Proteiner spelar en avgörande roll i människokroppen och det är därför viktigt att inta tillräckligt med essentiella aminosyror genom kosten. Tidigare har animaliska källor varit kända för att vara rika på dessa aminosyror, men nu förtiden räcker det inte för att tillfredsställa den globala behov. Dessutom väcker den animaliska proteiner allvarliga frågor om klimatförändringar och etik. Ärtor innehåller en essentiell profil av aminosyror och har många andra fördelar, såsom tillgänglighet, kostnadseffektivitet och förmåga att växa i låga temperaturer. Tidigare studier har visat att regelbunden intag av ärtprotein kan minska risken för hjärt-kärlsjukdomar och diabetes. Studier visade att Naturliga djupa eutektiska lösningsmedel (NADES) kan lösa upp proteiner utan att denaturera de, vilket är ett problem med traditionella lösningsmedel som kan påverka proteinstrukturer. Dessutom är NADES icke-toxiska och biologiskt nedbrytbara lösningar, vilket gör dem mer hållbara och miljövänliga än vanliga vätskor som används vid proteinextraktion. Syftet med studien var att använda NADES för att utveckla en ny metod för att extrahera proteiner från gula ärtor och jämföra denna med etablerade metoder. Dessutom skulle skillnader i proteinutbyte och strukturella förändringar uppskattas genom användning av Size Exclusion Chromatography (SEC) och Sodium Dodecyl Sulfate PolyAcrylamide Gel Elektrofores SDS-PAGE. Tre varianter av NADES framställdes från kolinklorid, citronsyra och olika vattenhalter. NADES 20% användes för extraktion, eftersom det har visat sig vara en lämplig metod för detta ändamål. Resultaten visade att extraktion med NADES är mer komplicerad än referensmetoderna för att proteinerna utfölls vid olika steg under extraktion process. SEC-kromatogram visade proteindenaturering där en tydlig topp erhölls vid senare elueringsvolym. SDS-PAGE analys genomfördes för att identifiera toppar men inge resultat kunde erhålla på grund av låg proteinkoncentration. Från ett optimistiskt perspektiv kan de denaturerade proteinerna fortfarande användas inom livsmedelsindustrin eller NADES kan användas som en pre-extraktionsmetod för att avlägsna föroreningar innan den verkliga extraktionsprocessen påbörjas. Det skulle vara användbart för framtida studier att prova att extrahera med en lägre vattenhalt i NADES eller med en annan typ av NADES. / Protein plays a crucial role in the human body, making it essential to consume sufficient amounts of essential amino acids through diet. Traditionally, animal sources have been known to be rich in these amino acids, but this is not sufficient to meet global demand. Additionally, this raises serious questions about climate change and ethics. Peas have been found to contain an essential profile of amino acids and have many other advantages, such as availability, cost-effectiveness, and the ability to grow in low temperatures. Previous studies have shown that regular consumption of pea protein can reduce the risk of cardiovascular disease and diabetes. Studies have shown that Natural Deep Eutectic Solvents (NADES) can dissolve proteins without denaturing them, which is a problem with traditional solvents that can affect protein structures. In addition, NADES are non-toxic and biodegradable solutions, making them more sustainable and environmentally friendly than common liquids used in protein extraction. The aim of the study was to use NADES to develop a new protein extraction method from yellow peas and compare this with established methods. In addition, differences in protein yield and structural changes were estimated using Size Exclusion Chromatography (SEC) and Sodium Dodecyl Sulfate PolyAcrylamide Gel Elektrofores SDS-PAGE. Three variants of NADES were prepared from choline chloride, citric acid, and different water contents. NADES 20% was used for extraction as it has been found to be a suitable method for this purpose. The results showed that extraction with NADES is more complicated than the reference methods because the proteins precipitated at different stages during the extraction process. SEC chromatogram showed protein denaturation where a clear peak was obtained at later elution volumes. SDS-PAGE analysis was performed to identify peaks, but no results could be obtained due to low protein concentration. From an optimistic perspective, the denatured proteins can still be used in the food industry, or NADES can be used as a pre-extraction method to remove impurities before the actual extraction process begins. It would be useful for future studies to try extracting with a lower water content in NADES or with a different type of NADES.

