Les désordres intellectuels (DI) comprennent une collection hétérogène de désordresneurodéveloppementaux qui émergent pendant l’enfance. Ils ont une incidence de 1 à 3% dansla population et sont associés avec des déficits dans les fonctions mentales et adaptives. Denombreuses mutations ont été identifiées dans des gènes codant pour des protéines quiremplissent des fonctions biologiques très diverses dans le cerveau. Parmi ces protéines,certaines sont enrichies à la synapse, supposant que les déficits cognitifs associés aux DIpourraient être reliés à des déficits synaptiques. L’objectif scientifique de notre équipe et decomprendre le rôle de certaines protéines dans la fonction synaptique et la cognition enutilisant des souris génétiquement modifiées portant des mutations dans le gènecorrespondant. Je me suis concentré sur Il1rapl1, un gène codant pour la protéine Interleukinreceptor-accessory-protein-like-1. Des mutations ou micro-délétions dans ce gène sont liés audéveloppement de DI chez l’homme. Dans les neurones, Il1rapl1 code pour une protéinetransmembranaire qui serait impliquée dans la formation et/ou la stabilisation de synapsesexcitatrices. Les conséquences de l’absence d’IL1RAPL1 à des niveaux plus intégrés restaientpeu étudiées lors du début de ma thèse. J’ai utilisé une souris déficiente pour IL1RAPL1 (KO) afinde comprendre le lien entre les déficits comportementaux et la fonction synaptique. Pour cela,j’ai soumis des souris KO à des taches comportementales de peur conditionnée. J’ai ensuiteutilisé une combinaison d’approches in vitro, ex vivo et in vivo afin de caractériser la fonctionsynaptique dans les circuits neuronaux dédiés : l’amygdale latérale et basolatérale. Desenregistrements electrophysiologiques ont montré une dérégulation de la balance entre latransmission inhibitrice et excitatrice (I/E) dans l’amygdale de souris Il1rapl1 KO, causant ainsides déficits dans la capacité d’acquérir et d’exprimer la mémoire de peur conditionnée. Lacorrection de ce déficit synaptique in vivo par pharmacologie ou par optogénétique a permis derestaurer le comportement chez les souris KO. / Intellectual disability (ID) comprises a highly heterogeneous collection of neurodevelopmentaldisorders that arise during childhood. They have an incidence of 1-3% in the population withimpairments in mental and adaptive functions. While the etiologies of IDs are thought to bevery heterogeneous, a significant proportion of ID has genetic origins. Mutations in single IDgenes lead to dysfunctions in proteins that fulfill highly different biological functions in thebrain. Interestingly, ID-related proteins are often found enriched at synapses, suggesting thatcognitive impairments defining ID could be related to alterations of synaptic function. The maingoal of our research team is to understand the role of ID-related proteins in synaptic functionand cognition using mouse models bearing gene mutations associated to ID in humans. Myresearch focused on the study of Il1rapl1, a gene coding for the Interleukin-receptor-accessoryprotein-like-1 protein. Micro-deletions or point mutations in this gene are directly linked to thedevelopment of ID and autism spectrum disorder in humans. In neurons, Il1rapl1 encodes atrans-membrane protein and several in vitro experiments point to its important role in thedifferentiation and formation/stabilization of excitatory synapses trough interactions withpresynaptic, trans-synaptic or postsynaptic partners. However, the consequences of Il1rapl1deficiency at more integrated levels remains poorly understood. The principal objective of mythesis is to explore the link between synaptic deficits and behavioral impairments in Il1rapl1-deficient mice. To achieve that, wild-type and mutant animals were first submitted to fearlearning tasks. I then used a combination of in vivo, ex vivo and in vitro functional essays tocharacterize synaptic functions in behaviorally relevant neuronal circuits. Ultimately, ourworking hypothesis were challenged in vivo by pharmacological and optogenetic approaches tonormalize behavioral deficits in Il1rapl1 KO mice. Altogether my work demonstrates thatInhibitory/Excitatory imbalances associated with the absence of Il1rapl1 impaired both thecapacity to form new memories as well as the expression of previously formed memories.

