Implication of DNA methylation, CTCF and BORIS factors in the transcriptional regulation of the human telomerase catalytic subunit hTERT

RESUME La télomérase confère une durée de vie illimitée et est réactivée dans la plupart des cellules tumorales. Sa sous-unité catalytique hTERT est définie comme le facteur limitant pour son activation. De l'identification de facteurs liant la région régulatrice d'hTERT, au rôle de la méthylation de l'ADN et de la modification des histones, de nombreux modèles de régulation ont été suggérés. Cependant, aucun de ces modèles n'a pu expliquer l'inactivation de la télomérase dans la plupart des cellules somatiques et sa réactivation dans la majorité des cellules tumorales. De plus, les observations contradictoires entre le faible niveau d'expression d'ARN messager d'hTERT dans les cellules télomérase-positives et la très forte activité transcriptionnelle du promoteur d'hTERT en transfection restent incomprises. Dans cette étude, nous avons montré que la région proximale du gène hTERT (exon 1 et 2) était impliquée dans la répression de l'activité de son promoteur. Nous avons identifié le facteur CTCF comme étant un inhibiteur du promoteur d'hTERT, en se liant au niveau de son premier exon. La méthylation de l'exon 1 du gène hTERT, couramment observée dans les tumeurs mais pas dans les cellules normales, empêcherait la liaison de CTCF. L'étude du profil de méthylation du promoteur d'hTERT indique qu'une partie du promoteur reste déméthylée et qu'elle semble suffisante pour permettre une faible activité transcriptionnelle du gène hTERT. Ainsi, la méthylation particulière des régions régulatrices d'hTERT inhibe la liaison de CTCF tout en permettant une faible transcription du gène. Cependant, dans certaines cellules tumorales, le promoteur et la région proximale du gène hTERT ne sont pas méthylés. Dans les lignées cellulaires tumorales de tesitcules et d'ovaires, l'inhibition de CTCF est contrée par son paralogue BORIS, qui se lie aussi au niveau de l'exon 1 d'hTERT, mais permet ainsi l'activation du promoteur. L'étude de l'expression du gène BORIS montre qu'il est exclusivement exprimé dans les tissus normaux de testicules et d'ovaires jeunes, ainsi qu'à différents niveaux dans la plupart des tumeurs. Sa transcription est sous le contrôle de deux promoteurs. Le promoteur proximal est régulé par méthylation et un transcrit alternatif majoritaire, délété de l'exon 6, est trouvé lorsque ce promoteur est actif. Tous ces résultats conduisent à un modèle de régulation du gène hTERT qui tient compte du profil épigénétique du gène et qui permet d'expliquer le faible taux de transcription observé in vivo. De plus, l'expression de BORIS dans les cancers et son implication dans l'activation du gène hTERT pourrait permettre de comprendre les phénomènes de dérégulation épigénétique et d'immortalisation qui ont lieu durant la tumorigenèse. SUMMARY Telomerase confers an unlimited lifespan, and is reactivated in most tumor cells. The catalytic subunit of telomerase, hTERT, is defined as the limiting factor for telomerase activity. Between activators and repressors that bind to the hTERT 5' regulatory region, and the role of CpG methylation and histone acetylation, an abundance of regulatory models have been suggested. None of these models can explain the silence of telomerase in most somatic cells and its reactivation in tumor cells. Moreover, the contradictory observations of the low level of hTERT mRNA in telomerase-positive cells and the high transcriptional activity of the hTERT promoter in transfection experiments remain unresolved. In this study, we demonstrated that the proximal exonic region of the hTERT gene (exon 1 and 2) is involved in the inhibition of its promoter. We identified the protein CTCF as the inhibitor of the hTERT promoter, through its binding to the first exon. The methylation of the first exon region, which is often observed in cancer cells but not in noimal cells, represses CTCF binding. Study of hTERT promoter methylation shows a partial demethylation sufficient to activate the transcription of the hTERT gene. Therefore, we demonstrated that the particular methylation profile of the hTERT regulatory sequences inhibits the binding of CTCF, while it allows a low transcription of the gene. Nevertheless, in some tumor cells, the promoter and the proximal exonic region of hTERT are unmethylated. In testicular and ovarian cancer cell lines, CTCF inhibition is counteracted by its BORIS paralogue that also binds the hTERT first exon but allows the promoter activation. The study of BORIS gene regulation showed that this factor is exclusively expressed in normal tissue of testis and ovary of young woman, as well as in almost all tumors with different levels. Two promoters were found to induce its transcription. The proximal promoter was regulated by methylation. Moreover, a major alternative transcript, deleted of the exon 6, is detected when this promoter is active. All these results lead to a model for hTERT regulation that takes into account the epigenetic profile of the gene and provides an explanation for the low transcriptional level observed in vivo. BORIS expression in cancers and its implication in hTERT activation might also permit the understanding of epigenetic deregulation and immortalization phenomena that occur during tumorigenesis.

