This thesis is inspired by a persistent limitation in the use of composite biomaterials for orthopaedic applications, namely the agglomeration of reinforcing particles in these composites, which has resulted in poor mechanical properties. The use of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and hydroxyapatite (HA) nanoparticles to produce biodegradable nanocomposites is investigated. More specifically, the thesis investigates different methods of composite processing, and interfacial modifying agents and the effect that these have on the nano- and micro- scale structure of composites and their mechanical properties. PHBV and HA were chosen because PHBV is a biodegradable/biocompatible polymer and it has a relatively high stiffness when compared to other biodegradable polymers frequently used in orthopaedic applications. HA is chemically similar to ceramic phase found in bones and hard tissues and the inclusion of HA into biomedical materials has been shown to enhance the rate of osteoconduction. HA/PHBV composites were produced using different dispersing agents including poly(acrylic acid) (PAA), a model dispersing agent, polyethyleneimine (PEI) which allowed for the development of a single solvent system for composite preparation, and heparin (Hep), a macromolecule which is produced in vivo. Additionally, HA/PAA/PHBV composites were prepared from both sonicated and non-sonicated HA/PAA suspensions up to approximately 17% by weight (wt %) of HA content. Attempts to prepare composites with higher HA loadings led to inhomogeneous composite mixtures, which were caused by the dual solvent system used for the composite preparation. The HA/PEI/PHBV and HA/Hep/PHBV composites were produced up to approximately 75 wt % of HA content. It was found that the HA/PEI/PHBV and HA/Hep/PHBV composites could be prepared at higher loadings than HA/PAA/PHBV composites due to the single solvent system used for the preparation of the HA/PEI/PHBV composites and the better dispersion of HA/Hep particles in precursor suspensions. Finally, selected HA/PEI/PHBV composites were further processed using a twin screw extruder. All of the composites were characterised in terms of their dispersion levels as well as their compressive mechanical properties. In addition, HA/PEI/PHBV composite reinforced with 20 wt % of HA content was also tested for its mechanical properties using three different test types; compression, three-point bending, and tensile tests. Finally, the HA/PAA/PHBV, HA/PEI/PHBV, and HA/Hep/PHBV composites were tested their compressive mechanical properties in wet state. It was found that the sonicated HA/PAA suspensions in general had better colloidal stability than non-sonicated ones and that this yielded composites with superior compressive moduli than those prepared from non-sonicated suspensions. In addition, the better dispersion of the particles in the composites prepared from the sonicated suspensions, as confirmed by transmission electron microscopic (TEM) images, led to higher percentage crystallinities when compared to the composites prepared from non-sonicated suspensions. It is likely that the greater number of individual HA particles and smaller HA agglomerates observed in the composites prepared from sonication treatment are acting as nuclei for crystal growth more effectively than large HA agglomerates. The largest modulus and yield strength that could be achieved with this system were approximately 1.45 GPa and 80 MPa, respectively. Composites of HA/PEI/PHBV and HA/Hep/PHBV with approximately 55 wt % of HA content were found to exhibit the largest compressive moduli of approximately 2.5 and 2.8 GPa, respectively. Moreover, the yield strengths for the same materials were found to be approximately 123 and 120 MPa, respectively. This was found to correlate with the better levels of dispersion within the nanocomposites that could be achieved using these stabilisers. The extruded samples were found to have an even greater degree of particle dispersion when compared to the unextruded ones. This improved degree of particle dispersion of the extruded samples resulted in higher moduli in comparison to unextruded samples. The largest compressive modulus and yield strength of the extruded samples were found to be approximately 3.2 GPa and 125 MPa, respectively. The compressive moduli of the composites produced in this thesis are significantly greater than that of cancellous bone (0.4 GPa), but significantly lower than that of cortical bone (12.8–17.7 GPa). However, maximum yield strengths of the HA/PEI/PHBV and HA/Hep/PHBV composites match to cortical bone (120–180 MPa), which is a noteworthy finding in this thesis. The wet mechanical results of all composites as well as pure PHBV polymer showed a reduction in both moduli and yield strengths when compared to dry state. In addition, after 2 weeks in wet state both moduli and yield strengths of the composites and pure polymer converged to approximately the same values. Finally, the HA/PEI/PHBV composite samples tested by tensile testing showed the highest Young’s modulus and those tested by compression testing possessed the lowest Young’s modulus. This resulted from the difference in periods of time for heating exposure and void contents of the tested samples, which were prepared by different methods. However, toughness values obtained from the samples tested using three-point bending and tensile tests, was not significantly different.

