IRF-1 (Interferon Regulatory Factor 1) is a transcription factor first identified as a regulator of Interferon expression. Two decades after its discovery, IRF-1 has been shown to be involved in numerous other pathways including apoptosis, cell cycle regulation, DNA damage/repair, immune cell development and inflammation. Transcriptional regulation of IRF-1 by a number of external agents has been extensively studied, however almost nothing is known about the posttranslational regulation of IRF-1 activity. In this study IRF-1 is shown to be phosphorylated at Thr180 by GSK3β (Glycogen Synthase Kinase 3β). Phosphorylated Thr180 promotes interaction with the ubiquitin E3 ligase SCFFbxw7u, (Skp1-Cu11-Fbxw7α) which increases turnover of IRF-1 protein. Phosphorylation dependent ubiquitination of IRF-1 was confirmed, as substitution of Thr180 to alanine reduced IRF-1 ubiquitination and increased stability. Enhanced phosphorylation of IRF-1 (by increasing GSK3β expression) promotes increased ubiquitination/degradation. Transactivation of the TRAIL (TNFα Related Apoptosis Inducing Ligand) promoter by IRF-1 was found to be dependent on GSK3β phosphorylation of Thr180 by use of reporter assays and inducible expression of IRF-1 in breast cancer cell lines. Importantly IRF-1 activity on the TRAIL promoter is dependent on proper turnover by the UPS (Ubiquitin Proteasome System), as chemical inhibition of the proteasome, or reduction in IRF-1 ubiquitination reduced activity in reporter assays. This suggests that phosphorylation of IRF-1 by GSK3β acts as a destruction signal through association with SCFFbxw7a. This signal dependent turnover of IRF-1 is required for proper transcriptional activation of the TRAIL promoter.

Co-crystals are multi-component, single phase materials in which the co-crystal formers exist in an uncharged state. This interaction between components occurs without affecting the intra-molecular covalent bonding of the involved components or altering their chemical integrity. This class of materials has recently gained the interest in the pharmaceutical industry for modifying the physico-chemical properties of some active pharmaceutical ingredients (APIs) such as solubility, hygroscopicity, and mechanical properties. However like all multi-component systems, there are some analytical challenges associated with using traditional quality control (QC) tools that involve sample preparation steps prior to the analysis. This can sometimes have an impact on the physical state of such systems and therefore affect the outcomes of analysis correlated with the actual materials. Flurbiprofen (FBP) and nicotinamide (NCT) in this work were selected to form a model co-crystal system. FBP falls in the carboxylated group of non steroidal anti-inflammatory drugs (NSAIDs), used herein as an API. It belongs to class II of the Biopharmaceutics Classification System (BCS), therefore its low aqueous solubility and dissolution rate affect its bioavailability. NCT is a vitamin B3 derivative and generally regarded as safe (GRAS) substance. It is used as co-crystallising agent due to its solubility enhancing property. FBP-NCT co-crystal system was previously studied by Berry et al., where a FBP-NCT co-crystal was prepared using Kofler method and screened by means of the hot-stage microscopy (HSM). FBP-NCT co-crystal was recently prepared via rapid evaporation from ethanol solution, and screened for its physicochemical and mechanical properties by Shing et al. who found an improvement in such properties compared to the pure API (FBP). There is a wide range of methods for preparing co-crystals, of which methodologies with minimal environment impact were mainly adopted in this work i.e., Ko er fusion and co-grinding methods. Within the Kofler fusion method, it was feasible to identify and screen simultaneously parent components as well as the emerging material at room temperature (RT) by means of confocal Raman microscopy. Detailed information from Raman mapping on the investigated phases were achieved using statistical analysis. The analysis resulted in a refinement regarding the traditional assumption about the binary phase diagram. It also revealed the presence of two forms of FBP-NCT co-crystals which were consequently explored by coupling thermal analysis with Raman spectroscopy and X-ray diffraction. Using co-grinding preparation, FBP-NCT co-crystals were prepared on a large scale; enough for solid dosage formulation. Subsequent crystallisation of the resulting compound using a solution method generated single crystals suitable for x-ray crystal structure determination. Moreover in this work, a solid-state dosage form of FBP-NCT co-crystal was prepared for the first time, then investigated using transmission Raman spectroscopy. The investigation included the presence of drug and excipient, and their composition in tablets. This procedure can be considered as a platform for studying the QC of drug preparation, using a reliable, non-destructive, non-invasive, and very rapid analytical tool. Such type of study complied well with the food and drug administrative (FDA) outlines on employing process analytical technology (PAT) protocol for analysing and controlling pharmaceutical manufacturing processes.

