During the time I've attended the programme to become a Transportation manager, I've been presented with quite a large number of different words. My purpose with this paper is to take a closer look at them, and see if they're sustainable concepts. This paper will be a literary study, and be on a wholly theoretical level. After a thorough examination of the various words, I've come to the conclusion that these words that I've chosen aren't just fad-words. They're going to be around for quite a while, this due to the fact that they're based on good ideas from the get-go. Many of them represent simple but functional solutions, that every one at work or at home that may partake in and thus create a better environment / Under den tiden jag har gått på Transportation manager programmet har jag fått ta del av en mängd ord. Dessa tänker jag gå in lite närmare på och se ifall de kan vara hållbara koncept. Detta arbetet kommer vara en litteraturstudie, och kommer att vara på en rent teoretiskt nivå. Efter att ha gått igenom de olika orden har jag kommit fram till att detta inte är några modeflugor jag har valt. De kommer att finnas kvar en lång tid, mest för att de är baserade på goda idéer från början. Många av dem representerar enkla men funktionella lösningar som var och en på företaget och hemma kan ta del av för att skapa en bättre miljö.

<p>The lateral superior olive (LSO) is a binaural nucleus that is critical for azimuthal sound localization. Bipolar principal cells of the LSO compute interaural level differences (ILDs) by comparing converging excitatory and inhibitory inputs driven by either ear. More specifically, this computation relies on integrating excitatory inputs from the ipsilateral cochlear nucleus with inhibitory, GABA/glycinergic inputs from the medial nucleus of the trapezoid body (MNTB), which are driven by sound originating at the contralateral ear. In order to reliably compute ILDs, the converging inputs must represent sounds of the same frequency. This specificity emerges during the first few weeks of postnatal life in rats as a result of functional and anatomical refinement. Interestingly, significant refinement of this auditory circuit occurs in the absence auditory experience. We focused on changes in the subcellular location of MNTB inputs and the expression of vesicular proteins before hearing onset.</p> <p>The subcellular distribution of inputs onto a neuron heavily influences synaptic integration and the mature distribution likely emerges during a period of circuit refinement. Little is known about how the inputs are distributed onto LSO principal cells and how the mature distribution is achieved. We studied the distribution of inhibitory inputs onto LSO neurons and found that significant re- distribution occurs before hearing onset. The mechanisms underlying the refinement of the inhibitory MNTB projection are not well understood but could be related to the transient co-transmission of the excitatory neurotransmitter glutamate. We studied the expression of vesicular proteins that may regulate the release of GABA/glycine and glutamate at the immature MNTB terminal. We found that MNTB terminals transiently express two Ca++ sensors, which may be associated with the different release properties for GABA/glycine and glutamate. Lastly, we asked one specific example of what controls the expression and sorting of vesicular proteins at the immature MNTB terminal.</p> / Doctor of Philosophy (PhD)

McMaster University MASTER OF SCIENCE (2019) Hamilton, Ontario (Medical Sciences) TITLE: Modulation of cyclic adenosine monophosphate for potentiation of long-acting β2-agonist and glucocorticoids in human airway epithelial cells AUTHOR: Yechan Kim, B.HSc. (McMaster University) SUPERVISOR: Dr. Jeremy Alexander Hirota NUMBER OF PAGES: xiv, 81 / In Canada, asthma is the third most common chronic disease resulting in 250 premature deaths annually and related healthcare expenses exceeding $2.1 billion/year. It is estimated that around 50-80% of asthma exacerbations are due to viral infections. Despite an advanced understanding on how to treat and manage the symptoms of asthma, current therapy is sub-optimal in 35-50% of moderate-severe asthmatics around the world resulting in lung inflammation, persistent impairment of lung function, and increased risk of mortality. Combination of long-acting β2 agonists (LABA) for bronchodilation and glucocorticoids (GCS) to control lung inflammation represent the dominant strategy for the management of asthma. Increasing intracellular cyclic adenosine monophosphate (cAMP) beyond existing combination LABA/GCS are likely to be beneficial for the management of difficult to control asthmatics that are hypo-responsive to mainstay therapy. In human airway epithelial cells (HAEC), cAMP is either exported by transporters or broken down by enzymes, such as phosphodiesterase 4 (PDE4). We have demonstrated that HAEC express ATP Binding Cassette Transporter C4 (ABCC4), an extracellular cAMP transporter. We also show that ABCC4 and PDE4 inhibition can potentiate LABA/GCS anti-inflammatory responses in a human epithelial cell line in a cAMP-dependent mechanism validating the pursuit of novel ABCC4 inhibitors as a cAMP elevating agent for asthma. / Thesis / Master of Science in Medical Sciences (MSMS) / Asthma is a common chronic lung disease characterized by narrow and inflamed airways that cause breathing difficulties. Current management includes the combination of bronchodilators, to relax the airway, and steroids, to decrease inflammation. Unfortunately, this combination therapy is suboptimal in 35-50% of users, increasing the risk of asthma attacks, hospitalization rate, and health care costs. Recently, there have been studies theorizing that we can improve the therapy’s ability to decrease inflammation by increasing cAMP, an important molecule for biological activities. We tested this claim by blocking the breakdown and export of cAMP to increase its levels and measured inflammatory cytokines, molecules that direct the action of immune cells. Our results show that in a model of viral infection, administering the combination therapy while increasing cAMP levels can further decrease inflammatory cytokines prompting further investigation for its potential implication in the clinic.

