How to improve supply chain performance of a specialized retailer: Managing landed costs of Zippy’s global supply chain

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Novel insights into the action of antimicrobial agents against human pathogens

Dissertation presented to obtain the Ph.D degree in Biochemistry

Electron Transfer and Ligand Binding Properties of Cytochromes

Dissertation presented to obtain the Ph.D degree in Biochemistry

Experimental Gravimetric Adsorption Equilibrium of n-Alkanes and Alkenes, Carbon Dioxide and Nitrogen in MIL-53(Al) and Zeolite 5A

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

Adsorption equilibria of flue gas components on activated carbon

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

Optimization of randomly amplified polymorphic DNA-polymerase chain reaction for molecular typing of Salmonella enterica serovar Typhi

Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.

Phenetic studies on randomly amplified polymorphic DNA-polymerase chain reaction-variability of four geographical populations of Lutzomyia whitmani (Diptera: Psychodidae) in Brazil

Previous evaluation of the genetic variability of four biogeographical populations of Lutzomyia whitmani from known foci of cutaneous leishmaniasis in Brazil demonstrated two main spatial clusters: Corte de Pedra-BA, Ilhéus-BA and Serra de Baturité-CE in the first cluster, and Martinho Campos-MG in the second. Further analysis showed a high degree of homogeneity in Corte de Pedra population but not in the others, which presented a significant percentage of specimens displaced from their phenon of origin (discrepant individuals). In the present work we analyzed the frequencies of association coefficients in the matrixes of similarity per population of Lutzomyia whitmani from both sexes and the general phenograms obtained, in a more detailed study of those discrepant specimens. Populational stability was observed for Corte de Pedra population, whereas the three remaining populations showed varying degrees of heterogeneity and different displacements according to sex. Our results strongly suggested the existence of a genetic flow between the lineages North-South/North-East and Ilhéus/Serra do Baturité of Lutzomyia whitmani.

The use of qualitative and quantitative polymerase chain reactions for diagnosis of cytomegalovirus infections in bone marrow and kidney transplant recipients

The purpose of this work was to test a cytomegalovirus qualitative PCR and a semi-quantitative PCR on the determination of CMV load in leukocytes of bone marrow and kidney transplanted (RT) patients. Thirty three BMT and 35 RT patients participated of the study. The DNA was subjected to a qualitative PCR using primers that amplify part of CMV gB gene. CMV load of positive samples was determined by a semi-quantitative PCR using quantified plasmids inserted with part of the gB gene of CMV as controls. The sensitivity of the test was determined to be 867 plasmid copies/µg DNA. CMV loads between 2,118 and 72,443 copies/µg DNA were observed in 12.1% BMT recipients and between 1,246 and 58,613 copies/µg DNA in 22.9% RT recipients. Further studies are necessary to confirm the usefulness of this CMV semi-quantitative PCR in transplanted patients.

Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differatiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni

We sequenced the internal transcribed spacer 2 of the ribosomal DNA (ITS2-DNAr) from the three Schistosoma mansoni intermediate hosts in Brazil: Biomphalaria glabrata, Biomphalaria tenagophila and Biomphalaria straminea. Analysis of a restriction map from those sequences allowed us to select putative restriction enzymes able to identify the snail species under study. Four restriction enzymes were used and HpaII provided simple species-specific profiles easily visualized in polyacrylamide gels. The use of ITS2 is advantageous as it provides a small fragment of 460 bp which may be easily amplified by PCR. In the current work, we showed that the amplification of ITS2-DNAr together with HpaII enzyme restriction is an auxiliary molecular tool for the morphological identification of such snails as well as for taxonomic and phylogenetic studies of neotropical planorbids.

Evaluation of a commercial test based on ligase chain reaction for direct detection of Mycobacterium tuberculosis in respiratory specimens

A ligase chain reaction DNA amplification method for direct detection of Mycobacterium tuberculosis (Abbott LCx MTB) in respiratory specimens was evaluated. Results from LCx MTB Assay were compared with those from acid fast bacilli smear, culture, and final clinical diagnosis for each patient. A total of 297 respiratory specimens (sputum and bronchial lavage) from 193 patients were tested. The sensitivity, specificity, positive predictive value and negative predictive value of LCx vs culture were 92.7%, 93%, 67.8% and 98.7%, respectively. When compared to the clinical final diagnosis, the sensitivity, specificity, PPV and NPV for LCx were 88.9%, 96.8%, 86.5% and 97.4%, respectively. The sensitivity of LCx MTB assay was 75% for smear-negative, culture positive samples. The results indicate that LCx MTB assay is a rapid, simple and valuable technique as a complementary tool for the diagnosis of tuberculosis.

