Bringing it home: the inter-American system and state obligations - using a gender approach regionally to address women's rights violations domestically

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Rate-limiting Steps in Transcription Initiation are Key Regulatory Mechanisms of Escherichia coli Gene Expression Dynamics

In all living organisms, the “blueprints of life” are documented in the genetic material. This material is composed of genes, which are regions of DNA coding for proteins. To produce proteins, cells read the information on the DNA with the help of molecular machines, such as RNAp holoenzymes and a factors. Proteins carry out the cellular functions required for survival and, as such, cells deal with challenging environments by adjusting their gene expression pattern. For this, cells constantly perform decision- making processes of whether or not to actively express a protein, based on intracellular and environmental cues. In Escherichia coli, gene expression is mostly regulated at the stage of transcription initiation. Although most of its regulatory molecules have been identified, the dynamics and regulation of this step remain elusive. Due to a limited number of specific regulatory molecules in the cells, the stochastic fluctuations of these molecular numbers can result in a sizeable temporal change in the numbers of transcription outputs (RNA and proteins) and have consequences on the phenotype of the cells. To understand the dynamics of this process, one should study the activity of the gene by tracking mRNA and protein production events at a detailed level. Recent advancements in single-molecule detection techniques have been used to image and track individually labeled fluorescent macromolecules of living cells. This allows investigating the intermolecular dynamics under any given condition. In this thesis, by using in vivo, single-RNA time-lapse microscopy techniques along with stochastic modelling techniques, we studied the kinetics of multi-rate limiting steps in the transcription process of multiple promoters, in various conditions. Specifically, first, we established a novel method of dissecting transcription in Escherichia coli that combines state-of-the-art microscopy measurements and model fitting techniques to construct detailed models of the rate-limiting steps governing the in vivo transcription initiation of a synthetic Lac-ara-1 promoter. After that, we estimated the duration of the closed and open complex formation, accounting for the rate of reversibility of the first step. From this, we also estimated the duration of periods of promoter inactivity, from which we were able to determine the contribution from each step to the distribution of intervals between consecutive RNA productions in individual cells. Second, using the above method, we studied the a factor selective mechanisms for indirect regulation of promoters whose transcription is primarily initiated by RNAp holoenzymes carrying a70. From the analysis, we concluded that, in E. coli, a promoter’s responsiveness to indirect regulation by a factor competition is determined by its sequence-dependent, dynamically regulated multi-step initiation kinetics. Third, we investigated the effects of extrinsic noise, arising from cell-to-cell variability in cellular components, on the single-cell distribution of RNA numbers, in the context of cell lineages. For this, first, we used stochastic models to predict the variability in the numbers of molecules involved in upstream processes. The models account for the intake of inducers from the environment, which acts as a transient source of variability in RNA production numbers, as well as for the variability in the numbers of molecular species controlling transcription of an active promoter, which acts as a constant source of variability in RNA numbers. From measurement analysis, we demonstrated the existence of lineage-to-lineage variability in gene activation times and mean transcription rates. Finally, we provided evidence that this can be explained by differences in the kinetics of the rate-limiting steps in transcription and of the induction scheme, from which it is possible to conclude that these variabilities differ between promoters and inducers used. Finally, we studied how the multi-rate limiting steps in the transcription initiation are capable of tuning the asymmetry and tailedness of the distribution of time intervals between consecutive RNA production events in individual cells. For this, first, we considered a stochastic model of transcription initiation and predicted that the asymmetry and tailedness in the distribution of intervals between consecutive RNA production events can differ by tuning the rate-limiting steps in transcription. Second, we validated the model with measurements from single-molecule RNA microscopy of transcription kinetics of multiple promoters in multiple conditions. Finally, from our results, we concluded that the skewness and kurtosis in RNA and protein production kinetics are subject to regulation by the kinetics of the steps in transcription initiation and affect the single-cell distributions of RNAs and, thus, proteins. We further showed that this regulation can significantly affect the probability of RNA and protein numbers to cross specific thresholds. Overall, the studies conducted in this thesis are expected to contribute to a better understanding of the dynamic process of bacterial gene expression. The advanced data and image analysis techniques and novel stochastic modeling approaches that we developed during the course of these studies, will allow studying in detail the in vivo regulation of multi-rate limiting steps of transcription initiation of any given promoter. In addition, by tuning the kinetics of the rate-limiting steps in the transcription initiation as executed here should allow engineering new promoters, with predefined RNA and, thus, protein production dynamics in Escherichia coli