In this dissertation I am using M. truncatula as a model legume that forms indeterminate nodules with rhizobia under limited nitrogen conditions. I take advantage of an M. truncatula Tnt1 mutant population that provides a useful resource to uncover and characterize novel genes. Here, I focused on several objectives. First, I carried out forward and reverse genetic screening of M. truncatula Tnt1 mutant populations to uncover novel genes involved in symbiotic nitrogen fixation. Second, I focused on reverse genetic screening of two genes, identified as encoding blue copper proteins, and characterization of their mutants' potential phenotypes. Third, I further characterized a nodule essential gene, M. truncatula vacuolar iron transporter like 8 (MtVTL8), which encodes a nodule specific iron transporter. I characterized the expression pattern, expression localization and function of MtVTL8. Additionally, I characterized several residues predicted to be essential to function using a model based on the known crystal structure of Eucalyptus grandis vacuolar iron transporter 1 (EgVIT1), a homologous protein to MtVTL8. I identified several potential essential residues of the MtVTL8 protein, mutagenized them, and through complementation experiments in planta and in yeast assessed functionality of the resulting protein. This helped us to better understand the potential mechanism by which MtVTL8 functions.

Utmaningar som ökad stillasittande, ohälsa och klimatförändringar kräver nya strategier. Vallastaden i Linköping exemplifieras som ett projekt som använt innovativa strategier för att möta globala utmaningar. Området betonar social hållbarhet men också ekologisk hållbarhet vilket integreras strategiskt i staden för att främja och uppmuntra till en hållbar livsstil och stad. Intresset riktas mot hur Vallastadens utformning påverkar invånarnas attityd till fysisk aktivitet och miljömedvetenhet. Fysisk aktivitet betonas som avgörande för hälsa och livslängd, och den fysiska miljöns utformning spelar en nyckelroll. Stadsidealet nyurbanism samt Edward Sojas teori, den socio-spatiala dialektiken, har applicerats för att undersöka uppsatsens syfte och besvara frågeställningarna. De teoretiska ramverken ger en djupare inblick i och förståelse för hur Vallastadens design och struktur påverkar invånarna och visar på samspelet mellan fysisk miljö och mänskligt beteende. För att undersöka Vallastadens påverkan på invånarnas fysiska aktivitet och hållbara val, samt bedöma hur väl kommunens mål uppnåtts, användes både kvalitativa och kvantitativa metoder. De kvalitativa metoderna – intervjuer, observation, enkät och litteraturstudier – gav djupare insikt i människors tankar och känslor medan de kvantitativa metoderna – enkät och statistik – möjliggjorde generalisering. Genom att kombinera båda metoderna strävade studien efter en mer omfattande och nyanserad förståelse av ämnet. Resultaten indikerar att Vallastaden, genom sin integrerade planering med grönområden och attraktiva offentliga platser, strävar efter att främja en aktiv livsstil och hållbara transportval. Dess strategi med parkeringshus och hastighetsbegränsningar framstår som en åtgärd för att skapa en säkrare miljö som främjar hälsa och gemenskap. Även om Vallastaden framhävs som ett framgångsrikt exempel, återges det att vissa invånare inte noterar en betydande förändring i sina aktivitetsnivåer. Detta pekar på en potentiell avvikelse från de större målen för projektet. Trots dessa avvikelser har Vallastadens resultat i huvudsak stämt överens med kommunens ambitioner.

