A targeting peptide at the N-terminus of a precursor protein usually directs the protein synthesized in the cytosol to a specific organelle in the cell. Interestingly, some targeting peptides, so-called dual targeting peptides (dTPs) can target their protein to both mitochondria and chloroplasts. In order to understand the mechanism of dual targeting, a dTP from threonyl tRNA synthetase (ThrRS-dTP) was investigated as a model dTP in this thesis work. The results suggest that ThrRS-dTP is intrinsically disordered in solution but has an α-helical propensity at the N-terminal part. Tom20 and Toc34 are the two primary receptors on the outer membranes of mitochondria and chloroplasts, respectively. We found that the N-terminal half of the ThrRS-dTP sequence, including an amphiphilic helix, is important for the interaction with Tom20. This part also contains a φχχφφ motif, where φ represents a hydrophobic/aromatic residue and χ represents any amino acid residue. In contrast, neither the amphiphilic helix nor φχχφφ motif in ThrRS-dTP has any special role for its interaction with Toc34. Instead, the entire sequence of ThrRS-dTP is important for Toc34 interaction, including the C-terminal part which is barely affected by Tom20 interaction. In addition, the role of lipids in the organelle membrane for the recognition of dual targeting peptides during protein import is also the focus of this thesis. The tendency to form α-helix in ThrRS-dTP, which is not observable in solution by CD, becomes obvious in the presence of lipids and DPC micelles. To be able to study such interactions, DMPC/DHPC isotropic bicelles under different conditions have also been characterized. These results demonstrate that bicelles with a long-chained/short-chained lipid ratio q = 0.5 and a concentration larger than 75 mM should be used to ensure that the classic bicelle morphology persists. Moreover, we developed a novel membrane mimetic system containing the galactolipids, MGDG or DGDG, which have been proposed to be important for protein import into chloroplasts. Up to 30% MGDG or DGDG lipids were able to be integrated into bicelles. The local dynamics of the galactolipids in bicelles displays two types of behavior: the sugar head-group and the glycerol part are rigid, and the acyl chains are flexible. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 2: In press.</p>

Context and rationale – This work originates from policy priorities established within Defra to manage exotic animal diseases (EAD); specifically to understand the causes of low probability events, and to establish contingencies to manage outbreak incidents. Outbreaks of exotic animal diseases, e.g. FMD, CSF and HPAI, can cause economic and social impacts of catastrophic proportions. The UK’s government develops and implements policies and controls to prevent EAD and thus minimise these impacts. Control policies to achieve this are designed to address the vulnerabilities within the control systems. However, data are limited for both the introduction of an EAD as well as its resurgence following the disposal of infected carcasses, i.e. the pre-outbreak and post-outbreak phases of an EAD event. These lack of data compromises the development of policy interventions to improve protection. To overcome these data limitations, predictive models are used to predict system vulnerabilities. Cont/d.

Lentiviruses can infect non-dividing cells, and various cellular nuclear transport proteins provide crucial functions for lentiviral nuclear entry and integration. Genome-wide small interfering RNA screens previously identified nuclear pore complex component nucleoporin 153 (NUP153) as being important for infection by human immunodeficiency virus type 1 (HIV-1). We found that HIV-1 infection of NUP153 depleted cells resulted in normal levels of reverse transcription, a moderate reduction of 2-long terminal repeat circles, and a relatively large reduction in integrated proviruses, consistent with a role for NUP153 during nuclear entry of the HIV-1 pre-integration complex. We ascertained the capsid (CA) to be the major viral determinant for NUP153 dependence during infection, and accordingly observed a direct interaction between the CA N-terminal domain and the phenylalanine-glycine (FG)-repeat enriched NUP153 C-terminal domain (NUP153C). NUP153C fused to the effector domains of the rhesus Trim5alpha restriction factor (Trim-NUP153C) potently restricted HIV-1, providing an intracellular readout for the NUP153C-CA interaction during retroviral infection. Primate lentiviruses and equine infectious anemia virus (EIAV) bound NUP153C under these conditions, results that correlated with direct binding between purified recombinant proteins in vitro. These binding phenotypes moreover correlated with the requirement for endogenous NUP153 function during infection. Mutagenesis experiments identified NUP153C and CA residues important for binding, and different FG motifs within NUP153C mediated binding to HIV-1 versus EIAV CA proteins. HIV-1 CA binding mapped to residues that line a common alpha helix 3/4 hydrophobic pocket that also mediates binding to the small molecule PF-3450074 (PF74) inhibitor and cleavage and polyadenylation specific factor 6 (CPSF6) protein, with Asn57 (Asp58 in EIAV) playing a particularly important role. PF74 and CPSF6 each competed with NUP153C for binding to HIV-1 CA, and significantly higher concentrations of PF74 were needed to inhibit HIV-1 infection in the face of Trim-NUP153C expression or NUP153 knockdown. Correlation between CA mutant viral cell cycle and NUP153 dependencies moreover indicated that the NUP153C-CA interaction underlies the ability of HIV-1 to infect non-dividing cells. We propose that HIV-1 CA binds NUP153 FG motifs to affect viral nuclear import, serving as a novel example of viral hijacking of a fundamental cellular process.

