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Pore forming toxins (PFTs) are a large group of proteins found mainly in bacteria with some exceptions found in animals. They bind and form pores in their target membranes and form pores, which leads to cell death. Among these are cholesterol-dependent cytolysins (CDC), which require the presence of cholesterol to bind target membranes. Mitilysin (Mly), a protein of interest in this project, belongs to the CDC group of pore forming toxins. It is produced by the bacterium Streptococcus mitis, a pathogen closely related to Streptococcus pneumoniae, found in human oral cavity, which causes several diseases such as Viridans Group Streptococcal (VGS) toxic shock syndrome and endocarditis. Mly is a homologue of the toxin Pneumolysin, which is produced by S. pneumoniae. However, the mechanism of pore formation is not well known. The purpose of this project is to understand the mechanism of CDC pore formation, focusing on the key amino acid residues that are responsible for transitioning from Mly pre-pore to pore state. The findings will aid in the design of inhibitors of pore formation as potential anti-bacterial drug candidates. The major goal of the project was to determine the 3-dimensional (3D) structure of assembled Mly pore. Mly is expressed in E.coli and purified by Ni-NTA affinity chromatography. Pore formation is confirmed by a hemolysis assay and negative stain-transmission electron microscopy. Mly pores are vitrified, analyzed and imaged in a cryo-electron microscope. 2D images were processed to generate a 3D density map. However, our Mly pore 3D map was incomplete due to lack of 2D projection angles resulting from preferred orientation of pore particles during sample preparation. To overcome this problem, we aim to use DNA origami, which requires His-tagged Mly. We were able to determine that His-tagged Mly retains its pore formation ability.

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政黨是我國政治中最重要的行動者，然而多數有關於政黨政治的研究多由大黨的角度出發，忽略了其他中、小規模的政黨在其中所能發揮的影響作用。本文以小黨為主要研究對象，由小黨的觀點解釋其如何在政治運作過程中發揮政治影響力，或是以哪些手段達成更多的政黨目的。本研究選擇的研究對象是民主化以來存續時間較長、規模較大的三個小黨：新黨、親民黨與台灣團結聯盟，研究時間從立法院第3屆起（1995年）至第6屆結束（2008年）為止，共12年的時間。 本文結合國內外國會、政黨研究的相關研究成果，建構一套小黨的行動理論，接著從結構性的制度規則、質性的立委深入訪談資料，以及量化的立法紀錄，檢視與解釋三個小黨在立法院的各種行動及背後的政治動機。在台灣，小黨的出現常常是因大黨在政黨立場上先出現變動，小黨有取代大黨原有的政黨立場與選票支持的企圖而興起。隨後在議事規則的政黨化改革過程中，又給予小黨透過參與選舉、晉身進入立法議事參與者的政治機會。小黨的興起及運作機會既與外在的政黨政治及制度條件密切相關，也即預示小黨在立法院的策略行動及政治結果，也受到現實的政黨政治關係、自身的政黨立場及議事制度等現實條件所限制框架。經由對於三小黨的質化訪談與量化立法紀錄資料的分析後發現，小黨的政黨行動兼具持續與彈性的雙重特徵，在不同的政治條件下，小黨的某些政黨行動並不因為政治條件的改變而轉變，如團結與妥協。但小黨又常隨著不同的大黨對立程度，調整其於政治過程中的議價方式，是其靈活彈性的另一面。

Purpose: The purpose of this thesis is to investigate how food start-ups (FS) can make their primary food packaging (PFP) more eco-friendly by identifying and evaluating the performance of suited packaging alternative. The purpose was fulfilled by answering the three research questions:  RQ1) How to assess the eco-friendliness of PFP? RQ2) Which are areas of improvement in environmental performance? RQ3) What are the differences in performance across similar PFP’s?  Methods: To answer the research questions both the literature review and empirical data was required. The literature study was conducted to gather relevant theories about primary food packaging in food start-ups. To get the required empirical data, a single case study was conducted at a case company that suited the subject. The case study consisted of multiple interviews and document study. This enabled for an analysis in the form of pattern matching in order to answer the research questions and achieve the purpose.  Findings: The Study found that to assess the PFP that have direct impact on the environment the functional features and the environmental framework play a central role in the eco- friendliness of PFPs which analyzed the requirements for the PFP and a multi criteria decision making approach for the environmental assessment for the Green-PE. The stakeholder expectations were found by analyzing the criterion for the PFP. In addition, a comparison for an eco-friendlier alternative was analyzed with the current Green-PE to justify the performance for the PFP in FS.  Implications: The study results present practical implications with assessing the current Green-PE and evaluating the gaps for improvement areas, while also comparing similar PFP which is an eco-friendlier option for food packaging start-ups. As there has been no general theoretical implications, the findings of the thesis can be used as a basis for deeper insights into the subject through more extensive research.  Delimitations & Scope: The focus was to identify and evaluate the current PFP environmental impact and not the other aspects of the life cycle assessment since the scope was limited. Also, a single case study was used rather than multiple case studies to analyze the eco-friendliness for PEPs.