To better understand the digestive and absorptive capacities of the chick intestine so that we may feed diets that better meet the nutritional needs of the chick, it is important to understand how expression of nutrient transporter genes changes in response to various factors. A series of feeding trials were conducted to evaluate the dietary and developmental regulation of nutrient transporter mRNA abundance in the small intestine of two lines of broilers selected on corn-based (Line A) or wheat-based (Line B) diets. Abundance of mRNA was quantified in all experiments using real time PCR and the absolute quantification method. The objective of the first study was to investigate intestinal nutrient transporter and enzyme mRNA in Line A and B broilers at embryo day 18 and 20, day of hatch, and d 1, 3, 7, and 14 posthatch. Genes evaluated included the peptide transporter, PepT1, 10 AA transporters (rBAT, bo,+AT, ATBo,+, CAT1, CAT2, LAT1, y+LAT1, y+LAT2, BoAT and EAAT3), four sugar transporters (SGLT1, SGLT5, GLUT5, and GLUT2), and a digestive enzyme, APN. For PepT1, Line B had greater quantities of mRNA compared with Line A (P = 0.001), suggesting a greater capacity for absorption of AA as peptides. Levels of PepT1 mRNA were greatest in the duodenum (P < 0.05), whereas the abundances of SGLT1, GLUT5 and GLUT2 mRNA were greatest in the jejunum (P < 0.05). Abundances of EAAT3, bo,+AT, rBAT, BoAT, LAT1, CAT2, SGLT5 and APN mRNA were greatest in the ileum (P < 0.05). Quantities of PepT1, EAAT3, BoAT, SGLT1, GLUT5, and GLUT2 mRNA increased linearly (P < 0.01), while CAT1, CAT2, y+LAT1, and LAT1 mRNA decreased linearly (P < 0.05) with age. The objective of the second study was to evaluate the effect of dietary protein quality on intestinal peptide, AA, and glucose transporter, and digestive enzyme mRNA abundance in Line A and B broilers. At day of hatch (doh), chicks from both lines were randomly assigned to corn-based diets containing 24% crude protein (CP) with either soybean meal (SBM) or corn gluten meal (CGM) as the supplemental protein source, ad libitum. Groups of chicks from both lines were also assigned to the SBM diet at a quantity restricted to that consumed by the CGM group (SBM-RT). Abundance of PepT1, EAAT3, and GLUT2 mRNA was greater in Line B (P < 0.03), while APN and SGLT1 were greater in Line A (P < 0.04). When feed intake was equal (CGM vs restricted SBM), a greater abundance of PepT1 and bo,+AT mRNA was associated with the higher quality SBM (P < 0.04), while a greater abundance of EAAT3 and GLUT2 mRNA was associated with the lower quality CGM (P < 0.01). When feed intake was restricted (SBM vs SBM-RT), a greater abundance of PepT1 mRNA was associated with the restricted intake (P < 0.04). The objective of the third study was to determine the effect of dietary protein composition on mRNA abundance of peptide and AA transporters, and a digestive enzyme. From day 8 to day 15 posthatch, Line A and B broilers were fed equal amounts of 1 of 3 diets (24% CP). Dietary protein sources included whey protein concentrate (whey), a partial whey hydrolysate (hydro), or a mixture of free amino acids (AA) similar to the composition of whey. Intestine was collected at days 8, 9, 11, 13, and 15. Expression of all genes except LAT1 was greater (P < 0.05) in Line B compared with A. Abundance of PepT1, EAAT3, y+LAT2, CAT1, bo,+AT, and APN mRNA varied little across diets in Line A but for CAT1 mRNA was greatest (P = 0.005) in Line A birds that consumed the AA diet. Expression of these genes was greatest (P < 0.006) in Line B birds consuming the hydro diet. A greater (P < 0.05) age response of bo,+AT, EAAT3, CAT1, and APN mRNA was observed in birds consuming the hydro or AA diets relative to the whey diet. Results from these studies collectively demonstrate that nutrient transporter gene expression is responsive to a variety of factors, including developmental stage, dietary manipulation, and genetic selection. Information from these studies can be used to improve dietary formulation so that nutrient utilization is enhanced, resulting in improved growth of the broiler. / Ph. D.

