921 resultados para self-organized critical
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Full text: The idea of producing proteins from recombinant DNA hatched almost half a century ago. In his PhD thesis, Peter Lobban foresaw the prospect of inserting foreign DNA (from any source, including mammalian cells) into the genome of a λ phage in order to detect and recover protein products from Escherichia coli [ 1 and 2]. Only a few years later, in 1977, Herbert Boyer and his colleagues succeeded in the first ever expression of a peptide-coding gene in E. coli — they produced recombinant somatostatin [ 3] followed shortly after by human insulin. The field has advanced enormously since those early days and today recombinant proteins have become indispensable in advancing research and development in all fields of the life sciences. Structural biology, in particular, has benefitted tremendously from recombinant protein biotechnology, and an overwhelming proportion of the entries in the Protein Data Bank (PDB) are based on heterologously expressed proteins. Nonetheless, synthesizing, purifying and stabilizing recombinant proteins can still be thoroughly challenging. For example, the soluble proteome is organized to a large part into multicomponent complexes (in humans often comprising ten or more subunits), posing critical challenges for recombinant production. A third of all proteins in cells are located in the membrane, and pose special challenges that require a more bespoke approach. Recent advances may now mean that even these most recalcitrant of proteins could become tenable structural biology targets on a more routine basis. In this special issue, we examine progress in key areas that suggests this is indeed the case. Our first contribution examines the importance of understanding quality control in the host cell during recombinant protein production, and pays particular attention to the synthesis of recombinant membrane proteins. A major challenge faced by any host cell factory is the balance it must strike between its own requirements for growth and the fact that its cellular machinery has essentially been hijacked by an expression construct. In this context, Bill and von der Haar examine emerging insights into the role of the dependent pathways of translation and protein folding in defining high-yielding recombinant membrane protein production experiments for the common prokaryotic and eukaryotic expression hosts. Rather than acting as isolated entities, many membrane proteins form complexes to carry out their functions. To understand their biological mechanisms, it is essential to study the molecular structure of the intact membrane protein assemblies. Recombinant production of membrane protein complexes is still a formidable, at times insurmountable, challenge. In these cases, extraction from natural sources is the only option to prepare samples for structural and functional studies. Zorman and co-workers, in our second contribution, provide an overview of recent advances in the production of multi-subunit membrane protein complexes and highlight recent achievements in membrane protein structural research brought about by state-of-the-art near-atomic resolution cryo-electron microscopy techniques. E. coli has been the dominant host cell for recombinant protein production. Nonetheless, eukaryotic expression systems, including yeasts, insect cells and mammalian cells, are increasingly gaining prominence in the field. The yeast species Pichia pastoris, is a well-established recombinant expression system for a number of applications, including the production of a range of different membrane proteins. Byrne reviews high-resolution structures that have been determined using this methylotroph as an expression host. Although it is not yet clear why P. pastoris is suited to producing such a wide range of membrane proteins, its ease of use and the availability of diverse tools that can be readily implemented in standard bioscience laboratories mean that it is likely to become an increasingly popular option in structural biology pipelines. The contribution by Columbus concludes the membrane protein section of this volume. In her overview of post-expression strategies, Columbus surveys the four most common biochemical approaches for the structural investigation of membrane proteins. Limited proteolysis has successfully aided structure determination of membrane proteins in many cases. Deglycosylation of membrane proteins following production and purification analysis has also facilitated membrane protein structure analysis. Moreover, chemical modifications, such as lysine methylation and cysteine alkylation, have proven their worth to facilitate crystallization of membrane proteins, as well as NMR investigations of membrane protein conformational sampling. Together these approaches have greatly facilitated the structure determination of more than 40 membrane proteins to date. It may be an advantage to produce a target protein in mammalian cells, especially if authentic post-translational modifications such as glycosylation are required for proper activity. Chinese Hamster Ovary (CHO) cells and Human Embryonic Kidney (HEK) 293 cell lines have emerged as excellent hosts for heterologous production. The generation