Парадигма юридичного ризику в системі національної безпеки України (прикладний аспект)

Розглядається загальна модель системи національної безпеки. Визначаються місце та роль юридичного ризику в функціонуванні системи національної безпеки. Аналізується категорія «юридичний ризик» та його основні складові, умови виникнення та функціонування. Пропонуються умови протидії ризикам та подається модель зниження юридичних ризиків у системі національної безпеки . Ключові слова: національна безпека, юридичний ризик.Рассматривается общая модель системы национальной безопасности. Определяется место и роль юридического риска при функционировании системы национальной безопасности. Анализируется категория «юридический риск» и его основные составляющие, условия возникновения и функционирования. Предлагаются условия противодействия рискам и подается модель снижения юридических рисков в системе национальной безопасности Ключевые слова: национальная безопасность, юридический риск.The problems of forming and development of the system of national safety are examined in the article. The general model of the system of national safety is offered and the role of legal risk is determined in this system. The location and role of legal risk is determined at functioning of the system of national safety. A category is analysed «legal risk» and his basic component elements, terms of origin and functioning. The terms of counteraction risks are offered and the model of decline of legal risks is given in the system of national safety. Key words: national safety, legal risk

Correction:How the COVID-19 pandemic highlights the necessity of animal research (vol 30, pg R1014, 2020)

(Current Biology 30, R1014–R1018; September 21, 2020) As a result of an author oversight in the originally published version of this article, a number of errors were introduced in the author list and affiliations. First, the middle initials were omitted from the names of several authors. Second, the surname of Dr. van Dam was mistakenly written as “Dam.” Third, the first name of author Bernhard Englitz was misspelled as “Bernard” and the surname of author B.J.A. Pollux was misspelled as “Pullox.” Finally, Dr. Keijer's first name was abbreviated rather than written in full. These errors, as well as various errors in the author affiliations, have now been corrected online

Chemical synthesis of lipopolysaccharide derived from klebsiella pneumoniae for conjugate vaccine development

Klebsiella pneumoniae (K. pneumoniae), a member of Gram-negative bacterium (GNB) belonging to Enterobacteriaceae family, is a commonly isolated nosocomial pathogens that can cause pneumonia, bloodstream infections and sepsis. Infections caused by carbapenem-resistant K. pneumoniae (CR-Kp) are particularly problematic due to a lack of therapeutic options leading to a 50% mortality. Capsular polysaccharide (CPS) and lipopolysaccharide (LPS) are two virulence factors of K. pneumoniae that provide attractive starting points for vaccine development. The large number (79) of serotypes, however, has complicated CPS vaccine development. In contrast, only nine serotypes of LPS have been identified to date. In addition, only two core types have been identified making the LPS core an attractive target for the development of a broad acting vaccine for K. pneumoniae. The chemical synthesis of an outer core tetrasaccharide is described in Chapter 2. The compound contains three amino groups in different forms, a Kdo residue which is a common component of inner core oligosaccharides, and a thiol-containing linker at the reducing end for protein conjugation. A two-carbon extension strategy was employed to synthesize a Kdo building block. The installation of challenging α-galactoside/α-Kdo linkages was accomplished by remote neighboring group participation. The subsequent Chapter 3 describes the synthesis of a pentasaccharide derived from the outer core of K. pneumoniae containing a nonstoichiometric L,D-heptose substituent. A crowded 4,5-branched Kdo residue was installed by a block synthetic strategy, and a heptosyl donor was prepared from D-mannose with excellent L-diastereoselectivity by using a one-carbon elongation approach. The immunological evaluation of the two conjugates is ongoing and will reveal the role of the heptoside in eliciting relevant immune responses. In Chapter 4, the synthesis of an inner core pentasaccharide derived from K. pneumoniae is described. Compared to the diversity of CPS, the inner core has only one type among K. pneumoniae making it attractive for the development of a broad acting vaccine. Galacturonic acid is expressed by the bacteria to provide negative charge since the phosphoryl modifications are absent in the inner core. To reveal the effects of the acidic substituent for eliciting immune response, a substructure lacking galacturonic acid was also synthesized containing a crowded 3,4-branched L,D-heptoside. The tetra- and pentasaccharide have been conjugated to carrier proteins, and future studies will focus on examining antigenic properties by mouse immunizations

Quantitative mass spectrometry-based proteomics in the study of iPSC-derived neurons and disease modeling: A study in neurodevelopment and neurological disorders