External urethral sphincter electromyography in asymptomatic women and the influence of the menstrual cycle.

OBJECTIVE: To investigate by electromyography (EMG), the presence of complex repetitive discharges (CRDs) and decelerating bursts (DBs) in the striated external urethral sphincter during the menstrual cycle in female volunteers with no urinary symptoms and complete bladder emptying. SUBJECTS AND METHODS: Healthy female volunteers aged 20-40 years, with regular menstrual cycles and no urinary symptoms were recruited. Volunteers completed a menstruation chart, urinary symptom questionnaires, pregnancy test, urine dipstick, urinary free flow and post-void ultrasound bladder scan. Exclusion criteria included current pregnancy, use of hormonal medication or contraception, body mass index of >35 kg/m(2) , incomplete voiding and a history of pelvic surgery. Eligible participants underwent an external urethral sphincter EMG, using a needle electrode in the early follicular phase and the mid-luteal phase of their menstrual cycles. Serum oestradiol and progesterone were measured at each EMG test. RESULTS: In all, 119 women enquired about the research and following screening, 18 were eligible to enter the study phase. Complete results were obtained in 15 women. In all, 30 EMG tests were undertaken in the 15 asymptomatic women. Sphincter EMG was positive for CRDs and DBs at one or both phases of the menstrual cycle in eight (53%) of the women. Three had CRDs and DBs in both early follicular and mid-luteal phases. Five had normal EMG activity in the early follicular phase and CRDs and DBs in the mid-luteal phase. No woman had abnormal EMG activity in the early follicular phase and normal activity in the luteal phase. There was no relationship between EMG activity and age, parity or serum levels of oestradiol and progesterone. CONCLUSIONS: CRDs and DB activity in the external striated urethral sphincter is present in a high proportion of asymptomatic young women. This abnormal EMG activity has been shown for the first time to change during the menstrual cycle in individual women. CRDs and DBs are more commonly found in the luteal phase of the menstrual cycle. The importance of CRDs and DBs in the aetiology of urinary retention in young women remains uncertain. The distribution and or quantity of abnormal EMG activity in the external urethral sphincter may be important. In a woman with urinary retention the finding of CRDs and DBs by needle EMG does not automatically establish Fowler's syndrome as the explanation for the bladder dysfunction. Urethral pressure profilometry may be helpful in establishing a diagnosis. Opiate use and psychological stress should be considered in young women with urinary retention.

The Evolution of the European Regulatory Framework for Electronic Communications

The paper reviews the historical transformation of the European regulatory framework for electronic communications from the era dominated by state-owned enterprises to the presence of regulated competition. In the course of these developments, the vision of the roles of the public and private sectors in electronic communications changed in expected and unexpected ways. While the period is characterized by a shift toward less direct state intervention, the intensity of regulation has increased in many areas. Most recently, in the wake of the financial crisis, new forms of state intervention can be observed, including public investment in communications infrastructure and public-private partnerships. As a result of the reforms, Europe has been able to achieve major successes but it also suffered unanticipated setbacks compared to other regions. The European Union emerged as the global leader in mobile communications during the 1990s and was able to roll-out first-generation broadband access networks more rapidly than many of its peers. Recently, however, Europe as a whole has not performed as well in deploying next-generation networks and advanced mobile communications services. The paper offers a political-economic explanation for these developments and assesses their effects on the performance of the European electronic communications sector and the economy. From this analysis, the European model emerges as a unique institutional arrangement with peculiar advantages and disadvantages. Once these are recognized, sensible next steps to build the strengths while avoiding the weaknesses of the model can be seen more clearly.

