In the first chapter is defined the fundamental concept of low cost airlines. Second and third section is dedicaded to history, creation and sequential developement of theese carriers and as well their characteristics. Following part is attended to airline traffic and its importance for tourism. In the last most extensive section, is analysed the influence of airline industry on tousim in the Czech Republic, at firts in general view of low cost carriers and tourism and further is this influence searched and analysed through three concrete airlines and four chosen regions.

The goal of my thesis is to analyze the current situation in the market of rail freight transport and to find the potential of the international railway transport of goods in the Czech Republic and Slovakia. It is focused on the comparison of these countries. Main topics of this thesis are i. a. the competitive environment in the Czech and Slovak railway, combined transport, system trains and the expected future progress.

The saliva of hematophagous ectoparasites contains a cocktail of vasodilators, anticoagulants and immunosuppressors that maintain blood in a liquid state at the site of the lesion and evade the host's defense mechanisms in suppressing the immune response. Since ticks have evolved to utilize mammals as a source of food, our understanding of the tick material, especially the salivary glands will enhance the control of tick infestation and allow the exploitation of the tick's natural resources. SGE protease activity was determined by measuring the degradation of azocasein. Proteolytic activity was found in the pH range of 3 to 11 with the highest activity at pH 9 followed by pH 7. At pH 3-5 the activity was mainly due to aspartic proteases, whereas at pH 7-9 the activity was due to the action of metallo- and serine proteases. At pH 11, the activity was mainly ascribed to metallo- and aspartic proteinase activity The fibrinogenolytic activity was determined by incubating human fibrinogen in the presence of SGE and monitoring the fibrinogen degradation by SDS-PAGE. SGE degraded the Au-chain of fibrinogen within 2 hours of incubation and even after 24 hours incubation there was no hydrolysis of the Bβ and γ-chains of fibrinogen. Characterization of the fibrinogenolytic activity revealed that metalloprotease activity was present over pH range of 3-9 and at pH 3-5, the cysteine proteases were active. No serine protease activity was found under similar experimental conditions. CE-HPLC separation of the SGE revealed three regions of proteolytic activity. Further characterization of the activity containing fractions using protease inhibitors at various pH values showed that the activity associated with region A is mainly due to the presence of aspartic and cysteine proteases in the lower pH range (< 5). Region B was mainly due to the activity of the metallo- and serine proteases, while the activity in region C was mainly due to the metalloproteinases which were more active in the higher pH range (> 9). CE-HPLC separation of SGE resulted in three regions exhibiting fibrinogenolytic activity at pH 7-9. In region A all four enzyme classes were found while in regions B and C, serine, cysteine and metalloproteinases were found to be responsible for the activity. Region A was further purified on the HIC-column and activity eluted in several peaks which after individual application on SE-HPLC column had similar retention times. The pooled samples were analyzed for purity using C5 RP-HPLC and reducing tricine SDS-PAGE and three bands of relative molecular masses 15, 22 and 12 kDa, respectively were found. In an attempt to purify the proteins in region C, four individual CE-HPLC runs were combined and applied to a fibrinogen affinity column. Reducing SDS-PAGE analysis of bound material showed two bands of relative molecular masses of 31 and 39 kDa, respectively. CE-HPLC region C as well as the SGE control was found to disaggregate platelets aggregated by ADP, epinephrine, collagen as well as TRAP. No disaggregation was observed for the saline negative control. The disaggregation is most probably due to the hydrolysis of the fibrinogen cross-linking platelets by the metalloproteinase activity in region C. Understanding of the proteolytic activities present in the salivary gland and therefore identifying molecules crucial for tick feeding could aid in the development of experimental vaccines. Even though the fibrinogenolytic activity was not purified to homogeneity, this study has laid the groundwork for further experiments in this field. / Dissertation (MSc (Biochemistry))--University of Pretoria, 2006. / Biochemistry / unrestricted