TRIM21 is a cytosolic ubiquitin ligase and an antibody receptor that providesa last line of defense against invading pathogens. By utilizing the diversity ofantibody repertoire to identify pathogens, TRIM21 serves as a link betweenintrinsic cellular defense and adaptive immunity. A variety of diseases havebeen linked to mutations of the TRIM family, including cancer, inflammatorydiseases, and autoimmune diseases. In this project, TRIM21 was producedand purified from Escherichia coli, (E.coli). Protein characterization wasperformed with SDS-PAGE, size exclusion chromatography and cryo-electronmicroscopy (cryo-EM). Previously TRIM21 has been shown to form a dimerwhen produced in SF9. Results from size exclusion chromatography show thatTRIM21 form a larger complex when expressed in E.coli. Cryo-EM resultsshow that the complex structure is more globular than previously thought.Purified TRIM21 was bound to the antibody IC100. SDS-PAGE and sizeexclusion chromatography results show much lower affinity to antibodies thanexpected. / TRIM21 är en cytosolisk ubiquitinligas- och antikroppsreceptor som ger ensista försvarslinje mot invaderande virus. Genom att använda mångfalden avantikroppsrepertoar för att identifiera patogener, fungerar TRIM21 som enlänk mellan inre cellulärt försvar och adaptiv immunitet. En mängd olikasjukdomar har kopplats till mutationer i TRIM-familjen, inklusive cancer,inflammatoriska sjukdomar och autoimmuna sjukdomar. I detta projekt produceradesoch renades TRIM21 från Escherichia coli, (E.coli). Proteinkarakteriseringutfördes med SDS-PAGE, gelfiltreringskromatografi och kryo-elektronmikroskopi(cryo-EM). Tidigare har TRIM21 visat sig bilda en dimer när den producerasi SF9. Resultat från gelfiltrering visar att TRIM21 bildar ett större komplexnär det uttrycks i E.coli. Cryo-EM-resultat visar att den komplexa strukturenär mer klotformig än man tidigare trott. Renad TRIM21 bands till antikroppenIC100. SDS-PAGE och gel-filtrerings resultat visar mycket lägre affinitet tillantikroppar än förväntat.

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Winter rye produces ice-modifying antifreeze proteins upon cold treatment. Two of these antifreeze proteins are members of the large, highly conserved, ??-1,3-endoglucanase family. This project was designed to identify glucanase genes that are expressed during cold acclimation, wounding, pathogen infection, drought or treatment with the phytohormones ethylene and MeJa. Additionally, a more detailed proteomic analysis was to be carried out to evaluate the glucanase content of the apoplast of cold-acclimated (CA) winter rye. Results of 2D SDS-PAGE analysis revealed that non-acclimated whole leaf protein extracts contain at least two ??-1,3-endoglucanses while CA whole leaf protein extracts contain at least three ??-1,3-endoglucanses. Subsequent 2D SDS-PAGE analysis was conducted on the apoplast extracts of NA and CA winter rye plants revealed the limitations of standard 1D SDS-PAGE. The 2-dimensional gel analysis revealed that there is a minimum of 25 proteins within the apoplast of CA winter rye, including at least 5 ??-1,3-endoglucanases. Genome walking was used to isolate cold-responsive glucanase genes. The five genes isolated were designated scGlu6, scGlu9, scGlu10, scGlu11 and scGlu12. The cis-element pattern within the promoter of each gene was evaluated using online databases of documented plant cis elements. As expected, all of the promoters contained elements associated with cold, biotic and abiotic stresses, light regulation, and development. The expression patterns predicted by the cis elements in each promoter were compared to the mRNA abundance produced by each gene as detected by semi-quantitative reverse transcriptase PCR. In most cases, the abundance of transcripts arising from each gene loosely corresponded to the expression pattern predicted by the cis elements the corresponding promoter. Transcripts of scGlu9, 10 and 11 were present in cold-treated tissues and are candidates for ??-1,3-endoglucanases with antifreeze activity. The results presented in this thesis provide additional insight into the apoplast proteome of CA winter rye plants as well as the complexity of the signals controlling the proteins that reside there. Although there are still a number of unresolved questions, this research opens new directions for future studies in the cold acclimation process in winter rye and specifically for the contribution of ?? -1,3-endoglucanses.

