Masters of Science / Invasive pneumococcal disease (IPD), which includes potentially fatal conditions such as meningitis, septicaemia and pneumonia poses a threat in children aged <5 years, pneumonia being the leading cause of child mortality worldwide. Even though capsular polysaccharides are the main antigens involved in the immunity to encapsulated bacteria, it was found that in children in that age group, the immune system was unresponsive. Conjugate vaccines however induce immunologic memory and provide long-term protective immunity. Therefore the aim of this project was to develop novel conjugation strategies towards a pneumococcal conjugate vaccines and focuses mainly on the serotypes that are a burden to the African continent. The chemistry involved in developing a conjugate vaccine is of importance beacuse while some polysaccharides contain chemical grouping which can be conveniently utilized for conjugation, many medically important ones require derivatization before they can be coupled to protein. Derivatization of which can be achieved through various strategies, important to note is through hypervalent iodine oxidants. Two hypervalent iodine reagents, O-Methyl substituted-1-hydroxy-1,2-benziodoxol-3(1H)-one 1-oxide (Me-IBX)and modified 1-hydroxy-1,2-benziodoxol-3(1H)-one 1-oxide (mIBX)were successfully synthesized in preparation for the use in polysaccharide, polyribitol phosphate, (PRP) oxidation. The polysaccharide to be oxidised was first size reduced by microfluidisation to allow maximum oxidation. However, the extent to which oxidisation was achieved was not enough to conjugate the polysaccharide to the protein of preference, Bovine Serum Albumin, (BSA).

The main objective of this work is to critically evaluate a sufficient portion of the circuit problem of software piracy and to propose solutions of sub-problems, to use the interpretation of all relevant laws and all other available resources and create a comprehensive picture of the case. The practical part should be primarily a manual for ordinary computer users, which should help them. At the same time I present a preview of the methods of the executive law enforcement authority - Police of the Czech Republic, which deals with the investigation of software piracy. Range of the methods used in my work is not wide, which is partly due to reluctance of companies to cooperate because it is a relatively sensitive information, but it fully covers the needs of work and make it possible to have a final vision of the subject.

Ce travail est une étude expérimentale et de modélisation d’une configuration de puce microfluidique réalisée par photoablation laser d’un polymère diélectrique, le polyéthylène téréphtalate (PET) pour des applications en bioanalyse. C’est une configuration dite « non-contact » dans laquelle deux microélectrodes sont galvaniquement isolées dans le PET sans contact avec l’électrolyte du microcanal. La caractérisation par spectroscopie d’impédance de la réponse diélectrique du PET seul a permis de conclure que le film de PET peut être considéré comme un élément à phase constante (CPE). Cela a permis la compréhension des impédances mesurées sur puce microfluidique suivant une perturbation alternative de 100 mV dans la gamme de fréquence comprise entre 1MHz à 1Hz est effectuée pour : un microcanal vide, et un microcanal rempli avec des solutions de différentes conductivités ioniques. Les résultats obtenus ont montré une grande sensibilité des mesures d’impédances par rapport aux variations de la conductivité électrique de la solution électrolytique. Un des résultats surprenant concernant les impédances identifiées a été le comportement CPE observé avec un exposant de 0,5 que nous avons relié à l’impédance de surface rugueuse due la microstructuration en surface du PET photoablaté. Tous ces résultats expérimentaux ont été confortés par la modélisation en utilisant les circuits électriques équivalents puis par les éléments finis. Ce qui a permis de modéliser à la fois les phénomènes physiques dans le microcanal et à l’interface PET/µ-canal photoablaté. Enfin, deux applications capteur puis biocapteur ont été mises en œuvre en suivant le module de l’impédance à fréquence fixe et en fonction du temps. Comme exemple ; l’étude de la cinétique d’adsorption de la BSA sur le PET et l’étude de la cinétique d’association/ dissociation entre la BSA et l’anti-BSA. / This work is an experimental study and modeling of a microfluidic chip configuration produced by laser photoablation process on a dielectric polymer, polyethylene terephthalate (PET) for new bioanalytical development. This configuration is named "non- contact" wherein two microelectrodes are galvanically isolated in the PET without contact with the streaming electrolyte. Characterization by impedance spectroscopy of the dielectric response of PET alone has concluded that the PET film can be regarded as a constant phase element (CPE). This has permitted a best understanding of the measured impedances in a microfluidic chip under a 100 mV AC excitation in the frequency range between 1 Hz to 1 MHz, which are performed for: an empty microchannel, and a filled microchannel with electrolytes at various ionic conductivities. The results obtained showed a high sensitivity of the impedance measurements in correlation with the electrical conductivity change of the electrolyte. One of the surprising results relating to the identified impedances was a CPE behavior observed with an exponent of 0.5. This latter has been connected to the impedance due to the PET surface roughness. All these experimental results were supported by modeling using the equivalent circuits and by the finite elements. This was used to model both the physical phenomena in the microchannel and at PET interface/? - channel photoablated. Finally, both sensor and biosensor applications have been implemented in the microchip by recording the impedance module at fixed frequency with time. As examples, the investigating of the kinetics of BSA adsorption on the PET and the kinetics of association / dissociation between BSA and anti?BSA have been demonstrated.