The bactericidal mechanism of hydrogen peroxide is poorly understood, with most evidence being obtained from studies involving greatly reduced concentrations aimed at investigating the effects of stress. Current theory suggests that this mechanism is based on the oxidation of protein, DNA and lipids within the cell by the production of free hydroxyl radicals through the interaction of hydrogen peroxide and intracellular iron. The mechanism of vapour phase hydrogen peroxide treatment remains unstudied, despite evidence that it may be different to the liquid phase. This study aimed to investigate the effects of bactericidal treatments of liquid and vapour phase hydrogen peroxide on the macromolecular components of a model organism, Escherichia coli strain K12. A set of treatment conditions producing a range of reductions in colony forming units was identified, and the effects of these conditions on the protein, DNA and lipid constituents of the cells assayed. No effect on the lipid contents and membrane integrity of treated cells was found. Liquid hydrogen peroxide was found to reduce the thiol content of cytoplasmic protein, but this was not found to be a major mechanism of bactericidal action. Extensive fragmentation of DNA was found to result by treatment with both phases, the degree of which was correlated with a reduction in colony forming unit counts. No effect on bactericidal action was found on addition of a hydroxyl radical scavenger or an inhibitor of protein synthesis, showing that DNA damage was due to the primary action of hydrogen peroxide, and that this damage was not caused by the production of free radicals within the cell. A modifed mechanism of hydrogen peroxide bactericidal action is proposed, whereby lethality is due solely to DNA damage caused by the production of ferryl radicals by the interaction of hydrogen peroxide and iron associated with the DNA backbone.

One third of the world’s population is estimated to be infected with M. tuberculosis, a pathogen which causes more human death and misery than any other bacterial disease. Whilst treatment is available, resistance to commonly used antimicrobials is a growing problem. Thus there is an urgent need to identify new compounds that can kill drug resistant isolates and are able to potentiate the activity of currently available antibiotics. The plant kingdom is a rich source of antibacterial compounds and a plant which has attracted particular interest is Humulus lupulus, more commonly known as the hop, which has been used as an antibacterial in beer for hundreds of years. Its antibacterial properties are thought to be due to the combined action of alpha and beta acids and polyphenols such as xanthohumol although the precise nature of their interactions and relative importance has yet to be determined. An optimised agar antimicrobial assay was developed and employed based on Mycobacterium smegmatis, to characterize the antibacterial activity of fifty commercially available hop varieties with a view to identifying novel antibacterial compounds. Surprisingly, no correlation was found between alpha and beta acid content and antibacterial activity. Chemical analysis of the most (Citra) and least (Galena) active hop variants using a combination of bioactivity based thin layer chromatography, mass spectrometry and HPLC revealed differences in the relative amounts of antimicrobial compounds such as humulone (alpha acid), lupulone (beta acid) and xanthohumol but failed to identify the presence of novel antibacterial compounds. Whilst no new antimicrobial compounds were identified, the Citra hop extract was able to potentiate the activity of the antibiotics imipenem and ciprofloxacin against clinical isolates of M. abscessus, a fast growing member of the mycobacterium family which infects individuals suffering from cystic fibrosis. The Citra hop extract also inhibited the growth drug resistant isolates of M. abscessus suggesting that it may have activity against other antibiotic resistant mycobacteria such as M. tuberculosis With regards to the mode of action, scanning electron microscopy revealed distinct changes to the outer cell structure of the bacteria, suggesting that hops contain compounds that interact with the bacterial cell membrane and/or cell wall. These changes were more profound in the presence of sub-inhibitory concentrations of imipenem, a compound which also targets the cell wall. Overall hops were shown to contain compounds which inhibited the growth of mycobacterium and were able to potentate the activity of antibiotics currently used to treat these pathogens. These findings suggest hops may be a fruitful source from which to isolate next generation compounds with which to treat increasingly drug resistant strains of mycobacteria.