Disponibiliza??o em conte?do parcial do trabalho, conforme Termo de Autoriza??o. / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-02-14T19:00:46Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lays_fernanda_nunes_dourado_parcial.pdf: 389665 bytes, checksum: 2289d8ac4ca389d5cdaf516878ddae78 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-03-06T12:24:50Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lays_fernanda_nunes_dourado_parcial.pdf: 389665 bytes, checksum: 2289d8ac4ca389d5cdaf516878ddae78 (MD5) / Made available in DSpace on 2017-03-06T12:24:50Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lays_fernanda_nunes_dourado_parcial.pdf: 389665 bytes, checksum: 2289d8ac4ca389d5cdaf516878ddae78 (MD5) Previous issue date: 2016 / Na ind?stria farmac?utica, os pol?meros s?o capazes de desempenhar a fun??o de matriz para libera??o controlada de f?rmacos. Isto tem como vantagem a minimiza??o dos efeitos adversos dos medicamentos e a otimiza??o do tratamento. Entre os pol?meros utilizados para tal finalidade, tem-se o poli(3-hidroxibutirato) (PHB) conhecido por sua origem natural, um material que se degrada a produtos n?o t?xicos para o organismo, o que garante o seu emprego como biomaterial. Quando utilizado em associa??o a outros pol?meros, s?o denominados Blendas, essas misturas modificam as caracter?sticas f?sico-qu?micas dos pol?meros para serem empregados em finalidades diversas. Desta forma este trabalho teve como objetivo o desenvolvimento de uma matriz polim?rica capaz de ser utilizada para libera??o controlada de f?rmacos. Para tal, foram produzidas duas membranas diferentes uma de PHB e outra de Blenda Poli (3-hidroxibutirato)/Polipropilenoglicol, PHB/PPG (90:10), com concentra??es diferentes de f?rmaco, contendo 5% e 25% em massa. O f?rmaco modelo foi a Sinvastatina. Estes dispositivos foram produzidos por dissolu??o seguida de evapora??o do solvente, formando filmes de 0,5 mm de di?metro que foram analisados quanto a sua degrada??o in vitro e in vivo, em implantes subcut?neos. Os materiais foram analisados utilizando espectroscopia na regi?o do infravermelho, termogravimetria, microscopia eletr?nica de varredura e por testes histol?gicos. Os filmes ? base de PHB, apresentaram degrada??o in vitro e in vivo mais lenta quando comparados ?s blendas, no entanto em ambos os casos, os materiais que continham maior percentual de Sinvastatina apresentaram maior degrada??o e libera??o do f?rmaco. As l?minas histol?gicas revelam a presen?a do tecido em torno dos dispositivos e aus?ncia de inflama??o o que comprova a biocompatibilidade dos materiais estudados. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / In the pharmaceutical industry, the polymers are able to drive the controlled release of drugs. Its brings the advantage of minimizing adverse effects of medicines and optimization of treatment. Among the polymers used for this purpose the poly (3-hydroxybutyrate) has known for their natural origin, its degradation to non-toxic effects to the life tissue which ensures their use as subcutaneous implants. A blend is a mixture of different polymers, these mixtures are able to modify the physicochemical characteristics of the polymers for several uses. This study aimed to develop a polymer matrix can be used as a way for controlled release of drugs. For this purpose, two different membranes were produced, the PHB and a Poly(3-hydroxybutyrate)/Polypropylene glycol blends, PHB/PPG (90:10), with different concentrations of drug with 5% and another 25%. The simvastatin was chosen as a model. These devices were manufactured by casting to form films of 0.5 mm diameter that were analyzed for in vitro and in vivo degradation tests using subcutaneous implants. The materials were analyzed using infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy and histological tests. The films based on PHB presented a slower degradation in vitro and in vivo degradation tests when compared to the blends, however in both cases those materials containing a higher percentage of simvastatin has shown the higher degradation and release of the drug. The histological sections showed a tissue colonization and absence of inflammation which demonstrates the biocompatibility of the implants.

