This work is will deal with the amount of costs and personal provisions with a view to the performance of regulations of independent regulatory authorities in the Czech Republic from 2001 until the present day. Independent regulatory authorities are characteristic for their specific placement within the hierarchy of state administration, their independence from the political will, and they are financed from the state budget. This work aims to find out what is the proportion of costs related to the output of regulations of independent regulatory authorities to the overall costs linked to the output of state administration, and analyze the development of this indicator in the aforesaid period. In the same way, the development in the personnel area will be analysed. As the source for data analysis will be primarily used annual reports of selected regulatory authorities, The Czech Statistical Authority and Ministry of Finance. The amount of regulations by the state and reinforcing or streamlining of the functions of the state administrations -- these are the topics of constant scholarly discussions both in the economic and political fields. At present, The European Commission requires that the Czech Republic establish two new independent regulatory authorities in the fields of railway transport and water management. The reason to establish an independent regulatory authority shall be to make sure of equal conditions of liberalized market of railway transportation and functioning of independent regulatory authority in the water-management field, as stated in the conditions of European Commission for subsidizing in the period between 2014 and 2020. The theoretical part will define the terms of regulations and describe the concepts of economic theories related to the regulations controlled by the state. Furthermore, the theoretical part will focus on definitions of independent regulatory authority and will describe the historical development of independent regulatory authorities. The practical part will concern itself with the institutional and legal framework, competency and activities of independent regulatory authorities. Furthermore, their budget and personnel aspects will be analysed. The goal of the Thesis is to analyse the development of costs and personnel scope of independent regulatory authorities with a view to the overall costs linked to the state administration and to the overall personnel provisions of the state administration in the mentioned period.

Over the years, the electronic letters of credit evolved as one of the developments to meet the international trade demands coupled with the exponential technology advancements of the current times which whetted an appetite for superfluous trade and competitiveness in the trade industry. Just like legal discrepancies pursuant to the use of the letter of credit in international trade, this too demanded some legal architecture to govern its utilization. However, unlike the traditional letters of credit, there are more legal stumbling blocks concerning this form of letters of credit. The primary legal constraints being, lack of legal recognition by the courts because of their nature (being data messages); lack of recognition in the laws of contracts (digital signatures, digital contracts), public perception more especially most of the developing countries, who because of lack of technology, resources and skilled man power, lacked knowledge on the advantages of technology advancement. So, the study interrogates the legal implications of an electronic letter of credit in the international trade transactions using Botswana as a case study. Importantly, it investigates the completeness and sufficiency of the legal regimes in Botswana to enable operation of the electronic letter of credit. The conclusions are that the Botswana e-legislation drafts so far are complete as regard to the legal principles enabling electronic transactions. It also argues that the laws are comprehensive enough, receptive to the electronic documents including the upcoming developments in technology and more importantly, the fact that it provides a level playing field for all the players by protecting the rights of the users of electronic transactions in general. / tm2017 / Centre for Human Rights / LLM / Unrestricted