A series of microgels have been prepared by a surfactant free emulsion polymerisation based on an N-isopropylacrylamide (NIPAM) monomer. Poly(NIPAM) is a thermo sensitive polymer which undergoes a conformational transition close to the human skin temperature. Poly(NIPAM) was co-polymerized with hydrophobic butyl acrylate (BA) to provide a more favourable environment for drug molecules to partition into within the particle. A second co-monomer, acrylic acid (AA) was used to prepare pH/temperature sensitive microgels. The coupling reactions between microgels and cotton cellulose are only feasible if they both have appropriate functionalities. For microgels, this was achieved by using different initiators which introduce different functional groups on the particle surfaces and different surface changes. Cotton samples were successfully modified by carboxymethylation, periodate oxidation, grafting of 1,2,3,4-butanetetracarboxylic acid, and chloroacetylation in order to target possible reactions with the terminal functional groups of the microgel particles. Microgels were attached to the cotton fabrics using different methods. The maximum weight increase of cotton samples due to the attached microgels was 23.51 ± 0.29% (w/w). Drug loading and drug release were studied for free (unattached) microgels and novel wound dressings. Methyl paraben (MP) was used as a model drug compound. The results of MP release from free microgels show no significant influence of surface charge on MP release. However, BA-containing microgel particles release less MP than 100% poly(NIPAM) microgels due to hydrophobic interactions between the MP and the BA. Temperature-sensitive dressings release up to 45% more MP at 40°C than at 30°C. The drug release for pH-sensitive dressings was up to 24% higher at pH 5 than at pH 8. This demonstrates that the novel dressings can be classed as “smart” materials as they can respond to subtle changes in simulated wound fluid temperature and pH.

Intravenous lipid emulsions are used ubiquitously through the medical field as a source of parenteral nutrition. Development of new formulations requires an understanding of the metabolism of the product. Current methods of rate determination and of metabolism analysis have several drawbacks. Radioactive labelled assays have lower biological relevance, are time consuming due to separation steps and require long substrate preparation. Existing fluorescence assays based around triglyceride hydrolysis are impractical in emulsion systems due to high signal-noise ratio as well as the use of non-specific fluorescent dyes. Colorimetric methods such as the non-esterified fatty acids (NEFA) assay is expensive, requires multiple steps and specialised machinery. Due to the limitations of these techniques we developed a novel fluorescent assay using a lipoprotein lipase specific substrate incorporated into lipid emulsions. The lipoprotein lipase substrate, EnzChek® Lipase Substrate, green fluorescent, 505/515, is based on a triglyceride structure with a fluorescent dye at the Sn1 position and a dark quencher at the Sn2 position. LPL cleaves preferentially at the Sn1 position of triglycerides, which separates the dye from the quencher creating a fluorescent signal. The signal can then be detected using a fluorescent plate reader. Both lipid emulsion particles and EnzChek are substrates for LPL, so the hydrolysis of EnzChek is analogous for native emulsions particles. Over time, in the presence of LPL, fluorescent signal increases as more EnzChek is hydrolysed in tandem with emulsion particles. We have designed a range of emulsions with varied oil and surfactant composition. Using the EnzChek emulsion assay detailed above we are able to follow the rate of metabolism in real-time. This assay has been tested and found to be robust and reproducible. It is able to investigate differences in metabolism between Soybean oil, medium-chain triglyceride and fish oil emulsions. As well as changes in surfactant type and concentration.