The web is the greatest information source in human history, yet finding all offers for flats with gardens in London, Paris, and Berlin or all restaurants open after a screening of the latest blockbuster remain hard tasks – as that data is not easily amenable to processing. Extracting web data into databases for easier processing has been a resource-intensive process, requiring human supervision for every source from which to extract. This has been changing with approaches that replace human annotators with automated annotations. Such approaches could be successfully applied to restricted settings such as single attribute extraction or for domains with significant redundancy among sources. Multi-attribute objects are often presented on (i) Result pages, where multiple objects are presented on a single page as lists, tables or grids, with most important attributes and a summary description, (ii) Detail pages, where each page provides a detailed list of attributes and long description for a single entity, often in rich format. Both result and detail pages are having their own advantages. Extracting objects from result pages is orders of magnitude faster than from detail pages, and the links to detail pages are often only accessible through result pages. Detail pages have a complete list of attributes and full description of the entity. Early web data extraction approaches requires manual annotations for each web site to reach high accuracy, while a number of domain independent approaches only focus on unsupervised repeated structure segmentation. The former is limited in scaling and automation, while the latter is lacked in accuracy. Recent automated data extraction systems are often informed with an ontology and a set of object and attribute recognizers, however they have focused on extracting simple objects with few attributes from single-entity pages and avoided result pages. We present an automatic ontology-based multi-attribute object extraction system AMBER, which deals with both result and detail pages, achieves very high accuracy (>96%) with zero site-specific supervision, and is able to solve practical issues that arise in real-life data extraction tasks. AMBER is also applied as an important component of DIADEM, the first automatic full-site extraction system that is able to extract structured data from different domains without site-specific supervision, and has been tested through a large-scale evaluation (>10, 000) sites. On the result page side, AMBER achieves high accuracy through a novel domain- aware, path-based template discovery algorithm, and integrates annotations for all parts of the extraction, from identifying the primary list of objects, over segment- ing the individual objects, to aligning the attributes. Yet, AMBER is able to tolerate significant noise in the annotations, by combining these annotations with a novel algorithm for finding regular structures based on XPATH expressions that capture regular tree structures. On the detail page side, AMBER integrates boilerplate removal, dynamic lists identification and page dissimilarity calculation seamlessly to identify data region, then employs a set of fairly simple and cheaply computable features for attribute extraction. Besides, AMBER is the first approach that combines result page extraction and detail page extraction by integrating attributes extracted from result pages and the attributes found on corresponding detail pages. AMBER is able to identify attributes of objects with near perfect accuracy and to extract dozens of attributes with > 96% across several domains, even in presence of significant noise. It outperforms uninformed, automated approaches by a wide margin if given an ontology. Even in absence of an ontology, AMBER outperforms most previous systems on record segmentation.

Introduction: Cellular membrane proteins, such as the cytochrome P450 enzyme superfamily (P450), have important roles in the physiology of the cell. P450s are important in metabolising endogenous molecules, as well as metabolising xenobiotic substances for detoxification and excretion. P450s are also implicated in cancer as they can act to 'negatively' de-activate or 'positively' activate cancer therapeutics. Identifying specific P450s that are highly up-regulated at the tumour site could be used to predict drug response and formulate targeted cancer therapy to help diminish systemic side-effects. Methods: Previous enrichment strategies have been unable to isolate the full complement of the P450 superfamily. To develop enrichment procedures for the P450s, a proteomic strategy was developed so that compounds could be screened for their effectiveness as general P450 probes. A standardised work-flow was created, encompassing affinity chromatography, protein concentration/desalting, followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and high performance liquid chromatography-mass spectrometry (HPLC-MS). A ketoconazole analogue and a 2-EN analogue, with known P450 inhibition, were immobilised on a solid support for comparison to immobilised histamine. Co-factor removal, competitive elution and DTT cleavage of disulfide bonds of probes were utilised to elute bound proteins. Results/Discussion: Inhibitor-beads bound a large range of proteins, including P450's, of which some were eluted by co-factor removal, some by competitive elution. Specificity of binding was improved by optimising buffer conditions and solid supports, however non-specific binding was not totally eradicated. All human P450s from spiked samples and 18 P450s from more complex mouse liver samples were recovered using one or more ligands.

Social networking sites are a major networking tool for consumer interactions as they provide a platform for communication, socialization, and learning activity. Subsequently, social media has become an important marketing tool for advertising companies’ messages. As a result, fashion brands such as H&M and Victoria’s Secret started to show more brand ads on Facebook. Facebook is one of the most powerful social networking sties due to its ability to reach to broad consumer groups through a brand page. However, research regarding this topic is limited as the social networking sites is relatively a new phenomenon. Therefore, the purpose of this research is to examine the effect of the brand’s comments in attenuating (enhancing) negative (positive) influence of other consumer’s postings on brand trust and purchase intention of other consumer’s postings on social media. Findings from this study revealed that there is no moderating effect of brand comments of the relationship between other consumer’s postings and brand trust, while positive other consumer’s postings has a significant effect on consumers’ brand trust. Also, there were no significant differences among other consumer’s postings, brand comments and purchase intention relationships. These findings add to the previous literature explains that brand should interact with consumers frequently in order to induce positive other consumer’s postings to develop brand trust. By using the consumer socialization theory to investigate Facebook marketing, this study provides insights and information on consumer attitudes and behaviors related to Facebook brand page.