of stable cell-lines is often an aspiration for synthesizing proteins expressed in mammalian cells, in particular if high volumetric yields are to be achieved. In his report, Buessow surveys recent structures of proteins produced using stable mammalian cells and summarizes both well-established and novel approaches to facilitate stable cell-line generation for structural biology applications. The ambition of many biologists is to observe a protein's structure in the native environment of the cell itself. Until recently, this seemed to be more of a dream than a reality. Advances in nuclear magnetic resonance (NMR) spectroscopy techniques, however, have now made possible the observation of mechanistic events at the molecular level of protein structure. Smith and colleagues, in an exciting contribution, review emerging ‘in-cell NMR’ techniques that demonstrate the potential to monitor biological activities by NMR in real time in native physiological environments. A current drawback of NMR as a structure determination tool derives from size limitations of the molecule under investigation and the structures of large proteins and their complexes are therefore typically intractable by NMR. A solution to this challenge is the use of selective isotope labeling of the target protein, which results in a marked reduction of the complexity of NMR spectra and allows dynamic processes even in very large proteins and even ribosomes to be investigated. Kerfah and co-workers introduce methyl-specific isotopic labeling as a molecular tool-box, and review its applications to the solution NMR analysis of large proteins. Tyagi and Lemke next examine single-molecule FRET and crosslinking following the co-translational incorporation of non-canonical amino acids (ncAAs); the goal here is to move beyond static snap-shots of proteins and their complexes and to observe them as dynamic entities. The encoding of ncAAs through codon-suppression technology allows biomolecules to be investigated with diverse structural biology methods. In their article, Tyagi and Lemke discuss these approaches and speculate on the design of improved host organisms for ‘integrative structural biology research’. Our volume concludes with two contributions that resolve particular bottlenecks in the protein structure determination pipeline. The contribution by Crepin and co-workers introduces the concept of polyproteins in contemporary structural biology. Polyproteins are widespread in nature. They represent long polypeptide chains in which individual smaller proteins with different biological function are covalently linked together. Highly specific proteases then tailor the polyprotein into its constituent proteins. Many viruses use polyproteins as a means of organizing their proteome. The concept of polyproteins has now been exploited successfully to produce hitherto inaccessible recombinant protein complexes. For instance, by means of a self-processing synthetic polyprotein, the influenza polymerase, a high-value drug target that had remained elusive for decades, has been produced, and its high-resolution structure determined. In the contribution by Desmyter and co-workers, a further, often imposing, bottleneck in high-resolution protein structure determination is addressed: The requirement to form stable three-dimensional crystal lattices that diffract incident X-ray radiation to high resolution. Nanobodies have proven to be uniquely useful as crystallization chaperones, to coax challenging targets into suitable crystal lattices. Desmyter and co-workers review the generation of nanobodies by immunization, and highlight the application of this powerful technology to the crystallography of important protein specimens including G protein-coupled receptors (GPCRs). Recombinant protein production has come a long way since Peter Lobban's hypothesis in the late 1960s, with recombinant proteins now a dominant force in structural biology. The contributions in this volume showcase an impressive array of inventive approaches that are being developed and implemented, ever increasing the scope of recombinant technology to facilitate the determination of elusive protein structures. Powerful new methods from synthetic biology are further accelerating progress. Structure determination is now reaching into the living cell with the ultimate goal of observing functional molecular architectures in action in their native physiological environment. We anticipate that even the most challenging protein assemblies will be tackled by recombinant technology in the near future.
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The Self-shrinking p-adic cryptographic generator (SSPCG) is a fast software stream cipher. Improved cryptoanalysis of the SSPCG is introduced. This cryptoanalysis makes more precise the length of the period of the generator. The linear complexity and the cryptography resistance against most recently used attacks are invesigated. Then we discuss how such attacks can be avoided. The results show that the sequence generated by a SSPCG has a large period, large linear complexity and is stable against the cryptographic attacks. This gives the reason to consider the SSPSG as suitable for critical cryptographic applications in stream cipher encryption algorithms.