Lentiviruses can infect postmitotic cells, indicative of a role for the nucleocytoplasmic transport machinery. Genome-wide RNA interference screens identified transportin 3 (TNPO3) that may regulate human immunodeficiency virus type 1 (HIV-1) preintegration complex (PIC) nuclear import but plays no role during murine leukemia virus (MLV) infection. Independently, TNPO3 was shown to bind HIV-1 integrase (IN), a PIC component, suggesting a potential mechanism for nuclear import. We demonstrated direct binding between TNPO3 and several retroviral INs, which did not correlate with TNPO3 dependency profiles of the respective retroviruses. Infectivity assays employing HIV-1/MLV chimeric viruses ascertained that the capsid (CA) domain, but not IN, was the functional determinant of TNPO3 dependence. A carboxy-terminal truncation mutant of the serine-arginine rich (SR) protein family member, cleavage and polyadenylation specific factor 6 (CPSF6), CPSF6-358, which lacks its RS domain, was shown to restrict HIV-1 PIC nuclear import. We demonstrated that CPSF6 interacts with HIV-1 CA, and a single point mutation in CA, Asn74Asp (N74D), abolished this interaction. N74D also rendered HIV-1 TNPO3-independent and impaired cyclophilin A (CypA) binding to CA. The CA:CPSF6 binding interface, as described in a partial co-crystal structure, defined a surface pocket on CA that faces the CA hexamer:hexamer interspace. Infectivities and CA binding profiles of CA mutants within this pocket or with aberrant CypA-related phenotypes were assessed to compare their CPSF6-358 sensitivity and TNPO3 dependence, which largely correlated. We showed an overall correlation between the CPSF6/CPSF6-358 binding profiles of these HIV-1 CA mutants and their CPSF6-358 sensitivity, whereas TNPO3 binding and TNPO3 dependence did not correlate. Based on similar infectivity profiles of CA mutants and the loss of the RS domain from CPSF6-358 we tested for a direct interaction between CPSF6 and TNPO3. We demonstrated specific binding between recombinant TNPO3 and the CPSF6.RS domain. Mutagenesis experiments suggested a multicontact binding interface. The interaction was downmodulated by Ras-related nuclear protein (Ran)-GTP, indicating that CPSF6 is a bona fide import substrate of TNPO3. Our results support a model where TNPO3 regulates nuclear CPSF6 localization and that in its absence CPSF6 may restrict infection by directly interacting with HIV-1 CA at the hexamer:hexamer interface.

