Characterization of polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 and relationship to the alcoholism in a Colombian population

Objective: Identify and characterize polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 in a Colombian population residing in the city of Bogotá and determine its possible relationship to the alcoholism. Methods: ADH2, ADH3, ALDH2, and CYP2E1 genotypes a population of 148 individuals with non-problematic alcohol and 65 individuals with alcoholism were determined with TaqMan probes and PCR-RFLP. DNA was obtained from peripheral blood white cells. Results: Significant difference was found in family history of alcoholism and use of other psychoactive substances to compare alcoholics with controls. When allelic frequencies for each category (gender) were considered, frequency of A2 allele carriers in ADH2 was found higher in male patients than controls. In women, the relative frequency for c1 allele in CYP2E1 was lower in controls than alcoholics. The ALDH2 locus is monomorphic. No significant differences in allele distributions of the loci examined to compare two populations were observed, however when stratifying the same trend was found that these differences tended to be significant. Conclusions: This study allows us to conclude the positive association between family history of alcoholism and alcoholism suggesting that there is a favorable hereditary predisposition. Since substance dependence requires interaction of multiple genes, the combination of genotypes ADH2*2, CYP2E1*1 combined with genotype homozygous ALDH2*1 found in this study could be leading to the population to a potential risk to alcoholism.

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.

Characterization of polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 and relationship to the alcoholism in a Colombian population

Objective: Identify and characterize polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 in a Colombian population residing in the city of Bogotá and determine its possible relationship to the alcoholism. Methods: ADH2, ADH3, ALDH2, and CYP2E1 genotypes a population of 148 individuals with non-problematic alcohol and 65 individuals with alcoholism were determined with TaqMan probes and PCR-RFLP. DNA was obtained from peripheral blood white cells. Results: Significant difference was found in family history of alcoholism and use of other psychoactive substances to compare alcoholics with controls. When allelic frequencies for each category (gender) were considered, frequency of A2 allele carriers in ADH2 was found higher in male patients than controls. In women, the relative frequency for c1 allele in CYP2E1 was lower in controls than alcoholics. The ALDH2 locus is monomorphic. No significant differences in allele distributions of the loci examined to compare two populations were observed, however when stratifying the same trend was found that these differences tended to be significant. Conclusions: This study allows us to conclude the positive association between family history of alcoholism and alcoholism suggesting that there is a favourable hereditary predisposition. Since substance dependence requires interaction of multiple genes, the combination of genotypes ADH2*2, CYP2E1*1 combined with genotype homozygous ALDH2*1 found in this study could be leading to the population to a potential risk to alcoholism.

Characterization of some bacterial strains isolated from animal clinical materials and identified as Corynebacterium xerosis by molecular biological techniques

Eighteen Corynebacterium xerosis strains isolated from different animal clinical specimens were subjected to phenotypic and molecular genetic studies. On the basis of the results of the biochemical characterization, the strains were tentatively identified as C. xerosis. Phylogenetic analysis based on comparative analysis of the sequences of 16S rRNA and rpoB genes revealed that the 18 strains were highly related to C. xerosis, C. amycolatum, C. freneyi, and C. hansenii. There was a good concordance between 16S rRNA and partial rpoB gene sequencing results, although partial rpoB gene sequencing allowed better differentiation of C. xerosis. Alternatively, C. xerosis was also differentiated from C. freneyi and C. amycolatum by restriction fragment length polymorphism analysis of the 16S-23S rRNA gene intergenic spacer region. Phenotypic characterization indicated that besides acid production from D-turanose and 5-ketogluconate, 90% of the strains were able to reduce nitrate. The absence of the fatty acids C(14:0), C(15:0), C(16:1)omega 7c, and C(17:1)omega 8c can also facilitate the differentiation of C. xerosis from closely related species. The results of the present investigation demonstrated that for reliable identification of C. xerosis strains from clinical samples, a combination of phenotypic and molecular-biology-based identification techniques is necessary.