1 Abstract BCR/ABL is a constitutively active tyrosine kinase that has been shown to be at the heart of the development of chronic myeloid leukemia (CML) and about 30% of acute lymphoblastic leukemia (ALL). With the recent advent of tyrosine kinase inhibitors (TKIs), exemplified by Imatinib, Nilotinib, Dasatinib and Bosutinib, patients with Ph+ CML or ALL are candidates for the therapy with these agents. From the available TKIs, Imatinib is considered as front-line therapy for CML patients in chronic phase, while for Ph+ ALL patients, 2nd generation TKIs (nilotinib, dasatinib, bosutinib) might be considered as more effective therapeutic option. Since the treatment with TKIs is a long-term affair, a substantial proportion of patients acquire some sort of mutation in kinase domain of BCR- ABL, which could be a reason of treatment failure. To date, over ninety BCR/ABL kinase domain mutations have been identified, affecting over 50 amino acids. Recurrent BCR/ABL kinase domain mutations have already been in vitro tested to approximate for their in vivo behavior. Our goal is to invent in vitro technique that would allow testing TKI sensitivity of novel BCR/ABL kinase domain mutations, identified at very low MRD levels. The technique makes use of site-directed mutagenesis to create the novel BCR/ABL kinase domain...

Compounds with carboxylic and amidic functions belong to basic structural blocks, which are used for construction of functional molecules in organic, organometallic and also in carborane chemistry. However, considering cobalt bis(dicarbollide)(1-) ion, the synthetic ways to these derivatives have been virtually unknown. A published procedure on lithiation in THF and reaction with CO2 leading to mono- and dicarboxylic acids had failed in our hands. Nevertheless, a detailed revision of the experimental conditions provided finally good yields of mixture of both acids, which could be separated by chromatography and crystallization, and compound of general formulation [(1-HOOC-1,2-C2B9H10)(1',2'-C2B9H10)-3,3'-Co(III)]- and stereoisomeric mixture of [(HOOC)2-(1,2-C2B9H10)2-3,3'-Co(III)]- were characterized for the first time by combination of NMR, MS and HPLC. Also, the carboxylic acid derivatives with methylene and ethylene connectors of the general formula [(1-HOOC-(CH2)n-1,2- C2B9H10)(1',2'-C2B9H10)-3,3'-Co(III)]- were prepared by lithiation of Cs1 in DME at low temperatures followed by reaction with BrCH2COOEt and subsequent hydrolysis of the resulting ester or by oxidation of the respective propylhydroxy derivative. The acids were converted to reactive p-nitrophenyl esters...

Hepatitis C Virus (HCV) is a small RNA virus that progresses to chronicity in 50-80% of infected individuals. Direct-acting antivirals (DAAs) are revolutionary treatments for HCV with 90-98% cure rates. However, over time, chronic HCV infections can result in advanced liver disease, including cirrhosis. Patients with advanced fibrosis experience a poor response to vaccination, recurrent infections and increased risk for hepatocellular carcinoma (HCC). These outcomes are, in part, a consequence of immune dysfunction. Increased inhibitory receptor and Galectin-9 (GAL-9) expression is a possible mechanism promoting lymphocyte dysfunction. In this study, blood samples were collected from chronic HCV patients with different degrees of liver fibrosis. I conducted a 13-parameter flow stain on the peripheral blood mononuclear cells (PBMC) of these patients. Next, I measured the expression of inhibitory receptors (PD-1, CTLA-4, LAG-3, TIGIT and TIM-3) and GAL-9 on bulk T cell and NK cells of 15 chronic HCV patients with no to moderate fibrosis (F0-F2) and 15 with advanced fibrosis (F3-F4). To analyze receptor co-expression, I employed t-distributed stochastic neighbor embedding (t-SNE) analysis to dimensionally reduce the multi-parametric data. Notably, I found that F3-F4 patients had higher frequencies of >3 inhibitory receptor co-expression on NK cells. Moreover, t-SNE analysis of bulk T cells revealed that F3-F4 patients manifest a higher frequency of cells in the clusters with CD25+TIGITmed-hi CD4+ T cells and PD-1medLAG-3med-hiGAL-9med-hi CD4+ T cells. t-SNE analysis of NK cells also showed that F3-F4 patients manifest a higher frequency of cells in the cluster with CD25+TIGITmed-hiTIM-3med-hi CD56Dim NK cells and CCR7+ PD-1medLAG-3med-hiGAL-9med-hi CD56Dim NK cells. Lastly, the frequency of cells in these clusters was found to positively correlate with patient’s extent of liver damage. In conclusion, I identified phenotypes of immune dysregulation that could explain the increased susceptibility to infection and HCC in chronic HCV patients with advanced fibrosis. These phenotypes could identify targets for combinatorial checkpoint blockade therapy to potentially improve immune function in these patients.