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The purpose of this study was to determine the knowledge and use of critical thinking teaching strategies by full-time and part-time faculty in Associate Degree Nursing (ADN) programs. ^ Sanders CTI (1992) instrument was adapted for this study and pilot-tested prior to the general administration to ADN faculty in Southeast Florida. This modified instrument, now termed the Burroughs Teaching Strategy Inventory (BTSI), returned reliability estimates (Cronbach alphas of .71, .74, and .82 for the three constructs) comparable to the original instrument. The BTSI was administered to 113 full-time and part-time nursing faculty in three community college nursing programs. The response rate was 92% for full-time faculty (n = 58) and 61% for part-time faculty (n = 55). ^ The majority of participants supported a combined definition of critical thinking in nursing which represented a composite of thinking skills that included reflective thinking, assessing alternative viewpoints, and the use of problem-solving. Full-time and part-time faculty used different teaching strategies. Full-time faculty most often used multiple-choice exams and lecture while part-time faculty most frequently used discussion within their classes. One possible explanation for specific strategy choices and differences might be that full-time faculty taught predominately in theory classes where certain strategies would be more appropriate and part-time faculty taught predominately clinical classes. Both faculty types selected written nursing care plans as the second most effective critical thinking strategy. ^ Faculty identified several strategies as being effective in teaching critical thinking. These strategies included discussion, case studies, higher order questioning, and concept analysis. These however, were not always the strategies that were used in either the classroom or clinical setting. ^ Based on this study, the author recommends that if the profession continues to stress critical thinking as a vital component of practice, nursing faculty should receive education in appropriate critical teaching strategies. Both in-service seminars and workshops could be used to further the knowledge and use of critical thinking strategies by faculty. Qualitative research should be done to determine why nursing faculty use self-selected teaching strategies. ^
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This study identifies and describes HIV Voluntary Counseling and Testing (VCT) of middle aged and older Latinas. The rate of new cases of HIV in people age 45 and older is rapidly increasing, with a 40.6% increase in the numbers of older Latinas infected with HIV between 1998 and 2002. Despite this increase, there is paucity of research on this population. This research seeks to address the gap through a secondary data analysis of Latina women. The aim of this study is twofold: (1) Develop and empirically test a multivariate model of VCT utilization for middle aged and older Latinas; (2) To test how the three individual components of the Andersen Behavioral Model impact VCT for middle aged and older Latinas. The study is organized around the three major domains of the Andersen Behavioral Model of service use that include: (a) predisposing factors; (b) enabling characteristics and (c) need. Logistic regression using structural equation modeling techniques were used to test multivariate relationships of variables on VCT for a sample of 135 middle age and older Latinas residing in Miami-Dade County, Florida. Over 60% of participants had been tested for HIV. Provider endorsement was found to he the strongest predictor of VCT (odds ration [OR] 6.38), followed by having a clinic as a regular source of healthcare (OR=3.88). Significant negative associations with VCT included self rated health status (OR=.592); Age (OR=.927); Spanish proficiency (OR=.927); number of sexual partners (OR=.613) and consumption of alcohol during sexual activity (.549). As this line of inquiry provides a critical glimpse into the VCT of older Latinas, recommendations for enhanced service provision and research will he offered.
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Some were born to do math, some persevered past fearful environments, while others withdrew. In this qualitative study, adults describe life with algebra and the meaning they sought. For all, pedagogy was critical, either positively or negatively; and all found salvation in intervention.
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As the nursing profession faces a shortage of nurses, workplace initiatives focused on retaining employees are critical to the United States healthcare industry (Sochalski, 2002). The purpose of this research was to determine whether self-reported intent to stay on the job was related to perceptions of workplace empowerment using Kanter's (1977) theory of organizational empowerment as a framework. ^ The sample consisted of 206 Florida registered nurses. Four self-report scales and a demographic questionnaire were administered by mail. The Conditions for Work Effectiveness Questionnaire (CWEQ; Chandler, 1987), Job Activity Scale (JAS; Laschinger, Kutzscher, & Sabiston, 1993), Organizational Relationships Scale (ORS; Laschinger, Sabiston, & Kutzscher, 1993) and an intent to stay instrument (Kim, Price, Mueller & Watson, 1996) were used to measure perceived access to empowerment structures, perceived formal power, perceived informal power, and intent to stay, respectively. The data were analyzed using descriptive statistics, correlational analysis, and hierarchical regression. ^ Twenty-eight percent of the variance of intent to stay was explained by perceived access to empowerment structures, perceived formal power, and perceived informal power when holding age, gender, education, overall nursing experience, and number of years on current job constant. Perceived access to empowerment structures (CWEQ total score) was the best predictor of self-reported intent to stay for this sample. Of the four components of perceived access to work empowerment structures, perceived access to opportunity and resources were the best predictors of nurses' intent to stay on the job. ^ This study was the first step in establishing the relationship between Kanter's full model and intent to remain on the job, which is a stepping stone for the development of effective retention strategies based on a workplace empowerment model. This knowledge is particularly important in today's healthcare industry where healthcare administrators and human resource development practitioners are ideally positioned to implement organizational strategies to enhance access to work empowerment structures and potentially reduce turnover and mitigate the effects of nursing shortage. ^