Characterization of medieval glass artefacts from Miranduolo site, Chiusdino, Italy

This archaeovitreological study deals with artefacts of Miranduolo site, Tuscany region (Italy), dated 1250-1350 AD. The Miranduolo site is a medieval hill-village dated from 7th to 14th century. The information obtained reveal that Miranduolo was under control of noble families, which displayed the social, economic and political power. It is marked by controlling the farmers and metal workers on the site, as well as having control over agricultural surpluses. No in situ glass workshop has been recovered, implying that the glass artefacts were imported. One aim of this work is application of SEM-EDS to visualize textural characteristics and thickness of the pristine glass and corrosion layers. Preliminary qualification and semi-quantification of major and minor chemical elements will provide the data on the glass group present and fluxes employed. The data obtained will be integrated with the one obtained by more sensitive techniques such as PIXE/PIGE and LA-ICP-MS. Twenty cross-sections of transparent glasses (colorless, azure, and different hues of yellow and) have been analyzed by VP-SEM. All the analyzed glasses display a homogenous matrix. Only four samples (MD 24, MD 139, MD 143, MD 259) show corrosion layers of various thickness with 2.25μm, 136-500 μm, 26.8 μm and 17.01 μm. EDS linescan analyses indicate strong depletion in the corrosion layers of Na and K, while Ca depletes to a minor extent. In general, both glass composition and the burial conditions were favorable for preservation. Samples can be classified as mainly plant ash Na-Ca-Si glasses made with both unpurified and purified Levantine ash. Only sample MD 243 is made from Barilla plant ash. Sample MD 139 cannot be classified into main compositional groups as K2O is 1.33 wt% and MgO 5.92 wt%. In 8 samples MnO content is lower than 0.8 wt%, meaning that in these samples MnO is naturally present. In other 12 samples, MnO above 0.8 wt% indicates deliberate addition as a decolorant agent to intentionally obtain different hues or the amount added was not successful in making the glass transparent. The results considering fluxes are compatible with archaeovitreological study from contemporary primary glass workshops in Tuscany. For determining the provenance of silica sources, further analysis with more sensitive techniques has to be carried out; Resumo: Este estudo “arqueovitreologia” lida com artefatos do local Miranduolo, região da Toscana (Itália), datados de 1250-1350 AD. O sitio de Miranduolo é uma colina vila medieval datada do séc.VII ao séc.XIV. As informações obtidas revelam que Miranduolo estava sob o controle de famílias nobres, que exibiu o poder social, económico e político. É marcado por controlar os agricultores e trabalhadores do metal no sitio, bem como ter controlo sobre os excedentes agrícolas. Não há na oficina de vidro in situ foi recuperado, o que implica que os artefactos de vidro foram importados. Um dos objetivos deste trabalho é a aplicação de SEM-EDS para visualizar características de textura e espessura das camadas de corrosão do vidro também como da áreas originais. qualificação preliminar e semi-quantificação de maiores e menores elementos químicos irá fornecer os dados sobre o grupo presente vidro e fluxos empregado. Os dados obtidos são integrados com os dados obtidos por meio de técnicas mais sensíveis, como PIXE / PIGE e LA-ICP-MS. Vinte secções transversais de vidros transparentes (incolor, azul celeste, e diferentes tons de amarelo) foram analisados por VP-SEM. Todos os vidros analisados exibir uma matriz homogénea. Apenas quatro amostras (MD 24, MD 139, MD 143, MD 259) mostram camadas de corrosão de várias espessuras com 2.25μm, 136-500μm, 26,8μm e 17,01μm. Análises Linescan EDS indicam forte esgotamento nas camadas de corrosão de Na e K, enquanto Ca esgota, em menor grau. Em geral, tanto a composição de vidro e as condições de depósito foram favoráveis para a preservação. As amostras podem ser classificados como vidros principalmente Na-Ca-Si feitas com cinzas de plantas do tipo levantino, não purificada e purificada. Apenas a amostra MD 243 é feita a partir de cinzas vegetais tipo “Barilla”. A amostra MD 139 não pode ser classificada em grupos principais de composição porque K2O é 1,33% em peso e MgO 5,92% em peso. Em 8 amostras, o teor de MnO é menor do que 0,8% em peso, o que significa que nestas amostras MnO está naturalmente presente. Em outras 12 amostras, MnO acima de 0,8% em peso indica adição intencional como um agente de colorante para obter intencionalmente diferentes matizes ou o valor acrescentado não foi bem sucedido em fazer o vidro transparente. Os resultados, considerando os fluxos são compatíveis com o estudo “arqueovitreologico” com as principais oficinas de vidro contemporâneos na Toscana. Para determinar a origem das fontes de sílica, uma análise mais aprofundada com técnicas mais sensíveis tem de ser levada a cabo.

Preparation and Characterization of Polymeric Beads and Membranes for Boron Removal From Water

Boron is an element essential for various biological processes, nevertheless at high concentration it can cause health issues in both plants and animals, thus making boron a pollutant element. Low cost and effective polymeric adsorbents capable of removing boron in aqueous solution at neutral pH were prepared for this purpose. The adsorbent selectivity towards boron was conferred taking advantage of the interaction between boric acid and the alcoholic groups of N-methyl-D-Glucamine, which are able to form specific complexes. Two different kinds of devices were produced and tested: cross-linked chitosan hydrogel beads (CCBMG) and PVA/chitosan membranes, the latter taking advantage of scCO2-assisted phase inversion technique. The capability of the adsorbents to be regenerated and to allow recovery of boric acid from a solution emulating the concentration of boric acid in seawater were evaluated.