As células T regulatórias (Tregs) são essenciais para a manutenção da tolerância periférica, prevenção de doenças autoimunes e limitantes nas doenças inflamatórias crônicas. Além disso, essas células exercem um papel fundamental no controle da rejeição de transplantes. Diferentes protocolos mostraram que é possível obter Tregs a partir de células T naive CD4+ in vitro. Para tal, é consenso que o TGF-? e a interleucina-2 (IL-2) são capazes de direcionar as células T naive CD4+ a se tornarem regulatórias após um estímulo antigênico (anti-CD3/CD28). Nosso grupo recentemente notou que, durante a imunomodulação de linfócitos T pelas células estromais mesenquimais (CTMs), estas eram capazes de produzir adenosina que, por sua vez, participa do processo de imunorregulação. Outros trabalhos indicam que as CTMs suprimem a proliferação dos linfócitos T pela geração de Tregs e que as CTMs induzem a geração de Tregs através da regulação negativa da via TCR e da via AKTmTOR. Evidências apontam que a adenosina pode atuar regulando negativamente a via mTOR. Portanto, acredita-se que a adenosina possa participar do processo de geração de Tregs através da modulação da via mTOR. Além disso, estudos recentes indicam que a ativação de receptores de adenosina, mais especificamente A2a, com agentes agonistas, leva ao aumento da produção de células Tregs, enquanto que a utilização de agentes antagonistas destes receptores leva à diminuição da diferenciação de Tregs. Porém, estes estudos mostram a geração de Tregs a partir de células T naive de camundongos. Visto a grande importância das Tregs no contexto imunológico, a produção eficiente de Tregs in vitro tem importância fundamental para o desenvolvimento de novos protocolos terapêuticos para o tratamento de doenças autoimunes e no combate à rejeição de transplantes. Assim, o objetivo central deste trabalho foi avaliar a participação de agonistas e antagonistas de receptores de adenosina na indução de células T regulatórias geradas in vitro (iTreg) pela ativação de células T CD4+ naive isoladas de sangue de cordão umbilical (SCU) humano. Para isso, células mononucleares foram isoladas de bolsas de SCU e as células T naive foram isoladas imunomagnéticamente. Essas células foram ativadas com beads ligadas a anticorpos anti-CD2/CD3/CD28 e cultivadas por cinco dias na presença de IL-2 e diferentes concentrações de drogas agonistas e antagonistas de receptores de adenosina. Em seguida, foram avaliados os principais marcadores de células T regulatorias por meio de citometria de fluxo e o meio de cultura foi coletado ao final da geração para quantificação de citocinas. Além disso, o RNA total foi extraído de todas as condições de cultivo para a análise da expressão de genes envolvidos na geração e desenvolvimento das Tregs, por PCR quantitativo. O potencial de supressão de células T efetoras também foi avaliado. / Regulatory T cells (Tregs) are essential for the maintenance of peripheral tolerance, prevention of autoimmune and limiting diseases in chronic inflammatory diseases. In addition, these cells play a key role in the control of transplant rejection. Different protocols have shown that it is possible to obtain Tregs from naive CD4+ T cells in vitro. To this end, there is consensus that TGF-? and interleukin-2 (IL-2) are capable of directing the naive CD4 + T cells to become regulatory following an antigenic stimulus (anti-CD3/CD28).. Our group recently noted that during the immunomodulation of T lymphocytes by mesenchymal stromal cells (MSCs), they were able to produce adenosine which in turn participates in the immunoregulation process. Other studies indicate that MSCs suppress the proliferation of T lymphocytes by generation of Tregs and that MSCs induce generation of Tregs by downregulation of the TCR pathway and the AKT-mTOR pathway. Evidence indicates that adenosine may act by downregulating the mTOR pathway. Therefore, it is believed that adenosine may participate in the generation of Tregs by modulating the mTOR pathway. In addition, recent studies indicate that activation of adenosine receptors, more specifically A2a, with agonist agents, leads to increased production of Treg cells, whereas the use of antagonistic agents of these receptors leads to a decrease in Treg differentiation.. However, these studies show the generation of Tregs from naive T cells of mice. In view of the great importance of Tregs in the immunological context, the efficient production of Tregs in vitro is of fundamental importance for the development of new therapeutic protocols for the treatment of autoimmune diseases and in the fight against transplant rejection. Thus, the central objective of this study was to evaluate the participation of adenosine receptor agonists and antagonists in induction of regulatory T cells generated in vitro (iTreg) by the activation of naive CD4+ T cells isolated from human umbilical cord blood (SCU). For this, mononuclear cells were isolated from SCU and naive T cells were immunomagnetic isolated. These cells were activated with beads bound to anti-CD2/CD3/CD28 antibodies and cultured for five days in the presence of IL-2 and different concentrations of agonist drugs and antagonists of adenosine receptors. Next, the major regulatory T-cell markers were assessed by flow cytometry and the culture medium was collected at the end of the generation for quantification of cytokines. In addition, total RNA was extracted from all culture conditions for the analysis of the expression of genes involved in the generation and development of Tregs by quantitative PCR. The potential for suppression of effector T cells was also evaluated.

