Development of methods for combinational approaches to cis-regulatory module interactions

The complexity and size of the higher animal genome and relative scarcity of DNA-binding factors with which to regulate it imply a complex and pleiotropic regulatory system. Cisregulatory modules (CRMs) are vitally important regulators of gene expression in higher animal cells, integrating external and internal information to determine an appropriate response in terms of gene expression by means of direct and indirect interactions with the transcriptional machinery. The interaction space available within systems of multiple CRMs, each containing several sites where one or more factors could be bound is huge. Current methods of investigation involve the removal of individual sites or factors and measuring the resulting effect on gene expression. The effects of investigations of this type may be masked by the functional redundancy present in some of these regulatory systems as a result of their evolutionary development. The investigation of CRM function is limited by a lack of technology to generate and analyse combinatorial mutation libraries of CRMs, where putative transcription factor binding sites are mutated in various combinations to achieve a holistic view of how the factors binding to those sites cooperate to bring about CRM function. The principle work of this thesis is the generation of such a library. This thesis presents the development of microstereolithography as a method for making microfluidic devices, both directly and indirectly. A microfluidic device was fabricated that was used to generate oligonucleotide mixtures necessary to synthesise combinatorial mutants of a CRM sequence from the muscle regulatory factor MyoD. In addition, this thesis presents the development of the optimisation algorithms and assembly processes necessary for successful sequence assembly. Furthermore, it was found that the CRM, in combination with other CRMs, is able to synergistically regulate gene expression in a position and orientation independent manner in three separate contexts. Finally, by testing a small portion of the available combinatorial mutant library it was shown that mutation of individual binding sites within of the CRM is not sufficient to show a significant change in the level of reporter gene expression

The 2nd International Electronic Conference on Applied Sciences

This book is focused on the works presented at the 2nd International Electronic Conference on Applied Sciences, organized by Applied Sciences from 15 to 31 October 2021 on the MDPI Sciforum platform. Two decades have passed since the start of the 21st century. The development of sciences and technologies is growing ever faster today than in the previous century. The field of science is expanding, and the structure of science is becoming ever richer. Because of this expansion and fine structure growth, researchers may lose themselves in the deep forest of the ever-increasing frontiers and sub-fields being created. This international conference on the Applied Sciences was started to help scientists conduct their own research into the growth of these frontiers by breaking down barriers and connecting the many sub-fields to cut through this vast forest. These functions will allow researchers to see these frontiers and their surrounding (or quite distant) fields and sub-fields, and give them the opportunity to incubate and develop their knowledge even further with the aid of this multi-dimensional network

Macrocriminology and Freedom

How can power over others be transformed to 'power with'? It is possible to transform many institutions to build societies with less predation and more freedom. These stretch from families and institutions of gender to the United Nations. Some societies, times and places have crime rates a hundred times higher than others. Some police forces kill at a hundred times the rate of others. Some criminal corporations kill thousands more than others. Micro variables fail to explain these patterns. Prevention principles for that challenge are macrocriminological. Freedom is conceived in a republican way as non-domination. Tempering domination prevents crime; crime prevention reduces domination. Many believe a high crime rate is a price of freedom. Not Braithwaite. His principles of crime control are to build freedom, temper power, lift people from poverty and reduce all forms of domination. Freedom requires a more just normative order. It requires cascading of peace by social movements for non-violence and non-domination. Periods of war, domination and anomie cascade with long lags to elevated crime, violence, inter-generational self-violence and ecocide. Cybercrime today poses risks of anomic nuclear wars. Braithwaite’s proposals refine some of criminology’s central theories and sharpen their relevance to all varieties of freedom. They can be reduced to one sentence. Strengthen freedom to prevent crime, prevent crime to strengthen freedom

Macrocriminology and Freedom

How can power over others be transformed to 'power with'? It is possible to transform many institutions to build societies with less predation and more freedom. These stretch from families and institutions of gender to the United Nations. Some societies, times and places have crime rates a hundred times higher than others. Some police forces kill at a hundred times the rate of others. Some criminal corporations kill thousands more than others. Micro variables fail to explain these patterns. Prevention principles for that challenge are macrocriminological. Freedom is conceived in a republican way as non-domination. Tempering domination prevents crime; crime prevention reduces domination. Many believe a high crime rate is a price of freedom. Not Braithwaite. His principles of crime control are to build freedom, temper power, lift people from poverty and reduce all forms of domination. Freedom requires a more just normative order. It requires cascading of peace by social movements for non-violence and non-domination. Periods of war, domination and anomie cascade with long lags to elevated crime, violence, inter-generational self-violence and ecocide. Cybercrime today poses risks of anomic nuclear wars. Braithwaite’s proposals refine some of criminology’s central theories and sharpen their relevance to all varieties of freedom. They can be reduced to one sentence. Strengthen freedom to prevent crime, prevent crime to strengthen freedom