Computational characterization of carbazole-benzonitrile derivatives for applications in Organic Light Emitting Diodes

The technology of Organic Light-Emitting Diodes has reached such a high level of reliability that it can be used in various applications. The required light emission efficiency can be achieved by transforming the triplet excitons into singlet states through Reverse InterSystem Crossing (RISC), which is the main process of a general mechanism called thermally activated delayed fluorescence (TADF). In this thesis, we theoretically analyzed two carbazole-benzonitrile (donor-acceptor) derivatives, 2,5-di(9H-carbazol-9-yl)benzonitrile (p-2CzBN) and 2,3,4,5,6-penta(9H-carbazol-9-yl)benzonitrile (5CzBN), and addressed the problem of how donor-acceptor (D-A) or donor-acceptor-donor (D-A-D) flexible molecular architectures influence the nature of the excited states and the emission intensity. Furthermore, we analyzed the RISC rates as a function of the conformation of the carbazole lateral groups, considering the first electronic states, S0, S1, T1 and T2, involved in TADF process. The two prototype molecules, p-2CzBN and 5CzBN, have a similar energy gap between the first singlet and triplet states (∆EST, a key parameter in the RISC rate), but different TADF performances. Therefore, other parameters must be considered to explain their different behavior. The oscillator strength of p-2CzBN, never tested as emitter in OLEDs, is similar to that of 5CzBN, which is an active TADF molecule. We also note that the presence of a second T2 triplet state, lower in energy than S1 only in 5CzBN, and the reorganization energies, associated with RISC processes involving T1 and T2, are important factors in differentiating the rates in p-2CzBN and 5CzBN. For p-2CzBN, the RISC rate from T2 to S1 is surprisingly higher than that from T1 to S1, in disagreement with El-Sayed rules, due to a large reorganization energy associated to the T1 to S1, process; while the contrary occurs for 5CzBN. These insights are important for designing new TADF emitters based on the benzo-carbazole architecture.

Structural and biophysical characterization of RAD52 as novel pharmacological target for synthetic lethality

RAD52 is a protein involved in various DNA reparation mechanisms. In the last few years, RAD52 has been proposed as a novel pharmacological target for cancer synthetic lethality strategies. Hence, this work has the purpose to investigate RAD52 protein, with biophysical and structural tools to shed light on proteins features and mechanistic details that are, up to now poorly described, and to design novel strategies for its inhibition. My PhD work had two goals: the structural and functional characterization of RAD52 and the identification of novel RAD52 inhibitors. For the first part, RAD52 was characterized both for its DNA interaction and oligomerization state together with its propensity to form high molecular weight superstructures. Moreover, using EM and Cryo-EM techniques, additional RAD52 structural hallmarks were obtained, valuable both for understanding protein mechanism of action and for drug discovery purpose. The second part of my PhD project focused on the design and characterization of novel RAD52 inhibitors to be potentially used in combination therapies with PARPi to achieve cancer cells synthetic lethality, avoiding resistance occurrence and side effects. With this aim we selected and characterized promising RAD52 inhibitors through three different approaches: 19F NMR fragment-based screening; virtual screening campaign; aptamers computational design. Selected hits (fragments, molecules and aptamers) were investigated for their binding to RAD52 and for their mechanism of inhibition. Collected data highlighted the identification of hits worthy to be developed into more potent and selective RAD52 inhibitors. Finally, a side project carried out during my PhD is reported. GSK-3β protein, an already validated pharmacological target was investigated using biophysical and structural biology tools. Here, an innovative and adaptable drug discovery screening pipeline able to directly identify selective compounds with binding affinities not higher than a reference binder was developed.

