SammanfattningUppsatsen är utformad efter ett förslag från Anders Ryberg vid Länsstyrelsen i Västra Götalands län. Förslaget var att lokalisera användningsområden för den ull som i dagsläget kasseras i Sverige. I början beskriver vi grundläggande fakta om ullfibern för att ge dig som läsare ett bättre begrepp om ullfiberns användningsområden som senare tas upp i arbetet. Detta följs av en beskrivning av de miljöaspekter som ullanvändningen medför, vilket dessutom är viktigt ur en marknadsföringssynpunkt. De ullkvalitéer vi har i Sverige beskrivs därefter kortfattat, följt av en presentation av svenska företag och projekt som arbetar med svensk ull. Därefter beskrivs Australiens, Storbritanniens och Norges ullindustrier. Deras marknadsfringsstrategier och infrastrukturer för ullhantering är viktiga att ta del av om den svenska ullindustrin ska kunna konkurrera i framtiden. Vi har även funnit många intressanta användningsområden för den svenska ullen. Däribland finner vi geotextil, isolering och ull i avfallshantering enklast att producera utifrån de förutsättningar vi har i Sverige. Våra slutsatser är att det krävs en hel del initiativ, framförallt från fårägarna i Sverige, för att skapa en större efterfrågan på svensk ull. Tillvägagångssättet är att skapa infrastruktur för uppsamling och sortering av ullen runtom i landet, sedan marknadsföra ullen med hjälp av information om dess unika egenskaper och positiva miljöaspekter. / <p>AbstractThe thesis is developed from a proposition by Anders Ryberg at the county administration board of Västra Götaland. The proposition was to evaluate new fields of utility for the Swedish wool which today is disregarded. First off we give a basic description of the facts concerning the wool fiber in order for you as a reader to better comprehend how the wool fiber applies in different fields of utility, which is brought up later in the thesis. This is followed by a description of the environmental aspects which comes with the usage of wool, which also is important from a marketing point of view. The different types of wool we have in Sweden are then briefly described, followed by a presentation of Swedish companies and projects who are working with Swedish wool. Thereafter we describe the wool industries of Australia, Great Britain and Norway. Their marketing strategies and infrastructures for wool handling are important to acknowledge if the Swedish wool industry are to be able to compete in the future. We have also found many interesting fields of utility for the Swedish wool. Among many we have found geotextile, isolation and wool in waste management the easiest ones to produce looking at the prerequisites we have in Sweden. Our conclusions are that a lot of initiative will be needed, especially from the Swedish sheep owners, to be able to create a larger demand for Swedish wool. The way to go about it is by creating an infrastructure for collecting and sorting of the wool around the country, then implicate marketing with information about the unique abilities of wool and its positive environmental aspects.</p><p>Program: Textil produktutveckling med entreprenörs- och affärsinriktning</p>

by Yam Hin Fai. / "June 1998." / Thesis (Ph.D.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (p. 118-151). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstract in Chinese.