Amino acids are necessary for protein synthesis and specialized metabolism in plants. Yet very little is known about how plants sense and regulate when and where to allocate amino acids to meet the demand for nitrogen in growing tissues. In particular, while characterized in yeast and mammals, no amino acid sensor has been identified in plants. Amino Acid Permease 1 (AAP1) has been previously characterized and was shown to mediate amino acid uptake from the soil. aap1 knockout plants and several EMS mutants affected in AAP1 sequence display enhanced tolerance to toxic concentrations of amino acids. Yet, two of the corresponding variant proteins appear to be functional transporters, effectively dissociating amino acid transport and phenotype. To understand this apparent discrepancy, I precisely studied AAP1 localization of expression at the plant and cellular level, and in specific tissue types of the root where AAP1 function is required for the tolerance phenotype and the amino acid uptake activity. I showed that AAP1 protein is present in the endoplasmic reticulum of the cortex in wild type plants Yet, its ectopic expression in root tip and phloem increased amino acid uptake, while expression in cortex could not. This and other of my results do not support the current model of AAP1 functioning in amino acid uptake by the root. I propose that the main effect of mutations in AAP1 is a disturbance in amino acid metabolism, possibly triggered by altered amino acid sensing. In this new model, AAP1 would be necessary for sensing amino acid status of cortex cells, possibly in the endoplasmic reticulum, and adjust amino acid metabolic activity and uptake to current availability. In effect, disruption of the sensing function, either by complete loss of AAP1 function (knockout) or by uncoupling the transport and sensing function (EMS mutants), would lead to the various characteristics of the phenotype of the aap1 mutants I observed. My main hypothesis is that AAP1 is a transporter endowed with sensing function, i.e., an amino acid transceptor. / Doctor of Philosophy / Changing environments create challenges for plants to grow under harsher, nutrient limiting conditions. Nitrogen is an essential nutrient for plant growth, used for the synthesis of amino acids and other nitrogen-containing metabolites. Amino acids are necessary for protein synthesis and other specialized metabolism – being targets for manipulation for improving agronomic traits. Protein content is a complex trait that involves many genes, possibly including amino acid transporters. In addition, the amount of nitrogen needed by and available to the plant increases or decreases depending on the environment conditions. How plants control nitrogen need and use at the molecular level is not well understood. The data presented here challenge a current model and I report how a protein (AAP1) involved in the acquisition of amino acids from the soil provides regulatory control over these processes. . This valuable information is useful for better understanding how plants use nitrogen and more precise breeding methods can be used to improve traits, such as protein content in agronomically important crops.

Generation and characterization of activation tagged potato mutants could aid in functional genomic studies. Morphological and molecular studies were conducted to compare potato cv. Bintje, its two mutants, underperformer (up), and nikku generated using the activation tagging vector pSKI074, and nikku revertant plants. Mutant up exhibited a dwarf phenotype (plant height 42 cm vs. 73 cm in cv. Bintje), abundant axillary shoot growth (3.1 shoots/plant compared to 0.7 shoots/plant in cv. Bintje; in vitro plants), greater tuber yield, altered tuber traits and early senescence compared to wild-type Bintje under in vitro conditions. Under in vivo conditions, the dwarf and early senescence phenotypes of the mutant were consistent, but the tuber yield of up was less (250 g/plant compared to 610 g/plant in wild-type Bintje) and had fewer axillary shoots compared to wild-type (1.9 shoots/plant in up vs. 4.7 shoots/plant in Bintje). Mutant nikku plants exhibited an extremely dwarf phenotype (plant height 2 cm in nikku vs. 6 cm in Bintje), had small hyponastic leaves, were rootless, and infrequently produced small tubers when compared to cv. Bintje. The overall nikku phenotype was suggestive of a constitutive stress response, which was further supported by the higher expression levels of several stress-responsive genes in nikku. The nikku revertant plants exhibited near normal stem elongation, larger leaves and consistent rooting, and it was a case of partial reversion. Southern blot analyses indicated the presence of single T-DNA insertions on chromosome 10 in the up and on chromosome 12 in the nikku mutant. The reversion in the nikku plants was not associated with the loss of enhancer copies from the original nikku mutant. Reverse transcriptase PCR analyses indicated transcriptional activation/repression of several genes in the up and nikku mutants, suggesting pleiotropic effects. In revertant, the expression levels of several genes which were differentially regulated in the nikku mutant were similar to Bintje. The gene immediately flanking the right border of the T-DNA insertion, which encoded a novel BTB/POZ (Broad complex, Tramtrac, Bric a brac; also known as Pox virus and Zinc finger) domain-containing protein, was highly up-regulated in the up mutant. This protein domain plays an important role in several important developmental, transcriptional and regulatory pathways. The mRNA-seq analyses resulted in 1,632 genes that were differentially expressed between mutant up and Bintje and the total number of up-regulated genes (661) were less than the number of genes down-regulated (971 genes) in the up mutant. Further analyses indicated that a variety of biological processes including decreased cell division, cell cycle activity, and abiotic stress responses were modified in the up mutant. In the nikku mutant, two potato genes, encoding an Acyl-CoA N-acyltransferases (NAT) superfamily protein, and a predicted major facilitator superfamily protein (MFS) were identified and overexpression lines Bintje/35S::NAT1 and Bintje/35S::PMT1 were created for recapitulation of the nikku mutant phenotype. Methylated DNA-PCR between the nikku and the revertant indicated a change in methylation status of the 35S enhancers, suggesting that the nikku revertant phenotype may be associated with some epigenetic modification. / Ph. D.

