An intercomparison exercise of the world-harmonized light-duty vehicle test procedure (WLTP) aiming at measuring ethanol, formaldehyde and acetaldehyde emissions from a flex-fuel light-duty vehicle using E85 was conducted in the Vehicle Emission Laboratory (VELA) at the European Commission Joint Research Centre (EC-JRC), Ispra, Italy. The instruments used during the intercomparison allowed online measurements of these compounds directly from the diluted exhaust. Measurements were done either in real time or immediately after the test. The measurement and analysis of exhaust emissions over the world-harmonized light-duty vehicle test cycle was done by means of Fourier transform infrared spectroscopy (FTIR), proton transfer reaction-mass spectrometry (PTR-Qi-ToF-MS), photoacoustic spectroscopy (PAS) and gas chromatography (GC). Results showed that online systems can perform measurements from the vehicle diluted exhaust assuring a good repeatability (within instrument variance) and reproducibility (between instrument variance) of the results. Measurements from all the instruments were in good agreement (|Z-score| < 2). Results showed that online systems can perform measurements from the vehicle diluted exhaust assuring the reproducibility and repeatability of the results. Results obtained measuring at the tailpipe using a FTIR were in good agreement with those acquired measuring at the constant volume sampler (CVS). Considering the low sensitivity of the current technique used to measure hydrocarbons emissions towards oxygenated compounds (flame ionization detector; FID), non-methane organic gases (NMOG) were calculated applying their FID response factors to the measured emissions of ethanol, acetaldehyde and formaldehyde. NMOG resulted to be up to 74% higher than measured non-methane hydrocarbons (NMHC).

The thesis deals with the issue of a teaching assistant aiding pupils with an autistic spectrum disorder. The aim is to uncover the specifics of teaching assistant's role in aiding pupils with an autistic spectrum disorder and enumerate the important aspects of their profession. The thesis is divided into a theoretical and a practical part. The theoretical parts summarise the findings regarding the discussed topic. The teaching assistant's institution is introduced here as it is included in the Czech school counselling system. After that, the autistic spectrum disorder is discussed. The attention is focused on the exhibition of this disorder within everyday schooling and the possibilities to eliminate its negative influence. The developmental milestones of pupils attending the elementary school are theoretically enumerated as well as the characteristics of each developmental period. The practical part introduces the reader to the experience of chosen teaching assistant aiding pupils with an autistic spectrum disorder. The methodology of the research is mentioned here. The research approach is qualitative. The data is picked employing partially structured interviews with five respondents. The analysis and the interpretation of the data are done via the grounded theory method. The characteristic...

The diploma thesis aims to map the lived reality of adult intact siblings of people with autism spectrum disorders in the Czech Republic. The theoretical part provides an insight into the current knowledge in the field of autism spectrum disorders and the manifestations thereof, sibling relationships and the impact of autism spectrum disorders on the family in general, and with respect to intact siblings, in particular. In the empirical part I focus on the description and analysis of qualitative research involving five intact siblings of people with autism spectrum disorders aged 28-29. For the purposes of the research, a semi-structured interview had been chosen, subsequently analyzed and interpreted. The interview topics concerned the past and present relationship of the informants to their siblings with ASD, ideas about their common future, the personality of an intact sibling or certain life choices of the informants. The results of the diploma thesis showed an interesting picture of the development of the sibling relationships with people with ASD, which are strongly affected by the nature of the disorder. The research confirmed a severe amount of stress in intact siblings of people with ASD, which stems from many different areas of their living together. It also turned out that living with...

Phosphate is an essential nutrient for all forms of life. Escherichia coli has a PhoR/PhoB two component regulatory system that controls the expression of various genes whose products allow the cell to thrive in low phosphate environments. The signaling mechanism of the PhoR/PhoB system has been studied and the phosphorylation cascade that controls gene expression is well understood. What is still unknown is how PhoR senses the phosphate level of the environment. The PstS, PstC, PstA, PstB, and PhoU proteins play a role in this signal sensing. This work confirms the hypothesis that the PstSCAB complex senses the environmental phosphate and that phosphate signal is passed through PhoU to PhoR. Further, this work characterizes residues important for interaction on PhoU and PhoR and identifies a structural model for interaction. This model points to a potential mechanism for PhoU mediated signaling to PhoR. We tested this model with direct coupling analysis and obtained further confirmation. Further use of these techniques may elucidate more of the interactions necessary for proper phosphate signaling.