O hidrogênio (H2) tem sido considerado uma fonte de energia limpa bastante promissora, pois sua combustão origina apenas moléculas de água, sendo uma alternativa ao uso de combustíveis fósseis. Entretanto, os métodos atuais de produção de H2 demandam matérias-primas finitas e uma grande quantidade de energia, tornando a sua obtenção não sustentável. Mais recentemente, a via fermentativa tem sido considerada para a produção de H2, utilizando como matérias-primas efluentes industriais, materiais lignocelulósicos e biomassa de algas, denominado de bio-hidrogênio de primeira, segunda e terceira geração, respectivamente. Neste trabalho foi isolada uma bactéria anaeróbia a partir de uma cultura mista (lodo) de um sistema de tratamento de vinhaça, após pré-tratamento do lodo a pH 3 por 12 horas. Este microrganismo foi identificado com 99% de similaridade como Clostridium beijerinckii com base na sequência do gene RNAr 16S denominado de C. beijerinckii Br21. A temperatura e o pH mais adequados para o crescimento e produção de H2 por esta cultura foi 35 °C e pH inicial 7,0. A bactéria possui a capacidade de utilizar ampla variedade de fontes de carbono para a produção de H2 por fermentação, especialmente, monossacarídeos resultantes da hidrólise de biomassa de algas, tais como glicose, galactose e manose. Foram realizados ensaios em batelada para a produção de H2 com a bactéria isolada empregando diferentes concentrações de glicose e galactose, visando a sua futura utilização em hidrolisados de alga. Os parâmetros cinéticos dos ensaios de fermentação estimados pelo modelo de Gompertz modificado, como a velocidade máxima de produção (Rm), a quantidade máxima de hidrogênio produzido (Hmáx) e o tempo necessário para o início da produção de hidrogênio (fase lag) para a glicose (15 g/L) foram de: 58,27 mL de H2/h, 57,68 mmol de H2 e 8,29 h, respectivamente. Para a galactose (15 g/L), a Rm, Hmáx e foram de 67,64 mL de H2/h, 47,61 mmol de H2 e 17,22 horas, respectivamente. O principal metabólito detectado ao final dos ensaios de fermentação, foi o ácido butírico, seguido pelo ácido acético e o etanol, tanto para os ensaios com glicose, como com galactose. C. beijerinckii é um candidato bastante promissor para a produção de H2 por fermentação a partir de glicose e galactose e, consequentemente, a partir de biomassa de algas como substratos. / Hydrogen (H2), considered an alternative to fossil fuels, is a promising source of clean energy because its combustion originates water molecules only. However, the current H2 production methods require finite raw materials and a large amount of energy, which makes them unsustainable. The fermentative pathway has been considered for H2 production from renewable raw materials such as industrial wastewater, lignocellulosic materials, and algal biomass, the so-called first, second, and third bio-hydrogen generation, respectively. In this work, after pre-treatment at pH 3 for 12 h, a H2-producing bacterium was isolated from a mixed culture (sludge) collected from an anaerobic bioreactor used to treat sugarcane vinasse. The microorganism was identified as Clostridium beijerinckii based on the sequence of the 16S rRNA gene; it was named C. beijerinckii Br21. The most appropriate temperature and initial pH to achieve H2 production by this strain was 35 °C and 7, respectively. The bacterium was able to use a wide variety of carbon sources, especially the monosaccharides glucose, galactose, and mannose resulting from hydrolysis of algal biomass. Batch assays using different concentrations of glucose and galactose were performed to produce H2. The kinetic parameters of the tests were estimated by the Gompertz modified model. The maximum production rate (Rm), the maximum amount of produced H2 (Hmáx), and the phase lag () for glucose and galactose, both at 15 g/L, were 58.27 and 67.64 mL of H2/h, 57.68 and 47.61 mmol of H2, and 8.29 and 17.22 h, respectively. The main metabolite detected at the end of fermentation tests was butyric acid, followed by acetic acid and ethanol. The results indicated that the new C. beijerinckii isolate is a promising candidate for fermentative H2 production from algal biomass.

One of the largest challenges humanity faces today is reducing CO₂-emissions to mitigate climate change. Part of the solution might be to increase the use of wood products. To do this, the efficiency of forestry has to be improved. In Sweden, a large obstacle for improving the efficiency of forestry is moose, or rather the damages its browsing causes on Scots pine. The aim of this study was to investigate how stand size and stand isolation affects the intensity of moose browsing damage on Scots pine. In addition, effects from tree density, stand age, and interactions between the different factors were investigated. This was done by using data from forest companies and field data collected from 29 Scots pine stands in central Sweden. The data were analysed in a multiple regression analysis, selecting the model that best explained variation in browsing damage. Results show that browsing damage was lower in more isolated stands. Stand size, however, did not have an effect on browsing damage. The factors stand age and Scots pine density had positive effects on the amount of browsing damage, but effects from stand age decreased with increasing pine density. In total, 77% of the variance in browsing damage was explained by the best model. In order to decrease browsing damage on Scots pine, I suggest that land owners take coordinated action to avoid creating large areas of young pine stands within the same area.