In this thesis we applied mass spectrometry-based proteomics to study neuronal differentiation derived from induced pluripotent stem cells (iPSCs) and to study neurodevelopmental disorders such as Spinal Muscular Atrophy (SMA) and Rett syndrome (RTT). In chapter one we give a general introduction to study human brain development. We introduce the generation of iPSCs that can be differentiated into neurons for several research areas. We give an overview of several neurological disorders that are being studied using this model system. Furthermore, we give an introduction to mass spectrometry-based proteomics describing general workflows and mass spectrometers. In addition, we describe several strategies used for quantitative mass spectrometry. In chapter two, we study the protein determinant during neuronal differentiation. Here, we used iPSCs and differentiated these cells into induced neurons (iN) and into small molecule-derived patterned motor neurons. We provide a mass spectrometry-based quantitative proteomic signature at temporat resolution. We identified key proteins showing significant expression changes during neuronal differentiation. Furthermore, we provide a rich source of information on several signaling pathways. Chapter three, describes the study of RTT syndrome using iPSC-based model with isogenic controls and a time-series of mass spectrometry-based proteomic analysis during early stages of neuronal development. We provide evidence of proteomic alterations long before symptoms of RTT syndrome become apparent. Such that proteins associated with dendrite morphology and synapses are differentially expressed at early neuronal stages of development. These changes increase from early to late neuronal phases giving awareness of the protein alterations at early onset of RTT syndrome. In chapter four, we again used quantitative mass spectrometry to study the proteomic changes during early stages of iPSC-derived motor neurons (MN) in SMA. Also here, we show altered proteins at early stages of MN differentiation that are associated with known SMA phenotypes such as defective ER to Golgi transport, mRNA splicing and protein ubiquitination. In addition, these proteins increase in differential expression in SMA towards later stages of MN differentiation. Furthermore, we highlight that differences in altered protein expression between SMA patients have similar biological functions. In this chapter we provide a rich source of molecular events during MN differentiation that might be interesting in the development of new biomarkers and therapeutic approaches. Chapter five is a future outlook describing the cross-talk between the multi-omic studies to better understand a biological mechanism. We introduce several challenges in the study of human brain 131 development associated with temporal and spatial resolution. Furthermore we discuss the latest studies done in SMA and RTT syndrome and provide a future perspective to further improve our understanding of these devastating disorders

UNRAVELING THE FUNCTIONS OF DETOXIFICATION ENZYMES AND DRUG TRANSPORTERS

A number of detoxification systems with broad substrate specificity, including drug transporters and drug-metabolizing enzymes, can protect the body from various toxins, and dramatically influence drug absorption, metabolism, distribution and elimination (ADME) processes, as well as in some cases physiological homeostasis. In Part I of this thesis, in view of the likely importance of the Carboxylesterase 2 enzyme in pharmacological and physiological processes, we generated and characterized several CES2-related genetically modified mouse models. We then demonstrated the value of these mouse models to investigate the pharmacological and physiological functions of the CES2 enzyme complex. In Part II of this thesis we explored and revealed the roles (especially in the blood-brain-barrier) of several drug transporters (ABCB1, ABCG2 and OATP1A/1B) and the drug-metabolizing CYP3A enzymes in pharmacokinetic behavior of several anti-tumor small-molecule inhibitors, as well as related drug-drug interaction effects. We believe that the obtained new mouse models and pharmacological and physiological insights will provide further useful information and support for improving drug discovery and development, and eventually clinical-therapeutic application regimens

The Effects of Mycotoxins on Pregnancy and Immune Development: (A Preclinical approach)