Cohabitation effect on the slowdown of the Neolithic expansion

We introduce the effect of cohabitation between generations to a previous model on the slowdown of the Neolithic transition in Europe. This effect consists on the fact that human beings do not leave their children alone when they migrate, but on the contrary they cohabit until their children reach adulthood. We also use archaeological data to estimate the variation of the Mesolithic population density with distance, and use this information to predict the slowdown of the Neolithic front speed. The new equation leads to a substantial correction, up to 37%, relative to previous results. The new model is able to provide a satisfactory explanation not only to the relative speed but also to the absolute speed of the Neolithic front obtained from archaeological data

Minimization of the experimental workload for the prediction of pollutants propagation in rivers. Mathematical modelling and knowledge re-use

The present thesis in focused on the minimization of experimental efforts for the prediction of pollutant propagation in rivers by mathematical modelling and knowledge re-use. Mathematical modelling is based on the well known advection-dispersion equation, while the knowledge re-use approach employs the methods of case based reasoning, graphical analysis and text mining. The thesis contribution to the pollutant transport research field consists of: (1) analytical and numerical models for pollutant transport prediction; (2) two novel techniques which enable the use of variable parameters along rivers in analytical models; (3) models for the estimation of pollutant transport characteristic parameters (velocity, dispersion coefficient and nutrient transformation rates) as functions of water flow, channel characteristics and/or seasonality; (4) the graphical analysis method to be used for the identification of pollution sources along rivers; (5) a case based reasoning tool for the identification of crucial information related to the pollutant transport modelling; (6) and the application of a software tool for the reuse of information during pollutants transport modelling research. These support tools are applicable in the water quality research field and in practice as well, as they can be involved in multiple activities. The models are capable of predicting pollutant propagation along rivers in case of both ordinary pollution and accidents. They can also be applied for other similar rivers in modelling of pollutant transport in rivers with low availability of experimental data concerning concentration. This is because models for parameter estimation developed in the present thesis enable the calculation of transport characteristic parameters as functions of river hydraulic parameters and/or seasonality. The similarity between rivers is assessed using case based reasoning tools, and additional necessary information can be identified by using the software for the information reuse. Such systems represent support for users and open up possibilities for new modelling methods, monitoring facilities and for better river water quality management tools. They are useful also for the estimation of environmental impact of possible technological changes and can be applied in the pre-design stage or/and in the practical use of processes as well.

The relativity of evaluative sentences: disagreeing over disagreement

Evaluative sentences (moral judgments, expressions of taste, epistemic modals) are relative to the speaker's standards. Lately, a phenomenon has challenged the traditional explanation of this relativity: whenever two speakers disagree over them they contradict each other without being at fault. Hence, it is thought that the correction of the assertions involved must be relative to an unprivileged standard not necessarily the speaker's. I will claim instead that so far, neither this nor any other proposal has provided an explanation of the phenomenon. I will point out several problems presented by them and I will hint to how this phenomenon could be explained by making minor adjustments to our semantic theory.

DEVELOPMENT OF LOGIC IN INDIA: SIGNIFICANCE OF 'THE DUOLOGUE BETWEEN PĀYĀSI AND KASSAPA' ("LONG DISCOURSES")

ABSTRACT 'The Duologue of King/Governor Pāyāsi' ("Long Discourses") has long been recognised as a source for the proto-materialism current at the time of the Buddha. What needs to be stressed is the significance of the text as a pointer to the development of Logic in India. Perception (observation and experiment employing the joint method of agreement and difference), which is an accepted method of experimental enquiry, and reasoning from analogy, which can lead at best to a probable conclusion - these two are the only means employed to settle the dispute concerning the existence of the other-world. The Jain version of the same duologue-cum-parable, though varying in minor details regarding the name and identity of the monk refuting the king/governor, contains the same contrast, namely, perception versus analogical reasoning. There can be little doubt that the original parable was conceived with a view to asserting the existence of the other-world. In the Kaṭha Upaniṣad (sixth century BCE), an earlier Brahmanical text, however, instead of argument by analogy, verbal testimony (śabda) was invoked to settle the same point. Naciketas is assailed by doubt about the existence of a person after his or her death. The authority of Yama, the Pluto of Indian mythology, is invoked to convince him that the other-world does exist. Thus, the three parables taken together exhibit three means of knowledge in operation: verbal testimony and argument by analogy pitted against perception.