Orientadores: João Vasconcellos Neto, Mário Eidi Sato / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-27T17:36:11Z (GMT). No. of bitstreams: 1 Mendonca_MarcioJoseCardoso_D.pdf: 1359711 bytes, checksum: 8fefaf30eddb842d45e30548a83edae1 (MD5) Previous issue date: 2015 / Resumo: Dinâmica populacional, preferência hospedeira e sensibilidade a agrotóxicos de Brevipalpus phoenicis (GEIJSKES 1939) e ácaros predadores em cafeeiro (Coffea spp.) Dentre as pragas que atacam o cafeeiro, destaca-se o ácaro Brevipalpus phoenicis (Geijskes, 1939) (Acari: Tenuipalpidae), que é o vetor do vírus da mancha-anular do cafeeiro. O estudo foi conduzido com o objetivo de se obter subsídios para o manejo da praga em cafeeiro no Brasil. Os objetivos específicos foram: 1) estudar a dinâmica populacional de B. phoenicis e das principais espécies de ácaros predadores (Phytoseiidae), incluindo análises sobre interação entre B. phoenicis e predadores, bem como a influência de fatores climáticos na população desses ácaros, em diferentes cultivares de café; 2) avaliar a sensibilidade a agrotóxicos em populações do ácaro-praga e ácaros predadores da espécie Euseius concordis (Chant, 1959), procedentes de diferentes regiões do estado de São Paulo. Um estudo sobre dinâmica populacional de ácaros foi conduzido em cafeeiros de diferentes cultivares (`Icatu Vermelho¿, `Catuaí Vermelho¿, `Mundo Novo¿, `Obatã¿, `Apoatã¿ (Robsuta), em Presidente Prudente, SP, onde foram realizadas coletas mensais de amostras de folhas de café, por um período de 19 meses. As espécies mais abundantes de ácaros em cafeeiro foram B. phoenicis, E. citrifolius Denmark & Muma e E. concordis. A espécie predominante de ácaro predador foi E. citrifolius, correspondendo a 92,4% dos fitoseídeos encontrados. Foram detectadas correlações significativas entre E. citrifolius e B. phoenicis em todos os cultivares de cafeeiro, sugerindo que esse ácaro fitoseídeo pode exercer controle biológico sobre o ácaro praga em cafeeiro. As maiores abundâncias do ácaro predador foram observadas para os cultivares `Apoatã¿ e `Catuaí Vermelho¿. Para o estudo de sensibilidade de ácaros a agrotóxicos, foram conduzidos dois experimentos em condições de laboratório. O experimento 1 foi sobre a influência de agrotóxicos sobre a sobrevivência de fêmeas adultas de B. phoenicis e E. concordis e sobre as taxas de crescimento populacional desses ácaros. Ciflumetofem, hexitiazox e fenpropatrina foram efetivos para o controle de B. phoenicis, porém, inócuos ao E. concordis, demonstrando bom potencial para uso em cafeeiro, visando ao manejo do ácaro-praga. Abamectina, cipermetrina+profenofós, deltametrina+triazofós e etoxazol foram altamente tóxicos para B. phoenicis e E. concordis. Oxicloreto de cobre afetou apenas o crescimento populacional de B. phoenicis, sem causar efeito significativo em E. concordis. Espirodiclofeno e malationa foram mais prejudiciais ao crescimento populacional de B. phoencis e E. concordis. O experimento 2 foi sobre toxicidade de agrotóxicos em populações de B. phoenicis e E. concordis de diferentes regiões do estado de São Paulo. Foram comparadas populações de B. phoenicis coletadas de cafeeiro nas regiões de Franca e Vera Cruz, e de E. concordis, originárias de cafeeiro de Franca, Marília e Vera Cruz. A população de B. phoenicis da região de Franca mostrou-se mais resistente a etoxazol a de Vera Cruz. A população de E. concordis de Franca mostrou-se mais resistente a abamectina e fenpropatrina, porém, mais suscetível a ciflumetofem, que Vera Cruz. Os ácaros E. concordis apresentaram maior tolerância aos agrotóxicos que os