Mycotoxins are among the most important and highly prevalent nutritional contaminants worldwide. They are naturally produced as secondary metabolites of different fungal species, which can contaminate a wide range of agricultural products, especially cereal and grain-based food. Fusarium species are amongst the most prevalent fungal contaminants of grains, and produce the two most important classes of mycotoxins, trichothecenes (including deoxynivalenol (DON), T-2 toxin and HT-2 toxin) and zearalenones (ZEN). In humans and animals, exposure to Fusarium mycotoxins is associated with mutagenic, teratogenic, estrogenic, hemorrhagic, neurotoxic, hepatotoxic, and immunotoxic effects. Considering the widespread occurrence of these food contaminants and inevitable human exposure, this thesis aimed to investigate the adverse effects of exposure to commonly occurring Fusarium mycotoxins and potential mechanisms involved, with a special focus on DON, as this is the most prevalently occurring food contaminant worldwide. In this thesis, we showed that exposure to the major Fusarium mycotoxin DON, can adversely affect innate and adaptive immune responses, placental barrier, fetal survival and development. We further demonstrated that consuming DON-contaminated diets during pregnancy and lactation, induces long-term effects on the developing immune system of the offspring and shifts the balance in immune cell populations, hence increasing the susceptibility to allergic reactions and reducing immune responses to vaccination in later stages of life. Some specific non-digestible oligosaccharides (NDOs) have prebiotic and immune- modulating properties comparable to those observed for certain human milk oligosaccharides (HMOs). Specific structures of NDOs and HMOs are confirmed to have positive effects on improving adaptive immune responses. In this thesis we showed that nutritional supplementation with a specific NDO mixture rich in 3’-GL, can be beneficial in preventing or neutralising the adverse effects of DON on immune system. Thus, NDOs might be a suitable candidate as a supplement in infant formulae for boosting the immune system and preventing the harmful effects of early-life exposure to DON on the intestinal and immune systems

Opwindend onderzoek : de ontwikkeling van de seksuele psychofarmacologie

Klinisch en dierexperimenteel seksueel psychofarmacologisch wetenschappelijk onderzoek vraagt tijd, en geduld. Het leidt echter tot nieuwe en vaak tot de verbeelding sprekende bevindingen. Maar wat is nu precies wetenschappelijk onderzoek? Aan het essay over wetenschapstheorie van de socioloog Prof. Dr. Johan Goudsblom ontleen ik graag de volgende kijk op wetenschappelijk onderzoek. Dit wordt volgens hem gekenmerkt door de vier relatief zelfstandige criteria: precisie, reikwijdte, systematiek, en relevantie [1]. De theorie kant van onderzoek heeft te maken met de wisselwerking tussen 1. relevantie en 2. reikwijdte. De empirische kant van wetenschappelijk onderzoek heeft daarentegen te maken met de wisselwerking tussen 3. precisie en 4. systematiek. Deze vier criteria liggen ten grondslag aan de tegenwoordig gangbare term “evidence based onderzoek” dat met name de vereisten precisie en systematiek verder uitwerkt. Tussen theorie en empirie bestaat een wisselwerking: beter empirisch onderzoek verbetert de onderliggende theorie. In het vervolg van mijn lezing zult u kunnen constateren dat seksueel psychofarmacologisch onderzoek, mits met precisie en systematiek uitgevoerd, bijdraagt aan relevantie en reikwijdte en daarmee tot een betere theorie over seksuele dysfuncties

Amyloidogenic peptides for design of inhibitors of peptide aggregation and as templates for new bionanomaterials

Misfolding of proteins from their soluble form into highly insoluble fibrillar deposits can lead to (non-)neurodegenerative disorders or systemic amyloidosis. This class of diseases (for which no therapy is available yet) is called amyloid diseases. Amyloid refers to the extracellular proteinaceous deposits consisting of fibrils. Fibrils are oligomers of misfolded peptides/proteins which are characterized by folding into beta-pleated sheets. A well accepted approach for the interference with beta-sheet formation is the design of soluble beta-sheet mimetics that are able to disrupt the hydrogen bonding network which ultimately leads to the disassembly of fibrils and aggregates. One prominent disease related to the formation of extracellular amyloid is type 2 diabetes. The western aging population shows a dramatic increase in the number of diabetic patients. Type 2 diabetes is characterized by the extracellular deposition of the misfolded human Islet Amyloid PolyPeptide (hIAPP or amylin) in the pancreatic islets of Langerhans and is found in 90% of the diabetic patients. The first part of this thesis describes the development of newly designed soluble beta-sheet breaker peptides based on human amylin. Several approaches towards such beta-sheet breaker peptides were investigated. In chapter 2, the synthesis, spectroscopic analysis and aggregation behaviour, of several backbone-modified amylin(20-29) derivatives is described. Three modified amide bonds at alternate positions were introduced such as ester bonds, N-butylated amides and peptoids using amylin(20-29) as core sequence. In chapter 3, the synthesis of the chiral peptoid building block of L-isoleucine, the solid phase synthesis of the peptoid and retropeptoid sequences of amylin(20-29), and the structural analysis of these amylin derivatives in solution by infrared spectroscopy, circular dichroism and transmission electron microscopy is described. Chapters 4 and 5 deal with the incorporation of a single beta-aminoethane sulfonyl amide moiety or an aminooxy moiety in the highly amyloidogenic peptide sequence amylin(20-29) and its subsequent benzylation featuring the Mitsunobu reaction. Chapter 6 deals with the synthesis of the enantiomer of the L-depsipeptide as was described in chapter 2. In chapter 7, aggregation studies of native amylin(1-37) and derivatives using NMR-spectroscopy are described. The role of histidine 18 in full-length amylin was studied to reveal the initial changes in secondary structure and fibril formation. In chapter 8, design of a covalent triangular shaped molecule with amyloidogenic Abeta(16-22) as a repeating unit and functioning as a cross recognition motif, is described. Identical triangular shaped molecules might be capable to induce spontaneous self-assembly into nanospheres like viral capsids or triangular shaped tubular nanostructures. Chapter 9 describes an alternative approach towards the synthesis of triangular shaped peptide constructs as described in chapter 8. Herein, an efficient and facile synthesis of an N-azido/C-alkynyl Abeta(16-22) derivative is described. Since peptides containing both an azide and an alkyne moiety can be polymerized using the microwave-assisted Cu(I)-catalyzed 1,3-dipolar cycloaddition reaction, this synthetic approach was chosen to obtain cyclic Abeta16-22) oligomers

