<p>The acceptance of blood contacting implants creating favorable conditions <i>in vivo</i> is decisively determined by their interaction with proteins that mediate inter cellular interactions with synthetic substrates. Adsorbed proteins can activate blood cascade systems like coagulation and complement that may result in serious blood clots, and/or immunological reactions. Poly (ethylene glycol) (PEG), heparin, and phosphoryl choline (PC) functional poly (methacrylates) are previously used polymers with known non-adhesive properties in blood contacting events.</p><p>This thesis contributes to this extensive research by introducing a novel type of biomaterial that equips biodegradable polymers with biomimetic functionalities. The phospholipid mimetic material is synthesized by combining biodegradable polymers with various functional polar end-groups consisting of zwitterionic phosphoryl choline (PC), anionic succinates, and cationic quaternary ammonium. The polymer backbone provides mechanical stability and biodegradability whilst the various head groups provide a variety of functions. The careful evaluation of the synthesis has allowed reaction conditions to be optimized leading to complete conversion at each step and subsequently high yields. Initially, poly (e-caprolactone) (PCL) was used since it provided a suitable synthetic starting point. However, the synthesis has also included poly (trimethylene carbonate) (PTMC) to provide a material that allows spontaneous surface enrichment of the polar PC group. This was achieved with an added hydrophilic environment. </p><p>Through the synthesis of multi PC functional PTMC, additional bulk organisation by the formation of zwitterionomers (PC ionomer) was achieved. Low modulus elasticity and water uptake were some of the properties of the formed material. As a result it was shown that the PC ionomer could be used for protein/drug loading and subsequent release. Furthermore, the material possessed non-adhesive properties in different biological environments.</p><p>Importantly, the result suggests that a versatile synthetic platform has been established that may provide a smorgasbord of different functional polymers, or combinations of such. This is indeed important since it was shown that the polymer in many ways dictates how the material may take advantage of an added functionality. </p><p>Such materials should be interesting for a variety of biomedical applications including the production of soft hemocompatible tissue.</p>

<p>The interactions between polymers and components of biological systems are an important area of interest within the fields of tissue engineering, polymer chemistry, medicine and biomaterials. In order to create such a biomimetic material, it must show the inherent ability to reproduce or elicit a biological function. How do we design synthetic materials in order to direct their interactions with biological systems?</p><p>This thesis contributes to this research with aspects of how polymers interact with biological materials with the help of ionic interactions. Polyesters, biodegradable or not, may after a hydrolytic cleavage interact ionically with protonated amines by the liberated carboxylate functions. Amines are found in proteins and this fact will help us to anchor proteins to polyester surfaces. Another type of interaction is to culture cells in polymeric materials, i.e. scaffolds. We have been working on compliant substrates, knitted structures, to allow cell culture in three dimensions. A problem that arises here is how to get a high cell seeding efficiency? By working on the interactions between polymers, proteins and finally cells, it is possible to create a polarized protein membrane that allows for very efficient cell seeding, and subsequent three dimensional cell cultures. Finally a synthetic route to taylor interaction was developed. Here a group of polymers known as ionomers were synthesized. In our case ionic end groups have been placed onto biodegradable polycarbonates, we have created amphiphilic telechelic ionomers. Functionalization, anionic or cationic, changes the properties of the material in many ways due to aggregation and surface enrichment of ionic groups. It is possible to add functional groups for a variety of different interactions, for example introducing ionic groups that interact and bind to the complementary charge of proteins or on the other hand one can chose groups to prevent protein interactions, like the phosphorylcholine zwitterionomers. Such interactions can be utilized to modulate the release of proteins from these materials when used in protein delivery applications. The swelling properties, Tg, degradation rate and mechanical properties are among other things that will easily be altered with the choice of functional groups or backbone polymer.</p>