Os líquidos iônicos, por geralmente apresentar, em sua estrutura, uma cadeia alquílica apolar e uma parte polar, como o cátion imidazólio, podem ser considerados estruturas anfifílicas, e apresentam semelhanças com os tensoativos. Tais estruturas, devido a este caráter anfifílico, podem interagir com estruturas do tipo Lipossomas ou até mesmo com biomoléculas, como a BSA. Neste trabalho, foram sintetizados três líquidos iônicos inéditos, com rendimentos de 52 a 66%, acoplados a sistemas quirais derivados de aminoácidos naturais e de fácil obtenção, como a L-Cisteína, a L-Serina e a L-Treonina. Estes foram caracterizados tanto do ponto de vista estrutural, quanto por suas propriedades fotofísicas. Como procedimento metodológico utilizou-se de ciclocondensação de aminoácidos, esterificação de Steglich e alquilação para inserção de cadeia carbônica ao anel metil-imidazol. Para investigação e confirmação dessas estruturas utilizou-se de estudos de Ressonância Magnética de 1H e de 13C, Infravermelho, além das espectroscopias de absorção na região do UV-Vis e de emissão de fluorescência. Por fim, estes novos líquidos iônicos foram testados com lipossomas e BSA, cujos resultados mostraram boa interação com essas biomoléculas. / Ionic liquids can be considered as amphiphilic structures and have similarities with the surfactants, once they generally have an apolar alkyl chain and a polar moiety, such as the imidazolium cation. Such structures, due to this amphiphilic character, may interact with liposome structures or even with biomolecules, such as BSA. In this work, three new ionic liquids containing chiral systems derived from naturally occurring amino acids, such as L-Cysteine, L-Serine and L-Threonine were synthesized, with yields ranging from 52 to 66%. They were fully characterized both from the structural point of view, and by its photophysical properties. As methodological procedures, amino acid cyclocondensation, Steglich esterification and alkylation were used. In order to investigate and confirm these structures, magnetic resonance studies of 1H and 13C, infrared, as well as absorption spectroscopies in the UV-Vis region and fluorescence emission were used. At last, these new ionic liquids were tested with liposomes and BSA, and the results showed good interaction with these biomolecules.

Manufactured oxide nanoparticles (NPs) have large production and widespread applications, which will inevitably enter the environment. NPs can interact with proteins in living beings due to the fact that NPs can transport into blood or across cell membranes into cells. Conformational change of protein molecules after sorption on oxide NPs has been reported. Therefore, it is important to understand the adsorption mechanism of protein onto oxide NPs surfaces. Although few works have reported protein adsorption behaviors, a general systematic comparison of the effects of particle size and surface groups on protein adsorption by widely studied NPs still needs to be made. Moreover, the relationship between adsorption maxima, which are related to protein conformational change and particle toxicity, and protein conformational change has not yet been studied. Therefore, in this work, the adsorption behavior of bovine serum albumin (BSA) protein on three types of nano oxide particles (viz., TiO2, SiO2, and Al2O3) was investigated in order to explore their interaction mechanisms, compared with that on regular bulk particles (BPs). The BSA adsorption maxima on oxide particles were regulated by the surface area of oxide particles. BSA adsorption was primarily induced by electrostatic attraction and ligand exchange between BSA and oxide surfaces. Surface hydrophilicity, surface charge and aggregation of oxide particles also affected their adsorption of BSA. Calculations suggested that a multilayer of BSA covered α-Al2O3, and single layer covered the other oxide particle surfaces. Primary structures of BSA molecules were adsorbed and changed on surfaces of oxide particles.