Quality of biopharmaceuticals: Comparability exercise and post-approval surveillance

Biopharmaceuticals (including biosimilars) are defined as a class of biologicals where the active substance is produced in living cells through recombinant DNA technology. When regulating biopharmaceuticals, among other things, the quality, where determining and measuring quality attributes (QAs), is considered a key regulatory aspect. A subset of QAs is considered critical QAs (CQAs) because small differences in these may potentially influence clinical outcomes of the therapy in patient care when they fall outside the acceptable range and limit. Therefore, the thesis aimed to study the quality of biopharmaceuticals (including biosimilars) by providing insight into (1) the comparability of QAs with emphasis on the CQAs, and (2) post-approval quality-related surveillance and actions by regulatory authorities after these medicines have been approved for use. The thesis provides learning regarding post-approval changes and defects in the quality of biopharmaceuticals that could potentially influence patient care. Moreover, the thesis reflects on the potential implications and future directions of comparability of CQAs and post-approval quality surveillance for regulation, policy-making, and patient care. The thesis concluded that the comparison of biopharmaceuticals relies significantly on the comparability of CQAs and also articulates that detecting small differences in and the criticality of QAs are crucial for demonstrating comparability. In the future, we will work on questions about the possible effects of small differences in clinical outcomes for patient care. The studies in the thesis have also shown that safety and efficacy of biopharmaceuticals must go hand in hand with ensuring pharmaceutical quality. Post-approval follow-up, surveillance, and regulatory action where necessary remain essential building blocks of a trusted biopharmaceutical system

To sugarcoat bacteria: Glycoengineering of lipooligosaccharides of bacteria with sialic acid derivatives

This thesis focuses on two approaches from the interdisciplinary field of chemical biology. The first approach is related to glycobiology. Glycans are present on the surface of all cells, also on bacteria. Since these molecules are on the cell surface, they are often the first to interact with the environment and therefore also play an important role in many host-microbe interactions. An illustrative example of their importance is glycan mimicry of host glycans by bacteria, by which bacteria can interact with host receptors or avoid immune recognition by the host. An overview of bacteria that display glycan mimicry and chemical biology techniques to study this occurrence are presented in this thesis. In general, the study of glycans is ideally suited for chemical biology. The first chemical biology approach of this thesis describes two techniques to introduce N-acetylneuraminic acid on bacterial glycans: selective exoenzymatic labeling (SEEL) and native sialyltransferases. These new techniques could contribute to elucidating the interaction between sialylated bacterial pathogens and the immune system. It also highlights the promising future of glycoengineering for the study of bacterial glycobiology. The second chemical biology approach of this thesis focuses on a screen of small molecules that induce a biological process somatic embryogenesis in plants. In this process, a somatic cell is reprogrammed and can grow into a new embryo after the addition of a chemical inducer, compound 2,4-D. A structure-activity relationship of 2,4-D was established based on a screen of a library of 2,4-D analogues. This research contributed to understanding the role of 2,4-D and stress in the process of somatic embryogenesis in plants. Both approaches described in this thesis show that chemical biology is a powerful approach to study and influence biological processes on the molecular level