Escherichia coli (E. coli) is an attractive host for production of recombinant proteins, since it generally provides a rapid and economical means to achieve high product quantities. In this thesis, the impact of the glucose uptake rate on the production of recombinant proteins was studied, aiming at improving and optimising production of recombinant proteins in E. coli. E. coli can be cultivated to high cell densities in bioreactors by applying the fed-batch technique, which offers a means to control the glucose uptake rate. One objective of this study was to find a method for control of the glucose uptake rate in small-scale cultivation, such as microtitre plates and shake flasks. Strains with mutations in the phosphotransferase system (PTS) where used for this purpose. The mutants had lower uptake rates of glucose, resulting in lower growth rates and lower accumulation of acetic acid in comparison to the wild type. By using the mutants in batch cultivations, the formation of acetic acid to levels detrimental to cell growth could be avoided, and ten times higher cell density was reached. Thus, the use of the mutant strains represent a novel, simple alternative to fed-batch cultures.   The PTS mutants were applied for production of integral membrane proteins in order to investigate if the reduced glucose uptake rate of the mutants was beneficial for their production. The mutants were able to produce three out of five integral membrane proteins that were not possible to produce by the wild-type strain. The expression level of one selected membrane protein was increased when using the mutants and the expression level appeared to be a function of strain, glucose uptake rate and acetic acid accumulation. For production purposes, it is not uncommon that the recombinant proteins are secreted to the E. coli periplasm. However, one drawback with secretion is the undesired leakage of periplasmic products to the medium. The leakage of the product to the medium was studied as a function of the feed rate of glucose in fed-batch cultivations and they were found to correlate. It was also shown that the amount of outer membrane proteins was affected by the feed rate of glucose and by secretion of a recombinant product to the periplasm. The cell surface is another compartment where recombinant proteins can be expressed. Surface display of proteins is a potentially attractive production strategy since it offers a simple purification scheme and possibilities for on-cell protein characterisation, and may in some cases also be the only viable option. The AIDA-autotransporter was applied for surface display of the Z domain of staphylococcal protein A under control of the aidA promoter. Z was expressed in an active form and was accessible to the medium. Expression was favoured by growth in minimal medium and it seemed likely that expression was higher at higher feed rates of glucose during fed-batch cultivation. A repetitive batch process was developed, where relatively high cell densities were achieved whilst maintaining a high expression level of Z. / QC 20110608

In Ethiopia industrial effluents containing high contents of organic matter, nitrogen and heavy metals are discharged into inland surface waters with little or no pre-treatment. Significant pollution concerns related to these effluents include dissolved oxygen depletion, toxicity and eutrophication of the receiving waters. This has not only forced the government to formulate regulations and standards for discharge limits but also resulted in an increasing interest and development of methods and systems by which wastewater can be recycled and used sustainably. The need for technologies for environmentally friendly treatment of industrial wastes such as tannery wastewaters is therefore obvious. Biological processes are not only cost effective but also environmentally sound alternatives to the chemical treatment of tannery wastewaters. The aim of the research presented in this thesis was to develop and optimize processes for biological nitrogen removal from tannery wastewaters and to identify the most efficient denitrifying organisms in tannery wastewaters laden with toxic substances. A pilot plant consisting of a predenitrification anoxic system, aerated nitrification compartment and a sedimentation tank (clarifier) all arranged in series was developed and installed on the premises of Addis Ababa University, Ethiopia. In spite of high influent chromium and sulphide perturbations over the successive feeding phases, the performance of the pilot plant was encouraging. The overall removal efficiency of the pilot plant over the experimental feeding phases varied between 82-98% for total nitrogen, 95-98% for COD, 96-98% for BOD5, 46-95% for ammonia nitrogen, 95-99% for sulphide and 93-99% for trivalent Chromium. Six isolates from over 1000 pure cultures were identified as the most efficient denitrifying bacteria. From both cellular fatty acid profiles and 16S rRNA gene sequencing, the six selected strains were phylogenetically identified as Brachymonas denitrificans in the β-subdivision of the Proteobacteria. All the six strains contain cd1-type nitrite reductase. The efficient isolates characterized in this study are of great value because of their excellent denitrifying properties and high tolerance to the concentrations of toxic compounds prevailing in tannery wastewaters. Bio-augmentation of the pilot plant with this bacterium showed a clear correlation between in situ denitrifying activities measured by nitrate uptake rate, population dynamics of the introduced B.denitrificans monitored by fluorescent in situ hybridization and the pilot plant performance, suggesting that the strategy of introducing this species for enhancing process performance has potential applications. Moreover, the nitrate-reducing, sulphur-oxidizing bacteria (NR-SOB) were also found in the pilot plant in abundance with steady sulphide removal efficiency during the study period. This could provide opportunities for the application of biologically mediated simultaneous removal of sulphide and nitrogen from tannery effluents. In addition to enriching high consortia of denitrifiers in the anoxic system to attain high denitrification efficiency and also improving the overall nitrification efficiency of the system, the predenitrification-nitrification pilot process plant stimulated the activity of indigenous NR-SOB to simultaneously remove sulphide from the system. Thus, the pilot plant was found to be operationally efficient for the removal of nitrogen, organic matter and other pollutants from tannery wastewaters. Keywords: Biological nitrogen and sulphide removal, denitrifying bacteria, nitrate-reducing, sulphur-oxidizing bacteria, nitrate uptake rate, fluorescent in situ hybridization, pollution, tannery effluents.