Estratégias para minimizar o efeito do balanço energético negativo (BEN) em vacas leiteiras vêm sendo estudadas constantemente no passar dos anos, pois o período de transição é caracterizado por grande estresse metabólico. O objetivo deste estudo foi avaliar parâmetros metabólicos, produtivos e reprodutivos em vacas da raça Holandesa após a aplicação de drench no pós-parto imediato e 24 h após, como alternativa para minimizar o BEN e proporcionar melhor desempenho produtivo e reprodutivo aos animais. Foram utilizadas 129 vacas, 64 no grupo 1 (1ª e 2ª lactação) e 65 no grupo 2 (≥ 3ª lactação), distribuídas em: verão (n= 63) e inverno (n= 66) e região da Serra Gaúcha (Fazenda A, n= 48) e Planato Médio (Fazenda B, n= 81). Foram coletadas amostras de sangue para obtenção de soro e plasma e determinação de β-hidroxibutirato (BHB) com sistema Abbot Precision Xtra. Foi medido o escore de condição corporal e a temperatura corporal, e realizado acompanhamento de controle leiteiro para avaliar produção e composição de leite. Foi realizada ultrassonografia ovariana para avaliação de ciclicidade ovárica e primeira ovulação pós-parto. O tratamento drench provocou uma tendência a diminuir os valores de BHB sérico no período experimental, sendo que, no período de verão essa diminuição foi significativa na Fazenda A. Na Fazenda B, nas semanas 3 e 4, o tratamento com drench aumentou significativamente a produção de leite nos grupos 1 e 2. A exceção foi no grupo 1 no período de verão, onde o efeito foi negativo. Também na Fazenda B, o tratamento com drench teve efeito positivo no percentual de gordura nas vacas do grupo 1 e efeito negativo nas vacas do grupo 2. Parâmetros reprodutivos não tiveram interações significativas. Em conclusão, o tratamento com drench preveniu a ocorrência de cetose subclínica, aumentou a produção leiteira no inverno e no verão em vacas de 3 ou mais lactações e também aumentou o percentual de gordura no leite em vacas de uma ou duas lactações. / Strategies to minimize the effect of negative energy balance (NEB) in dairy cows are being studied constantly over the years, since the transition period is characterized by excessive metabolic stress. The present study had the objective to evaluate metabolic, productive and reproductive parameters in dairy cows after application of drench in the immediate postpartum period and 24 hours after as an alternative to minimize NEB providing better productive and reproductive performance of animals. 129 cows, 64 were used in group 1 (1st and 2nd lactation ) and 65 in group 2 ( ≥ 3rd lactation ), distributed in: summer ( n = 63 ) and winter ( n = 66 ) and Serra Gaucha (Farm A, n = 48) and Planato Médio (Farm B, n = 81 ) . Blood samples were collected to obtain serum and plasma and determination of β-hydroxybutyrate (BHB) with the Abbott Precision Xtra system . Body condition score and body temperature, were also obtained by milk information system to evaluate production and milk composition was measured. Ovarian ultrasonography was performed for evaluation of cyclicity and days to first ovulation postpartum. The drench treatment had a trend towards significance in the reduction of mean serum BHB during the experimental period, whereas in the summer period this decrease was significant in farm A. On farm B at weeks 3 and 4, drench significantly increased milk production in groups 1 and 2. Except for group 1 during the summer, where the effect was negative. Also on farm B, drench had a positive effect on fat percentage in cows of group 1 and negative effect on cows in group 2. Reproductive parameters showed no significant interactions. In conclusion, drench prevented the occurrence of subclinical ketosis, increased milk production during winter and during summer in cows of third or more lactation and also increased of milk fat percentage in cows of first and second lactations.

