SUR2A-mediated regulation of KATP channels is affected by residues belonging to the C terminus of the first nucleotide binding domain (NBD1). We studied the C-terminal region of NBD1 by comparing experiments using NBD1 S615-D914 and NBD1 S615-K972 constructs to studies of NBD1 S615-L933 also performed in our laboratory. Our NMR data suggests that the C-terminal region of NBD1 from residues Q915 to L933 is disordered and transiently contacts the NBD1 core, which may affect NBD1 phosphorylation. Tryptophan quenching fluorescence experiments corroborate that the Q915-L933 C-terminal tail contacts the NBD1 core. Fluorescence thermal denaturation experiments suggest that NBD1 S615-D914 has a higher affinity for MgATP compared with NBD1 S615-L933, implying that the C-terminal tail varies MgATP binding. Additional experiments were performed to identify soluble constructs of hSUR1 NBD1 and rSUR2A NBD2 that would allow detailed biophysical studies of these domains. Some of the constructs studied showed improved solubility and stability.

<p style="text-align: justify;">U ovom radu sintetisani su hidrogelovi na osnovu akrilamida i akrilne kiseline, radikalnom polimerizacijom, primenom konvencionalne i mikrotalasne metode sinteze. Varirani su početni odnosi monomera i udeo umreživača, u cilju ispitivanja uticaja sastava reakcione sme&scaron;e na svojstva dobijenih hidrogelova. Optimizovani su uslovi sinteze u mikrotalasnom polju kao brže, jednostavnije i ekonomičnije metode. U cilju uspostavljanja korelacije između mehanizma sinteze, strukture i svojstava dobijenih hidrogelova, primenom relevantnih metoda karakterizacije, upoređena su apsorpciona, reolo&scaron;ka, toplotna i strukturna svojstva hidrogelova dobijenih dvema metodama.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Ustanovljeno je da se mikrotalasnom metodom sinteze na brži i jednostavniji način uz smanjen utro&scaron;ak vremena i energije dobijaju hidrogelovi konkurentni onima koji se dobijaju konvencionalnim zagrevanjem u vodenom rastvoru. Drugi deo doktorata obuhvata sintezu hidrofilnih polimernih mreža na osnovu natrijum karboksimetilceluloze (NaCMC) i karboksilnih kiselina, od kojih je jedna serija sintetisana prožimanjem linearnim kopolimerima akrilamida i akrilne kiseline u cilju povećanja potencijala primene u floku-lacione svrhe. Rezultati ispitivanja svojstava bubrenja, strukturnih toplotnih i flokulacionih svojstava pokazali su međusobno slaganje. Dobijeni rezultati pokazali su da od primenjenih karboksilnih kiselina, linunska kiselina u udelu od 15% u odnosu na masu NaCMC daje hidrogelove najboljih svojstava. Kombinacijom ove mreže sa kopolimerom akrilamida i akrilne kiseline u masenom odnosu 10/90, stvara se teorijska platforma za dobijanja flokulanta koji bi mogao da pokaže visoku efikasnost u preči&scaron;ćavanju vode u kojoj dominiraju pozitivno naelektrisane čestice, pravilnim izborom parametara flokulacije.</p> / <p>In this paper, hydrogels based on acrylamide and acrylic acid were synthesized using conventional and microwave synthetic methods via free-radical polymerization. The initial monomers ratio and amount of crosslinking agent were varied in order to investigate the effect of the composition of the reaction mixture on the properties of the obtained hydrogels. The conditions of synthesis in the microwave field as faster simpler and more economical method have been optimized. In order to establish a correlation between the mechanism of synthesis, structure and properties of the obtained hydrogels using the relevant methods of characterization, the absorption, rheological, thermal and structural properties of the hydrogels obtained by the two methods were compared. It has been found that the microwave synthesis is a faster and simpler method, which enables reduced consumption of time and energy and produces hydrogels competitive to those ones obtained by conventional heating in aqueous solution. The second part of the thesis includes the synthesis of hydrophilic polymer networks based on sodium carboxymethylcellulose (NaCMC) and carboxylic acids, whereby one series is synthesized by interpenetration of the network using the linear acrylamide and acrylic acid copolymers in order to increase the potential application of hydrogels for flocculation purposes. The results of measurements of swelling, structural, thermal and flocculation properites have shown mutual agreement. The obtained results have shown that among applied carboxylic acids, citric acid in the amount of 15% per mass of NaCMC, has given the hydrogels with the best properties. The Combination of this network with a copolymer of acrylamide and acrylic acid in a mass ratio of 10/90 has created a theoretical platform for the production of flocculant which could show high efficacy in purifying of water dominated by positively charged particles.</p>