During the transformation of the bacterial endosymbiont into current mitochondria the protein import apparatus had to be created de novo. The reduced mitochondria (mitosomes) of the parasitic protist Giardia intestinalis represent unique cellular model for the examination of these fundamental transport processes. As the main objective of this project I will try to characterize the motor complex, which propels the protein transport, and also the translocation channel in the inner mitosomal membrane. To this aim I will exploit the presence of two membrane components Pam16 and Pam18, which were discovered in our laboratory, and which constitute the functional core of the motor complex. Based on the information from the analogous systems of yeast and mammalian mitochondria, these two components should physically interact with so far unknown translocation channel. In all other eukaryotes this channel is formed by a conserved protein Tim23. The absence of this protein in the genome of G. intestinalis suggests presence of completely novel, or maybe the original-bacterial protein. Having in hand this simplified mitochodrial model the project has potential to bring not only new data in parasite biology but also generate new information on the function and evolution of mitochondrial protein import.

Memoria para optar al título de Ingeniero Civil en Computación / El presente documento describe la implementación de un sistema web bajo la arquitectura de aplicación de página única (en inglés, SPA: Single Page Application), con el objetivo de dar apoyo a la gestión interna de OTECs (Organismos Técnicos de Capacitación), dando una experiencia más fluida a los usuarios del sistema a través de una aplicación de escritorio. Las tecnologías escogidas en este proyecto permiten generar una arquitectura modular, que facilita la modificación de su estructura, con el objetivo de agregar o quitar servicios según los procesos internos que posea un OTEC en particular. Además, las funcionalidades desarrolladas en este sistema permiten apoyar procesos transversales de un OTEC, como por ejemplo: registrar usuarios, crear y corregir pruebas, y realizar encuestas de satisfacción, entre otras. Este sistema web permite al OTEC, registrar diversos tipos de usuario: internos, empresas y estudiantes regulares. Entre las principales funciones de los usuarios internos (empleados del OTEC), se encuentra: registrar estudiantes y empresas, generar y revisar pruebas para las capacitaciones, cargar documentos de apoyo, y generar credenciales, diplomas y actas de entrega. Por su parte, los usuarios empresa pueden: controlar el avance los alumnos, y descargar las credenciales, diplomas y actas de entrega. Finalmente, estudiantes regulares pueden ver su participación en las capacitaciones, y descargar sus credenciales, diplomas y responder pruebas. La solución desarrollada permite a la OTEC reducir el número de errores en la carga de información, y evitar la duplicidad de la misma. Por otro lado, al ser éste un sistema online, permite el trabajo en paralelo y distribuido, siempre que se cuente con acceso a internet. Este trabajo de memoria se desarrolló en el marco un OTEC particular, quien destacó el software como un elemento diferenciador respecto a otros OTEC. Aunque este proyecto será continuado con el objetivo de cubrir todas las necesidades de un OTEC, los resultados actuales del software han sido evaluados positivamente, y le permiten al OTEC operar con niveles de error muy por debajo de sus niveles históricos.

Arbetet undersöker huruvida laddtider för en single-page applikation av typen kundvagn påverkas beroende på vilket av JavaScript ramverken AngularJS och Vue som använts under utvecklingsfasen. Undersökningen görs mot bakgrunden att e-handeln hastigt växt på webben under det senaste decenniet till att bli globalt konkurrenskraftig. Den mest centrala delen all e-handel är kundvagnen och det finns en uppsjö av ramverk som kan förenkla utvecklandet som alla har olika styrkor och svagheter. Studien kommer genomföras som ett experiment där en applikation för respektive ramverk skapas på en lokal webbserver och jämförs mot varandra. Testerna kommer genomföras på en MacBook genom automatiserade script i Tampermonkey. Dessa skillnader framhäver behovet av att göra ett val. Resultatet visar tydligt att vilket ramverk som använts påverkar laddtider men till fördel för olika ramverk vid olika tillfällen. Arbetet kan ge en rad framtida arbeten härledas där resultat kan styrkas och utvidgas för att bli än mer givande.