Nuestro conocimiento de las interacciones moleculares nos ha conducido hoy hacia una perspectiva ingenieril, donde diseños e implementaciones de sistemas artificiales de regulación intentan proporcionar instrucciones fundamentales para la reprogramación celular. Nosotros aquí abordamos el diseño de redes de genes como una forma de profundizar en la comprensión de las regulaciones naturales. También abordamos el problema de la diseñabilidad dada una genoteca de elementos compatibles. Con este fin, aplicamos métodos heuríticos de optimización que implementan rutinas para resolver problemas inversos, así como herramientas de análisis matemático para estudiar la dinámica de la expresión genética. Debido a que la ingeniería de redes de transcripción se ha basado principalmente en el ensamblaje de unos pocos elementos regulatorios usando principios de diseño racional, desarrollamos un marco de diseño computacional para explotar este enfoque. Modelos asociados a genotecas fueron examinados para descubrir el espacio genotípico asociado a un cierto fenotipo. Además, desarrollamos un procedimiento completamente automatizado para diseñar moleculas de ARN no codificante con capacidad regulatoria, basándonos en un modelo fisicoquímico y aprovechando la regulación alostérica. Los circuitos de ARN resultantes implementaban un mecanismo de control post-transcripcional para la expresión de proteínas que podía ser combinado con elementos transcripcionales. También aplicamos los métodos heurísticos para analizar la diseñabilidad de rutas metabólicas. Ciertamente, los métodos de diseño computacional pueden al mismo tiempo aprender de los mecanismos naturales con el fin de explotar sus principios fundamentales. Así, los estudios de estos sistemas nos permiten profundizar en la ingeniería genética. De relevancia, el control integral y las regulaciones incoherentes son estrategias generales que los organismos emplean y que aquí analizamos. / Rodrigo Tarrega, G. (2011). Computational design and designability of gene regulatory networks [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/14179 / Palancia

The decline in the profitability of credit unions with less than $10 million in assets harms the number of small credit unions available to serve local communities. Grounded in the financial intermediation theory, the purpose of this quantitative correlational study was to examine the relationship between regulatory compliance costs, operation costs, and profitability. The population of this study consisted of federally insured credit unions with less than $10 million in assets and located in the state of Texas. Archival data from the National Credit Union Administration database were collected and analyzed. Multiple regression was used to identify a statistically significant predictive model, F (2, 49) = 3.834, p = .028, R2 =.135. The implications for positive social change include the potential for credit union managers to improve decision-making processes related to current and future operations and investments, which could increase profitability and contribute to the financial prosperity of employees, employees' families, communities, and local economies.