Estratégias para minimizar o efeito do balanço energético negativo (BEN) em vacas leiteiras vêm sendo estudadas constantemente no passar dos anos, pois o período de transição é caracterizado por grande estresse metabólico. O objetivo deste estudo foi avaliar parâmetros metabólicos, produtivos e reprodutivos em vacas da raça Holandesa após a aplicação de drench no pós-parto imediato e 24 h após, como alternativa para minimizar o BEN e proporcionar melhor desempenho produtivo e reprodutivo aos animais. Foram utilizadas 129 vacas, 64 no grupo 1 (1ª e 2ª lactação) e 65 no grupo 2 (≥ 3ª lactação), distribuídas em: verão (n= 63) e inverno (n= 66) e região da Serra Gaúcha (Fazenda A, n= 48) e Planato Médio (Fazenda B, n= 81). Foram coletadas amostras de sangue para obtenção de soro e plasma e determinação de β-hidroxibutirato (BHB) com sistema Abbot Precision Xtra. Foi medido o escore de condição corporal e a temperatura corporal, e realizado acompanhamento de controle leiteiro para avaliar produção e composição de leite. Foi realizada ultrassonografia ovariana para avaliação de ciclicidade ovárica e primeira ovulação pós-parto. O tratamento drench provocou uma tendência a diminuir os valores de BHB sérico no período experimental, sendo que, no período de verão essa diminuição foi significativa na Fazenda A. Na Fazenda B, nas semanas 3 e 4, o tratamento com drench aumentou significativamente a produção de leite nos grupos 1 e 2. A exceção foi no grupo 1 no período de verão, onde o efeito foi negativo. Também na Fazenda B, o tratamento com drench teve efeito positivo no percentual de gordura nas vacas do grupo 1 e efeito negativo nas vacas do grupo 2. Parâmetros reprodutivos não tiveram interações significativas. Em conclusão, o tratamento com drench preveniu a ocorrência de cetose subclínica, aumentou a produção leiteira no inverno e no verão em vacas de 3 ou mais lactações e também aumentou o percentual de gordura no leite em vacas de uma ou duas lactações. / Strategies to minimize the effect of negative energy balance (NEB) in dairy cows are being studied constantly over the years, since the transition period is characterized by excessive metabolic stress. The present study had the objective to evaluate metabolic, productive and reproductive parameters in dairy cows after application of drench in the immediate postpartum period and 24 hours after as an alternative to minimize NEB providing better productive and reproductive performance of animals. 129 cows, 64 were used in group 1 (1st and 2nd lactation ) and 65 in group 2 ( ≥ 3rd lactation ), distributed in: summer ( n = 63 ) and winter ( n = 66 ) and Serra Gaucha (Farm A, n = 48) and Planato Médio (Farm B, n = 81 ) . Blood samples were collected to obtain serum and plasma and determination of β-hydroxybutyrate (BHB) with the Abbott Precision Xtra system . Body condition score and body temperature, were also obtained by milk information system to evaluate production and milk composition was measured. Ovarian ultrasonography was performed for evaluation of cyclicity and days to first ovulation postpartum. The drench treatment had a trend towards significance in the reduction of mean serum BHB during the experimental period, whereas in the summer period this decrease was significant in farm A. On farm B at weeks 3 and 4, drench significantly increased milk production in groups 1 and 2. Except for group 1 during the summer, where the effect was negative. Also on farm B, drench had a positive effect on fat percentage in cows of group 1 and negative effect on cows in group 2. Reproductive parameters showed no significant interactions. In conclusion, drench prevented the occurrence of subclinical ketosis, increased milk production during winter and during summer in cows of third or more lactation and also increased of milk fat percentage in cows of first and second lactations.