<p>Degradable polymers have gained an increased attention in the field of biomedical applications over the past decades, for example in tissue engineering. One way of improving the biocompatibility of these polymers is by chemical surface modification, however the risk of degradation during the modification procedure is a limiting factor. In some biomedical applications, for example in nerve guides, a patterned surface is desired to improve the cell attachment and proliferation.</p><p>In this thesis a new non-destructive, single-step, and solvent free method for surface modification of degradable polymers is described. Poly(L-lactide) (PLLA) substrates have been functionalized with one of the following vinyl monomers; N-vinylpyrrolidone (VP), acrylamide (AAm), or maleic anhydride (MAH) grafts. The substrates were subjected to a vapor phase atmosphere constituted of a mixture of a vinyl monomer and a photoinitiator (benzophenone) in a closed chamber at very low pressure and under UV irradiation. Poly(ε-caprolactone) (PCL), poly(lactide-co-glycolide) (PLGA), and poly(trimethylene carbonate) (PTMC) have been surface modified with VP using the same procedure to show the versatility of the method. The wettability of all of the four substrates increased after grafting. The surface compositions were confirmed by ATR-FTIR and XPS. The VP grafted PLLA, PTMC and PLGA substrates have been shown to be good substrates for the normal human cells i.e. keratinocytes and fibroblasts, to adhere and proliferate on. The topography of substrates with well defined nano patterns was preserved during grafting, since the grafted layer is very thin. We have also shown that the method is useful for a simultaneous chemical and topographical modification of substrates by masked vapor phase grafting. The surface topography was determined with SEM and AFM.</p> / <p>Intresset för användningen av nedbrytbara polymerer till biomedicinska applikationer som till exempel vävnads rekonstruktion har ökat avsevärt de senaste decennierna. Ett sätt att öka biokompatibiliteten hos dessa polymerer är genom kemisk ytmodifiering, men risken för nedbrytning under själva modifieringen är en begränsande faktor. I vissa biomedicinska applikationer, till exempel nervguider, är det önskvärt att ha en väldefinierad ytstruktur för att öka vidhäftningen och tillväxten av celler.</p><p>I den här avhandlingen presenteras en ny ickeförstörande, lösningsmedelsfri enstegsprocess för ytmodifiering av nedbrytbara polymerer. Substrat av poly(L-laktid) (PLLA) har ytfunktionaliserats med var och en av följande vinylmonomerer, N-vinylpyrrolidon (VP), akrylamid (AAm) eller maleinsyraanhydrid (MAH). Substraten har exponerats för en gasfasatmosfär av en blandning av en vinylmonomer och en fotoinitiator (bensofenon) i en tillsluten reaktor vid mycket lågt tryck och under UV-strålning. Metodens mångsidighet har även påvisats genom att ytmodifiera substrat av poly(ε-kaprolakton) (PCL), poly(laktid-co-glykolid) (PLGA) och poly(trimetylen karbonat) (PTMC) med VP. Vätbarheten ökade för alla fyra materialen efter ympning med en vinylmonomer. Ytsammansättningen fastställdes med ATR-FTIR och XPS. De VP ympade filmerna av PLLA, PLGA och PTMC visade sig vara bra substrat för mänskliga celler, i detta fall keratinocyter och fibroblaster, att vidhäfta och växa på. Yttopografin hos filmer med väldefinierade nanomönstrade ytor kunde bevaras efter ympning, tack vare att det ympade lagret är så tunt. Gasfas metoden har också visat sig användbar för att simultant ytmodifiera både kemiskt och topografiskt genom maskad gasfasympning. Yttopografin bestämdes med SEM och AFM.</p>

Degradable polymers have gained an increased attention in the field of biomedical applications over the past decades, for example in tissue engineering. One way of improving the biocompatibility of these polymers is by chemical surface modification, however the risk of degradation during the modification procedure is a limiting factor. In some biomedical applications, for example in nerve guides, a patterned surface is desired to improve the cell attachment and proliferation. In this thesis a new non-destructive, single-step, and solvent free method for surface modification of degradable polymers is described. Poly(L-lactide) (PLLA) substrates have been functionalized with one of the following vinyl monomers; N-vinylpyrrolidone (VP), acrylamide (AAm), or maleic anhydride (MAH) grafts. The substrates were subjected to a vapor phase atmosphere constituted of a mixture of a vinyl monomer and a photoinitiator (benzophenone) in a closed chamber at very low pressure and under UV irradiation. Poly(ε-caprolactone) (PCL), poly(lactide-co-glycolide) (PLGA), and poly(trimethylene carbonate) (PTMC) have been surface modified with VP using the same procedure to show the versatility of the method. The wettability of all of the four substrates increased after grafting. The surface compositions were confirmed by ATR-FTIR and XPS. The VP grafted PLLA, PTMC and PLGA substrates have been shown to be good substrates for the normal human cells i.e. keratinocytes and fibroblasts, to adhere and proliferate on. The topography of substrates with well defined nano patterns was preserved during grafting, since the grafted layer is very thin. We have also shown that the method is useful for a simultaneous chemical and topographical modification of substrates by masked vapor phase grafting. The surface topography was determined with SEM and AFM. / Intresset för användningen av nedbrytbara polymerer till biomedicinska applikationer som till exempel vävnads rekonstruktion har ökat avsevärt de senaste decennierna. Ett sätt att öka biokompatibiliteten hos dessa polymerer är genom kemisk ytmodifiering, men risken för nedbrytning under själva modifieringen är en begränsande faktor. I vissa biomedicinska applikationer, till exempel nervguider, är det önskvärt att ha en väldefinierad ytstruktur för att öka vidhäftningen och tillväxten av celler. I den här avhandlingen presenteras en ny ickeförstörande, lösningsmedelsfri enstegsprocess för ytmodifiering av nedbrytbara polymerer. Substrat av poly(L-laktid) (PLLA) har ytfunktionaliserats med var och en av följande vinylmonomerer, N-vinylpyrrolidon (VP), akrylamid (AAm) eller maleinsyraanhydrid (MAH). Substraten har exponerats för en gasfasatmosfär av en blandning av en vinylmonomer och en fotoinitiator (bensofenon) i en tillsluten reaktor vid mycket lågt tryck och under UV-strålning. Metodens mångsidighet har även påvisats genom att ytmodifiera substrat av poly(ε-kaprolakton) (PCL), poly(laktid-co-glykolid) (PLGA) och poly(trimetylen karbonat) (PTMC) med VP. Vätbarheten ökade för alla fyra materialen efter ympning med en vinylmonomer. Ytsammansättningen fastställdes med ATR-FTIR och XPS. De VP ympade filmerna av PLLA, PLGA och PTMC visade sig vara bra substrat för mänskliga celler, i detta fall keratinocyter och fibroblaster, att vidhäfta och växa på. Yttopografin hos filmer med väldefinierade nanomönstrade ytor kunde bevaras efter ympning, tack vare att det ympade lagret är så tunt. Gasfas metoden har också visat sig användbar för att simultant ytmodifiera både kemiskt och topografiskt genom maskad gasfasympning. Yttopografin bestämdes med SEM och AFM. / QC 20101014

Acrylamide is a common dietary exposure that crosses the human placenta. It is classified as a probable human carcinogen, and developmental toxicity has been observed in rodents. OBJECTIVES: We examined the associations between prenatal exposure to acrylamide and birth outcomes in a prospective European mother-child study. METHODS: Hemoglobin (Hb) adducts of acrylamide and its metabolite glycidamide were measured in cord blood (reflecting cumulated exposure in the last months of pregnancy) from 1,101 singleton pregnant women recruited in Denmark, England, Greece, Norway, and Spain during 2006-2010. Maternal diet was estimated through food-frequency questionnaires. RESULTS: Both acrylamide and glycidamide Hb adducts were associated with a statistically significant reduction in birth weight and head circumference. The estimated difference in birth weight for infants in the highest versus lowest quartile of acrylamide Hb adduct levels after adjusting for gestational age and country was -132 g (95% CI: -207, -56); the corresponding difference for head circumference was -0.33 cm (95% CI: -0.61, -0.06). Findings were similar in infants of nonsmokers, were consistent across countries, and remained after adjustment for factors associated with reduced birth weight. Maternal consumption of foods rich in acrylamide, such as fried potatoes, was associated with cord blood acrylamide adduct levels and with reduced birth weight. CONCLUSIONS: Dietary exposure to acrylamide was associated with reduced birth weight and head circumference. Consumption of specific foods during pregnancy was associated with higher acrylamide exposure in utero. If confirmed, these findings suggest that dietary intake of acrylamide should be reduced among pregnant women.