Introdução: o aumento gradativo da resistência dos microrganismos aos antimicrobianos usados na prática clínica tem contribuído efetivamente para que as infecções hospitalares sejam consideradas um problema de saúde pública. Assim, diminuir a disseminação destes microrganismos no ambiente hospitalar tem se tornado um desafio aos profissionais que atuam nos serviços de controle de infecção hospitalar. Objetivo: identificar a sensibilidade e especificidade dos critérios para isolamento de pacientes admitidos num hospital especializado em oncologia. Material e Método: trata-se de um estudo de corte transversal de caráter prospectivo, aprovado pelo Comitê de Ética em Pesquisa de um hospital especializado em oncologia. A população do estudo foi composta por 61 pacientes admitidos no hospital, no período de 01 março a 31 de agosto de 2009, segundo o protocolo para isolamento de pacientes procedentes de outros hospitais. Os dados foram obtidos por meio de entrevista individual e consulta aos resultados de cultura provenientes de swab nasal e anal, coletados pela equipe de enfermagem no momento da admissão no hospital e disponibilizados no laboratório de microbiologia. Resultado: dos 61 pacientes admitidos no hospital, 56 preencheram os critérios para que as precauções de contato fossem instituídas. Destaca-se que 30(49,2%) pacientes tiveram culturas positivas para microrganismos multirresistentes, sendo o Staphylococcus aureus resistente à oxacilina o microrganismo mais freqüentemente isolado no swab nasal e anal. A sensibilidade dos critérios para isolamento de pacientes admitidos no referido hospital e procedentes de outros hospitais foi de 90%. Conclusão: foram altamente sensíveis os critérios utilizados para o isolamento de pacientes provenientes de outros hospitais, portanto a maioria dos pacientes colonizados por microrganismos multirresistentes foi isolada no momento da admissão. / Introduction: the gradual increase of microorganisms\' resistance to antimicrobials used in clinical practice has effectively contributed to hospital-acquired infections be considered a public health problem. Hence, reducing the dissemination of these microorganisms in the hospital setting has become a challenge for professionals working in hospital-acquired infection control services. Objective: to identify the sensitivity and specificity of criteria for isolation of patients admitted to a cancer specialized hospital. Material and method: this prospective cross-sectional study was approved by the Research Ethics Committee of a cancer specialized hospital. The study\'s population was composed of 61 patients, who were admitted to the hospital between March 1 and August 31 2009 according to the protocol for isolation of patients from other hospitals. Data were collected through individual interview and consultation of the results from nasal and anal swabs culture collected by the nursing team at the moment patients were admitted to the hospital and available in the microbiology laboratory. Results: 56 out of the 61 patients met the criteria establishing contact precautions. It is worthy noting that 30(49,2%) patients presented positive cultures for multi-resistant microorganisms, while the Staphylococcus aureus resistant to oxacillin was the microorganism most frequently isolated in nasal and anal swabs. The sensitivity of the criteria for isolation of patients from other hospitals admitted in the studied hospital was of 90%. Conclusions: criteria used for isolation of patients from other hospitals were highly sensitive, thereby the majority of patients colonized by multi-resistant microorganisms were isolated at the moment of admission.