da espécie B. phoenicis. Estudos dessa natureza são de grande relevância para o estabelecimento de programas de manejo integrado de ácaros-praga em cafeeiros no Brasil / Abstract: Population dynamics, host preference and agrochemicals susceptibility of Brevipalpus phoenicis (Geijskes, 1939) and predaceous mites on coffee (Coffea spp.) Among the pests that attack coffee, it stands out the mite Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae), stands out the vector of the coffee ringspot virus (CoRSV). This study was conducted in order to obtain basic information that management keeping of this pest coffee in Brazil.The objectives were: 1) to study the population dynamics of B. phoenicis and of the main species of predatory mites (Phytoseiidae), including analyzes of interactions between B. phoenicis and predators, as well as the influence of climatic factors on the population of these mites on different coffee cultivars; 2) to evaluate the pesticide susceptibility in populations of this mite and the predaceous mite Euseius concordis (Chant, 1959), from different regions of São Paulo state. A study on population dynamics of mites was conducted on different coffee cultivars (`Icatu Vermelho¿, `Catuaí Vermelho¿, `Mundo Novo¿, `Obatã¿, `Apoatã¿) in Presidente Prudente-SP, where samples of coffee leaves were taken monthly for a period of 19 months. The most abundant mite species on coffee were B. phoenicis, Euseius citrifolius Denmark & Muma and E. concordis. The predominant species of predatory mites was E. citrifolius, corresponding to 92.4% of the phytoseiids found on coffee plants. Significant correlations between E. citrifolius and B. phoenicis were detected in all the coffee cultivars studied, indicating that this phytoseiid mite exerts biological control of this coffee pest. The highest abundances of predaceous mites were observed for `Apoatã¿ and `Catuaí Vermelho¿ cultivars. To study the susceptibility of mites to agrochemicals, two experiments were conducted under laboratory conditions. Experiment 1 was about the influence of agrochemicals on adult female survival of B. phoenicis and E. concordis and on the population growth rates of the mites. Cyflumetofen, hexythiazox and fenpropathrin were effective for the control of B. phoenicis but harmless to the predator E. concordis, showing good potential for the use in coffee mite management program. Abamectin, cypermethrin + profenofos, deltamethrin + triazophos and etoxazole were highly toxic to B. phoenicis and E. concordis. Copper oxychloride only affected the population growth of B. phoenicis, without causing significant effects on E. concordis. Spirodiclofen and malathion were more harmful to population growth of B. phoencis than E. concordis. Experiment 2 was on agrochemical toxicity in populations of B. phoenicis and E. concordis from different regions of São Paulo state. Populations of B. phoenicis collected from coffee fields in the regions of Franca and Vera Cruz, were compared to populations of E. concordis, collected from coffee in Franca, Marilia and Vera Cruz. The population of B. phoenicis from Franca was more resistant to etoxazol than that from Vera Cruz. The population of E. concordis of Franca was more resistant to abamectin and fenpropathrin, but more susceptible to cyflumetofen, than that from Vera Cruz. Euseius concordis showed higher tolerance to acaricides than B. phoenicis. These kind of studies are greatest importance to establishment of integrated management programs for coffee pest mites in coffee in Brazil / Doutorado / Biodiversidade Animal / Doutor em Biologia Animal

