Zinc is a metal involved in structure of many enzymes, in the growth and differentiation of plants. Wheat is one of the most consumed cereals. Some wheat cultivars can&amp / #8217 / t deal with zinc deficiency and this situation not only reduces grain yield but also weakens the resistance of cereals to diseases and impairs the nutritional quality of the grain. Some wheat cultivars are not affected by zinc deficiency. In this study, &amp / #8216 / differential display&amp / #8217 / , used for determination differentially expressed genes between two samples, was performed. The most zinc efficient bread wheat cultivar Kira&ccedil / -66 was grown in hydroponics medium and samples were taken at different time periods. RNA isolations were done and differential display technique was performed. After examining the results, differentially expressed bands were selected and sequenced. DNA sequence analysis were done in available databases which showed that three of the bands were fragments of putative zinc transporters. In this study we have found threee putative gene fragments using differential display technique on zinc efficient plants grown under differeing zinc concentrations. These fragments showed homology with zinc transporter, ABC transporter and ADH (Alcohol Dehydrogenase). It is known that all of these three genes are involved in zinc efficiency mechanism. Further studies will be conducted on these gene fragments.

Depression is a major psychological disorder that affects a person's mental and physical abilities. The National Institute of Mental Health (NIMH) classified it as a serious medical illness. It causes huge economic, as well as financial impact on the people, and it is also becoming a major public health issue. Antidepressant drugs are prescribed to mitigate the suffering caused by this disorder. Different generations of antidepressants have been developed with dissimilar mechanisms of action. According to the Center for Disease Control, the usage of antidepressants has skyrocketed by 400 percent increase over 2005- 2008 survey period. This dramatic rise in usage indicates that these are the most prescribed drugs in the US. Even with the FDA mandated "black box" warning of increased suicidal thoughts upon use of selected antidepressants, these drugs are still being used at a higher rate. <br>All classes of antidepressants are plagued by side effects with mainly sexual dysfunction common among them. To avoid the adverse effects, an emphasis is to discover novel structural drug scaffolds that can be further developed as a new generation of antidepressants. The importance of this research is to discover structurally novel antidepressants by performing in silico virtual screening (VS) of chemical databases using the serotonin transporter (SERT). In the absence of a SERT crystal structure, a homology model was developed. The homology model was utilized to develop the first structure-based pharmacophore for the extracellular facing secondary ligand binding pocket. The pharmacophore captured the necessary drug-SERT interaction pattern for SERT inhibitory action. This pharmacophore was employed as one of the filters for VS of candidate ligands. The ten compounds identified were purchased and tested pharmacologically. Out of the ten hits, three structurally novel ligands were identified as lead compounds. Two of these compounds exhibited selectivity towards SERT; the remaining lead compound was selective towards the dopamine transporter and displayed cocaine inhibition. The two SERT selective compounds will provide new opportunities in the development of novel therapeutics to treat depression. <br>For dopamine transporter (DAT), the study was based on recently developed structurally diverse photo probes. In an effort to better understand the binding profile similarities among these different scaffolds, the photo probes were docked into DAT. The finger print analysis of the interaction pattern of docked poses was performed to identify the inhibitor-binding sites. <br>For mitogen activated protein kinase 5 (MEK5), given the lack of structural information, a homology model of MEK5 was developed to guide the rational design of inhibitors. Docking of known MEK5 inhibitors into the homology model was performed to understand the inhibitory interaction profile. Several series of analogues were designed utilizing the generated interaction profile. / Bayer School of Natural and Environmental Sciences / Chemistry and Biochemistry / PhD / Dissertation

Ischemic strokes are a major cause of death and disability, yet efficacious pharmacotherapies remain limited. Although neuronal cell death during stroke is primarily induced via excessive Ca2+ influx through NMDARs following overactivation by uncontrolled glutamate release, antagonism of these receptors has been shown to be ineffective due to intolerable side effects. This thesis highlights a novel therapeutic strategy for stroke wherein NMDAR-mediated excitotoxicity is temporarily and dynamically mitigated via the initiation of a process termed “glycine induced NMDAR internalization” (GINI). While GINI occurs in vitro following application of high doses of glycine, achieving these levels of glycine in vivo has long been thought impossible as glycine transporters (GlyT1) maintain synaptic glycine levels well below saturating concentrations. Here, we show that GINI can be triggered in vivo when mice are administered a glycine transporter-1 antagonist (GlyT1-A) prior to stroke and that this strategy provides neuroprotection. Mice pre-treated with a GlyT1-A, which elevates glycine levels, exhibited significantly smaller stroke volumes, reduced cell death, and significantly minimized behavioural deficits following stroke induction by either photothrombosis (PT) or endothelin-1 (ET-1). Moreover, we observed preservation of vasculature function and morphology in the peri-infarct area. These data strongly suggest that elevating brain glycine levels with GlyT1-As should be considered as a novel pharmacotherapy for ischemic stroke.