Qi Wei. / "November 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (p. 158-185). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

by Mock Wai Yin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 112-122). / ACKNOWLEDGEMENTS --- p.I / ABSTRACT --- p.II / TABLE OF CONTENT --- p.IV / Chapter CHAPTER 1: --- INTRODUCTION --- p.1 / Chapter CHAPTER 2: --- PURIFICATION OF α- AND β-MMCs --- p.29 / Chapter CHAPTER 3: --- RIBONUCLEASE ACTIVITY OF MMCs --- p.50 / Chapter CHAPTER 4: --- PURIFICATION AND RIBONUCLEASE ACTIVITY OF RNase-MC --- p.85 / Chapter CHAPTER 5: --- CONCLUSION --- p.109 / REFERENCES --- p.112

Lam Sze Kwan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 172-200). / Abstracts in English and Chinese. / Acknowledgements --- p.ii / Table of contents --- p.ii / Abstract --- p.xii / 撮要 --- p.xv / List of Abbreviations --- p.xvi / List of Tables --- p.xvii / List of Figures --- p.xix / TABLE OF CONTENTS / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1. --- Overview of Chitinases --- p.3 / Chapter 1.1.1. --- Classification of Chitinases --- p.7 / Chapter 1.1.1.1. --- Family 19 Chitinases --- p.7 / Chapter 1.1.1.1.1. --- Class I Chitinases --- p.9 / Chapter 1.1.1.1.2. --- Class II Chitinases --- p.10 / Chapter 1.1.1.1.3. --- Class IV Chitinases --- p.10 / Chapter 1.1.1.1.4. --- Class V Chitinases --- p.11 / Chapter 1.1.1.1.5. --- Class VI Chitinases --- p.11 / Chapter 1.1.1.2. --- Family 18 Chitinases --- p.12 / Chapter 1.1.1.2.1. --- PR-8/Class III Chitinases --- p.12 / Chapter 1.1.1.2.2. --- PR-11 Chitinases --- p.15 / Chapter 1.1.1.3. --- The PR-4 Family --- p.16 / Chapter 1.1.2. --- Catalytic Mechanism of Chitinases --- p.19 / Chapter 1.1.2.1. --- Catalytic Mechanism of Family 18 Chitinases --- p.20 / Chapter 1.1.2.2. --- Catalytic Mechanism of Family 19 Chitinases --- p.21 / Chapter 1.1.3. --- Biological Properties of Chitinases --- p.22 / Chapter 1.1.3.1. --- Antifungal Activity of Chitinases in vitro --- p.22 / Chapter 1.1.3.2. --- Antifungal Activity of Chitinases in vivo --- p.23 / Chapter 1.1.3.3. --- Other Functions --- p.23 / Chapter 1.2. --- Overview of Ribonucleases --- p.25 / Chapter 1.2.1. --- Classification of Ribonucleases --- p.26 / Chapter 1.2.1.1. --- RNase T1 Family --- p.26 / Chapter 1.2.1.1.1. --- Action Mechanism of RNase T1 Family --- p.32 / Chapter 1.2.1.2. --- RNase T2 Family --- p.34 / Chapter 1.2.1.2.1. --- Action Mechanism of RNase T2 Family --- p.36 / Chapter 1.2.2. --- Biological Activities of Plant Ribonucleases --- p.38 / Chapter 1.2.2.1. --- Phosphate Remobilization --- p.38 / Chapter 1.2.2.2. --- Senescence --- p.39 / Chapter 1.2.2.3. --- Programmed Cell Death --- p.40 / Chapter 1.2.2.4. --- Plant Defense --- p.41 / Chapter 1.2.2.5. --- RNA Processing and Decay --- p.43 / Chapter 1.2.2.6. --- Antitumor Activities --- p.43 / Chapter 1.3. --- Overview of plant ribosome-inactivating proteins (RIPs) --- p.45 / Chapter 1.3.1. --- General properties of RIPs --- p.46 / Chapter 1.3.1.1. --- Classification of RIPs --- p.46 / Chapter 1.3.2. --- Activities of Ribosome-inactivating Proteins --- p.52 / Chapter 1.3.2.1. --- RNA N-glycosidase activity --- p.52 / Chapter 1.3.2.2. --- Protein synthesis inhibitory activity --- p.58 / Chapter 1.3.2.3. --- Abortifacient activity --- p.59 / Chapter 1.3.2.4. --- Immunosuppressive activity --- p.60 / Chapter 1.3.2.5. --- Antiviral activity --- p.61 / Chapter 1.3.3. --- Roles of RIPs in plants --- p.63 / Chapter 1.3.3.1. --- Defensive