Malaysian Reference Network Modelling For Photovoltaic Impact Study

The burning of abundant fossil fuel amount to accommodate the energy demand causes emission of the greenhouse gas (GHG) which is contribute to the climate change. The Distributed Generation (DG) and renewable energy resources is one promising approach to reduce the emission, thereby mitigating climate change. However, most of the DG and renewable energy resources, especially the photovoltaic (PV) system is intermittent and often fluctuates. In this regard, this thesis focuses on the analysing the impact of PV integration at the medium voltage (MV) network. More precisely, the network losses and voltage issues are used to evaluate the effects of four variables; PV system installed locations, PV variability index (VI), different time resolution of PV generation profiles used and the PV penetration levels. The Malaysian Reference Network (RN) is modelled with the intention to analyse the impact of PV integration at the Medium Voltage (MV) network. Moreover, the breakdown of network losses for the Malaysian MV network is quantified by utilising the RN models. All case studies have been carried out for the urban, semi-urban and rural MV networks. The findings suggest that PV system installed at the end of the 11kV feeder for the rural and semi-urban produces significant network losses reduction than urban networks, which is driven by feeder length and load size. The findings also show that different categories of VI produce different impact on the networks. Voltage fluctuation and voltage step change are the two main concerns of various PV variability on the MV network. In addition, the different time resolution of real solar PV generation profile was collected to be used in the case study. By analysing different ranges of time resolution, it is suggested that a 15-minute time resolution PV generation profiles data are sufficient in network assessment study with approximately 5% error. The use of hourly PV generation data will cause up to 31% loss of accuracy. Furthermore, a case study was done to evaluate the impact of different PV penetration levels. The results show that PV integration above 60% and 100% penetration level will likely cause voltage rise violation in rural and urban networks, respectively. The research also suggests that the maximum PV penetration levels for a MV Malaysian network are based on several network performance indicators. In conclusion, the results show that location of PV system, PV variability, time resolution and PV penetration level are the key parameters to study the impact of PV in the MV networks

Determining the Level of Patient Satisfaction in a Dental Hygiene Setting.

Not many studies have been conducted in the past to determine patients\u27 level of satisfaction in academic dental hygiene settings. This patient satisfaction study analyzed the level of patient satisfaction with the dental hygiene clinic at East Tennessee State University, Johnson City, TN. The purpose of the study was to determine if there was a statistically significant influence of demographic characteristics of the patients and the affective behavior of the care providers on level of patient satisfaction. It was concluded that except for age, other demographic variables did not have any statistically significant influence on patient satisfaction. Also, care provider\u27s affective behavior significantly influenced patient satisfaction. Overall, the dental hygiene patients were satisfied with the clinic. As patients\u27 needs are prioritized in this customer-driven industry, such positive patient satisfaction data can be used for the welfare of the patients, the care providers, and the health care organization

T. Janakiraman’s ‘Uyirthen’ Novel in Feminism View

Feminism has branched out into a three-way approach to the concept of time. Despite the fundamental differences of opinion, the generality can also be seen in the feminist genres that developed into the three divisions of thought. Equality feminism appeared when the inadequacy of moderate feminism was realized. Extremist feminism and its consequent extreme feminism have emerged in the world as a result of the inadequacy of Egalitarian feminism. Today we can see that there is a tendency for women to choose one feminine genre as a thought-provoking guide for their life and ignore the other based on the land, family, social environment, education and economic status. In the pulsating literature of the community, one can find the people of that community and their thoughts. Feminist thought can also be seen in the works of T. Janakiraman, who creates female characters with the power of strength. The purpose of this article is to examine T. Janakiraman's novel 'Survived' by Feminist Thoughts. In T. Janakiraman 's novels, women are portrayed as a powerful personality. There are three types of feminism: T. Janakiraman; Sengamma, Renganayaki and Anusuya can be compared to the three characters in the novel 'Uyirthen'. This comparison will help us to identify the thinking resources of the three-fold feminism, to focus on the creative technique of the female characters and the social value of the woman