Rupture of the anterior cruciate ligament (ACL) is one of the most common ligamentous injuries of the knee. Post rupture, the ACL does not heal on itself due to poor vasculature and hence surgical intervention is required to treat the ACL. Current surgical management of ACL rupture consists of reconstruction with autografts or allografts. However, the limitations associated with these grafts have prompted interest in tissue engineered solutions that combine cells, scaffolds and stimuli to facilitate ACL regeneration. This thesis describes a ligament tissue engineering strategy that involves incorporating biomolecules within fibers-based electrospun meshes which mimics the extra-cellular matrix microarchitecture of ligament. However, challenges exist with incorporation of biomolecules. Therefore, the goal of this research project was to develop two techniques to incorporate biomolecules within electrospun meshes: (1) co-axially electrospinning fibers that support surface-grafting of biomolecules, and (2) co-axially electrospinning fibers decorated with biomolecule-loaded microspheres. In the first approach, chitosan was co-axially electrospun on the shell side of poly caprolactone (PCL) and arginine-glycine-aspartate (RGD) was attached to the electrospun meshes. Bone marrow stromal cells (BMSCs) attached, spread and proliferated on these meshes. In the second approach, fluorescein isothiocyanate labelled bovine serum albumin (FITC-BSA) loaded chitosan-alginate (CS-AL) microspheres were fabricated. The effects of cation to alginate ratio, type of alginate and concentration of CaCl2 on microsphere size, FITC-BSA loading and release were systematically evaluated. The CS-AL microspheres were then incorporated into the sheath phase of co-axially electrospun meshes to achieve microsphere-decorated fiber composite meshes. The results from these model study suggest that both approaches are tractable for incorporating biomolecules within fibers-based electrospun meshes. Both these approaches provide platform for future studies that can focus on ligament-relevant biomolecules such as FGF-2 and GDF-5. / Master of Science

Biological development and progression are managed by a diverse macromolecular group called proteins. Protein structure results from a complex folding process that leads to a final active form. This protein state is susceptible to changes in the surrounding environment and an incorrect structure can be produced. Changes in the protein conformation can lead to the formation of protein aggregates. Adsorption of proteins onto surfaces is utilized in many research analyses, but is capable of irreversibly changing the protein structure and causing aggregation. Albumin is a plasma protein that adsorbs on many different surfaces because the structure easily rearranges. The structure of albumin once adsorbed has been shown to deteriorate; however, outcomes of both stabilization and aggregation have been found. A dynamic laser light scattering instrument will be utilized to measure the differences in size and determine the amount of aggregation. Our lab has developed a z-axis translating laser light scattering device (ZATLLS) that has been used to measure the sedimentation velocity of several different materials in solution. In this case, bovine serum albumin (BSA) will be adsorbed onto polystyrene particles and the particle settling velocity determined. The settling solution viscosity and density will also be ascertained, so Stoke's law can infer the average aggregate size of each experiment. BSA-coated polystyrene particles displayed a more controlled settling behavior compared to non-coated polystyrene particles. Although the BSA-coated particles had a smaller sedimentation velocity, a larger aggregate size was found due to the greater solution viscosity. Therefore, the ZATLLS instrument can be employed to measure sedimentation velocities of multiple interactions and the aggregation level inferred. Although most albumin molecules are remarkably similar, there are subtle differences in amino acid residues, length, and charge. Sedimentation velocities for human serum albumin (HSA) coated polystyrene particles and BSA-coated polystyrene particles only had a small difference. However an almost 50% higher solution viscosity was measured in BSA experiment solutions, and resulted in the slower settling of the larger aggregates compared to HSA-coated particles. Viscosity calibration curves for each albumin species were used to determine the amount of protein desorbed from the particles during the settling process. The larger solution viscosity for BSA-coated particle experiments