Estratégias para minimizar o efeito do balanço energético negativo (BEN) em vacas leiteiras vêm sendo estudadas constantemente no passar dos anos, pois o período de transição é caracterizado por grande estresse metabólico. O objetivo deste estudo foi avaliar parâmetros metabólicos, produtivos e reprodutivos em vacas da raça Holandesa após a aplicação de drench no pós-parto imediato e 24 h após, como alternativa para minimizar o BEN e proporcionar melhor desempenho produtivo e reprodutivo aos animais. Foram utilizadas 129 vacas, 64 no grupo 1 (1ª e 2ª lactação) e 65 no grupo 2 (≥ 3ª lactação), distribuídas em: verão (n= 63) e inverno (n= 66) e região da Serra Gaúcha (Fazenda A, n= 48) e Planato Médio (Fazenda B, n= 81). Foram coletadas amostras de sangue para obtenção de soro e plasma e determinação de β-hidroxibutirato (BHB) com sistema Abbot Precision Xtra. Foi medido o escore de condição corporal e a temperatura corporal, e realizado acompanhamento de controle leiteiro para avaliar produção e composição de leite. Foi realizada ultrassonografia ovariana para avaliação de ciclicidade ovárica e primeira ovulação pós-parto. O tratamento drench provocou uma tendência a diminuir os valores de BHB sérico no período experimental, sendo que, no período de verão essa diminuição foi significativa na Fazenda A. Na Fazenda B, nas semanas 3 e 4, o tratamento com drench aumentou significativamente a produção de leite nos grupos 1 e 2. A exceção foi no grupo 1 no período de verão, onde o efeito foi negativo. Também na Fazenda B, o tratamento com drench teve efeito positivo no percentual de gordura nas vacas do grupo 1 e efeito negativo nas vacas do grupo 2. Parâmetros reprodutivos não tiveram interações significativas. Em conclusão, o tratamento com drench preveniu a ocorrência de cetose subclínica, aumentou a produção leiteira no inverno e no verão em vacas de 3 ou mais lactações e também aumentou o percentual de gordura no leite em vacas de uma ou duas lactações. / Strategies to minimize the effect of negative energy balance (NEB) in dairy cows are being studied constantly over the years, since the transition period is characterized by excessive metabolic stress. The present study had the objective to evaluate metabolic, productive and reproductive parameters in dairy cows after application of drench in the immediate postpartum period and 24 hours after as an alternative to minimize NEB providing better productive and reproductive performance of animals. 129 cows, 64 were used in group 1 (1st and 2nd lactation ) and 65 in group 2 ( ≥ 3rd lactation ), distributed in: summer ( n = 63 ) and winter ( n = 66 ) and Serra Gaucha (Farm A, n = 48) and Planato Médio (Farm B, n = 81 ) . Blood samples were collected to obtain serum and plasma and determination of β-hydroxybutyrate (BHB) with the Abbott Precision Xtra system . Body condition score and body temperature, were also obtained by milk information system to evaluate production and milk composition was measured. Ovarian ultrasonography was performed for evaluation of cyclicity and days to first ovulation postpartum. The drench treatment had a trend towards significance in the reduction of mean serum BHB during the experimental period, whereas in the summer period this decrease was significant in farm A. On farm B at weeks 3 and 4, drench significantly increased milk production in groups 1 and 2. Except for group 1 during the summer, where the effect was negative. Also on farm B, drench had a positive effect on fat percentage in cows of group 1 and negative effect on cows in group 2. Reproductive parameters showed no significant interactions. In conclusion, drench prevented the occurrence of subclinical ketosis, increased milk production during winter and during summer in cows of third or more lactation and also increased of milk fat percentage in cows of first and second lactations.

This master’s thesis deals with plasticization of poly(3-hydroxybutyrate), polylactid acid and their blend. It explores effect of chemical structure of plasticizer on mechanical properties of this polymer blend and on its diffusion from the polymer blend. Syntheses of plasticizers based on oligomeric polyadipates, citrates, lactate and esters of 2 ethylhexanoic acid with poly(ethyleneglycol) were carried out. Molecular weight distribution of synthesized plasticizers was determined using gel permeation chromatography. Poly(3-hydroxybutyrate), polylactid acid and their blend were plasticized with synthesized and commercial plasticizers. From commercial, chosed plasticizers were based on citrates and ester of 2-ethylhexanoic acid with poly(ethyleneglycol). Thermal stability of selected commercial plasticizers in polylactid acid was studied using thermogravimetry. Diffusion of plasticizers from poly(3-hydroxybutyrate), polylactid acid and their blend during exposure to 110 °C was also investigated. Mechanical properties of prepared blends were tested by tensile test. Almost all used plasticizers showed positive softening effect in blend. The highest elongation at break was detected for the blend with commercial acetyltributylcitrate, where elongation at break reached 328 % relative to 21 % for neat non-plasticized blend.