Nutrient sensing kinases sense available nutrients and regulate cell activity accordingly. Three of these enzymes are AMP regulated kinase (AMPK, or Snf1 in yeast), PAS kinase, and target of rapamycin (TOR), are conserved from yeast to man and have overlapping function. AMPK and Snf1 are important in sensing when nutrient status in the cell is low and down regulating energy consuming pathways. PAS kinase is required for glucose homeostasis in the cell, and responds to glucose levels. TOR senses nutrients such as amino acids and upregulates cell growth pathways primarily through protein synthesis. Due to the varying nature of these enzymes, cross talk is expected in order for the cell to properly regulate cellular metabolism and growth in response to energy and nutrient availability. Previous studies have shown that activation of yeast PAS kinase under nutrient stress conditions requires the presence of Snf1. The aim of this thesis is to determine whether Snf1 directly phosphorylates and activates PAS kinase through both in vivo and in vitro approaches. PAS kinase was found to require Snf1 for both activation and phosphorylation in vivo. In vitro kinase assays were also performed to confirm a direct phosphorylation event. The results from this study support the direct phosphorylation and activation of PAS kinase by Snf1, linking cellular energy status to glucose allocation.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Ataxin-2 is an RNA-binding protein that is involved in many crucial cellular processes such as R-loop regulation, mRNA stability, TOR signaling regulation, and stress granule formation. Ataxin-2 is highly conserved, found in organisms ranging from Saccharomyces cerevisiae to Caenorhabditis elegans and Homo sapiens. Recently, ataxin-2 has been linked to the neurodegenerative disease Amyotrophic Lateral Sclerosis (ALS). ALS is a fatal disease that causes loss of motor neurons. In addition to ataxin-2 interacting with known ALS risk factor proteins, research into the relationship between ataxin-2 and ALS shows that polyglutamine expansions in ataxin-2 are gain-of-function mutations that lead to overactivity of ataxin-2 and probable neurodegeneration. In fact, targeting ataxin-2 using gene silencing techniques dramatically slows the progression of ALS in both mice and man.The Grose laboratory has characterized a serine-threonine kinase, PAS kinase as upstream kinase and putative activator of ataxin-2. We hypothesize that knockdown of PAS kinase could, therefore, have similar effects to directly downregulating ataxin-2 and its cellular functions. Characterization of Ataxin-2 has revealed that its gain or loss of function lead to distinct cellular phenotypes. One study concluded that lowering ataxin-2 levels reduced the size and number of stress granules in mammalian cells, which was observed through microscopy. Another study found that activation and overexpression of ataxin-2 lead to reduced mTOR levels because of its sequestration to stress granules. Lastly, preliminary data obtained by the Grose laboratory suggests that yeast deficient in Pbp1 (the yeast homologue of ataxin-2) have altered cell cycles.This project describes the cellular readouts used to determine if PAS kinase downregulation confers the same cellular phenotypes as ataxin-2 downregulation. First, we found that PAS kinase does influence ataxin-2 abundance in mammalian cells. Using yeast as a model, we found that Pbp1 influences the cell cycle through its binding partners, causing a reduction in the percentage of cells in the G2 phase compared to the G1 phase. PAS kinase conferred an opposite change, most likely due to the activity of other PAS kinase substrates. Additionally, we found that Pbp1 deficiency is synthetically lethal when in conjunction with deficiency of any one of its cell cycle-related binding partners. The cellular changes cause by Pbp1 deficiency highlight not only the importance of ataxin-2 in the cell, but also the importance of understanding the effects of downregulation of ataxin-2.

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For over 20 years, the phenomenon of parental alienation and its related characteristics has been described in literature. Various clinicians independently researched and described the pathological alignment of a child within the context of divorce. In the 1980’s Richard Gardner coined this “Parental alienation syndrome” (PAS). According to Gardner PAS is characterised by a child’s profound preoccupation with criticism against a parent. This criticism is overstated and ungrounded, and it is caused by both conscious and unconscious behaviour by the alienating parent, which influences the child negatively against the alienated parent. In essence, PAS is the subjective contamination of the child’s understanding and beliefs about his/her environment. The alienating parent gives the impression that the targeted parent is unworthy, dangerous, unloving and deserving of the child’s rejection. This is done by a series of alienation strategies like bad-mouthing, blaming, limiting contact and belittling. The alienated child, as a result responds with unjustified expressions of hate and discontent towards the targeted parent. The experience of divorce can be very challenging to children. Research about the relationship between divorce and child adjustment holds that a child’s exposure to inter parental conflict and the quality of the parent-child relationship are the two major predictors of children’s adjustment during divorce. Research suggests that the negative effects of PAS may include guilt, self-hatred, distortion of reality testing, and general emotional and psychological problems. The aim of this study was to provide an in-depth exploration and description of how PAS is experienced, and the possible effect it has on children from the perspective of young adults who was possible exposed to PAS as children. This was done by exploring their memories and their recalled experiences of their parents’ divorce and the possible effect on their current lives. In this study a collective exploratory/descriptive case study design was used. Nine voluntary participants, between 18 and 28 years of age, were chosen for this study by means of purposeful sampling strategies. Data were collected through in-depth semi-structured interviews that were audio taped and transcribed verbatim. Transcribed data were analysed by means of thematic analysis from which themes and sub-themes were derived. Two main themes with sub-themes were identified. It was found that some of the parental behaviour evident in PAS cases may fall under specific subtypes of psychological maltreatment and leave children feeling angry, worthless, flawed, unloved, unwanted, incompetent and sceptical about trusting other individuals. It was found that parents who engage in alienation strategies are likely to discourage autonomous behaviour and lack nurturance and emotional responsiveness towards their children’s needs. As result children exposed to PAS learn parental love and acceptance is conditional and is based needs fulfilment of the alienating parent. These expectations are brought forward from the early relationship into adulthood and influences individual’s expectations, behaviour, and beliefs about relationships across the lifespan. The results indicate that the lack parental support, encouragement, and responsiveness may negatively influence the self-esteem, autonomy, competence, and relatedness of individuals exposed to PAS. They reported difficulties with trust, intimacy and social skills and depression as adults. / MA (Clinical Psychology), North-West University, Potchefstroom Campus, 2014