Micro-isolation of PCR-ready from fresh and dried legumes seeds and food products containing legumes (hummus) was tested. For optimization process magnetic microparticles PGMAox was used. Optimum weight plants and food material was 200 mg. For isolation of DNA from fresh legumes, mixture containing 500 L of CTAB extraction buffer, 1 L -mercaptoethanol and 500 L chloroform-octanol was used. For isolation of DNA from dried legumes seeds and food products volume of CTAB extraction buffer was increased to 1 000 L. To achieve higher purity of DNA some prepared homogenates was precipitate by isopropylalcohol. The optimized process of DNA isolation was used to prepare a homogenate from food products which DNA was isolated by different types of magnetic carriers. For comparison, non-porous magnetic carriers poly(glycidyl methacrylate) PGMAox, poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) P(HEMA-co-GMA)ox covered by carboxyl groups and porous fully crosslinked microparticles poly(styrene-co-divinylbenzene) HPS-B-M-22-NH2, magnetic porous glass MPG and nanoparticles of iron oxides covered by poly(L-lysine) PLL were used. In average the highest concentration and the best purity of DNA was isolated by magnetic carriers P(HEMA-co-GMA)ox a PGMAox.

Acid-sensitive liposome has drawn much interest as drug and gene carriers that release payloads specifically at the low-pH target sites, such as in solid tumors, tissues with inflammation, and ischemia sites. Also, it helps drug/gene to escape endosome trapping and followed lysosome degradation. The goal of this thesis research is to develop novel trans-2-aminocyclohexanols based lipids and their liposome that can be switched by mildly acidic pH. NMR study · show that in certain acidic medium, the amine group on cyclohexane will attract proton and form hydrogen bond with the neighboring -OH. This change will force the bonds switch to from equatorial conformation to axial conformation. This conformational change is transmitted by the structure of the molecular, and induces consequently dramatic conformational change of the two long lipid tails. Fluorescence leakage assay was conducted on liposomes that encapsulated with ANTs/DPX fluorescence dyes. For certain special designed cyclohexane compounds, the pH triggered lipid conformation change will rupture liposome membrane, release the encapsulated content, and thus help them escape lysosome degradation. This would in tum improve the efficiency of liposome drug delivery and gene transfection. Luciferase gene transfection was conducted on B16F10 cultured cells. The lipoplex comprising trans-2-aminocyclohexanollipid 1 significantly enhanced the Luciferase gene expression. The gene transfection efficiency correlated well with the pH-triggered membrane-rupture in the trans-aminocyclohexanol-based lipoplexes.

The aim of the work was the selective isolation of bacterial DNA with help of magnetic carriers covered by streptavidine (PGMA-NH2-STV, MPG® Streptavidin). Conditions of functionalisation of carriers using two biotinylated probes were optimized: the amount of carrier, the amount of probe, binding of biotinyled probe to streptavidine. Purified DNA Lactobacillus was used for hybridization. DNA binding to the probe (DNA/DNA hybridization) and nospecific adsorption of DNA to the carrier were tested. Target DNA eluted from the carrier was identified using PCR with primers R16-1 and LbLMA1-rev and with primers P_eub and F_eub. The amount of probe bound to the carrier was estimated using UV spectrophotometry. It was estimated that biotinyled probe can be used for functionalisation in concentration 5 pmol/µl added to the carrier in the ration carrier : probe 1:1. It was shown that nonspecific DNA adsorption to the MPG® Streptavidin is significantly lower than to the carrier PGMA-NH2-STV.Using DNA/DNA hybridization and the MPG® Streptavidin, DNA from pure culture Lactobacillus was isolated. Procedure was applicated for DNA isolation from milk products.

