Crystal structures of anhydrous pinacol, the hexagonal pinacol, pinacol monohydrate, and pinacol hexahydrate were studied. In all the structures crystal packing is unusual and complicated. The origin of the complexity may be the difficulty in filling space densely and while also satisfying the H-bonding requirements when the molecule has few internal degrees of freedom. Five 15-crown-5 complexes of M(NO3)2 (M = Cu, Zn, Mg, Co, Mn) were synthesized and characterized using X-ray diffraction and differential scanning calorimetry. The system is rich in polymorphs. Nine definite solid-state phases were identified. More phases probably exist in the solid state at temperatures slightly above the room temperature. Most phase transformations in this system take place in single crystals without the loss of crystallinity. The nine phases crystallize in five crystal structures. The crown ether ligands have very similar conformation in all the structures. The asymmetric units in all the structures are complicated and pseudosymmetric, which is the consequence of the presence of the packing problem. The origin of the packing problem that leads to the complicated phase behavior is the odd number of -CH2-O-CH2- units in the crown ether ligand.

Since apoE allele status is the predominant Alzheimers disease (AD) genetic risk factor, functional single nucleotide polymorphisms (SNPs) in brain apoE receptors represent excellent candidates for association with AD. Therefore, three low density lipoprotein receptor (LDLR) SNPs were evaluated by TaqMan allelic discrimination assays for association with AD and I found that certain haplotypes alter the odds of AD. A SNP within LDLR exon 12, rs688, was identified in silico as neutralizing a putative exon splicing enhancer (ESE). Since LDLR is a major apoE receptor in the brain, I hypothesized that rs688 modulates LDLR splicing in neural tissues and associates with AD. To evaluate this hypothesis, I analyzed splicing patterns in human hippocampus samples and established that this SNP was associated with significantly decreased LDLR exon 12 splicing efficiency when the minor allele T is present in vivo. Lastly, I evaluated whether rs688 associates with AD by genotyping DNA from the Religious Orders Study (ROS) series. The rs688T/T genotype was associated with increased AD odds in males, but not in females, in a dataset consisting of 1,457 men and 2,055 women drawn from three case-control series. The rs688T/T genotype was associated with increased AD odds in males (recessive model, odds ratio (OR) of 1.49, 95% confidence interval (CI) of 1.13- 1.97, uncorrected p=0.005), but not in females. In summary, these studies identify a functional apoE receptor SNP that is associated with AD in a sex-dependent fashion.

Colorectal cancer is estimated to cause approximately 50,000 deaths each year in the United States. Epidemiological studies have demonstrated an inverse association between sunlight exposure, which stimulates the formation of vitamin D in the skin, and colorectal carcinoma. Laboratory studies report that metabolites of vitamin D, acting through the vitamin D receptor (VDR), regulate cellular proliferation, differentiation and apoptosis. In addition, VDR contains a polymorphic variant, FokI, which results in two different isoforms of VDR. We have demonstrated a differential suppression of β-catenin transcriptional activity by these isoforms in the presence of 1,25(OH)₂D₃ (1,25D). Epidemiological evaluation of metachronous colorectal adenoma formation indicates that VDR includes several single nucleotide polymorphisms (SNPs) which influence the odds of developing colorectal adenoma. In addition, we have found full length Adenomatous Polyposis Coli (APC), a frequently mutated tumor suppressor gene in colorectal cancer, augments both the interaction of VDR and β-catenin as well as the suppression of β-catenin transcriptional activity in the presence of 1,25D. We have also demonstrated in epidemiological studies that the presence of a T-A haplotype in APC codons 486 and 1822, respectively, reduces the odds of any metachronous adenoma by 27% [odds ratio (OR), 0.73; 95% confidence interval (95% CI), 0.59 – 0.91]. Taken together, these data support not only a protective role for vitamin D acting through the VDR, but also for an important role of heritable polymorphic variation in VDR and APC in carcinogenesis.

<p>The aim of this study was to investigate the forensic usefulness of various Y-chromosome short tandem repeat loci among South African sub-populations. Three different sets of Y-chromosome short tandem repeat loci were chosen for investigation.</p>