The role of warehouse in the value stream, plays a significant part in how well a company can satisfy the requirements of a customer. To be able to support the demand of the customers, production companies are forced to produce a bigger variety of models in the product range, which results in an increasing assortment. With an effective stock keeping, there is a potential to reduce waste and increase profitability. In the choice of warehouse-design, warehouse-operations have the opportunity of choosing between a number of different designs containing manual and automated put away.It has become common that companies are choosing a combination of automated and manual processes. With this combination of processes, the companies are faced with choices about stock locations. With the help of well developed criterias about stock locations, there is a potential to achieve a more effective put away. The purpose of the study has been formulated as ”Investigate when automation in warehouse management makes the put away process in a central warehouse more efficient”.   Through a case study on a warehouse, located in Nässjö, this report will answer the research questions regarding which of the manual and automated put away processes to use. The research questions are: Which criteria affect the decision of automated or manual put away? Based on the criteria that are raised in research question 1, when is it more efficient to store carton in AS/RS in comparison with manual picking with truck in pallet racking? The research questions wish to lift criteria as a basis for decision making and when it is more effective to use automated put away. The warehouse that this study is based on is using both automated and manual materials handling. The specific automation system that has been the basis for this study’s result is AutoStore. This report has a qualitative approach, the methods that has been used for collecting data are interviews, observations and document studies.   Dimension, process time and utilization were the three criteria that are presented in the result. Based on the pallets limit of space in the automated system, the dimension of the article is the first criteria. In addition to the article’s measurements, the articles shape and material of the package had an impact if the article would fit in the load carrier. The two put away processes include different operations that differ in how long they take to execute, which has an impact on the company's efficiency. One reason for the time consuming difference, turned out to be how the quantity affects the time for put away in AutoStore comparing with the constant time for manual put away. A time consuming operation in the process with AutoStore was the one with repacking for the systems pallet. The operation also turned out to affect the utilization rate, due to its measurement of a pallets fill rate.  The quantity of put away in combination with the dimensions of the article, affects the total utilization rate.   With the result, the aim of the study is to contribute to companies that contains a combination of warehouse-design with a basis for the decision making about stock locations.

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Nos últimos dez anos têm ocorrido uma intensificação da produção vitícola na região da Campanha Gaúcha, mas ainda existem deficiências nas principais técnicas de manejo dos vinhedos. Os objetivos deste trabalho foram avaliar a antecipação da poda hibernal e testar os efeitos de sistemas de poda e estimuladores de brotação sobre o potencial de produção e qualidade enológica da uva ‘Merlot’ enxertada em SO4. O experimento foi realizado nos ciclos vegetativos de 2013/2014 e 2014/2015 em vinhedo comercial da Cooperativa Nova Aliança, situada em Santana do Livramento-RS (30º 44’ 53,031” Sul, 55º 23’ 49,445” Oeste e 180 m de altitude). As plantas tinham 8 anos, conduzidas em espaldeira e espaçadas de 1,0 x 2,80 m. Foram realizados dois estudos principais: no primeiro, em 2013/2014 foram testadas três épocas de poda e cinco concentrações de cianamida hidrogenada (0; 1,0; 1,75; 2,5 e 3,25) nos sistemas de poda em Cordão Esporonado e Guyot Duplo, em parcelas sub-subdivididas. No segundo ciclo produtivo, o estudo foi repetido com a adição de mais uma época de poda. No segundo estudo, foi realizada apenas a poda de agosto no sistema Guyot Duplo e testado o efeito de concentrações de Erger® testemunha, Erger® 2,5 % + nitrato de cálcio 5,0 %; Erger® 5,0 % + nitrato de cálcio 5,0 %, Erger® 7,5 % + nitrato de cálcio 5,0 % e nitrato de cálcio 5,0 %, organizados em blocos casualizados. Nas avaliações foram coletados dados referentes à brotação, fenologia e variáveis produtivas. Os resultados mostraram interação significativa entre sistema de poda e concentração de cianamida hidrogenada, sendo que o Cordão Esporonado apresenta melhor brotação e proporcionou maior produção que o sistema de poda em Guyot Duplo. A antecipação da