this case the Lee form is also parallel with respect to the Levi-Civita and to the torsion connections. In particular, the Lee form is harmonic.Comment: 14 pages, no figures. arXiv admin note: substantial text overlap with arXiv:2307.05619, arXiv:2307.0500

The Fitness Landscape Metaphor: Dead but Not Gone

Dans cet article, je présente une approche sémantique de l’analyse de la fonction de la métaphore du paysage dans la biologie de l’évolution. Le concept de paysage adaptatif a suscité une attention considérable dans la philosophie de la biologie récente. La plupart des auteurs ont considéré ce concept de l’une des deux manières suivantes: en tant qu’outil heuristique, comme partie intrinsèque de modèles mathématiques robustes, ou comme un ensemble définissable d’analogies sur lesquelles les modèles sont basés et testés. Chacune de ces visions conduit à la conclusion que la valeur de la métaphore du paysage dépend seulement du succès des modèles que sous-tend la métaphore, en vue de représenter adéquatement la dynamique de l’évolution. J’essaie de montrer que cette conclusion, ainsi que les visions qui y conduisent, ne tiennent pas compte d’épisodes importants dans l’histoire de la métaphore du langage. Ces visions proviennent plutôt de thèses générales, en philosophie des sciences, quant au rôle des métaphores dans les théories scientifiques. L’analyse sémantique que je propose met en lumière le fait que la fonction première du concept de relief adaptatif, au cours de la synthèse évolutionniste, a été de servir de cadre général d’unification conceptuelle, qui a rendu possible la conciliation de phénomènes empiriques hétérogènes. De ce point de vue, la métaphore du paysage est un outil linguistique-théorique qui ne doit pas être abandonné (et qui ne l’est de fait pas) suite à la falsification des modèles construits et interprétés au moyen de la métaphore.In this paper I present a semantic approach to the analysis of the function of the landscape metaphor within evolutionary biology. The concept of adaptive landscape has drawn a considerable attention in recent philosophy of biology. Most writers have treated the concept in one of the following ways: as a heuristic tool, as an intrinsic part of robustly defined mathematical models, or as a definable set of analogies on which models are based and tested. All these views lead to the conclusion that the value of the landscape metaphor depends only on the success of the models, which the metaphor underlies, to adequately represent evolutionary dynamics. I have tried to show that this conclusion, and respectively, the views which imply it, do not account for important episodes from the history of the landscape metaphor. These views rather stem from the general views, in philosophy of science, about the role of metaphors in scientific theories. The semantic analysis which I propose reveals that the concept of adaptive reliefs’ primary function during the evolutionary synthesis has been to serve as a general unifying conceptual framework which has made possible the theoretical reconciliation of heterogeneous empirical phenomena. From this perspective, the landscape metaphor is a linguistic theoretical tool which should not be abandoned (and in fact has not been abandoned) after the falsification of the models which have been built and interpreted by means of the metaphor