Fysisk planering har möjligheten att påverka hurstaden utvecklas och med det hur invånarnautvecklas med staden. För att kunna påverkautvecklingen behövs kunskapen om vilkafaktorer som styr utvecklingen i önskad riktning.För att människor ska resa mer med hållbaratransportmedel krävs förutsättningarna för det istaden. Det är planerarens jobb att ha kunskapenom vilka faktorer som påverkar valet till hållbaratransportmedel och hur de ska implementeras. I den här uppsatsen analyseras hur Kumla stadoch sex kvarter i Kumla kan omgestaltas för attfrämja hållbara transporter. Uppsatsen visar påhur en omgestaltning kan se ut i en liten svenskpendlingskommun. Förslaget på omgestaltningkan appliceras i generella drag i andra liknandekommuner.

Syfte och frågeställningar Studiens syfte var att ta reda på om pojkar respektive flickor som bedriver aktiva transporter är fysiskt aktiva på annat sätt och vilka faktorer som påverkar val av aktivpendling samt om årstiden och plats i landet har någon avgörande betydelse. - Hur kommer eleverna till skolan? - Vilka faktorer kan förklara aktiv pendling och finns det samband mellan aktiva transporter och årstider?  Metod En kvantitativ enkät användes som metod för att undersöka aktiva transporter och fysisk aktivitet. En anonym enkätundersökning genomfördes med 125 högstadieelever. Urvalet bestod av samtliga elever i årskurs nio i Kalix Kommun, Norrbotten. Resultaten analyserades och bearbetades via korstabeller. Studiens teoretiska utgångspunkter var befintlig forskning om ungas fysiska aktivitet samt pågående forskning om aktiva transporter. Resultat Resultatet visar på att eleverna i årskurs nio i Kalix Kommun bedriver aktiva transporter till en liten del, pojkar 27% under den kalla årstiden och 33% under den varma årstiden. Motsvarande resultat för flickor är 26% under den kalla årstiden och 23% under den varma årstiden. Resultatet visar också att många elever åker buss eller bil till och från skolan. De elever (92 stycken) som uppgett annat färdsätt uppger att de har färdats med något slags fordon (90%). Det verkar som att färdvägens längd och det kalla klimatet i Norrbotten kan vara avgörande faktorer som påverkar valet av aktiva transporter för både pojkar och flickor. Få elever bedriver aktiva transporter till och från skolan jämfört med dem som använder fordon. Enligt korstabellerna är det de flickor och pojkar som bedriver aktiva transporter som också är mer aktiva både inom föreningsidrott och på annat sätt på fritiden. Pojkarna inom denna grupp är också de som är mest aktiva i föreningsidrott eller liknande jämfört med flickor. Det verkar även som att det är dessa pojkar som i större utsträckning än flickorna, kommer upp till miniminivå i tid för daglig fysisk aktivitet . Slutsats Färdvägens längd i glesbygd och det kalla klimatet i Norrbotten kan vara avgörande faktorer som påverkar valet av aktiva transporter och är en trolig orsak till den låga andelen av elever som bedriver aktiva transporter. Flickorna verkar vara mer fysiskt aktiva på fritiden än pojkarna. Det kan ha att göra med att fysisk aktivitet har uppmärksammats under det senaste årtiondet. Pojkar som bedriver aktiva transporter är mer aktiva både i föreningsidrott och på fritiden jämfört med flickor som bedriver aktiva transporter. Denna grupp av pojkar når upp till miniminivån i tid för fysisk aktivitet per dag i större utsträckning än motsvarande grupp flickor.