led to a much larger degree of desorption. HSA was shown to be the more stable albumin species when adsorbed onto polystyrene particles. Temperature denaturing was performed to aid in the determination of the stability of BSA. Reversible and irreversible conformational changes in BSA were produced at 46ºC and 76ºC respectively. The solutions were cooled to room temperature before adsorption ontopolystyrene particles and the sedimentation velocities measured. A 50% difference in average viscosity between the reversibly and irreversibly changed BSA was found. This caused the larger aggregates formed in the 76ºC BSA experiments to have an almost equivalent sedimentation velocity to those in the reversibly denatured BSA experiments. Average aggregate size for reversibly denatured BSA was well within the ranges found for non-denatured BSA. In conclusion, irreversibly denatured BSA formed larger aggregates and was more likely to desorb from the polystyrene particles than reversibly changed BSA. / Master of Science

Verankerte Polymere können die Funktionalität einer Oberfläche beeinflussen. Der Schwerpunkt dieser Arbeit liegt auf der Untersuchung von Polymerbürsten aus Polyacrylsäure (PAA) und Poly(N,N-dimethylaminoethyl methacrylat) (PDMAEMA). Auf Oberflächen, die mit PAA Bürsten beschichtet sind, können Proteine im nativen Zustand immobilisiert werden. Für PDMAEMA ist eine Reaktion auf externe Reize bekannt. So kann dessen untere kritische Lösungstemperatur (LCST) zur Einstellung der Hydrophobizität von Oberflächen verwendet werden. Erstmalig im Rahmen dieser Arbeit wurde untersucht, wie sich hydrostatischer Druck von bis zu 1000 bar auf die Funktionalität der verwendeten Polymerbürsten auswirkt. Aus Diblock-Kopolymeren wurden Langmuir-Filme unterschiedlicher Ankerdichte mit der Langmuir-Schäfer Technik auf feste Substrate übertragen. Die Funktionalität der PAA Bürsten wurde vor und nach der Adsorption von Rinderserumalbumin (BSA) in gepufferter D2O-Lösung mit Hilfe der Neutronenreflektometrie (NR) bei 1 bar und 900 bar an der fest-flüssig Grenzfläche untersucht. Es wurden Volumenfraktionsprofile der PAA Bürste und adsorbierten BSA extrahiert, woraus sich eine lineare Abhängigkeit zwischen Ankerdichte und Menge an adsorbiertem Protein feststellen ließ. Erhöhung des hydrostatischen Druckes auf 900 bar veränderte weder die PAA Volumenprofile noch die Immobilisierung von BSA. Die PDMAEMA Bürsten wurden mittels NR bei Temperaturen von 20-60 °C und Drücken von 1-1000 bar untersucht. Zur Analyse der Daten wurde ein neuartiges Dichteprofil-Modell verwendet. Temperaturerhöhung führt zur stetigen Abnahme der Bürstendicke. Dies lässt sich durch den LCST induzierten Phasenübergang der Polymere vom hydrophilen in einen hydrophoben Zustand erklären. Es wurde gefunden, dass eine Erhöhung des hydrostatischen Druckes diesem Prozess entgegenwirkt. Strukturänderungen der Polymerbürsten bei Erhöhung der Temperatur um 10 K ließen sich durch Erhöhung des Druckes um 1000 bar rückgängig machen. / The functionality of an interface can be modified by polymer brushes. The focus of this work is on brushes of either polyacrylic acid (PAA) or poly(N,N-dimethylaminoethyl methacrylate) (PDMAEMA). PAA brushes provide a soft interface that prevents the denaturation of adsorbed proteins. PDMAEMA is known to respond to external stimuli. The lower critical solution temperature (LCST) of PDMAEMA can be used to tune the hydrophobicity of the interface with temperature. For the first time, the effect of elevated hydrostatic pressure, up to 1000 bar, on the functionality of these systems is investigated. Planar PAA and PDMAEMA brushes are prepared from precursor diblock copolymer Langmuir layers with varied grafting density utilizing the Langmuir-Schäfer transfer technique. For solvent-swollen PAA brushes, neutron reflectivity (NR) measurements are conducted at the solid-liquid interface after incubation in buffered D2O and after the adsorption of bovine serum albumin (BSA) from the aqueous liquid phase at 1 bar and 900 bar. Detailed volume fraction profiles of the PAA brush and adsorbed BSA proteins are extracted. The amount of adsorbed BSA is found to scale linearly with grafting density. An elevated hydrostatic pressure of 900 bar is found to have no impact on the structure of the PAA brush and its capability to bind BSA proteins. The PDMAEMA brushes are investigated by NR at the solid-liquid interface in a temperature range of 20 to 60 °C for hydrostatic pressures from 1 to 1000 bar. A novel theoretical model of the brush density profile is used to fit the experimental NR data. Increasing the temperature causes a continuous decrease of the polymer brush thickness due to a hydrophobic coil to globule transition of the polymer chains when crossing the LCST. Hydrostatic pressure is found to act antagonistic to temperature. The hydrophobic collapse of the PDMAEMA brush caused by a temperature increase of 10 K is counterbalanced by a pressure increase of 1000 bar.