Billions of cells die everyday as a result of normal tissue turnover, infection, trauma or injury. These dead cells are taken up, processed and presented to T cells by antigen presenting cells resulting in tolerance or immunity. Apoptotic cells induce tolerance; however, the precise mechanisms are still unknown. Previous studies have shown that direct infusion of apoptotic cells induce tolerance mediated by TRAIL-expressing CD8+ T cells. We hypothesized that immunologic tolerance induced by apoptotic cells is dependent on the activation status of apoptotic cells and mediated by direct killing of target cells by TRAIL-expressing CD8+ T cells. Three different experimental systems were used to elucidate the mechanisms by which apoptotic cells regulate immune responses. Using a classical system of tolerance induction, we examined the immunological consequence of intravenous (i.v.) delivery of ex vivo-generated naïve or activated apoptotic cells. Naïve apoptotic cells induced tolerance when injected i.v.; however, previously activated apoptotic cells induced immunity. Further analysis revealed a key role for CD154 in the tolerogenic or immunogenic nature of the naïve or activated apoptotic cells, respectively, as tolerance resulted after i.v. injection of either naïve or activated apoptotic CD154-/- cells, while co-injection of an agonistic anti-CD40 mAb with naïve apoptotic T cells induced robust immunity. The infusion of large numbers of apoptotic cells has limited physiological relevance, so the investigation of the influence of apoptotic cells on the immune system turned to another experimental tolerance model where soluble peptide antigen is injected systemically to induce the peripheral deletion of a population of antigen-specific T cells. Using this system, we investigated how apoptotic cells generated in vivo leads to T cell tolerance. Following adoptive transfer of OT-II cells, wild-type mice injected with soluble OVA323-339 became unresponsive to subsequent CFA/OVA immunization. Interestingly, Trail-/- or Dr5-/- mice developed robust immunity; even though all strains displayed peripheral deletion of OVA-specific T cells. Subsequent investigation found the mechanism of action of the CD8+ T cells was TRAIL-mediated deletion of the OVA-responsive T cells in a TCR-specific manner. The experimental systems used above have some clinical relevance but are still not physiologic. To study the impact of apoptotic cells in a physiologic setting, we took advantage of the medical condition sepsis, which is accompanied by massive apoptosis of multiple immune cell populations. Thus, the final set of experiments in this thesis examined the tolerance induced during sepsis using a clinically-relevant cecal-ligation and puncture (CLP) model that included a secondary bacterial infection. CLP-treated WT mice had a reduced ability to control the secondary bacterial infection, which was paralleled by suppressed T cell responses, compared to sham-treated WT mice. In contrast, CLP- and sham-treated Trail-/- and Dr5-/- mice were able to similarly control the bacterial infection and generated bacterial antigen-specific T cell responses. The ability of CLP-treated wild-type mice to control the secondary infection and generate T cell immunity could be restored by the administration of a blocking anti-TRAIL mAb. These results suggest the importance of TRAIL in the induction of sepsis-induced immune suppression, such that TRAIL neutralization may be a potential therapeutic target to restore cellular immunity in septic patients.

Maintaining cells in an undifferentiated totipotent state is essential for initiating developmental programs that lead to a fully formed organism in each generation and for maintaining immortal germ cells across generations. Caenorhabditis elegans is a powerful genetic model organism to study early germ cell development due to the animal’s transparency and the ability to screen for mutant phenotypes. However, our ability to use standard techniques to study gene expression using fluorescent reporter genes has been limited due to germline-specific silencing mechanisms that repress transgenes. Therefore, we lack even basic knowledge of how expression is regulated in C. elegans germ cells. In this study, we develop methods to overcome these silencing mechanisms by using a class of noncoding DNA, called Periodic An/Tn Clusters (PATCs), to prevent transgene silencing in the germline. We use these improved tools to test the proposed role of putative germline-specific regulatory DNA motifs and the role a periodic TT signal within germline promoters. We fused GFP to the promoter of a germline expressed gene (pcn-1), which is enriched for PATCs and contains a germline-specific motif (TTAAAG). Our results show that despite enrichment and phylogenetic conservation, the TTAAAG motif is not required for germline expression. To test additional motifs and periodic TTs, we have designed a system that will allow us to test synthetic gene fragments for bi-directional germline expression. These tools will allow us to rapidly test motif redundancy, motif spacing, and TT periodicity using gfp and rfp signals in the germline and will enable experiments aimed at understanding the role of germline regulatory elements.