Half of the cholinergic neurons of human and primate intrinsic cardiac ganglia (ICG) have a dual cholinergic/noradrenergic phenotype. Likewise, a large subpopulation of cholinergic neurons of the mouse heart expresses enzymes needed for synthesis of norepinephrine (NE), but they lack the vesicular monoamine transporter type 2 (VMAT2) required for catecholamine storage. In the present study, we determined the full scope of noradrenergic properties (i.e. synthetic enzymes and transporters) expressed by cholinergic neurons of mouse ICG, estimated the relative abundance of neurons expressing different elements of the noradrenergic phenotype, and evaluated the colocalization of cholinergic and noradrenergic markers in atrial nerve fibers. Stellate ganglia were used as a positive control for noradrenergic markers. Using fluorescence immunohistochemistry and confocal microscopy, we found that about 30% of cholinergic cell bodies contained tyrosine hydroxylase (TH), including the activated form that is phosphorylated at Ser-40 (pSer40 TH). Dopamine β-hydroxylase (DBH) and norepinephrine transporter (NET) were present in all cholinergic somata, indicating a wider capability for dopamine metabolism and catecholamine uptake. Yet, cholinergic somata lacked VMAT2, precluding the potential for NE storage and vesicular release. In contrast to cholinergic somata, cardiac nerve fibers rarely showed colocalization of cholinergic and noradrenergic markers. Instead, these labels were closely apposed but clearly distinct from each other. Since cholinergic somata expressed several noradrenergic proteins, we questioned whether these neurons might also contain trophic factor receptors typical of noradrenergic neurons. Indeed, we found that all cholinergic cell bodies of mouse ICG, like noradrenergic cell bodies of the stellate ganglia, contained both tropomyosin-related kinase A (TrkA) and p75 neurotrophin receptors. Collectively, these findings demonstrate that mouse intrinsic cardiac neurons (ICNs), like those of humans, have a complex neurochemical phenotype that goes beyond the classical view of cardiac parasympathetic neurons. They also suggest that neurotrophins and local NE synthesis might have important effects on neurons of the mouse ICG.

System L (SL), a basolateral amino acid transporter, transports large neutral amino acids (LNAAs) in a Na+-independent manner. Previously, we identified two isoforms of transporters: L-type amino acid transporter 1 (LAT1) and 2 (LAT2) and revealed their distinct substrate selectivity and transport properties. In this study, to establish more stable human LAT1 (hLAT1) and LAT2 (hLAT2) in vitro assay systems, we established mouse cell lines stably expressing hLAT1 (S2-LAT1) and hLAT2 (S2-LAT2). Real-time quantitative RT-PCR analysis revealed that S2-LAT1 and S2-LAT2 cells express hLAT1 and hLAT2 mRNAs at 20 - 1000-fold higher levels than those of endogenous mouse Lat1 and Lat2. S2-LAT1 and S2-LAT2 mediated [14C]L-leucine transport properties were measured and corresponded to results observed via Xenopus oocytes. Using these cells, the data demonstrate that hLAT1 and hLAT2 exhibit different characters in the acceptance of α-methyl amino acids and amino acid-related compounds with bulky side chains such as thyroid hormones and melphalan. S2-LAT1 and S2-LAT2 cells are expected to facilitate hLAT1 and hLAT2 substrate recognition research and contribute to drug development by providing an efficient assay system to screen for chemical compounds that interact with hLAT1 and hLAT2.