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This study documented differences between substance using adolescent participants who either completed or dropped out of a brief motivational intervention. Therapeutic alliance, working alliance and patient involvement were used to describe differences in treatment process ratings in a sample of majority Latino males who either (a) completed a adolescent substance abuse intervention called Alcohol Treatment Targeting Adolescents In Need (ATTAIN) or (b) dropped out after the first or second Guided Self-Change therapy session. Fifteen-minute segments were copied from the midpoint of previously recorded audio-tapes of Guided Self-Change therapy sessions. Raters were trained to a criterion level of interrater reliability for both the Working Alliance Inventory-Short and Vanderbilt Psychotherapy Process Scale. Correlations among Working Alliance Inventory- Short and Vanderbilt Psychotherapy Process Scale subscales reflected a general similarity in the assignment of ratings to client-therapist dyads. Findings underscore why these concepts are often used interchangeably in the treatment process literature. The Vanderbilt Psychotherapy Process Scale patient participation subscale demonstrated substantial empirical differentiation from overall therapeutic alliance. Discriminant function analysis demonstrated the Working Alliance Inventory-Short goal subscale and the Vanderbilt Psychotherapy Process Scale patient participation and therapist warmth and friendliness subscales as successful classifiers of groups of mostly Latino youth based on completion status. Follow-up logistic regression analyses confirmed major findings and successfully predicted group membership. Treatment process constructs can be used as clinical tools to identify participants who may be susceptible to dropping out of treatment services. Further investigation of treatment process may enhance understanding of the influence of alliance between clients and Guided Self-Change therapists. Investigating the role of treatment process as a critical component of brief motivational interventions for substance-using adolescents will inform both practitioners and researchers regarding the effectiveness of community-based substance abuse interventions for adolescents.
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As researchers and practitioners move towards a vision of software systems that configure, optimize, protect, and heal themselves, they must also consider the implications of such self-management activities on software reliability. Autonomic computing (AC) describes a new generation of software systems that are characterized by dynamically adaptive self-management features. During dynamic adaptation, autonomic systems modify their own structure and/or behavior in response to environmental changes. Adaptation can result in new system configurations and capabilities, which need to be validated at runtime to prevent costly system failures. However, although the pioneers of AC recognize that validating autonomic systems is critical to the success of the paradigm, the architectural blueprint for AC does not provide a workflow or supporting design models for runtime testing. ^ This dissertation presents a novel approach for seamlessly integrating runtime testing into autonomic software. The approach introduces an implicit self-test feature into autonomic software by tailoring the existing self-management infrastructure to runtime testing. Autonomic self-testing facilitates activities such as test execution, code coverage analysis, timed test performance, and post-test evaluation. In addition, the approach is supported by automated testing tools, and a detailed design methodology. A case study that incorporates self-testing into three autonomic applications is also presented. The findings of the study reveal that autonomic self-testing provides a flexible approach for building safe, reliable autonomic software, while limiting the development and performance overhead through software reuse. ^
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Favelas are Brazilian informal housing settlements that are areas of concentrated poverty. In Rio de Janeiro, favelas are perceived as areas of heightened criminal activity and violence, and residents experience discrimination, and little access to quality education and employment opportunities. In this context, hundreds of non-formal educational arts and leisure programs work to build the self-esteem and identity of youth in Rio's favelas as a way of preventing the youth from negative local influences. The Morrinho organization, located in the Pereira da Silva favela in Rio, uses art as a way for the local male youth to communicate their lived reality. This study used a visual critical ethnographic methodology to describe the way in which the Morrinho participants interpret living in a favela. Seventeen semi-structured interviews with young men aged 15 to 29, the feature-length documentary film on the organization, 206 researcher produced documentary style photographs of the Morrinho artwork, and the researcher's field notes were analyzed. Truth claims, ways of seeing as communicated through words and actions, were induced through a cyclical process of reconstructive horizon analysis that incorporated the societal context and critical theory. The participants communicated their concerns about life in a favela; however, they did not describe their societal positions in terms of complete marginalization. They named multiple benefits of living in Pereira da Silva, discussed positive and negative experiences in school, and described ways they circumvented discrimination. Morrinho as an organization was described as an enthralling game and a social project that benefited dozens of local youth. Character development was a valuable result of participation at Morrinho. The Morrinho artwork communicates a nuanced vision of both benevolent and violent social actors, and counters the overwhelmingly negative dominant characterization of Rio de Janeiro's favelas. This study has implications for an inclusive critical pedagogy and the use of art as a means to facilitate a transformative education. Further research is recommended to explore terminology used to refer to favelas, and perceptions that favela residents have of their experiences in public education.