poda é viável, podendo esta atividade ser antecipada em até dois meses, sem influenciar a brotação e a qualidade das uvas. A concentração de 2,0 % de cianamida é a mais adequada para o sistema de poda em Guyot. No entanto, não se justifica o uso em Cordão Esporonado, pois não foram obervados efeitos sobre a produção. O fertilizante Erger® apresentou resultados satisfatórios de brotação nas plantas podadas em agosto em Santana do Livramento. / In the last ten years there has been an intensification of grape growing and wine production in the Campanha Gaúcha Region, but deficiencies still exist regarding vineyards main management techniques. The objectives of this study were to evaluate the anticipation of winter pruning and to test the effect of pruning systems and bud break stimulants on the production potential and enological quality of Merlot grape grafted in SO4. The experiment was conducted in the 2013/2014 and 2014/2015 vegetative cycles in the Nova Aliança Cooperative commercial vineyard, located in Santana do Livramento-RS (30º 44’ 53,031” South, 55º 23’ 49,445” West and 180 m altitude). The vines were 8 years-old, trained into VSP system (vertical shoot positioned) and spaced 1.0 m among vines and 2.80 m among rows. Two main studies were conducted: in the first one, in the 2013/2014 cycle, three pruning times and five hydrogen cyanamide concentrations were tested (0; 1.0; 1.75; 2.5 e 3.25 %) on the Cordon and Double Guyot pruning systems in splitplot design. In the second cycle, this study was repeated with an additional pruning time. In the second study, pruning was performed only in August, adopting Double Guyot system and three Erger® concentrations were tested control, 2.5 % Erger® + 5.0 % calcium nitrate; 5.0 % Erger® + 5.0 % calcium nitrate; 7.5 % Erger® + 5.0 % calcium nitrate and 5.0 % calcium nitrate, arranged in randomized blocks. Data related to bud break, phenology and production variables were collected. Results showed a significant interaction between pruning system and hydrogen cyanamide concentration. The Cordon system presented better bud break and production than the Double Guyot system. The pruning anticipation is viable, and a period of up to two months does not influence bud break and grape quality. The 2.0 % hydrogen cyanamide concentration is the most suitable for the Double Guyot pruning system, however, its use in Cordon system is not recommended because no effects were observed on the production. Erger® fertilizer showed satisfactory results over bud break of vines pruned in August at Santana do Livramento.

A fragmentação e a redução das Florestas naturais vem ocorrendo em muitas partes do mundo. Enquanto isso, as plantações de árvores para fins de silvicultura aumentam em extensão, muitas vezes substituindo áreas anteriormente cobertas pela vegetação natural. Considerando a situação florestal atual, as tentativas de aumentar a cobertura florestal natural são metas importantes no mundo todo. Este trabalho tem como principais objetivos: Avaliar a estrutura arbórea e regenerativa em monoculturas de espécies arbóreas (sem manejo) e áreas nativas. Avaliar a semelhança da regeneração nativa com as áreas de plantio. Estimar a influência de fatores ambientais sobre a regeneração de espécies nativas. Descrever a estrutura funcional do componente arbóreo adulto e regenerativo. Avaliar a diferenciação dos atributos funcionais entre as áreas e a influência dos fatores ambientais sobre esta variação. Este estudo foi realizado na Floresta Nacional de Passo Fundo, localizada no município de Mato Castelhano, Rio Grande do Sul, Brasil. Para tanto, foram selecionadas aleatoriamente cinco unidades amostrais para cada área. Dentro de cada uma das unidades amostrais foram demarcadas 10 subunidades para o levantamento do componente arbóreo adulto e regenerativo. O levantamento da vegetação mostrou que, além das áreas de Floresta Nativa, os plantios apresentaram um recrutamento de espécies tanto em fase regenerativa quanto adulta. No entanto, os plantios, no decorrer do tempo, causaram diferenças nas características do solo. Isto implicou em diferenças notáveis em termos de composição. Apesar destas diferenças, o estabelecimento de espécies nas áreas de plantio foi abundante, apresentando diversas famílias ocorrentes na Floresta Nativa. O componente adulto apresentou diferenças significativas para os traits entre as comunidades, enquanto o regenerativo apresentou apenas pequenas diferenças. A análise RLQ demonstrou a separação das áreas para ambos os componentes indicando diferenças na composição específica, funcional, edáfica e de incidência de luz. Nosso estudo mostra que devido ao componente regenerativo bem desenvolvido das áreas de plantios é possível a transformação dessas áreas em florestas naturais. Também podemos concluir que a ação das plantações como filtros ambientais influenciam diretamente o recrutamento e a riqueza de espécies assim como a diversidade funcional.