role of RIPs in plants --- p.63 / Chapter 1.3.3.2. --- Role of RIPs in stress adaptation in plants --- p.66 / Chapter 1.3.4. --- Possible application of RIPs --- p.67 / Chapter 1.3.4.1. --- Use of RIPs in therapies --- p.67 / Chapter 1.3.4.1.1. --- Antiviral agents --- p.67 / Chapter 1.3.4.1.2. --- Immunotoxins --- p.68 / Chapter 1.3.4.1.3. --- Anti-HIV drugs --- p.69 / Chapter 1.3.4.2. --- Use of RIPs in agriculture --- p.71 / Chapter 1.4. --- Overview of the PR-5 Family: Thaumatin-Like Proteins (TLPs) --- p.72 / Chapter 1.4.1. --- Occurrence of Thaumatin-Like Proteins --- p.76 / Chapter 1.4.2. --- Biological properties of TLPs --- p.77 / Chapter 1.4.2.1. --- Antifungal Activity --- p.77 / Chapter 1.4.2.2. --- TLPs as Anti-Freeze Protein --- p.78 / Chapter 1.4.3. --- Biotechnological Application ´ؤ Transgenic Plants --- p.79 / Chapter Chapter 2 --- Materials and Methods --- p.81 / Chapter 2.1. --- Materials --- p.81 / Chapter 2.2. --- Preparation of Crude Extract --- p.82 / Chapter 2.3. --- Purification --- p.83 / Chapter 2.4. --- Chromatography --- p.84 / Chapter 2.4.1. --- CM-Cellulose Chromatography --- p.84 / Chapter 2.4.2. --- Mono S® HR 5/5 and Mono Q® HR 5/5 --- p.85 / Chapter 2.4.3. --- Affi-gel Blue gel --- p.86 / Chapter 2.4.4. --- Superdex75 --- p.87 / Chapter 2.5. --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.88 / Chapter 2.6. --- Protein Concentration Determination --- p.89 / Chapter 2.7. --- Preparation of Rabbit Reticulocyte Lysate --- p.90 / Chapter 2.8. --- Determination of N-terminal Amino Acid Sequence --- p.91 / Chapter 2.9. --- Biological Activity Assays --- p.92 / Chapter 2.9.1. --- Assay for Antifungal Activity --- p.92 / Chapter 2.9.2. --- Assay for Cell-Free Translation Inhibitory Activity --- p.93 / Chapter 2.9.3. --- Assay of Cytotoxic Activity on Cancer Cell Lines --- p.94 / Chapter 2.9.4. --- Assay for HIV-1 Reverse Transcriptase (RT) Inhibitory Activity --- p.95 / Chapter 2.9.5. --- Assay of Mitogenic Activity --- p.97 / Chapter 2.9.6. --- Assay for N-Glycosidase Activity --- p.98 / Chapter 2.9.6.1. --- RNA Extraction --- p.98 / Chapter 2.9.6.2. --- Aniline Treatment --- p.99 / Chapter 2.9.6.3. --- Formaldehyde Gel Electrophoresis --- p.99 / Chapter 2.9.7. --- Assay of Ribonuclease Activity --- p.100 / Chapter 2.9.7.1. --- Assay for Yeast tRNA --- p.100 / Chapter 2.9.7.2. --- Activity toward Polyhomoribonucleotides --- p.100 / Chapter Chapter 3 --- Purification and Characterization of Pathogenesis-Related Proteins from their Respective Sources --- p.101 / Chapter 3.1. --- Purification and Characterization of Chitinase and Ribonuclease from the Roots of Panax notoginseng --- p.102 / Chapter 3.1.1. --- Introduction --- p.102 / Chapter 3.1.2. --- Results --- p.104 / Chapter 3.1.3. --- Purification --- p.107 / Chapter 3.1.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.108 / Chapter 3.1.3.2. --- Affinity Chromatography on Affi-gel Blue gel --- p.111 / Chapter 3.1.3.3. --- Cation-Exchange Chromatography on Mono S Column --- p.114 / Chapter 3.1.3.4. --- Gel Filtration on Superdex 75 Column --- p.115 / Chapter 3.1.4. --- Characterization of Chitinase --- p.117 / Chapter 3.1.4.1. --- N-terminal Amino Acid Sequence --- p.117 / Chapter 3.1.4.2. --- Assay for Antifungal Activity --- p.118 / Chapter 3.1.4.3. --- Assay for Cell-Free Translation-inhibitory Activity --- p.120 / Chapter 3.1.4.4. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.120 / Chapter 3.1.5. --- Characterization of Ribonuclease --- p.121 / Chapter 3.1.5.1. --- N-terminal Amino Acid Sequence --- p.121 / Chapter 3.1.5.2. --- Assay for Ribonuclease Activity --- p.122 / Chapter 3.1.5.3. --- Assay for Cell-Free Translation-inhibitory Activity --- p.125 / Chapter 3.1.5.4. --- Assay for Antifungal Activity --- p.125 / Chapter 3.1.5.5. --- Assay for Antiproliferative Activity --- p.126 / Chapter 3.1.6. --- Discussion --- p.127 / Chapter 3.2. --- Purification and Characterization of Ribosome-Inactivating Protein and Antifungal Protein from the mushroom Lyophyllum shimeji --- p.131 / Chapter 3.2.1. --- Introduction --- p.131 / Chapter 3.2.2. --- Results --- p.132 / Chapter 3.2.3. --- Purification --- p.134 / Chapter 3.2.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.135 / Chapter 3.2.3.2. --- Affinity Chromatography on Affi-gel Blue Gel --- p.137 / Chapter 3.2.3.3. --- Cation-Exchange Chromatography on Mono S --- p.140 / Chapter 3.2.4. --- Characterization of Ribosome-Inactivating Protein and Antifungal Protein from Lyophyllum shimeji --- p.142 / Chapter 3.2.4.1. --- N-terminal Amino Acid Sequence --- p.142 / Chapter 3.2.4.2. --- Assay for Antifungal Activity --- p.144 / Chapter 3.2.4.3. --- Assay for N-glycosidase Activity --- p.147 / Chapter 3.2.4.4. --- Assay for Mitogenic Activity --- p.147 / Chapter 3.2.4.5. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.148 / Chapter 3.2.5. --- Discussion --- p.150 / Chapter 3.3. --- Purification and Characterization of Ribosome-inactivating Protein from the Hypsizigus marmoreus --- p.153 / Chapter 3.3.1. --- Introduction --- p.153 / Chapter 3.3.2. --- Result --- p.154 / Chapter 3.3.3. --- Purification --- p.155 / Chapter 3.3.3.1. --- Cation-Exchange Chromatography on CM-Cellulose --- p.156 / Chapter 3.3.3.2. --- Affinity-Chromatography on Affi-gel Blue Gel --- p.158 / Chapter 3.3.3.3. --- Anion-Exchange Chromatography on Mono Q Column --- p.160 / Chapter 3.3.4. --- Characterization of Ribosome-inactivating Protein from Hypsizigus marmoreus --- p.162 / Chapter 3.3.4.1. --- N-terminal Amino Acid Sequence --- p.162 / Chapter 3.3.4.2. --- Assay for Cell-Free Translation-Inhibiting Activity --- p.163 / Chapter 3.3.4.3. --- Assay for Antifungal Activity --- p.164 / Chapter 3.3.4.4. --- Assay for N-glycosidase Activity --- p.166 / Chapter 3.3.4.5. --- Assay for HIV-1 Reverse Transcriptase Inhibitory Activity --- p.166 / Chapter 3.3.4.6. --- Assay for mitogenic Activity --- p.167 / Chapter 3.3.4.7. --- Assay for Antiproliferative Activity --- p.167 / Chapter 3.3.5. --- Discussion --- p.159 / Chapter Chapter 4 --- General Discussion --- p.170 / References --- p.172

Ip Lai Fong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 81-92). / Abstracts in English and Chinese. / Table of Contents --- p.1 / Abbreviations --- p.4 / Chapter Chapter 1 --- Introduction and Background --- p.5 / Chapter 1.1 --- Endothelin system --- p.5 / Chapter 1.1.1 --- Endothelins --- p.5 / Chapter 1.1.2 --- Endothelin converting enzyme (ECE) isoforms --- p.12 / Chapter 1.1.3 --- Endothelin receptors --- p.24 / Chapter 1.2 --- Signal-transduction mechanisms in ET system --- p.27 / Chapter 1.3 --- The aim of the present thesis --- p.31 / Chapter Chapter 2 --- Materials and Methods --- p.32 / Chapter 2.1 --- Primer Design --- p.32 / Chapter 2.2 --- Total RNA Isolation --- p.33 / Chapter 2.3 --- Reverse transcriptase polymerase chain reaction (RT-PCR) --- p.34 / Chapter 2.3.1 --- First Strand cDNA Synthesis --- p.34 / Chapter 