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-07-25T13:32:13Z No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-26T12:05:00Z (GMT) No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-26T12:05:00Z (GMT). No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2002-12-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The main function of the most superficial layer of the epidermis, the Stratum Corneum (SC), is to provide a physical barrier that controls the transepidermal water loss as well as the permeation of another substances in both directions across the skin. The SC is formed by anabolically dead cells, the terminally differentiated corneocyte, and its function is essentially accomplished by forming a highly insoluble protein structure on the surface of the corneocytes, termed the cornified cell envelope, and by impeding water diffusion across the SC by mortaring the corneocytes together by layers of skin-specific lipids, essentially ceramide, cholesterol and fatty acid. In this work the cell envelope of the SC was spin labeled with a sulfhydryl-specific nitroxide reagent to investigate the water content effects upon the protein dynamics directly in the intact tissue. A two-state model for the nitroxide side chain described the coexistence of two spectral components in the electron paramagnetic resonance (EPR) spectra. The so-called strongly immobilized component, S, is associated with the EPR signal of a motionally restricted nitroxide fraction having its N-O group hydrogen bonded to protein (rigid structure) while the weakly immobilized component, W, corresponds to the signal provided by the spin labels with higher mobility (~10 times greater) exposed to the aqueous environment. The relative populations between these two mobility states, S and W, are in thermodynamic equilibrium. The standard Gibbs free energy, enthalpy and entropy changes for transferring the nitroxide side chain from the state contacting the solvent, W, to the one contacting protein, S, indicated that the reduction of the SC water content to below ~h 0.69, g H2O per g dry SC, stabilizes the protein interacting state, S. Upon decreasing the SC hydration level below ~h 0.69 the segmental motion of the polypeptide chains and the rotational motion of the spin-labeled side chain were also constrained. To test our methodology in a pure and very well known protein, we also studied the effects of two types of detergents on the bovine serum albumin (BSA). Both detergents, the anionic sodium dodecyl sulfate (SDS) and the zwitterionic N-hexadecyl-N,N-dimethyl-3-ammonium-1-propanesulfonate (HPS) increase the mobility of the protein backbone and of the nitroxide side chain. The thermodynamic parameters indicated that these detergents destabilize the protein favoring less compact conformations. This work can also be useful to improve the spectral analysis of site-directed spin labeling, especially for a more quantitative description in terms of thermodynamic parameters. / A camada mais superficial da epiderme, o Estrato Córneo (EC), tem como função principal a formação de uma barreira física que controla a perda de água do corpo bem como a permeação de outras substâncias em ambas as direções da pele. O EC é formado por células anabolicamente mortas, os corneócitos, os quais sofreram diferenciação celular terminal, e sua função é realizada formando uma estrutura de proteínas altamente insolúveis na superfície do corneócito, chamada de envelope celular, e também uma matriz lipídica, essencialmente ceramídios, colesterol e ácidos graxos, que dificultam a difusão da água. Neste trabalho, o EC foi marcado com marcadores de spin específicos para reagir com os grupos sulfidrilas das proteínas, para investigar os efeitos do conteúdo de água na dinâmica de proteínas diretamente no tecido intacto. Um modelo de dois estados para a cadeia lateral do nitróxido descreveu a coexistência de duas componentes espectrais de ressonância paramagnética eletrônica (RPE). A componente denominada fortemente imobilizada (S), surge de uma fração de marcadores com o átomo de oxigênio do nitróxido ligado à proteína (estrutura rígida) enquanto a componente fracamente imobilizada é gerada pelos marcadores com mobilidade mais alta (~10 vezes maior) e expostos ao ambiente aquoso. As