Aspergillus fumigatus is a ubiquitous fungus associated with several airway complications and diseases including asthma, allergies, cystic fibrosis, and most commonly invasive aspergillosis. The airway epithelium, a protective barrier, is the first anatomical site to interact with A. fumigatus. Although this host-fungal interaction is often asymptomatic for immunocompetent individuals, for immunocompromised persons, due to a weakened competence of the immune system, they have an increased likelihood of fungal infection. This dissertation aims to investigate the effect of A. fumigatus on the transcriptional response of human airway epithelial cells, focusing on the relationship between innate immunity and iron regulation from the host perspective. The trace element iron is needed by both the fungus and the host for cellular maintenance and survival, but tightly controlled iron regulation in the host is required to prevent oxidative stress and cell death. The research methods in this dissertation employ a systems biology approach, by incorporating mathematical modeling, RNA-seq analysis, and experimental biology techniques to assess the role of airway epithelial cells in the host-fungal interaction. Both the quantitative and qualitative research design allows for characterization of airway epithelial cells and the downstream changes in iron importer genes. This study addresses literature gaps through analysis of the host transcriptome using multiple time points, by performing an extensive evaluation of the effect of cytokines on iron importer genes, and conceptualization of a comprehensive mathematical model of the airway epithelial cell. The major findings suggest the following: 1) airway epithelial cells avidly respond to A. fumigatus through modification of the expression of immune response related genes at different infection stages, 2) during A. fumigatus co-incubation with airway epithelial cells, the iron importers genes respond in strikingly different ways, and 3) cytokines have a significant effect on the increase in expression of an iron importer gene. We illuminated the role of airway epithelial cells in fungal recognition and activation of the immune response in signaling cascades that consequently modify iron importer genes and hope to use this information as a platform to discover potential therapeutic targets. / Ph. D.

The mesolimbic dopamine (DA) system originates in the ventral tegmental area (VTA) and projects to the nucleus accumbens (NAc) and other areas including the basolateral amygdala (BLA), prefrontal cortex, and the hippocampus. Drug use induces reward and leads to dysregulation in these brain areas and eventually to substance use disorders (SUDs). Chapter 1 introduces the mesolimbic DA system and its relationship to drug use and their relevance to each chapter. Chapter 2 explores opioid effects on BLA circuitry which is known to play a role in the emotional response including anxiety and stress in SUDs. We showed that morphine induced an inhibitory effect on GABAergic lateral paracapsular cells (LPCs). These cells inhibit BLA principal neuron output and are influenced by opioids. Opioid activation in LPCs leads to upregulated BLA output, and activation in the NAc and central amygdala which may have important implications for stress/anxiety response for patients with SUDs. Chapter 3 explores the effect of interleukin-10 on the mesolimbic DA system. Specifically, cell-attached recordings of VTA DA neurons increase their firing rate in the presence of IL-10, and in vivo studies showed increased DA release in the NAc. Interleukin-10 receptors were expressed in VTA DA neurons and signals through the phosphoinositide 3-kinase. Surprisingly, IL-10 induced conditioned place aversion in mice which may be related to depression- and anxiety-like behaviors reported by others. Thus, IL-10 appears to be regulating the mesolimbic DA system and its association with reward which may be important in understanding the relationship between inflammation and SUDs. Chapter 4 explores the DA transporter (DAT) kinetics in the presence of psychostimulants using DA iontophoresis. We showed that iontophoretic DA delivery increased DA concentration and clearance rates compared to evoked release making it an important tool in measuring DAT kinetics. Cocaine was bath applied and slowed DAT reuptake at high concentrations and D2 stimulant quinpirole slowed the reuptake process but did not show any effect on DAT trafficking, and D2 antagonist eticlopride showed no change in reuptake or DAT trafficking. Cocaine-injected mice increased locomotion and reduced anxiety-like behavior, and iontophoresis experiments slowed reuptake with bath-applied cocaine. Thus, DA iontophoresis is useful in studying DAT blocker kinetics but has limitations in studying the effects of DAT trafficking. Chapter 5 discusses the impact these studies have on society, the limitations of each chapter, and future directions for this dissertation. Together these studies explore the reward system and its relationship with SUDs. The overarching aim has been to understand the involvement of DA in motivation and reward in the context of SUDs and the influence of opioids, cytokines, and psychostimulants.