abstract: This study investigated low regulatory flexibility as a mechanism of the associations of rumination with affect, internalizing symptoms, and substance use and problems. 403 first-year college students completed an online baseline survey assessing rumination, regulatory flexibility, internalizing symptoms, alcohol use, cannabis use, alcohol problems, and cannabis problems. Roughly 2.67 months later, 261 of these participants completed a follow-up survey assessing internalizing symptoms and substance use and problems. Additionally, 71 of the 403 participants completed an experimental study. Thirty-three participants were randomly assigned to undergo a rumination induction, and 38 were assigned to a control condition. All lab participants underwent an interpersonal stress task during which regulatory flexibility was observed and completed pre-test and post-role-play measures of positive and negative affect. Experimental study results showed regulatory flexibility did not mediate effects of rumination induction on positive (indirect effect: standardized beta (β)=-0.01, unstandardized beta (b)=-0.12, 95% Confidence Interval (CI) [-0.64, 0.41], p=.66) or negative affect (indirect effect: β=0.01, b=0.17, 95% CI [-0.29, 0.63], p=.48). Longitudinal study results showed regulatory flexibility did not mediate associations between baseline rumination and follow-up internalizing symptoms (indirect effect: b=0.01, 95% CI [-0.03, 0.05], p=.57), alcohol use (indirect effect: b=-0.03, 95% CI [-0.09, 0.04], p=.39), cannabis use (indirect effect: b=0.10, 95% CI [-0.06, 0.26], p=.21), alcohol problems (indirect effect: b=-0.05, 95% CI [-0.18, 0.07], p=.40), or cannabis problems (indirect effect: b=-0.10, 95% CI [-0.36, 0.16], p=.43). However, rumination predicted greater internalizing symptoms (Incidence Rate Ratio (IRR)=1.26, b=0.23, 95% CI [0.08, 0.37], p=.003) and cannabis problems (IRR=1.73, b=0.55, 95% CI [0.23, 0.87], p=.001). Regulatory flexibility predicted fewer alcohol use days (IRR=0.76, b=-0.27, 95% CI [-0.49, -0.05], p=.015) and problems (IRR=0.58, b=-0.55, 95% CI [-0.95, -0.15], p=.007), and less cannabis use for women (IRR=0.59, b=-0.53, 95% CI [-0.92, -0.14], p=.007) and fewer cannabis problems for men (IRR=0.21, b=-1.55, 95% CI [-2.50, -0.60], p=.001). Lack of agreement about how best to measure regulatory flexibility makes it unclear whether null associations were due to measurement problems or actual null effects. Research on how best to measure this construct is a priority. Findings indicate rumination and regulatory flexibility may be promising intervention targets. / Dissertation/Thesis / Doctoral Dissertation Psychology 2020

The overall need for the Kudu Gas Project is for Namibia to meet its projected electricity demand and to export excess power to neighboring countries. As the Namibian population grows and the economy develops so does the increase in the demand for electricity and the current generation capacity is below the demand needed and this has led to Namibia importing approximately over 50% of its electricity demands from other utilities in the region, primarily Eskom of South Africa. The development of operations of the Kudu gas field would therefore be imperative for the energy supply goals of the country as it is also in line with the main targets of the Namibia white paper on energy policy aiming towards reducing electricity imports and achieving security of electricity supply in Namibia. Although the Kudu gas field was discovered in 1974, today in 2018 which is 44 years later, gas operations have still not commenced. This raises a point of concern as to what the possible delay to the project is, accordingly this paper focused on establishing how legal risks may have contributed thereto by assessing how legal risks affect offshore upstream gas operations. This was done by conducting a legal risk analysis of the legal framework that governs legislative and contractual upstream gas risk and determining what the consequences of these risks are towards the development of the Kudu gas project. It was established that that legal risk can hamper the development of offshore upstream operations quite negatively if legal compliance to the regulatory requirements are not adhered to, however, from the information available at the time this study was done, the Kudu gas project complies very well with regulatory requirements and the project delay is not linked to legal risks. The paper observed that other risks to the project, namely market and financial risk may be the underlying cause of the project delay. As oil and gas operations are plagued with various risk and constant increases in regulatory pressure the recommendations of this paper were directed towards the establishment of a risk management process for the Kudu gas project which will identify and monitor risks and implement risk responses that have been established by internal risks controls to mitigate and avoid the stern consequences that come with no- compliance with legal and regulatory requirements. / Mini Dissertation (LLM)--University of Pretoria, 2019. / Public Law / LLM / Unrestricted

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