populações relativas entre estes dois estados de mobilidade, S e W, estão em equilíbrio termodinâmico. Os parâmetros da termodinâmica: energia livre padrão de Gibbs, entalpia e entropia, envolvidos na transferência da cadeia lateral do nitróxido do estado W, contatando ao solvente, para o estado S, contatando a proteína, indicaram que a redução do conteúdo de água para abaixo de ~0.69g de H2O por g de EC seco, estabiliza o estado S (cadeia lateral do nitróxido dobrada sobre a cadeia principal da proteína). Ao diminuir o nível de hidratação para abaixo de ~ h 0.69 (g H2o/g EC seco) o movimento local da cadeia polipeptídica e o movimento rotacional da cadeia lateral do marcador de spin foram ambos reduzidos. Para testar nossa metodologia em uma proteína pura e bem conhecida, estudamos os efeitos de dois tipos de detergentes sobre a albumina do soro bovino (BSA). Ambos os detergentes, o aniônico dodecil sulfato de sódio (SDS) e o ziteriônico N-hexadecil-N,N-dimetil-3-amônio-1-propanosulfonato (HPS) aumentaram a mobilidade da cadeia principal da proteína e da cadeia lateral do nitróxido. Os parâmetros termodinâmicos indicaram que estes detergentes desestabilizam a proteína favorecendo conformações menos compactas. Os resultados do presente trabalho também podem contribuir para aprimorar a

<p>I ett samhälle där upphovsrätten och immaterialrätten blir allt mer uppmärksammad är det viktigt för företag att vara medvetna om sitt innehav av licenser för mjukvaror. I detta arbete kommer företags medvetenhet om deras licenshanteringssituation att undersökas. I den här rapporten kommer problemställningen att undersökas med hjälp av intervjuer med företag. Dessa intervjuer kommer att ske via telefon och per e-post. Ett liknande arbete har tidigare inte gjorts och därför känns det relevant samt intressant att göra denna undersökning.</p><p>Arbetet resulterade i intervjusvar från 11 företag. Det visade sig att företagens medvetenhet om deras licenshantering varierade stort, vilket även att storleken på företagen och innehav av en policy för licenshantering bland företagen gjorde. Metoden för licenshantering och licensinventering skiljde sig åt bland företagen.</p><p>Ett flertal program, tillammans med pärmar och Excel-listor var de dominerande licenshanteringsmetoderna bland företagen. Mjukvarutillverkarna och deras intresseorganisationer däribland BSA ställer krav på att företag som använder deras licensierade programvaror skall ha en strukturerad licenshantering. De ser gärna att företagen följer någon vedertagen metod för detta, en sådan metod är Software Asset Management, SAM.</p><p>Det är svårt att rekommendera någon licenshanteringsmetod och program som fungerar för samtliga företag, då användandet av dessa är väldigt företags- och organisationsspecifikt. De flesta större mjukvaror som finns i dagsläget följer en vedertagen metod för licenshantering och fungerar därmed bra att tillförlitligt inventera licenser med.</p> / <p>In a community where copyright and intellectual property law is getting more attention in the media, it is important for companies to be aware of their possession of software licenses. In this thesis we are going to investigate the awareness of the software licensing situation in some companies. To collect data that can be analyzed, we are interviewing companies. The interviews are performed by phone or e-mail. A similar report hasn’t been done before therefore it’s an interesting subject to look at.</p><p>The interviews resulted in answers from 11 companies. The answers showed that the awareness about software licensing among the companies had big variations. Some other things that varied among the companies were the size of the companies and the usage of software licensing policy within the companies. The way and method of handling software licensing also varied among the companies.</p><p>A lot of software programs, along with folders and Excel-lists dominated the method of handling software licenses among the companies. Software developing companies together with their interest organization for example BSA has demands on companies using their software to have a foreseeable documentation for the software licenses. They gladly see that the companies uses a best practice method to manage their license possessions, one best practice method is Software Asset Management, SAM.</p><p>It is hard to recommend a method for software license management to a non-specific company, because every company has a unique organization and computer network structure. Most of the big license management software is following a best practice method for software license management.</p>