In this work two overall subjects were addressed. 1. In recent years high pressure perturbance has become a tool to investigate the folding energy landscape, the volumetric properties and the conformational equilibria of proteins. Conformational states which are not populated at ambient conditions thus become accessible to spectroscopic characterization. In this work a high pressure application was combined with EPR spectroscopy to investigate three spin labeled proteins, BSA from Bos taurus, HisJ from Salmonella enterica serovar Typhimurium and α-synuclein from Homo sapiens. The goal of these studies was to comprehend the influence of pressure on the respective EPR spectra and to identify changes in conformational equilibria and volumetric properties of the investigated proteins. Studies on BSA revealed a negative activation volume for rotational diffusion of the spin labeled site. Moreover, a rotameric equilibrium was derived from the pressure-dependent side chain dynamics and a correlating negative partial molar volume was observed, indicating a shift of the rotameric equilibrium to lesser order. In this regard it was also shown that a chaotropic medium (guanidine hydrochloride) supports the pressure-dependent effect. Spin labeled sites in the substrate binding protein HisJ revealed to be highly influenceable by low pressures between ambient conditions and 200 bar. Pressurization induced oligomerization and precipitation of the protein. Substrate binding revealed differences in pressure-dependence with regard to a decreased precipitation effect but not in relation to oligomerization. The natively unfolded protein α-synuclein plays a key role in Parkinson´s disease and is known for forming β-sheet rich aggregates, so called amyloid fibrils. The experimental data of this work revealed that hydrostatic pressure can induce a non-amyloid aggregation of monomeric α-synuclein which produces an unspecific oligomer. Furthermore, it was shown that α-synuclein amyloid fibrils can be dissolved by hydrostatic pressure. From the pressure dependent conformational equilibrium between the monomer and the fibril form the change of the partial molar volume of the investigated site was determined. 2. The second subject of this work was focused on different import systems, ATP-binding cassette (ABC) transporters and Energy-Coupling-Factor (ECF) transporters, for amino acids, vitamins and metal ions in prokaryotes. Studies on one bacterial ABC and two ECF transporter systems from two different organisms, the histidine ABC-type transporter HisQMP2 from Salmonella enterica serovar Typhimurium, the biotin ECF-type importer BioMNY from Rhodobacter capsulatus and the cobalt-specific ECF-type transporter CbiMNQO from Rhodobacter capsulatus, were performed using DEER and cw EPR spectroscopy. The goal of the studies on HisQMP2 and BioMNY was to shed light on conformations and dynamics connected to their transporter function. Studies on CbiMNQO aimed at the detection of the substrate in the transporter´s substrate binding unit. For HisQMP2 transport cycle dependent conformational changes and interactions with the substrate binding protein HisJ were revealed. Three different distance values between sites H101R1 and H101’R1 in the transporter´s nucleotide binding domains were assigned to the apo-, the ATP-bound and the posthydrolysis state. It was shown that the closed conformation of the nucleotide binding domains is achieved only in the presence of the ligand-bound HisJ which indicates a transmembrane communication of the association of HisJ to the transporter. Furthermore, interspin distances were determined between sites D86R1-A96R1, C197R1-C104R1 and A118R1-G123R1 in the transmembrane domains HisQ and HisM revealing distinguishable conformational states which correlate to the different states of the nucleotide binding sites during the hydrolysis cycle. Measured interspin distances between HisJ and HisM in the HisQMP2 complex showed that interaction only occurred in the closed state of the HisP2 dimer, the nucleotide bound state. Two different, substrate-dependent interactions between site G24R1 in HisJ and site A96R1 in HisQMP2 were observed, revealing that the substrate-free and substrate-bound form of HisJ both associate with HisQMP2. Distance measurements between sites G24R1 and T151R1 in HisJ in the presence and absence of its substrate revealed interspin distance changes that correlate with the proteins open and closed conformation. Investigations on the ECF transporter BioMNY, reconstituted into nanodiscs, revealed a closure and reopening of the nucleotide binding domains between sites H87R1 and H87’R1 using DEER spectroscopy which delivered interspin distance values that correlate with the apo-, the ATP-bound and the posthydrolysis state of the transporter. Further experiments were aimed to shed light on the transporters substrate-translocation mechanism with regard to the so called toppling over mechanism. Unfortunately, the experiments of this work were not able to give a distinct answer with respect to the proposed model because of the transmembrane domains tendency to oligomerize when reconstituted into nanodiscs. In this work we showed that substrate uptake by the substrate binding unit CbiM of the cobalt-specific ECF transporter CbiMNQO depends on the presence of the small transmembrane protein CbiN. Measurements of spin labeled CbiMN in detergent showed oligomerization of CbiM.

Sweden has ambitious climate goals, such as the overarching goal, the 2045 goal. The goal is for Sweden to have zero net emissions of greenhouse gases by 2045 at the latest. In order to achieve the overall climate goal and the interim targets, electrification of society is an important component. The industrial sector and the transport sector each account for about a third of Sweden’s emissions, where the conversion to electricity is an important solution. The electrification of passenger cars and buses has meant that emissions from domestic transport have decreased every year, and in order to achieve the interim target of 70 percent lower emissions of greenhouse gases in 2030 compared to 2010, the electrification of heavy duty trucks is one of the key components. However, there are long-term challenges with the power grid and already today grid owners have capacity challenges. Regional grid owners cannot increase their power subscription, while local grid owners cannot grant new connections. Regarding the electrification of long-haul transportation, there are challenges in terms of charging infrastructure. Truck drivers operate on a strict schedule, and minimizing down time is crucial to keep costs down. By law, truck drivers must take a 45-minute break after 4.5 hours of driving time, which means that during this break it is desirable to recharge the vehicle before departure. This means that high power demands are placed on the charging infrastructure that exists to be able to transmit the desired energy.   A new standard, the Megawatt Charging System (MCS), which meets the high power requirements has been developed and is included in pilot projects. The maximum power that the charging standard can deliver is 3.75 MW. The results of the thesis show that these high-power chargers place high demands on thepower grid. When connecting a charging station with MCS charging points to the grid, it may require local upgrading of lines and transformers, but also upgrading in otherparts of the network. Examples of such upgrades are reactive power compensation to be able to support the network locally at peak loads to obtain voltage levels within stable voltage ranges, but also upgrades of lines and transformers to be able to deliver the desired power. Integration of a battery storage in connection with a charging station relieves the powergrid and its components. However, it is important to highlight that since the battery needs to be recharged, this means that a more even power requirement is needed. However, the  maximum load on both transformers and lines is reduced, which can be a desirable effect when a charging station of the same nature is put into operation.

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