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This study investigated how ethnicity, perceived family/friend social support (FSS), and health behaviors are associated with diabetes self-management (DSM) in minorities. The participants were recruited by community outreach methods and included 174 Cuban-, 121 Haitian- and 110 African-Americans with type 2 diabetes. The results indicated that ethnicity and FSS were associated with DSM. Higher FSS scores were associated with higher DSM scores, independent of ethnicity. There were ethnic differences in several elements of FSS. DSM was highest in Haitian- as compared to African-Americans; yet Haitian Americans had poorer glycemic control. The findings suggest FSS together with ethnicity may influence critical health practices. Studies are needed that further investigate the relationships among minorities with diabetes, their intimate network (family and friends) and the diabetes care process.
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The purpose of this study was to determine the knowledge and use of critical thinking teaching strategies by full-time and part-time faculty in Associate Degree Nursing (ADN) programs. Sander's CTI (1992) instrument was adapted for this study and pilottested prior to the general administration to ADN faculty in Southeast Florida. This modified instrument, now termed the Burroughs Teaching Strategy Inventory (BTSI), returned reliability estimates (Cronbach alphas of .71, .74, and .82 for the three constructs) comparable to the original instrument. The BTSI was administered to 113 full-time and part-time nursing faculty in three community college nursing programs. The response rate was 92% for full-time faculty (n = 58) and 61 % for part-time faculty (n = 55). The majority of participants supported a combined definition of critical thinking in nursing which represented a composite of thinking skills that included reflective thinking, assessing alternative viewpoints, and the use of problem-solving. Full-time and part-time faculty used different teaching strategies. Fulltime faculty most often used multiple-choice exams and lecture while part-time faculty most frequently used discussion within their classes. One possible explanation for specific strategy choices and differences might be that full-time faculty taught predominately in theory classes where certain strategies would be more appropriate and part-time faculty taught predominately clinical classes. Both faculty types selected written nursing care plans as the second most effective critical thinking strategy. Faculty identified several strategies as being effective in teaching critical thinking. These strategies included discussion, case studies, higher order questioning, and concept analysis. These however, were not always the strategies that were used in either the classroom or clinical setting. Based on this study, the author recommends that if the profession continues to stress critical thinking as a vital component of practice, nursing faculty should receive education in appropriate critical teaching strategies. Both in-service seminars and workshops could be used to further the knowledge and use of critical thinking strategies by faculty. Qualitative research should be done to determine why nursing faculty use self-selected teaching strategies.
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This thesis extends previous research on critical decision making and problem-solving by refining and validating a self-report measure designed to assess the use of critical decision making and problem solving in making life choices. The analysis was conducted by performing two studies, and therefore collecting two sets of data on the psychometric properties of the measure. Psychometric analyses included: item analysis, internal consistency reliability, interrater reliability, and an exploratory factor analysis. This study also included regression analysis with the Wonderlic, an established measure of general intelligence, to provide preliminary evidence for the construct validity of the measure.