2.3.2 --- PCR reaction --- p.34 / Chapter 2.4 --- Agarose gel electrophoresis --- p.35 / Chapter 2.5 --- Ligation of PCR inserts to cloning vector by TA cloning method --- p.35 / Chapter 2.6 --- Competent cell preparation --- p.36 / Chapter 2.7 --- Transformation and Screening --- p.37 / Chapter 2.8 --- Plasmid DNA Extraction --- p.38 / Chapter 2.9 --- DNA sequencing --- p.38 / Chapter 2.10 --- DIG RNA Labeling --- p.38 / Chapter 2.10.1 --- Plasmid Linearization --- p.38 / Chapter 2.10.2 --- Transcription --- p.39 / Chapter 2.10.3 --- Probe purification --- p.39 / Chapter 2.11 --- In situ hybridizaion --- p.40 / Chapter 2.11.1 --- Tissue preparation and slide mounting --- p.40 / Chapter 2.11.2 --- Non-radioactive in situ hybridization --- p.41 / Chapter 2.12 --- Whole Mount non-radioactive in situ hybridization --- p.42 / Chapter 2.12.1 --- Dissection and fixation --- p.42 / Chapter 2.12.2 --- Hybridization --- p.43 / Chapter 2.12.3 --- Antibody incubation --- p.43 / Chapter 2.12.4 --- Histochemistry --- p.44 / Chapter Chapter 3 --- Results --- p.46 / Chapter 3.1 --- The molecular cloning of ECE-2 from rat brain --- p.46 / Chapter 3.2 --- Sequence characteristics of rat ECE-2 --- p.52 / Chapter 3.3 --- Comparison of rat ECE-2 with bovine and human ECE-2 and with the rat ECE-1 --- p.53 / Chapter 3.4 --- Tissue distribution of ECE-2 in rat and localization in C6 glial cells by RT-PCR --- p.60 / Chapter 3.5 --- ECE-2 in rat embryos at different gestation stages by RT-PCR --- p.60 / Chapter 3.6 --- ECE-2 distribution in C6 glioma cells --- p.63 / Chapter 3.7 --- ECE-2 distribution in rat embryo E15.5 --- p.63 / Chapter 3.8 --- ECE-2 distribution in rat brain sections --- p.63 / Chapter Chapter 4 --- p.74 / Discussion --- p.74 / References --- p.81

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-10-17T12:13:57Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_carlos_eduardo_pouey_da_cunha (2).pdf: 934243 bytes, checksum: 07247a5a64f709717d8bc06370d25dc7 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-10-23T11:07:13Z (GMT) No. of bitstreams: 2 dissertacao_carlos_eduardo_pouey_da_cunha (2).pdf: 934243 bytes, checksum: 07247a5a64f709717d8bc06370d25dc7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-10-23T11:07:32Z (GMT) No. of bitstreams: 2 dissertacao_carlos_eduardo_pouey_da_cunha (2).pdf: 934243 bytes, checksum: 07247a5a64f709717d8bc06370d25dc7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-10-23T11:07:43Z (GMT). No. of bitstreams: 2 dissertacao_carlos_eduardo_pouey_da_cunha (2).pdf: 934243 bytes, checksum: 07247a5a64f709717d8bc06370d25dc7 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-10-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A leptospirose é uma zoonose negligenciada difundida mundialmente com mais de 800.000 casos em humanos por ano. A doença é causada por espiroquetas patogênicas do gênero Leptospira, classificadas em 10 espécies e mais de 260 sorovares, agrupados em mais de 30 sorogrupos. As principais espécies responsáveis por casos de leptospirose humana são L. interrogans, L. borgpetersenii e L. kirschneri. No Brasil, onde os climas tropical e subtropical favorecem a disseminação da doença, a maioria dos casos é causada por L. interrogans sorogrupo Icterohaemorrhagiae. Apesar de Pelotas apresentar um clima temperado, a taxa de infecções por Leptospira (10 casos por 100 mil habitantes) é superior a regiões com climas tropical e subtropical (3.5 casos por 100 mil habitantes). Trabalhos