published_or_final_version / Economics / Master / Master of Philosophy

Bakgrund och problem: Inköpsfunktionen ökar i betydelse i takt med att konkurrensen från andra företag ökar. Inköpsfunktionen ser olika ut i olika företag. För att hålla låga priser är det många företag som väljer att importera produkter från lågkostnadsländer. För textilföretag är Indien ett bra alternativ då de på grund av långa traditioner anses vara duktiga på textilproduktion. De har billig arbetskraft och stora råvarutillgångar. Stora företag har mer resurser än små och det påverkar inköpsprocessen. Därför har följande tre problemfrågor formulerats: 1. Hur ser inköpsprocessen ut hos mindre svenska företag inom textilbranschen vid inköp från Indien? 2. Vilka problem stöter mindre svenska företag inom textilbranschen på vid inköp från Indien? 3. Varför uppstår dessa problem? Syfte: Syftet med denna studie är att identifiera problem som uppstår i inköpsprocessen vid textilinköp från Indien. Vidare är syftet att ge en förklaring till varför dessa problem uppstår. Metod: Studien är av kvalitativ art och insamling av uppsatsens empiriska material har skett genom en flerfallstudie och med semi-strukturerade intervjuer. Respondenterna har varit personer som företagen anser mest lämpliga. Fallföretagen i studien är Elfsborg Interiör Aktiebolag, Polytuft of Sweden Aktiebolag och Värnamo Sängkläder Aktiebolag. Slutsatser: Svenska mindre företag inom textilbranschen har en inköpsprocess som innehåller många steg som återfinns i van Weeles inköpsprocess men kan inte fullt ut beskrivas som likadana eller lika detaljerade. Samtliga tre företag upplever problem längs inköpsprocessen. Gemensamt för alla tre är att de upplever det svårt att lokalisera kvalificerade leverantörer med avseende på kvalitet och inköpspris när val av leverantör i Indien ska göras. Det geografiska avståndet ses, av samtliga fallföretag, som en stor anledning till varför undersökning innan och bevakning under produktionen är svår att genomföra. Tydlig dokumentation och detaljerad förteckning över leverantörer i Indien är till stor hjälp då uppföljning och utvärdering genomförs.

New technologies gives us possibility to step to new, more automated, level and to guarantee bigger efficiency. It requires comprehensive perfection and stepping to higher level. The ability of human being to analyze and process information flows is quite limited and requires big time recourses, that’s why complex of technical and programmable instrumentality would be needed to guarantee exactness of information, timeliness, automated information processing. In these days companies more are discussing about the possibilities how to computerize the activity process than about the need of this process generally. In any activity important is exact, timeliness and automated information. By the way, we must not forget that any program equipment will not fulfill all the job: to fulfill the tasks we need to have human recourses who will be able to use this program equipment as a tool, who will have ability to analyze and estimate received results. Only in this way there is possibility to increase efficiency with the help of program equipment. There are four production lines connected in one mill, that can work separately according their production capacity. Many big companies in their activity already long time are using mobile telecommunications. But only in these years, when the amount of jobs and speed of life is increasing and after new technologies came in to the life, mobile technologies are more and more entering to the business of big or middle companies as well as... [to full text]

Darbe analizuojama įvairių veiksnių bei priežastinių ryšių įtaką Lietuvos maisto produktų gamybos įrenginių rinkos parametrų pokyčiams – įrenginių importo, eksporto rinkoje apimtims bei jų vertei. Tyrimų tikslas - išanalizuoti Lietuvos maisto produktų gamybos įrenginių rinkos kiekybinius pokyčius, įvertinant makroaplinkos įtaką. Darbo uždaviniai – nustatyti Lietuvos maisto produktų gamybos įrenginių importo, eksporto kiekybinius pokyčius šalyje; paklausos vertinimo ir prognozavimo ekonominių aspektų analizė bei prognozavimo metodų parinkimo kriterijai; parinkti prognozavimo metodą maisto gamybos įrenginių importo prognozei sudaryti; įvertinti gautas pirmines prognozes. Darbe yra 60 teksto puslapių, 8 lentelės, 13 paveikslų, 53 priedų puslapiai, naudota 40 literatūros šaltinių. / In this work there is tried to analyse of the various factors and causalities to the rates of market alterations of the food equipment in Lithuania – to the quantity and to the value of the equipment import and export in the market. The object of this research is to analyse the market quantitative alterations of the food equipment in Lithuania and to evaluate macroeconomic influence. Main tasks are to clarify the market quantitative alterations of the Lithuanian food equipment import and export in this country; the economic dimensions analyse of the demand evaluation and forecasting and the selection parameters of the forecasting methods; to select the forecasting method to make the forecast for the food equipment import; to evaluate forthcoming primary forecasts. There are 60 text pages, 8 tables, 13 pictures, 53 extras pages used 40 sources of literature.