Objective: The aim was to summarize the clinical literature on interactions between common illicit drugs and prescription therapies. Methods: Medline, Iowa Drug Information Service, International Pharmaceutical Abstracts, EBSCO Academic Search Premier, and Google Scholar were searched from date of origin of database to March 2011. Search terms were cocaine, marijuana, cannabis, methamphetamine, amphetamine, ecstasy, N-methyl-3,4- methylenedioxymethamphetamine, methylenedioxymethamphetamine, heroin, gamma-hydroxybutyrate, sodium oxybate, and combined with interactions, drug interactions, and drugdrug interactions. This review focuses on established clinical evidence. All applicable full-text English language articles and abstracts found were evaluated and included in the review as appropriate. Results: The interactions of illicit drugs with prescription therapies have the ability to potentiate or attenuate the effects of both the illicit agent and/or the prescription therapeutic agent, which can lead to toxic effects or a reduction in the prescription agent's therapeutic activity. Most texts and databases focus on theoretical or probable interactions due to the kinetic properties of the drugs and do not fully explore the pharmacodynamic and clinical implications of these interactions. Clinical trials with coadministration of illicit drugs and prescription drugs are discussed along with case reports that demonstrate a potential interaction between agents. The illicit drugs discussed are cocaine, marijuana, amphetamines, methylenedioxymethamphetamine, heroin, and sodium oxybate. Conclusion: Although the use of illicit drugs is widespread, there are little experimental or clinical data regarding the effects of these agents on common prescription therapies. Scientific Significance: Potential drug interactions between illicit drugs and prescription drugs are described and evaluated on the Drug Interaction Probability Scale by Horn and Hansten.

Growing concerns of the negative effects on the environment and dependency of fossil fuelsare major driving forces for finding novel sustainable production pathways for plastic.Metabolic engineering has emerged as a powerful tool to enable microorganisms to producenon-native metabolites. The aim of this project was recombinant production of 4-hydroxybutyrate (4-HB) by expressing two enzymes in the model organism Escherichia coli.α-ketoglutarate decarboxylase (SucA) from Mycobacterium smegmatis followed by 4-hydroxybutyrate dehydrogenase (4-HBd) from Clostridium kluyveri was expressed inEscherichia coli. Results showed that the genes were successfully transformed and expressedin E. coli and after protein purification a concentration of 0.9 g/L SucA and 9.8 g/L 4-HBdwas achieved. Furthermore, some protein activity was detected by a coupled reaction withSucA and 4-HBd. When the enzymes got coupled together a change in NADH concentrationcould be detected spectrophotometrically. The enzymes were also tested for substratespecificity by using substrates with various carbon chain lengths and a decrease in NADHconcentration was seen. However, a decrease in the negative control for the experiments wasalso seen indicating a breakdown of NADH over time rather than consumption. Therefore, noconclusion could be drawn about the promiscuity of the enzymes. Lastly a single plasmidssystem was tested where both the genes were ligated on the same plasmid (pCDF duet) andexpressed successfully in E. coli Bl21DE3. / Ökad oro för miljön samt behovet av fossila resurser för produktion av plaster har gjort detnödvändigt att skapa nya och mer hållbara produktions vägar. Genetisk modifikation av olikaorganismer har utvecklats som ett starkt redskap för att få mikroorganismer att framställametaboliter som de normalt inte producerar. Målet med detta projekt var rekombinantproduktion av gamma hydroxibutansyra (4-HB) genom att uttrycka två enzym i modellorganismen Escherichia coli. Dessa enzym bestod av α-ketoglutarat dekarboxylas (SucA) frånMycobacterium smegmatis samt 4-hydroxybutyrate dehydrogenas (4-HBd) från Clostridiumkluyveri. Resultaten visade att proteinerna lyckades utryckas i E. coli med en koncentration av0,9 g/L SucA och 9,8 g/L 4-HBd som uppnåddes efter rening. Utöver detta detekterades ävenviss enzymaktivitet genom att kopplad enzymreaktion mellan 4-HBd och SucA och mätakonsumtionen av NADH spektrofotometriskt över tid. Enzymen testades även försubstratspecificitet genom att köra reaktionen med substrat med olika längd på kolkedjan. Dåkunde en minskning i NADH koncentrationen ses men det gjordes det även för de negativakontrollerna vilket indikerar nedbrytning av NADH och inte konsumtion av NADH. Ingaslutsatser angående enzymens substratspecificitet kunde därför dras. Det sista som gjordes varatt sätta in båda generna i ett en plasmidsystem där båda generna sattes in på samma plasmid(pCDF duet) och uttrycktes framgångsrikt i E. coli Bl21DE3.