A case control study was performed in the conservancy area of the Kruger National Park (KNP), South Africa to find out whether the faeces of impala (Aepyceros melampus) were more likely to contain tetracycline resistant Escherichia coli (TREC) when they drank from rivers that contained these bacteria, compared to rivers that were uncontaminated with TREC. Five perennial rivers (Crocodile, Letaba, Olifants, Sabie and Sand) were selected. A total of 11 points in these rivers were sampled on three separate occasions and cultured for E. coli. Impala herds within 5 kilometres of each water collection site were identified and between 5 and 10 fresh faeces were collected for each collection period (n=209 faecal specimens). Selective culturing of E. coli was done and the resulting colonies were screened for tetracycline resistance by using the Lederberg Replica Plating (LRP) method. Resistant colonies were those that grew in the presence of 4 mg/L of tetracycline. Both a resistant and/or a susceptible isolate were then selected from each specimen, and subjected to the minimum inhibitory concentration (MIC) micro-broth dilution test for tetracyclines. The breakpoint for the MIC method was considered ≥ 8 mg/L (which is higher than for the LRP method). Twenty one of the 33 water specimens examined were found to be contaminated by E. coli. Among them (n=21), 76.19% (n=16) were resistant to tetracycline using the LRP method, although using the MIC method only 19.05% (n=4) were resistant. All of the resistant strains originated from the Letaba, Olifants and Crocodile rivers (TRECpos rivers). Among the 209 impala faeces sampled, 191 were positive for the presence of E. coli (91.38%). Within these (n=191), 36.64% (n=70) showed TREC using the RPL method, while using the MIC, 9.95% (n=19) were found to be TREC. The RPL and MIC methods showed a concordance of only 48%. Resistant isolates screened by PCR for tet(A) and tet(B) genes were found to be negative and it was concluded that other resistant genes were responsible. The odds ratios (OR) showed that impala drinking from TRECpos rivers were 19.3 (2. 63-141.68) times more likely to be infected with TREC than unexposed impala / Dissertation (MSc)--University of Pretoria, 2007. / Paraclinical Sciences / unrestricted

The thermophilic Campylobacters, Campylobacter jejuni and Campylobacter coli are found as commensals in the intestinal tract of healthy mammals and birds. Campylobacter jejuni is one of the leading causes of sporadic food-borne bacterial disease in humans which is predominantly contracted from poultry products. Although the vast majority of these infections are mild, life-threatening complications should be treated with antimicrobials. Patients are usually treated with either macrolides of fluoroquinolones. However, globally there is an increased trend in the development of resistance to these antibiotics. This trend has also been observed in infection of poultry and pigs. The aim of this investigation was to determine antimicrobial sensitivity of thermophilic Campylobacters isolated from pigs and poultry by broth microdilution minimum inhibitory concentration testing. A total of 482 samples of the small intestinal content from poultry and pigs from the Western Cape and Gauteng Provinces were collected and analysed. Thirty-eight Campylobacter isolates were obtained. Statistical analyses included percentage resistance, minimum inhibitory concentrations (MIC50 and MIC90) as well as the distribution percentages of the MICs. The non-parametric Mann-Whitney U test was used to establish any significant differences at an interspecies, interhost and interprovincial level. Analyses of the data obtained revealed indications of decreasing susceptibility to several antibiotic groups including the tetracyclines, macrolides, erythromycin and tylosin, as well as the lincoasamides, and fluoroquinolones. It was found that isolates from the Western Cape were more likely to be resistant to the fluoroquinolones (p = 0.0392), macrolides (p = 0.0262), and lincoasmides (p = 0.0001) and, as well as to a certain extent the pleuromutulins (p = 0.0985), whereas isolates from Gauteng were more resistant to the tetracycyclines (p = <.0001). Poultry Campylobacter spp. were more prone to be resistant to enrofloxacin (p = 0.0021). Campylobacter jejuni, mainly isolated from poultry, was more liable to be resistant to the tetracyclines (chlrotetracycline p= 0.0307), whereas C. coli, predominatly isolated from pigs was more likely to be resistant to the macrolides (tylosin p= 0.063). Four of the bacteria isolated from the Western Cape were resistant to three or more antibiotic classes, namely; tetracyclines, macrolides, lincosamides, pleuromutulins and fluoroquinolones. No multi-resistant Campylobacter spp. were isolated from the flocks in Gauteng. With the exception of tiamulin, the bacterial populations could clearly be divided into resistant and susceptible populations. As consequence of the increased resistance to the antimicrobial classes used for human therapy and the geographical differences in antimicrobial susceptibility, it is recommended that an antimicrobial resistance monitoring system for the thermophilic Campylobacter spp. be initiated in the South Africa National Veterinary Surveillance and Monitoring Programme for Resistance to Antimicorbial Drugs (SANVAD) Copyright / Dissertation (MSc)--University of Pretoria, 2009. / Veterinary Tropical Diseases / unrestricted