In vivo bioluminescence imaging in preclinical trials of genetic vaccines

DNA immunization is a rapidly developing vaccine platform for cancer, infectious disease, and allergies. The efficiency of DNA vaccination is largely determined by the efficiency of delivery and subsequent expression of the HIV-1 genes in the cells. DNA immunogens are generally administered by intramuscular or intradermal injections, followed by electroporation to enhance the DNA uptake into the cells. An intense debate on the pros and cons of different routes of DNA delivery is still ongoing. A number of studies have compared the effect of the delivery methods on the amount and quality of DNA-directed immunogen expression, as well as on the magnitude and specificity of the immune response they generate. Several studies were based on post mortem studies of the tissues, or on indirect expression monitored by techniques such as in vivo imaging of reporter genes co-delivered or fused to immunogens. The aim of this work was to develop in vivo imaging applications for DNA immunization. The first aim was to optimize delivery techniques in order to increase the efficacy and immunogenicity of DNA vaccines. Furthermore we set out to use the differences in the strength and type of immune response induced by DNA immunogens administered by intradermal (ID) or intramuscular (IM) injection routes, each followed by electroporation. In particular, the task was to determine the extent to which the method of DNA delivery influences the immune response to Th1 and Th2 type immunogens, represented by the viral protease (PR) and reverse transcriptase (RT) of HIV-1, respectively. Our final objective was to use the acquired results in an attempt to model immune responses induced by DNA immunogens in silicon. BALB/c mice were immunized with DNA immunogens mixed with a gene encoding a bioluminescent reporter. We used bioluminescence imaging (BLI) as a tool to monitor the expression of delivered reporter genes in vivo. By combining the readouts form BLI and immunoassays we were able to produce a set of delivery parameters that result in the best immunization outcome in terms of expression and immunogenicity. Upon the optimization of delivery conditions we exploited different immunization routes to determine the one that is best suited and providing maximal immunogenicity for DNA vaccines. Here we show that ID administration of DNA immunogens results in a significant enhancement of both cellular and humoral immune responses in mice as compared to IM. The increase in the magnitude of immune responses was evident regardless of the nature of the immunogen (Th1 vs. Th2). The kinetics of the loss of co-delivered reporter gene expression was found to correlate with the antigen-specific production of IFN-γ and IL-2 and could thus be used to characterize the strength of specific immune responses against the delivered immunogen. Thus, we were able to assess the immunogenicity of a DNA vaccine by non-invasive imaging of bioluminescence from the co-delivered reporters. The use of bioluminescent reporters is a new strategy to assess the delivery of DNA immunogens and their expression from the start to the completion of the immunization experiment. The level of reporter expression in the presence of the DNA immunogens reflects the in vivo immunogenicity of the construct, presenting anon-invasive method (technique) to assess the dynamics of the immune responses in individual DNA immunogen recipients useful for determination of the study end-points. The application of this technique allows us to significantly refine and reduce animal experimentation in gene vaccine development

In vivo bioluminescence imaging in preclinical trials of genetic vaccines

DNA immunization is a rapidly developing vaccine platform for infectious diseases, cancer and allergies. The efficiency of DNA vaccination is largely determined by the efficiency of delivery and subsequent expression of genes encoding microbial and tumor antigens or allergens in the cells of vaccine recipients. DNA immunogens are generally administered by intramuscular or intradermal injections, followed by electroporation to enhance the DNA uptake into the cells. An intense debate on the pros and cons of different routes of DNA delivery is still ongoing. The aim of this work was to develop in vivo imaging applications for improvement of DNA immunization. The first aim was to optimize delivery techniques in order to increase the efficacy of in vivo delivery of DNA vaccines and subsequent immune response. Using model DNA immunogens encoding luciferase, and HIV-derived immunogens encoding protease (PR) and reverse transcriptase (RT), we defined the differences in the strength and type of immune responses induced by them when administered by intradermal or intramuscular injection routes followed by electroporation. Furthermore, we determined the extent to which the method of DNA delivery influences the immune response to Th1 and Th2 type immunogens, represented by plasmids encoding PR and RT of HIV-1. Finally, we developed imaging applications for the in vivo assessment of the effector/lytic potential of the immune response in tumor and surrogate pathogen challenge models. We immunized mice with DNA immunogens mixed with a gene encoding a bioluminescent reporter. Bioluminescence imaging (BLI) served as a tool to monitor the expression of delivered reporter genes in vivo. By combining the readouts form BLI and immunoassays we defined a set of delivery parameters that led to the best immunization outcome in terms of both immunogen expression and subsequent immune response. After optimizing the delivery conditions we tested different immunization routes to determine the one that ensures maximal immunogenicity of DNA immunogen. Here we show that intradermal administration resulted in a significant enhancement of both cellular and humoral immune responses as compared to intramuscular delivery. This was evident regardless of the nature of the immunogen (Th1 vs. Th2). The kinetics of the loss of co-delivered reporter gene expression was found to correlate with the antigen-specific production of IFN-γ and IL-2 and could thus be used as in vivo correlate of the strength of specific immune responses. Thus, non-invasive imaging allowed to assess the immunogenicity of DNA vaccines in vivo. Using the same parameters we developed a surrogate method that could assess effector memory responses. Finally, we applied BLI to study the growth of luciferase-labeled tumors in luciferase-immunized animals, which provided a functional measure of vaccine efficacy. Overall, the use of BLI allowed us to establish a methodology to increase the efficacy of delivery, define optimal regimens and test the effector capacity of the immune response induced by DNA vaccination. The application of this technique made it possible to significantly refine and reduce animal experimentation in gene vaccine development