In most cases, organic materials exist in the solid phase as polymorphs, solvatomorphs or amorphous forms. Physico-chemical properties in the solid-state are all affected primarily in terms of dissolution, solubility, bioavailability, stability and processability. Therefore investigation into the polymorphic behaviour of APIs has become a mandatory part of drug characterisation studies by pharmaceutical companies (Giron, 2001). The influence polymorphism has on bioavailability and the need for the development of drugs in the amorphous form have instigated regulatory bodies such as the FDA to require solid-state characterisation of pharmaceuticals (Strachan et al., 2005). Subsequently a study was conducted to determine the physico-chemical properties of two poorly watersoluble macrolides; clarithromycin and spiramycin. Characterisation methods included: XRPD, IR, TGA, DSC, SEM, Karl Fischer titration, solubility and stability studies. Recrystallisations of spiramycin from various solvents indicated that this API mainly exists in the amorphous form. The DSC proved to be of little value in the characterisation of this particular macromolecular antibiotic, since wide inter-sample variations were mostly obtained. TGA results showed higher solvent uptake than expected. This was ascribed to the amorphous, sponge-like character of this drug. For the sake of reproducibility and quality of the results, characterisation of spiramycin was more reliant on spectroscopic and crystallographic methods. Samples generated from 2- butanol, chloroform, ethyl acetate, 1.4-dioxane, methanol, n-propanol, iso-propanol and tetrahydrofuran showed characteristic peaks in the range of 2000-2400 cm-1 that were not present in the IR spectrum of the raw material. Conversely, the XRPD patterns were all identical, exhibiting a characteristic “halo” pattern with no detectable Bragg diffraction peaks. A solubility assessment showed no significant differences between the raw material and the recrystallisation products. In fact the raw material seemed to be the form with the highest solubility, albeit it only by a small margin. According to the literature, clarithromycin exists in five forms. Form 0 exists as a solvate, form I is a metastable form, form II is the stable form (Liu & Riley 1998; Deshpande et al., 2006), form III is a solvate of acetonitrile (Liu et al., 2003; Liang & Yao, 2008) and form IV is a hydrate (Avrutov et al., 2003). The stable form II is used in formulations currently on the market. A follow-up study was done relating to a study performed by De Jager (2005). The raw material (form II) was recrystallised from acetonitrile, chloroform and ethyl acetate. Two new crystal forms were prepared from chloroform and acetonitrile. With the necessary driving force, both of these crystals forms are able to convert to the thermodynamically stable form II. In addition, a solvate recrystallised from chloroform together with its corresponding desolvate, showed a 4 and 1.5 fold respective increase in solubility when compared to the raw material. The recrystallisations from ethyl acetate delivered crystals with an XRPD pattern similar to form II. This proved that clarithromycin can be recrystallised directly from this solvent without the need of an additional conversion step, as was the case in the study done by De Jager (2005). / MSc (Pharmaceutics), North-West University, Potchefstroom Campus, 2013

Pollinator-driven selection is thought to drive much of the extraordinary diversity of flowering plants. Plants that produce floral traits preferred by particular pollinators are more likely to receive conspecific pollen and to evolve further adaptations to those pollinators that enhance pollination and ultimately generate floral diversity. Two mechanisms in particular, sensory bias and learning, are thought to explain how pollinator preference can contribute to divergence and speciation in flowering plants. While the preferences of pollinators, such as bees, flies, and birds, are frequently implicated in patterns of floral trait evolution, the role of learning in generating reproductive isolation and trait divergence for different floral types within plant populations is not well understood. Floral color polymorphism in particular provides an excellent opportunity to examine how pollinator behavior and learning might maintain the different floral morphs. In this study we asked if bumble bees showed innate preferences for different color morphs of the pollen-only plant Solanum tridynamum, whether bees formed preferences for the morphs with which they had experience collecting pollen from, and the strength of those learned preferences. Using an absolute conditioning protocol, we gave bees experience collecting pollen from a color polymorphic plant species that offered only pollen rewards. Despite initially-naïve bees showing no apparent innate bias toward human-white versus human-purple flower morphs, we did find evidence of a bias in learning. Specifically, bees learned strong preferences for purple corollas, but learned only weak preferences for hypochromic (human-white) corollas. We discuss how our results might explain patterns of floral display evolution, particularly as they relate to color polymorphisms. Additionally, we propose that the ease with which floral visual traits are learned—i.e., biases in learning—can influence the evolution of floral color as a signal to pollinators.

The metabolic expense of producing and operating neural tissue required for adaptive behaviour is considered a significant selective force in brain evolution. In primates, brain size correlates positively with group size, presumably owing to the greater cognitive demands of complex social relationships in large societies. Social complexity in eusocial insects is also associated with large groups, as well as collective intelligence and division of labour among sterile workers. However, superorganism phenotypes may lower cognitive demands on behaviourally specialized workers resulting in selection for decreased brain size and/or energetic costs of brain metabolism. To test this hypothesis, we compared brain investment patterns and cytochrome oxidase (COX) activity, a proxy for ATP usage, in two ant species contrasting in social organization. Socially complex Oecophylla smaragdina workers had larger brain size and relative investment in the mushroom bodies (MBs)-higher order sensory processing compartments-than the more socially basic Formica subsericea workers. Oecophylla smaragdina workers, however, had reduced COX activity in the MBs. Our results suggest that as in primates, ant group size is associated with large brain size. The elevated costs of investment in metabolically expensive brain tissue in the socially complex O. smaragdina, however, appear to be offset by decreased energetic costs.