In schizophrenia, subtle aberrations in the brain cause functional disturbances like psychotic symptoms and social disability. There are, however, also disturbances outside the CNS indicating a systemic manifestation in the disease. The aim of the present thesis was to gain deeper understanding of the pathophysiological mechanisms underlying schizophrenia with a particular interest in peripheral and systemic manifestations with relevance for the increased risk of obesity and metabolic complications seen in the disease. Therefore, resting energy expenditure (REE), physical capacity, and relevant body composition variables were measured in patients with schizophrenia and in healthy controls. Also niacin skin flush response and electrodermal activity (EDA) were studied. Patients with schizophrenia exhibited significantly lower REE expressed as kJ/kg, and also lower values compared with predicted levels than the controls. The difference could not be attributed to medication or variations in body composition between the two groups. There was a gender difference with the lowest levels found in male patients. Male patients exhibited significantly lower physical capacity in terms of predicted maximal oxygen uptake capacity and faster increase in respiratory quotient than male controls. The oral niacin test revealed a significantly delayed skin flush reaction in patients compared with controls. The patients also exhibited lower EDA response. There was a significant association in response patterns for the niacin and the EDA tests in the patients, but not in controls. In a test-retest study in patients there was acceptable stability for EDA measures but low test-retest stability for niacin variables. The previously found association in responses for the two tests was, however, replicated. The results gain support for the concept of schizophrenia as a disease with systemic manifestations including metabolic dysregulation. The findings add to the understanding of the weight gain and the increased risk for cardiovascular morbidity seen in this condition.

<p><font face="Times New Roman" size="3"><font face="Times New Roman" size="3"> <p align="left">The aim of the study is to explore the barriers associated with low VCT uptake by the TB patients attending primary health care clinics within the Buffalo City municipality. <font face="Times New Roman" size="3"><font face="Times New Roman" size="3">The study population was drawn from TB patients attending the primary health care facilities in Buffalo city municipality in the Eastern Cape Province. Eight participants were purposively selected to include those who had accepted VCT as well as those who did not.</font></font></p> </font></font></p>

<p>Aim: To establish whether a family centered approach to HIV care in which HIV positive adults are counseled on the importance of having their children tested results in the adults bringing their children under the age of five years for testing and or accessing HIV care, and to explore challenges faced by caregivers in bringing children for testing and care.</p>

Lichens are fascinating symbiotic systems, where a fungus and a unicellular alga, most often green (bipartite green algal lichens; 90% of all lichens), or a fi lamentous cyanobacterium (bipartite cyanobacterial lichens; 10% of all lichens) form a new entity (a thallus) appearing as a new and integrated organism: in about 500 lichens the fungus is associated with both a cyanobacterium and an alga (tripartite lichens). In the thallus, the lichen bionts function both as individual organisms, and as a symbiont partner. Hence, in lichens, the participating partners must both be able to receive and acquire resources from the other partner(s) in a controlled way. Lichens are particularly successful in harsh terrestrial environments. In part this is related to their poikilohydric nature and subsequent ability to repeatedly become desiccated and hydrated. Metabolic activity, i.e. photosynthesis, respiration, and for cyanobacterial lichens N2-fixation, is limited to periods when the thallus is suffi ciently hydrated. Mineral nutrients are mainly acquired from dry or wet deposition directly on the thallus. Taken together it then appears that lichens are to a large extent passively controlled by their environment, making their control over resource allocation and acquisition particularly challenging. The aim of this thesis was to investigate resource acquisition and allocation processes in different lichens, and to see how these respond to changes in resource availability. This was done by following lichen growth in the fi eld during manipulation of water, light, and nutrient supply, and by assessing the responses of both the integrated thallus as well as the individual bionts. As a fi rst step, resource allocation and acquisition was investigated for a broad range of lichens aiming to determine the magnitude of metabolic variation across lichens. Seventy-fi ve lichen species were selected to cover as broad a spectrum as possible regarding taxonomy, morphology, habitat, and nitrogen requirements. The lichens had invested their nitrogen resources so that photosynthetic capacity matched respiratory carbon demand around a similar equilibrium across the contrasting species. Regulation of lichen growth was investigated in another study, using the two tripartite species Nephroma arcticum and Peltigera aphthosa, emphasizing the contribution of both internal and external factors. The empirical growth models for the two lichens were similar, showing that weight gain is to a higher extent dependent on those external factors that regulate their photosynthesis, whilst area gain is more controlled by internal factors, such as their nitrogen metabolism. This might be inferred from another study of the same species, where nitrogen manipulations resulted in an undisturbed weight gain, a similar resource allocation pattern between the bionts, but a distorted area gain. Aiming to investigate lichen nitrogen relations even further, lichens’ capacities to assimilate combined nitrogen in the form of ammonium, nitrate and amino acids were assessed using 14 contrasting boreal species. All these had the capacity to assimilate all the three nitrogen forms, with ammonium absorption being more passive, and nitrate uptake being low in bipartite cyanobacterial lichens. Differences in uptake capacities between species were more correlated to photobiont than to morphology or substrate preferences. Finally, to investigate intra-specifi c plasticity in relation to altered nutrient supply, resource investments between photo- and mycobiont were investigated in the two bipartite green algal lichens Hypogymnia physodes and and Platismatia glauca in a low and a high nutrient environ- in a low and a high nutrient environ- ment. In both species, more of the resources had been directed to the photobiont in the high nutrient environment also increasing their overall carbon status. Taken together, my studies indicate that in spite of the apparent passive environmental control on lichen metabolism, these symbiotic organisms are able to both optimize and control their resource acquisition and allocation processes.