de vigilância em leptospirose desempenham papel fundamental no combate à doença identificado espécies e sorovares endêmicos a determinada região. Este trabalho descreve o isolamento e caracterização de uma cepa de Leptospira de uma paciente da zona rural de Pelotas que relatou contato com coleções hídricas, bovinos, caninos e roedores, e apresentou sintomas clássicos da fase aguda da doença. O isolado foi caracterizado como L. kirschneri sorogrupo Pomona sorovar Mozdok, conforme determinado por multilocus sequence typing (MLST). O genoma do isolado possui 3.398 sequencias codificadoras, 39 tRNAs, 1 região codificadora de rRNA e conteúdo G+C de 34,6%. O cromossomo I tem 3.738.643 pares de base, e o segundo, 335.634, totalizando 4,07 Mb. Imunofluorescência indireta mostrou a expressão dos fatores de virulência LipL32, LigA e LigB. O isolado foi capaz de causar danos severos aos tecidos renal, hepático e pulmonar, como revelado por histopatologia e a presença de leptospiras nesses tecidos confirmada pela técnica de imprint. Este é o primeiro relato de isolamento de L. kirschneri sorovar Mozdok de caso clínico humano no Hemisfério Sul, de acordo com a literatura disponível. Os resultados aqui apresentados são de grande importância epidemiológica, bem como para o desenvolvimento de novas vacinas e testes rápidos de diagnóstico. / Leptospirosis is a neglected zoonosis spread worldwide with more than 800,000 human cases each year. The disease is caused by pathogenic members of the genus Leptospira, which is classified in 10 species and more than 260 serovars, grouped in more than 30 serogroups. The main species associated with human cases of leptospirosis are L. interrogans, L. borgpetersenii and L. kirschneri. L. interrogans serogroup Icterohaemorrhagiae causes the majority of human leptospirosis cases in Brazil, where the tropical and subtropical climates favour the spread of the disease. Even though the climate in Pelotas is temperate, Leptospira infection rates (10 cases per 100 thousand inhabitants) are higher than other regions with tropical and subtropical climates (3.5 cases per 100 thousand inhabitants). Leptospirosis surveillance studies play a vital role against the disease through the identification of endemic species and sorovars to a given region. This work describes the characterization of a Leptospira strain isolated from an inhabitant of the rural area of Pelotas. The patient reported contact with water, cattle, dogs and rodents and presented classical symptomatology for leptospirosis. Multilocus sequence typing (MLST) determined the isolate to be a L. kirschneri serogroup Pomona serovar Mozdok. The genome of the isolate includes of 3,398 coding sequences, 39 tRNAs, 1 rRNA coding region and 34.6% G+C content. Chromosome I has 3,738,643 base pairs, and chromosome II has 335,634 base pairs, totalizing 4.07 Mb. Indirect immunofluorescence showed the expression of LigA and LigB virulence factors as well as the LipL32 protein. The isolate was capable of causing severe renal, hepatic and lung damage, as revealed by histopathological analysis and the presence of leptospiras in these tissues was confirmed by the imprint technique. This is the first report of isolation of L. kirschneri serovar Mozdok from a human patient in the Southern Hemisphere, according to the literature. Results presented herein are important for epidemiologic studies, as well as for the development of new vaccines and rapid diagnosis tests.