Le paludisme est une cause majeure de morbidité et de mortalité, plus particulièrement en Afrique sub-saharienne. De très nombreuses observations sont en faveur de l'existence de facteurs génétiques contrôlant le devenir de l'infection palustre. Il est très probable que certains variants génétiques de gènes candidats du paludisme affectent la résistance du paludisme à travers leur effet sur la réponse immunitaire acquise. Afin de vérifier cette hypothèse, nous avons étudié, dans une population vivant au Burkina Faso, des variants génétiques de HBB, IL4, IL12B, TNF, LTA, FCGR2A et NCR3 dont l'association avec des phénotypes liés à la résistance au paludisme a été publiée; nous avons évalué leur influence sur les niveaux d'IgG dirigés contre les antigènes de Plasmodium falciparum en utilisant un test d'association basé sur les familles. Ainsi, nous avons détecté, l'effet de l'hémoglobine C, FCGR2A-H131, le TNF-857T, et TNF1304A sur les niveaux des sous-classes d'IgG anti-P. falciparum. Ces résultats constituent une base utile pour des études ultérieures du contrôle génétique de la réponse immunitaire chez des individus vivant dans une zone d'endémie. Un autre projet a porté sur l'étude fonctionnelle du polymorphisme NCR3-412, dont nous avions montré l'association avec les accès palustres simples. Nos résultats basés sur des techniques moléculaires montrent l'effet de ce polymorphisme situé dans le promoteur sur la liaison de protéines nucléaires. / Malaria is the major cause of morbidity and mortality especially in the Sub-Saharan Africa. There is a growing body of evidence for genetic factors controlling the outcome of malaria infection. It is thought that some genetic variants of malaria candidate genes affect malaria resistance through their effect on the acquired immune response. In order to verify this hypothesis, we worked on genetic variants of HBB, IL4, IL12B, TNF, LTA, FCGR2A and NCR3, which have been associated with malaria resistance phenotypes, to determine their influence on levels of anti-P. falciparum IgG in urban population of Burkina Faso. Using family-based association analysis, we detected the effect of Hemoglobin C, FCGR2A-H131, TNF-857T, and TNF1304A on the levels of anti-P. falciparum IgG. This study can pave the way towards further comprehension of genetic control of an individual's immune response against malaria. Another project focused on functional study of polymorphism NCR3-412, which has already been associated to mild malaria. We investigated the functional effect of this polymorphism located in the promoter by using molecular techniques and showed the effect of this polymorphism on the binding of nuclear proteins.

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michal Rataj Supervisor: Doc. PharmDr. Martin Beránek Ph.D. Title of diploma thesis: Genetic polymorphism in the NBS1 gene for diagnosis and treatment of patients with cervical carcinoma The aim of this diploma thesis is to find optimal methods for screening of mutation 657del5 and estimate frequency of heterozygotes and homozygotes for the mutation 657del5 in population of the Czech republic. In the first section of the theoretical part is comprehensively pointed out the effect of factors affecting the integrity of genetic information and the formation of mutations in DNA. On the contrary, the second section devotes to the ability of cells to respond to this damage. In detail, the thesis devotes to the NBS1 gene and its product nibrin. In the complex MRE11/Rad50/NBN nibrin is an important member of the mechanisms of repair of double strand breaks NHEJ (non-homologous end joining) and HR (homologous recombination). The thesis is focused on nibrin and its functions, but also to mutations that prevent these functions and causes genetic disease Nijmegen breakage syndrome. Nibrin is translated from the sequence of the NBS1 gene. Gene NBS1 appears in population with several various...

1 Abstract Charles University in Prague, Faculty of Pharmacy in Hradci Králové Department of: Pharmaceutical technology Consultant: PharmDr. Ondřej Holas, Ph.D. Student: Tomáš Máčalík Title of Thesis: Problematics of active compounds recrystalisation in pharmaceutical preparations The topic of the thesis is the recrystallization of active substances in pharmaceutical preparations. The aim of this work is to demonstrate this phenomenon on a real particular pharmaceutical product, in which it has occurred. It includes a detailed description of the manufacturing process including all mandatory conditions, possible critical points, investigations and measures taken. The thesis consists of two parts. The first describes in details the current state of knowledge. It is based on the findings of the available literature on the subject. The basic concepts of crystallization and polymorphism are explained in detail, which form the basis for the subsequent case study. The case study includes a detailed description of a specific medicinal product. Subsequently, it's production, good manufacturing practice and gradual investigation of the discovered technological incompatibility are explained step by step. The result of the work points out that the issue of recrystallization and polymorphism is very closely related to...