High housing- and construction costs is a fact in Sweden today. Prices on building materials more rapidly than producer price index for industrial goods. Prices tend to rise during market boom but does not adapt during diminishing demand. The effects of fading demand can rather be deduced through discounts and quotations. During 1992-1998 when the Swedish construction sector found it self in severe crisis the prices of construction materials kept rising more rapidly than PPI for industrial merchandise. The development of cost for construction materials has been significant. Subsequently, report published by the building commission 2002:115 presents that the market structure of the Swedish supply market of construction materials often can be likened with oligopoly. Since 1994 to 2006 the cost of production for one square meter building area has practically doubled. The cost of production for apartment buildings from 1996 to 2001 was just about 50 % while at the same time the inflation was only 4,3 % according to SCB. This report primarily intends to study the possibilities and actions necessary to increase the use of foreign suppliers of construction materials at the larger construction contractors. The survey shows that the use of foreign suppliers is highly limited at 46 of the 50 largest building companies in Sweden. Results show that 90 % of the respondents has had totally 5 or less purchases from foreign sellers during 2012 till April 2013. At the same time the report shows that international trade frequently has a positive competition effect on the Swedish supplier market. Leading actors in international trade of construction goods states that they have succeeded in breaking the interacting price controlling at the Swedish suppliers of construction materials. The study further shows that knowledge and competence within the companies is missing in order to obtain a successful international procurement. The person accountable of purchases in the companies emphasizes that more resources within the business is fundamental to incept such a purchasing method. The survey confirms that a service that offers knowledge and information about foreign suppliers would ease the international purchases for Swedish contractors.

Baigiamajame darbe atlikta importo, eksporto ir tranzito informacinių srautų Lietuvoje analizė per 2006 m. sausio 1 d. - 2007 m. birželio 30 d laikotarpį. Išanalizuoti kreipimaisi į Informacinių technologijų paslaugų centrą dėl minėtus srautus aptarnaujančių sistemų problemų. Pateiktos išvados ir pasiūlymai. Efektyvus informacinių technologijų panaudojimas atlieka svarbų vaidmenį Lietuvos muitinės sistemos veikloje. Pastaruoju metu tiek mūsų šalyje, tiek ir tarptautiniu mastu aktyviai diskutuojama apie kuo spartesnį elektroninės muitinės aplinkos sukūrimą Europos Sąjungoje, dar veiksmingesnį informacinių technologijų pritaikymą atliekant prekių importo, eksporto bei tranzito srautų statistinę analizę, vykdant muitinio įforminimo dokumentų apdorojimą. Šio magistro baigiamojo darbo tikslas – atlikti muitinio įforminimo proceso informacinių srautų Lietuvoje analizę ir identifikuoti pagrindines problemas bei pateikti pasiūlymų dėl šių problemų sprendimo. Magistro baigiamąjį darbą sudaro trys dalys. Pirmoje darbo dalyje trumpai pristatoma Lietuvos muitinės istorija, aptariamos pagrindinės šiuolaikinės muitinės veiklos kryptys ir funkcijos, ypač akcentuotas Muitinės informacinių sistemų centro vaidmuo Lietuvos muitinės sistemoje. Antroje dalyje apžvelgiami prekių importo, eksporto bei tranzito procesai bei informacinių technologijų panaudojimas muitinio įforminimo dokumentų apdorojime. Trečioji dalis skirta muitinio įforminimo proceso informacinių srautų statistinei analizei... [toliau žr. visą tekstą] / In the Master’s paper the analysis of the import, export and transit flows of goods in Lithuania during the period from 1 January 2006 till 30 June 2007 is performed. The calls to Information Technology Service Centre regarding the problems of the systems servicing these flows is analysed. Some conclusions and proposals are submitted. The effective use of information technology (IT) plays an important role in the activities of the Lithuania’s customs system. Currently the issues of a faster creation of an EU-wide e-Customs, more efficient use of IT in performing the statistical analysis of the import, export and transit flows of goods or in processing customs documents have been seriously considered both nationally and internationally. The goal of this Master’s paper is to perform the analysis of information flows of the customs documents processing, to identify the problems as well as to present some proposals for the solution to these problems. The Master’s paper consists of three parts. In the first part a brief overview of the customs’ history in Lithuania is presented, the main current functions and activities of the Lithuanian customs are described and much attention is paid to the role of the Customs Information System Centre in the Lithuanian customs system. The second part is devoted to the survey of the import, export and transit processes of goods as well as the use of IT in the Lithuanian customs. In the third part a thorough statistical analysis of... [to full text]