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucia Dziaková Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Study on interactions of PARP inhibitors with ABC drug efflux transporters. ATP-binding cassette (ABC) transporters are integral membrane proteins that use the energy obtained from ATP to carry transport of numerous endogenous substrances out of the cells, but attention is drawn primarily to the fact that they transfer also xenobiotics. Their overexpression in tumor tissue contributes to multidrug resistance (MDR), which in most cases leads to therapy failure. Poly(ADP-ribose)polymerase inhibitors (PARPi) represent a promising therapeutic approach in the treatment of cancers that exhibit defects in homologous recombination (HR). This work focuses on four selected PARPi (olaparib, rucaparib, niraparib, veliparib) and their interaction potential towards ABC drug efflux transporters (ABCB, ABCC1, ABCG2). In our work, we worked with MDCKII cells (parent, transduced by the transporters of interest) and utilized the principle of accumulation studies based on the measurement of fluorescence intensity of specific model substrates (hoechst33342, calcein AM, daunorubicin, mitoxantrone). We used established inhibitors of studied...

Title: Neurological deficit after focal cerebral ischemia in rats - pharmacological intervention Objectives: The goal of the thesis was to determine the effect of 7-nitroindazole, a selective inhibitor of neuronal nitric oxide synthase, after focal ischemic stroke in rats. Methods: Twenty adult male Wistar rats were used in this experiment. The rats were randomly divided into four groups: ischemic stroke was given to half of them, the rest were sham operated. 10 animals were given 7-nitroindazole (25mg/kg) to protect neuronal ischemic brain damage. After a few weeks the rats were tested with a set of behavioral tests: Ladder rung walking test, Bar holding test, Rotarod test and Open field test. To evaluate the volume of brain damage the stereotactic method was used. The brain sections were cut and compared with atlas. This study was supported by Institute of Physiology, Academy of Sciences ČR in Prague. Results: The present results show that the 7-nitroindazole has no side effects on healthy rats. The long-term effect on rats after ischemic stroke was not proved. There were a few positive trends observed such as an increase of locomotor speed, increased explorative behaviour and better coordination outcome on RotaRod. On the other hand the brain tissue damage was bigger and the time of hanging in...

Institution/department: Charles University, Faculty of Pharmacy in Hradec Kralove, Department of Social and Clinical Pharmacy Title of diploma thesis: The rational use of proton pump inhibitors among geriatric patients in the Czech Republic Supervisor: Assoc. Prof. Daniela Fialova, PharmDr. Ph.D. Author: Ivana Geletova Introduction: The dynamic process of ageing requires adequate attention at a global level, mainly in the area of providing a proper healthcare. Ageing process is specific by significant involutional changes and is often complicated by higher polymorbidity and polypharmacotherapy. Therefore, it is necessary to eliminate the negative impact of possible drug-related and nondrug-related risk factors and to ensure the rational use of medicines. Proton pump inhibitors (PPIs) are one of the most frequently prescribed classes of drugs in older adults, administered especially for the treatment of gastrointestinal tract (GIT) disorders associated with increased secretion of gastric acid and/or impairment of GIT or as a preventive treatment of potential drug-related and other gastropathies. This diploma thesis focused on evaluating the prevalence of use of PPIs among geriatric patients in acute, ambulatory and pharmacy settings of healthcare in the Czech Republic, in the EUROAGEISM H2020...

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