Cognitive impairments are an increasing health problem worldwide. In the developed countries, the average life expectancy has dramatically increased over the last decades, and with an elderly population more cases of cognitive impairments appear. Age, genetics, and different medical conditions such as diabetes mellitus, and substance use disorders may all contribute to declined cognitive ability. Physiological functions also decrease with increasing age, as does the activity of the growth hormone (GH)/insulin-like growth factor-1 (IGF-1) axis. Interestingly, both GH and IGF-1 are recognized for their neuroprotective effects and cognitive enhancement. The overall aim of this thesis was to investigate the impact of the somatotrophic axis (i.e. GH/IGF-1 axis) in rodents with cognitive deficiencies induced by diabetes or long-term drug exposure. For the first time cognitive impairments were characterized in diabetic mice using a spatial learning and memory task called the Barnes maze (BM). In diabetic mice, impaired learning in the BM was associated with decreased expression of the GH receptor (GHR) in the frontal cortex, a region important for e.g. working memory. Treatment with GH reversed certain cognitive impairments seen in diabetic animals. In rats treated with gamma-hydroxybutyrate (GHB), a significant decrease of Igf1 mRNA expression in the frontal cortex was observed. This observation may explain the impaired cognitive function previously seen following GHB administration. Furthermore, rats exposed to chronic morphine delivered in mini-osmotic pumps displayed memory impairments in the Morris water maze (MWM), an effect that seems to be associated with the composition of the N-methyl-d-aspartate (NMDA) receptor complex in the frontal cortex. In conclusion, the result strengthens the evidence for GH being a cognitive enhancer. Moreover, the result within this thesis identifies the frontal cortex as an important brain region, where gene expression related to the somatotrophic system is affected in rodents with cognitive impairments. The thesis especially emphasizes the importance of the local somatotrophic system in the brain with regard to cognitive function.

O objetivo deste trabalho foi o estudo das propriedades mecânicas e térmicas do PHB e de compósitos de PHB/pó de madeira processados, reprocessados e irradiados. Foram preparados compósitos com PHB e pó de madeira com concentrações de PHB/pó de madeira de 90/10, 80/20 e 70/30 (m/m). Foram realizados ensaios de propriedades mecânicas, HDT, ponto de amolecimento Vicat, TGA e DSC. A incorporação do pó de madeira aumentou o grau de cristalinidade e a temperatura de cristalização do polímero, e nos compósitos PHB/pó de madeira 80/20 e 70/30 a rigidez do material aumentou em relação ao polímero puro. O compósito PHB/pó de madeira 70/30 apresentou os melhores resultados em termos econômicos, de processamento e de estabilidade em relação ao reprocessamento. A irradiação após o processamento, em dose de 30 kGy, provocou aumento da rigidez do PHB puro e dos compósitos PHB/pó de madeira 90/10 e 80/20, embora as outras propriedades tenham decrescido. O compósito PHB/pó de madeira 70/30 apresentou queda de todas as propriedades com a irradiação. / The purpose of this work was the study of the mechanical and thermal properties of processed, reprocessed and irradiated PHB, as well as PHB/wood flour composites. PHB/wood flour composites were prepared with PHB/wood relation of 90/10, 80/20 e 70/30 (w/w). Mechanical properties, HDT, Vicat melting point and thermo analyses (TGA and DSC) tests were carried out. The introduction of the wood flour increased both polymer crystallinity and crystallization temperature. The material stiffness increased in PHB/wood flour composites (80/20 and 70/30) resulting in higher values in some properties when compared with pure PHB. The PHB/wood flour composite 70/30 showed the best results in terms of economy, processing and stability during reprocessing. The irradiation after processing in 30kGy doses led to a stiffness increasing of pure PHB and PHB/wood flour composites (90/10 and 80/20) while other properties have decreased. The PHB/wood flour 70/30 showed decreasing in all properties after irradiation.