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Presently, there are numerous Native English Teacher (NETs) teaching in Korean post-secondary educational (PSE) institutions. The aim of this thesis is to explore the views held by NETs with regards to their self-perceived teaching perspectives while working in a Korean PSE setting. The thesis also aims to answer the assertion made in the literature that English as Foreign Language (EFL) teachers are "acritical and atheoretical". To this end, the thesis intends to identify the extent of the NETs’ preference for social reform as a teaching perspective, the NETs stated reasons for identifying with roles as social reformers, how these views are reflected in the NETs’ practice (praxis), what the barriers impeding the adoption and enactment of social reform are, and how the NETs’ perspectives relate to critical pedagogy. The results reveal that NETs in Korean PSE do not align themselves with social reform, yet categorizing NETS as "acritical and atheoretical" may be overly-simplistic. The results show that there are three kinds of obstacles that prevent NETs from engaging more with social reform and being less acritical and atheoretical: 1) NETs teaching in Korean EFL are conflicted and/or confused about their roles as English teachers; 2) there are significant cultural constraints to teaching in Korean EFL as a NET; 3) there are significant pedagogical constraints to teaching in Korean EFL as a NET.
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The Mermaid Series (1887-1909) edited by Havelock Ellis was a major watershed in appreciation of Elizabethan and Jacobean drama. Before it appeared plays were available to general readers in scattered anthologies, large expensive collected editions or in expurgated selections which included only the more lyrical speeches and memorable scenes. Criticism of the drama followed suit; the majority of critics concentrated on the sections which appealed to the romantic and sentimental tastes of nineteenthcentury readers. The two men who conceived the Mermaid Series, John Addington Symonds and Havelock Ellis, approached the drama differently from their contemporaries; Symonds studied a play as a whole work of art and Ellis concentrated on its view of life. Both were unsatisfied with the "select beauties", fragmented approach and wanted readers to have the best plays in their entirety easily available in handy, inexpensive editions. Symonds's awareness of the drama as theatre was combined with a historical perspective allowing him to judge the drama in relation to its own time. He made a lasting but hitherto underestimated contribution to study of Beaumont and Fletcher, Dekker, Marlowe, and Ford. Ellis's work on the drama is overshadowed today by his studies of sex but his concentration on ideas and appreciation of unconventional behaviour enabled him to formulate new views on Ford, Middleton and Chapman. The two other major editors to work on the series, A. C. Swinburne and Arthur Symons had more conventional nineteenth-century approaches. Both were impressionistic critics who were most attracted to the l~nguage of the drama. Swinburne, however, occasionally transcended his fragmented approach and offered significant interpretations of Tourneur, Massinger; 'The .Changeling, Heywood. Symons's range was more limited but his form of impressionism was valuable for its concentration on the aesthetic experience at the heart of a work of art. His most important contributions were the study of Middleton and Massinger. Besides these four major critics numerous lesser writers worked on the series. Their editorial work was valuable and some, notably Ernest Rhys, c. H. Herford and Thomas Dickinson offered criticism of enduring importance. In my first chapter I consider the general availability of texts of the Elizabethan and Jacobean drama in the nineteenth century, the general attitudes towards the drama, and the critical approaches of each of the editors. The subsequent chapters are organized around the volumes of the series. I consider the climate of opinion in which each appeared, assess its critical and editorial contribution and evaluate the work of the other Mermaid editors on the dramatist included in the volume. My study shows that the concept of the Mermaid Series and the work of its editors helped to revolutionize study of the Elizabethan and Jacobean dramatists by providing good texts and by pointing the way to our present view of the plays as whole works of art.
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We analyze a recent proposal for spontaneous mirror symmetry breaking based on the coupling of first-order enantioselective autocatalysis and direct production of the enantiomers that invokes a critical role for intrinsic reaction noise. For isolated systems, the racemic state is the unique stable outcome for both stochastic and deterministic dynamics when the system is in compliance with the constraints dictated by the thermodynamics of chemical reaction processes. In open systems, the racemic outcome also results for both stochastic and deterministic dynamics when driving the auto-catalysis unidirectionally by external reagents. Nonracemic states can result in the latter only if the reverse reactions are strictly zero: these are kinetically controlled outcomes for small populations and volumes, and can be simulated by stochastic dynamics. However, the stability of the thermodynamic limit proves that the racemic outcome is the unique stable state for strictly irreversible externally driven autocatalysis. These findings contradict the suggestion that the inhibition requirement of the Frank autocatalytic model for the emergence of homochirality may be relaxed in a noise-induced mechanism.