Carbon prices. Dynamic analysis of European and Californian markets

Dissertação para obtenção do Grau de Doutor em Alterações Climáticas e Políticas de Desenvolvimento Sustentável

Application of NMR RDC’s, relaxation and self-diffusion for the study of dynamic processes of small molecules in solution

Spin-lattice Relaxation, self-Diffusion coefficients and Residual Dipolar Couplings (RDC’s) are the basis of well established Nuclear Magnetic Resonance techniques for the physicochemical study of small molecules (typically organic compounds and natural products with MW < 1000 Da), as they proved to be a powerful and complementary source of information about structural dynamic processes in solution. The work developed in this thesis consists in the application of the earlier-mentioned NMR techniques to explore, analyze and systematize patterns of the molecular dynamic behavior of selected small molecules in particular experimental conditions. Two systems were chosen to investigate molecular dynamic behavior by these techniques: the dynamics of ion-pair formation and ion interaction in ionic liquids (IL) and the dynamics of molecular reorientation when molecules are placed in oriented phases (alignment media). The application of NMR spin-lattice relaxation and self-diffusion measurements was applied to study the rotational and translational molecular dynamics of the IL: 1-butyl-3-methylimidazolium tetrafluoroborate [BMIM][BF4]. The study of the cation-anion dynamics in neat and IL-water mixtures was systematically investigated by a combination of multinuclear NMR relaxation techniques with diffusion data (using by H1, C13 and F19 NMR spectroscopy). Spin-lattice relaxation time (T1), self-diffusion coefficients and nuclear Overhauser effect experiments were combined to determine the conditions that favor the formation of long lived [BMIM][BF4] ion-pairs in water. For this purpose and using the self-diffusion coefficients of cation and anion as a probe, different IL-water compositions were screened (from neat IL to infinite dilution) to find the conditions where both cation and anion present equal diffusion coefficients (8% water fraction at 25 ºC). This condition as well as the neat IL and the infinite dilution were then further studied by 13C NMR relaxation in order to determine correlation times (c) for the molecular reorientational motion using a mathematical iterative procedure and experimental data obtained in a temperature range between 273 and 353 K. The behavior of self-diffusion and relaxation data obtained in our experiments point at the combining parameters of molar fraction 8 % and temperature 298 K as the most favorable condition for the formation of long lived ion-pairs. When molecules are subjected to soft anisotropic motion by being placed in some special media, Residual Dipolar Couplings (RDCs), can be measured, because of the partial alignment induced by this media. RDCs are emerging as a powerful routine tool employed in conformational analysis, as it complements and even outperforms the approaches based on the classical NMR NOE or J3 couplings. In this work, three different alignment media have been characterized and evaluated in terms of integrity using 2H and 1H 1D-NMR spectroscopy, namely the stretched and compressed gel PMMA, and the lyotropic liquid crystals CpCl/n-hexanol/brine and cromolyn/water. The influence that different media and degrees of alignment have on the dynamic properties of several molecules was explored. Different sized sugars were used and their self-diffusion was determined as well as conformation features using RDCs. The results obtained indicate that no influence is felt by the small molecules diffusion and conformational features studied within the alignment degree range studied, which was the 3, 5 and 6 % CpCl/n-hexanol/brine for diffusion, and 5 and 7.5 % CpCl/n-hexanol/brine for conformation. It was also possible to determine that the small molecules diffusion verified in the alignment media presented close values to the ones observed in water, reinforcing the idea of no conditioning of molecular properties in such media.

Development of novel ionic liquids based on biological molecules

Ionic Liquids (ILs) belong to a class of compounds with unusual properties: very low vapour pressure; high chemical and thermal stability and the ability to dissolve a wide range of substances. A new field in research is evaluating the possibility to use natural chiral biomolecules for the preparation of chiral ionic liquids (CILs). This important challenge in synthetic chemistry can open new avenues of research in order to avoid some problems related with the intrinsic biodegradability and toxicity associated to conventional ILs. The research work developed aimed for the synthesis of CILs, their characterization and possible applications, based on biological moieties used either as chiral cations or anions, depending on the synthetic manipulation of the derivatives. Overall, a total of 28 organic salts, including CILs were synthesized: 9 based on L-cysteine derivatives, 12 based on L-proline, 3 based on nucleosides and 4 based on nucleotides. All these new CILs were completely characterized and their chemical and physical properties were evaluated. Some CILs based on L-cysteine have been applied for discrimination processes, including resolution of racemates and as a chiral catalyst for asymmetric Aldol condensation. L-proline derived CILs were also studied as chiral catalysts for Michael reaction. In parallel, the interactions of macrocyclic oligosugars called cyclodextrins (CDs) with several ILs were studied. It was possible to improve the solubility of CDs in water and serum. Additionally, fatty acids and steroids showed an increase in water solubility when ILs-CDs systems were used. The development of efficient and selective ILs-CDs systems is indispensable to expand the range of their applications in host-guest interactions, drug delivery systems or catalytic reactions. Novel salts derived from nucleobases were used in order to enhance the fluorescence in aqueous solution. Additionally, preliminary studies regarding ethyl lactate as an alternative solvent for asymmetric organocatalysis were performed.