Social ventures : the development of the theory of sustainable contributive advantage and initial empirical tests

This thesis attempts to fill gaps in both a theoretical basis and an operational and strategic understanding in the areas of social ventures, social entrepreneurship and nonprofit business models. This study also attempts to bridge the gap in strategic and economic theory between social and commercial ventures. More specifically, this thesis explores sustainable competitive advantage from a resource-based theory perspective and explores how it may be applied to the nonmarket situation of nonprofit organizations and social ventures. It is proposed that a social value-orientation of sustainable competitive advantage, called sustainable contributive advantage, provides a more realistic depiction of what is necessary in order for a social venture to perform better than its competitors over time. In addition to providing this realistic depiction, this research provides a substantial theoretical contribution in the area of economics, social ventures, and strategy research, specifically in regards to resource-based theory. The proposed model for sustainable contributive advantage uses resource-based theory and competitive advantage in order to be applicable to social ventures. This model proposes an explanation of a social venture’s ability to demonstrate consistently superior performance. In order to determine whether sustainable competitive advantage is in fact, appropriate to apply to both social and economic environments, quantitative analyses are conducted on a large sample of nonprofit organizations in a single industry and then compared to similar quantitative analyses conducted on commercial ventures. In comparing the trends and strategies between the two types of entities from a quantitative perspective, propositions are developed regarding a social venture’s resource utilization strategies and their possible impact on performance. Evidence is found to support the necessity of adjusting existing models in resource-based theory in order to apply them to social ventures. Additionally supported is the proposed theory of sustainable contributive advantage. The thesis concludes with recommendations for practitioners, researchers and policy makers as well as suggestions for future research paths.

Marketers’ perception of the implementation of social media marketing: Evidence from large Finnish companies’ marketing campaigns

Social media is a multidimensional marketing and communications channel which can support and enhance a business’ reputation, sales and even longevity. Social media as a business tool encourages an interaction between customers and companies which gives opportunities for a company to better understand their customers, to target them more effectively and to collaborate and create dialogues with them which is not possible through traditional media channels. The aim of a social media strategy is to increase brand awareness, image, loyalty and recognition. The peer networks that social media creates allows a company to disseminate information through loyal customers to new and prospective customers to ultimately increase reach. The purpose of the study is to understand the marketer’s perspective of social media marketing use and how it is currently utilized in marketing and communications activities in Finland. Three companies were interviewed covering fourteen different implementations of social media marketing campaigns. These were then analysed to ascertain the utilization methods and experience gained on recent campaigns in the Finnish market The utilization of social media marketing was analysed using the methods of thematic analysis and inductive and abductive reasoning. Elements and themes were drawn out of the separate interviews to create a framework with which to explore, evaluate and match theories that define social media usage by companies. It became clear from all of the interviews that social media as a tool is most effective when it captures the viewer’s interest through rich and entertaining content. This directed the theoretical research towards Engagement Theory and Content Marketing which look to emphasize the importance of communities, collaboration, interaction, and peer-sharing as the key drivers of a social media marketing campaign.

Comparison of scientists of the Brazilian Academy of Sciences and of the National Academy of Sciences of the USA on the basis of the h-index

A new scientometric indicator, the h-index, has been recently proposed (Hirsch JE. Proc Natl Acad Sci 2005; 102: 16569-16572). The index avoids some shortcomings of the calculation of the total number of citations as a parameter to evaluate scientific performance. Although it has become known only recently, it has had widespread acceptance. A comparison of the average h-index of members of the Brazilian Academy of Sciences (BAS) and of the National Academy of Sciences of the USA (NAS-USA) was carried out for 10 different areas of science. Although, as expected, the comparison was unfavorable to the members of the BAS, the imbalance was distinct in different areas. Since these two academies represent, to a significant extent, the science of top quality produced in each country, the comparison allows the identification of the areas in Brazil that are closer to the international stakeholders of scientific excellence. The areas of Physics and Mathematics stand out in this context. The heterogeneity of the h-index in the different areas, estimated by the median dispersion of the index, is significantly higher in the BAS than in the NAS-USA. No elements have been collected in the present study to provide an explanation for this fact.