The acceptance of blood contacting implants creating favorable conditions in vivo is decisively determined by their interaction with proteins that mediate inter cellular interactions with synthetic substrates. Adsorbed proteins can activate blood cascade systems like coagulation and complement that may result in serious blood clots, and/or immunological reactions. Poly (ethylene glycol) (PEG), heparin, and phosphoryl choline (PC) functional poly (methacrylates) are previously used polymers with known non-adhesive properties in blood contacting events. This thesis contributes to this extensive research by introducing a novel type of biomaterial that equips biodegradable polymers with biomimetic functionalities. The phospholipid mimetic material is synthesized by combining biodegradable polymers with various functional polar end-groups consisting of zwitterionic phosphoryl choline (PC), anionic succinates, and cationic quaternary ammonium. The polymer backbone provides mechanical stability and biodegradability whilst the various head groups provide a variety of functions. The careful evaluation of the synthesis has allowed reaction conditions to be optimized leading to complete conversion at each step and subsequently high yields. Initially, poly (e-caprolactone) (PCL) was used since it provided a suitable synthetic starting point. However, the synthesis has also included poly (trimethylene carbonate) (PTMC) to provide a material that allows spontaneous surface enrichment of the polar PC group. This was achieved with an added hydrophilic environment. Through the synthesis of multi PC functional PTMC, additional bulk organisation by the formation of zwitterionomers (PC ionomer) was achieved. Low modulus elasticity and water uptake were some of the properties of the formed material. As a result it was shown that the PC ionomer could be used for protein/drug loading and subsequent release. Furthermore, the material possessed non-adhesive properties in different biological environments. Importantly, the result suggests that a versatile synthetic platform has been established that may provide a smorgasbord of different functional polymers, or combinations of such. This is indeed important since it was shown that the polymer in many ways dictates how the material may take advantage of an added functionality. Such materials should be interesting for a variety of biomedical applications including the production of soft hemocompatible tissue.

Purpose: The purpose of this study was to investigate if there was any difference in energy expenditure (kcal/min) and oxygen consumption (VO2) between subjects walking with Masai Barefoot Technology ® (MBT) shoes and regular orthopedic shoes. The research hypothesis was that MBT shoes demand more energy expenditure than regular orthopedic shoes. Methods: Seven women aged 49-65 were recruited for the study. The subjects were tested in two sessions, with a minimum of two weeks in between each sesssion. On each test session the subjects walked with both MBT shoes and orthopedic shoes which were adjusted in mass (g) to match the mass of the MBT shoes. While the subjects walked on a treadmill, the oxygen consumption (VO2), heart rate (min-1) and self selected velocity (m/s) for each of the shoe types was measured. Results: Results showed that there is no significant difference in oxygen consumption (VO2) between the MBT and orthopedic shoes. Energy expenditure (kcal/min) was also calculated from the data and the results revealed that there is no significant differ-ence between MBT and orthopedic shoes in energy expenditure (kcal/min) either. The self selected velocity (m/s) between the two shoe types was also found to be insignificant. Conclusion: The results showed no significant difference between the shoes. This could indicate that the specific construction of the MBT shoe has no effect on the energy expenditure (kcal/min) of its user. This lack of difference may be due to the equal mass of the shoes, but since oxygen consumption (VO2) was not investigated in orthopedic shoes with different shoe masses, this conclusion cannot be confirmed. The self selected velocity (m/s) was found to be insignificant and this finding could suggest to that prolonged usage of the MBT shoe may diminish gait parameters dissimilarities during ambulation. This study should therefore be seen as a pilot study and further investigation in this area should be pursued.