Carbon dots: nanopartículas de carbono fluorescentes para marcação e visualização de células

A imagiologia por fluorescência é uma técnica extremamente útil em investigação biomédica. Actualmente existe uma vasta gama de fluoróforos disponíveis para marcação por fluorescência. Contudo estes marcadores possuem limitações que condicionam a sua aplicação em sistemas biológicos. As nanopartículas de carbono fluorescentes (CNPs) constituem uma recente classe de marcadores fluorescentes com elevada biocompatibilidade. O objectivo deste trabalho consistiu em produzir de CNPs através de métodos simples, a sua caracterização e aplicação como marcadores celulares para visualização de células em microscopia de fluorescência. Inicialmente foram produzidas nanopartículas (NPs) seguindo métodos mencionados na literatura. Seguidamente foram produzidas CNPs a partir de PAA, por via hidrotérmica, e a partir da carbonização de grãos de cortiça para as quais foi feito um estudo do efeito da variável temperatura de carbonização. Das amostras produzidas, nove foram devidamente estudadas. A espectroscopia de absorção no UV-Vis revelou perfis de absorção característicos para este tipo de NPs. A emissão de fluorescência das CNPs caracterizadada por espectroscopia de fluorescência evidenciou comportamentos emissivos típicos destas NPs tais como dependência do máximo de emissão com o comprimento de onda de excitação. A intensidade da fluorescência das CNPs sintetizadas por via hidrotérmica é, em geral, maior com rendimentos quânticos de fluorescência a variar entre 4 e 11%. Os rendimentos quânticos das CNPs produzidas por carbonização variam entre 2 e 5%. As imagens de microscopia electrónica demonstram que as CNPs possuíam formas esféricas. Os tamanhos determinados por SEM, TEM e DLS revelaram que as dimensões das NPs caem entre os 2 e 150nm. Por DRX constatou-se que as CNPs possuem uma estrutura atómica desorganizada. A análise FTIR mostrou que as amostras de CNPs produzidas a partir de macromoléculas pelo método hidrotérmico possuíam uma grande quantidade de precursor não degradado. Para as restantes CNPs foi verificada a presença de grupos funcionais polares que lhes conferem solubilidade em meio aquoso. Com 1H-RMN verificou-se uma diminuição de grupos alifáticos e aumento de grupos aromáticos nas CNPs de cortiça carbonizada, com o aumento da temperatura de carbonização. O potencial ζ da amostra obtida com maior temperatura de carbonização foi -25,7mV. Nos estudos in vitro realizados apenas as NPs produzidas a partir de ácido cítrico e etilenodiamina por via hidrotérmica marcaram eficazmente as linhas celulares de osteoblastos e de fibroblastos. A eficiência da marcação aparenta ser dependente do tempo de incubação com CNPs.

Influence of Lean and Green on supply chain performance: an interpretive structural modelling model

This work aims to identify and rank a set of Lean and Green practices and supply chain performance measures on which managers should focus to achieve competitiveness and improve the performance of automotive supply chains. The identification of the contextual relationships among the suggested practices and measures, was performed through literature review. Their ranking was done by interviews with professionals from the automotive industry and academics with wide knowledge on the subject. The methodology of interpretive structural modelling (ISM) is a useful methodology to identify inter relationships among Lean and Green practices and supply chain performance measures and to support the evaluation of automotive supply chain performance. Using the ISM methodology, the variables under study were clustered according to their driving power and dependence power. The ISM methodology was proposed to be used in this work. The model intends to provide a better understanding of the variables that have more influence (driving variables), the others and those which are most influenced (dependent variables) by others. The information provided by this model is strategic for managers who can use it to identify which variables they should focus on in order to have competitive supply chains.