Orientador: Renato Grillo / Resumo: Inúmeros sistemas nanoestruturados vêm sendo desenvolvidos para o transporte e liberação modificada de defensivos agrícolas nos últimos anos. Tais sistemas vêm se mostrando uma potencial ferramenta para reduzir a toxicidade destes ingredientes ativos no ambiente, bem como vêm melhorando a eficiência de pesticidas no campo. Entretanto pouco se conhece sobre o destino, toxicidade e mecanismos de ação destes nanocarreadores no ambiente. Neste sentido, o objetivo do presente trabalho foi desenvolver potenciais sistemas de liberação modificada, encapsulando o herbicida atrazina (ATZ) em carreadores lipídicos nanoestruturados (CLNs) sem e contendo nanopartículas inorgânicas (magnéticas (Fe3O4@AO) e plasmônicas (Au@CTAB)), a fim de construir um sistema de liberação marcado que possa ser rastreado, auxiliando assim em futuras análises destes nanocarreadores no ambiente. Para o desenvolvimento das nanoformulações a técnica de emulsificação/evaporação solvente foi aplicada e diversos métodos físico-químicos foram empregados para a sua caracterização. As nanoformulações permaneceram estáveis por mais de 60 dias à temperatura ambiente. Além disso, os CLNs demostraram ter índice de polidispersão (PDI) < 0,2, potencial zeta entre -5,38 ± 0,27 a -23,5 ± 1,25 mV e uma eficiência de encapsulação maior que 90 % para a ATZ. As análises de microscopia eletrônica de varredura (MEV) revelaram ausência de agregados e morfologia esférica ao sistema. Além disso, as técnicas de espectroscopia de Infra... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Numerous nanostructured systems have been developed for the transportation and modified release of crop protection products in recent years. Such systems have been shown to be a potential tool to reduce the toxicity of these active ingredients in the environment, as well as improving the efficiency of pesticides in the field. However, little is known about the fate, toxicity and mechanisms of action of these nanocarriers in the environment. In this sense, the objective of the present work was to develop potential modified release systems, encapsulating the herbicide atrazine (ATZ) in nanostructured lipid carriers (CLNs) without and containing inorganic (magnetic (Fe3O4 @ AO) and plasma (Au @ CTAB)) nanoparticles) , in order to build a marked release system that can be tracked, thus assisting in future analyzes of these nanocarriers in the environment. For the development of nanoformulations, the solvent emulsification / evaporation technique was applied and several physical-chemical methods were used for its characterization. The nanoformulations remained stable for more than 60 days at room temperature. In addition, CLNs have been shown to have a polydispersity index (PDI) <0.2, zeta potential between -5.38 ± 0.27 to -23.5 ± 1.25 mV and an encapsulation efficiency greater than 90% for the ATZ. Scanning electron microscopy (SEM) analyzes revealed the absence of aggregates and spherical morphology to the system. In addition, Infrared spectroscopy (FTIR) and X-ray (DRX) techniq... (Complete abstract click electronic access below) / Mestre

in English Plants with their sessile life-style are exposed to many stimuli from environment. They have developed mechanisms how to coordinate their growth and development, which allows them to survive sometimes in very difficult conditions. Plant hormones are one of the most important regulators of this signal transduction. Auxins, as the oldest known group of plant hormones, play important role in many physiological processes in plants. To allow perceiving the information in every single cell, auxin molecule is transported by cell-to-cell manner. Auxin molecules enter the cell by passive diffusion or by active uptake by auxin influx carriers. To reach the next cell, they are transported actively out of the cell by auxin efflux carriers. Athough active auxin transport has been studied for almost four decades, past two decades contributed to the identification and characterization of particular auxin carriers remarkably. This thesis contributes to the knowledge on the auxin efflux and influx carriers and their involvement in the processes occurring from a single cell level to the level of a whole plant. Firstly, it brings detailed description of auxin transport characteristics in Arabidopsis thaliana cell suspension cells (Ath cells). Secondly, it shows that the overproduction of PIN1 auxin efflux...

The non-uniform distribution of the plant growth regulator (phytohormone) auxin is known to mediate many fundamental processes in plant development. Auxin is transported through the plant body either via vascular pathways or from cell to cell by specialized polar auxin transport machinery. This machinery consists of a balanced system of passive diffusion combined with the activities of auxin influx and efflux carriers. This work is focused on the processes that are involved in the uptake of auxin into plant cells. On the basis of molecular-biological and biochemical characterization, the function as an auxin influx carrier was confirmed for PaLAX1 protein from wild cherry (Prunus avium). The sequences of isolated cDNA of the PaLAX1 gene and of its protein product are highly similar to both the cDNAs and the corresponding protein products of the AUX1/LAX-type genes, coding for putative auxin influx carriers in model plant A. thaliana. On the level of organs and single cells, we have shown that the overproduction of PaLAX1 in transgenic lines resulted in an increase of the content of native auxin indole-3-acetic acid as well as of the uptake of synthetic auxin, 2,4-dichlorophenoxyacetic acid. Further, the mechanism of action of putative auxin influx inhibitors 1-naphthoxyacetic acid (1-NOA),...