Synthesis and characterization of prussian blue analogue materials for post-Li ion batteries

Energy issues have always been a subject of concern to people. During the past 30 years, rechargeable Li-ion batteries (LIBs) have been widely used in portable electronic devices and power tools because of their high energy density and efficiency among practical secondary batteries. While the unevenly distribution of Lithium sources and the increasing cost of lithium-raw material can not satisfy the requirement for further cost reduction, especially for the grid-scale energy storage. Post-lithium ion batteries as promising replacement for LIBs have attracted wide attention, owing to their high abundant resources and adequate insertion potential. Similar with Li-ion batteries, finding a suitable electrode material is the key for the research and application of the post-Li ion batteries. In our project, we focus our study on Prussian blue analogues (PBAs), with formula AxM[M’(CN)6]1-y□y•zH2O (0≤x≤2, 0of metal-organic framework, with 3D frameworks which allow for a facile insertion/ extraction of ions with negligible lattice strain. By substituting the metal sites with different transition metals, we can get a series of compounds that can be used as both cathode and anode material for both Li-ion and post-Li batteries. The most commonly studied PBAs are metal haxacyanoferrate, with the carbon-sites of -CN- ligands fix connected with Fe. Here, we synthesized three different PBAs: manganese hexacynoferrate (MnHCF), zinc hexacynoferrate (ZnHCF) and titanium hexacynoferrate (TiHCF), using co-precipitation method, and their electrochemical properties were tested in both aqueous Na+, K+, Mg2+, Zn2+ and organic Li+, Na+ electrolytes. Various X-ray techniques were employed to study their electronic and structural properties of electrodes and electrochemical reaction mechanism during cycling.

Design and characterization of automated agricultural vehicle systems

Nowadays, technological advancements have brought industry and research towards the automation of various processes. Automation brings a reduction in costs and an improvement in product quality. For this reason, companies are pushing research to investigate new technologies. The agriculture industry has always looked towards automating various processes, from product processing to storage. In the last years, the automation of harvest and cultivation phases also has become attractive, pushed by the advancement of autonomous driving. Nevertheless, ADAS systems are not enough. Merging different technologies will be the solution to obtain total automation of agriculture processes. For example, sensors that estimate products' physical and chemical properties can be used to evaluate the maturation level of fruit. Therefore, the fusion of these technologies has a key role in industrial process automation. In this dissertation, ADAS systems and sensors for precision agriculture will be both treated. Several measurement procedures for characterizing commercial 3D LiDARs will be proposed and tested to cope with the growing need for comparison tools. Axial errors and transversal errors have been investigated. Moreover, a measurement method and setup for evaluating the fog effect on 3D LiDARs will be proposed. Each presented measurement procedure has been tested. The obtained results highlight the versatility and the goodness of the proposed approaches. Regarding the precision agriculture sensors, a measurement approach for the Moisture Content and density estimation of crop directly on the field is presented. The approach regards the employment of a Near Infrared spectrometer jointly with Partial Least Square statistical analysis. The approach and the model will be described together with a first laboratory prototype used to evaluate the NIRS approach. Finally, a prototype for on the field analysis is realized and tested. The test results are promising, evidencing that the proposed approach is suitable for Moisture Content and density estimation.

Design, fabrication and mechanical characterization of a structural node made using Wire and Arc Additive Manufacturing (WAAM) technology

Additive Manufacturing (AM), also known as “3D printing”, is a recent production technique that allows the creation of three-dimensional elements by depositing multiple layers of material. This technology is widely used in various industrial sectors, such as automotive, aerospace and aviation. With AM, it is possible to produce particularly complex elements for which traditional techniques cannot be used. These technologies are not yet widespread in the civil engineering sector, which is slowly changing thanks to the advantages of AM, such as the possibility of realizing elements without geometric restrictions, with less material usage and a higher efficiency, in particular employing Wire-and-Arc Additive Manufacturing (WAAM) technology. Buildings that benefit most from AM are all those structures designed using form-finding and free-form techniques. These include gridshells, where joints are the most critical and difficult elements to design, as the overall behaviour of the structure depends on them. It must also be considered that, during the design, the engineer must try to minimize the structure's own weight. Self-weight reductions can be achieved by Topological Optimization (TO) of the joint itself, which generates complex geometries that could not be made using traditional techniques. To sum up, weight reductions through TO combined with AM allow for several potential benefits, including economic ones. In this thesis, the roof of the British Museum is considered as a case study, analysing the gridshell structure of which a joint will be chosen to be designed and manufactured, using TO and WAAM techniques. Then, the designed joint will be studied in order to understand its structural behaviour in terms of stiffness and strength. Finally, a printing test will be performed to assess the production feasibility using WAAM technology. The computational design and fabrication stages were carried out at Technische Universität Braunschweig in Germany.

Characterization Of Single Use Reactor Used For The Manufacturing Of Formulated Products Within The Pharmaceutical Industry

Mixing is a fundamental unit operation in the pharmaceutical industry to ensure consistent product quality across different batches. It is usually carried out in mechanically stirred tanks, with a large variety of designs according to the process requirements. A key aspect of pharmaceutical manufacturing is the extensive and meticulous cleaning of the vessels between runs to prevent the risk of contamination. Single-use reactors represent an increasing trend in the industry since they do not require cleaning and sterilization, reducing the need for utilities such as steam to sterilize equipment and the time between production batches. In contrast to traditional stainless steel vessels, single-use reactors consist of a plastic bag used as a vessel and disposed of after use. This thesis aims to characterize the fluid dynamics features and the mixing performance of a commercially available single-use reactor. The characterization employs a combination of various experimental techniques. The analysis starts with the visual observation of the liquid behavior inside the vessel, focusing on the vortex shape evolution at different impeller speeds. The power consumption is then measured using a torque meter to quantify the power number. Particle Image Velocimetry (PIV) is employed to investigate local fluid dynamics properties such as mean flow field and mean and rms velocity profiles. The same experimental setup of PIV is exploited for another optical measurement technique, the Planar Laser-Induced Fluorescence (PLIF). The PLIF measurements complete the characterization of the reactor with the qualitative visualization of the turbulent flow and the quantitative assessment of the system performance through the mixing time. The results confirm good mixing performances for the single-use reactor over the investigated impeller speeds and reveal that the filling volume plays a significant role in the fluid dynamics of the system.

Characterization Of Microsatellite Markers Developed From Prosopis Rubriflora And Prosopis Ruscifolia (leguminosae - Mimosoideae), Legume Species That Are Used As Models For Genetic Diversity Studies In Chaquenian Areas Under Anthropization In South America.

Prosopis rubriflora and Prosopis ruscifolia are important species in the Chaquenian regions of Brazil. Because of the restriction and frequency of their physiognomy, they are excellent models for conservation genetics studies. The use of microsatellite markers (Simple Sequence Repeats, SSRs) has become increasingly important in recent years and has proven to be a powerful tool for both ecological and molecular studies. In this study, we present the development and characterization of 10 new markers for P. rubriflora and 13 new markers for P. ruscifolia. The genotyping was performed using 40 P. rubriflora samples and 48 P. ruscifolia samples from the Chaquenian remnants in Brazil. The polymorphism information content (PIC) of the P. rubriflora markers ranged from 0.073 to 0.791, and no null alleles or deviation from Hardy-Weinberg equilibrium (HW) were detected. The PIC values for the P. ruscifolia markers ranged from 0.289 to 0.883, but a departure from HW and null alleles were detected for certain loci; however, this departure may have resulted from anthropic activities, such as the presence of livestock, which is very common in the remnant areas. In this study, we describe novel SSR polymorphic markers that may be helpful in future genetic studies of P. rubriflora and P. ruscifolia.

Development And Characterization Of Microsatellite Loci For Ocotea Species (lauraceae) Threatened With Extinction.

The Atlantic rainforest species Ocotea catharinensis, Ocotea odorifera, and Ocotea porosa have been extensively harvested in the past for timber and oil extraction and are currently listed as threatened due to overexploitation. To investigate the genetic diversity and population structure of these species, we developed 8 polymorphic microsatellite markers for O. odorifera from an enriched microsatellite library by using 2 dinucleotide repeats. The microsatellite markers were tested for cross-amplification in O. catharinensis and O. porosa. The average number of alleles per locus was 10.2, considering all loci over 2 populations of O. odorifera. Observed and expected heterozygosities for O. odorifera ranged from 0.39 to 0.93 and 0.41 to 0.92 across populations, respectively. Cross-amplification of all loci was successfully observed in O. catharinensis and O. porosa except 1 locus that was found to lack polymorphism in O. porosa. Combined probabilities of identity in the studied Ocotea species were very low ranging from 1.0 x 10-24 to 7.7 x 10-24. The probability of exclusion over all loci estimated for O. odorifera indicated a 99.9% chance of correctly excluding a random nonparent individual. The microsatellite markers described in this study have high information content and will be useful for further investigations on genetic diversity within these species and for subsequent conservation purposes.

Development And Characterization Of Microsatellite Loci For Tabebuia Cassinoides (bignoniaceae).

Tabebuia cassinoides (Lam.) DC., popularly known as caxeta, is a tree species that belongs to the plant family Bignoniaceae. This species is endemic to the Brazilian Atlantic Forest and is widely exploited commercially. To date, little is known about its genetic structure, preventing the establishment of adequate management plans for this taxon. The objective of this study was to construct a microsatellite-enriched genomic library for T. cassinoides to select polymorphic loci, and standardize polymerase chain reaction amplification conditions. Of the 15 loci examined, 5 were polymorphic. The number of alleles per locus ranged from 2 to 8, with a mean of 4.4. The microsatellite loci described here represent the basis for detailed population genetic studies of this species, which will greatly contribute for the development of better conservation strategies for this taxon.