Changes in sodium appetite evoked by lesions of the commissural nucleus of the tractus solitarius

Ablation of the area postrema/caudal nucleus of the tractus solitarius (NTS) complex increases sodium intake, but the effect of selective lesions of the caudal NTS is not known. We measured depletion-induced sodium intake in rats with electrolytic lesions of the commissural NTS that spared the area postrema. One day after the lesion, rats were depleted of sodium with furosemide (10 mg/kg body weight, sc) and then had access to water and a sodium-deficient diet for 24 h when 1.8% NaCl was offered. Water and saline intakes were measured for 2 h. Saline intake was higher in lesioned than in sham-lesioned rats (mean ± SEM: 20 ± 2 vs 11 ± 3 mL/2 h, P < 0.05, N = 6-7). Saline intake remained elevated in lesioned rats when the tests were repeated 6 and 14 days after the lesion, and water intake in these two tests was increased as well. Water intake seemed to be secondary to saline intake both in lesioned and in sham-lesioned rats. A second group of rats was offered 10% sucrose for 2 h/day before and 2, 7, and 15 days after lesion. Sucrose intake in lesioned rats was higher than in sham-lesioned rats only 7 days after lesioning. A possible explanation for the increased saline intake in rats with commissural NTS lesions could be a reduced gastrointestinal feedback inhibition. The commissural NTS is probably part of a pathway for inhibitory control of sodium intake that also involves the area postrema and the parabrachial nucleus.

Hypertensive effects of the iv administration of picomoles of ouabain

Ouabain, an endogenous digitalis compound, has been detected in nanomolar concentrations in the plasma of several mammals and is associated with the development of hypertension. In addition, plasma ouabain is increased in several hypertension models, and the acute or chronic administration of ouabain increases blood pressure in rodents. These results suggest a possible association between ouabain and the genesis or development and maintenance of arterial hypertension. One explanation for this association is that ouabain binds to the α-subunit of the Na+ pump, inhibiting its activity. Inhibition of this pump increases intracellular Na+, which reduces the activity of the sarcolemmal Na+/Ca2+ exchanger and thereby reduces Ca2+ extrusion. Consequently, intracellular Ca2+ increases and is taken up by the sarcoplasmic reticulum, which, upon activation, releases more calcium and increases the vascular smooth muscle tone. In fact, acute treatment with ouabain enhances the vascular reactivity to vasopressor agents, increases the release of norepinephrine from the perivascular adrenergic nerve endings and promotes increases in the activity of endothelial angiotensin-converting enzyme and the local synthesis of angiotensin II in the tail vascular bed. Additionally, the hypertension induced by ouabain has been associated with central mechanisms that increase sympathetic tone, subsequent to the activation of the cerebral renin-angiotensin system. Thus, the association with peripheral mechanisms and central mechanisms, mainly involving the renin-angiotensin system, may contribute to the acute effects of ouabain-induced elevation of arterial blood pressure.

Explaining the intention to play and behavior of playing sports betting games of young Finnish adults : an application of the theory of planned behavior

The aim of this Master’s Thesis was to examine the determinants of intention and behavior of playing sports betting games in order to explain the intention to play in a more precise way and to be able to understand the behavior of playing. The theory of planned behavior was applied in explaining the intention of young Finnish adults aged 18 to 34. A quantitative research method was applied and an online survey was sent to the students of Lappeenranta University of Technology and to the subscribers of Urheilulehti in order to reach a sample that present the young population of Finland. The theory of the study focused on the theory of planned behavior and its antecedents, attitude towards behavior, subjective norms, perceived behavioral control as well as motivation. By analyzing the data, causal relationships were found through which the explanation of intention was possible. The results showed that attitude towards playing, subjective norms, perceived behavioral control and motivation impact the formation of intention significantly. The results also indicated that intention impacts significantly to the playing frequency.

Academics as parents--parents as academics : a study of the integration of intellectual and child-care labours /

Discussions concerning the challenges of combining work and family are certainly not new, and still actively continue. There is, however, a silence in the related literature regarding a comprehensive description of integrating specifically university academic ~. work and family responsibilities. This silence is especially evident for men who are parents as well as academics. With the participation of 4 key informants, this qualitative research study gave voice to men and women who participate in the academic labour of a Canadian university as professors, and as graduate students, along with the parenting labour of at least 1 child under the age of7. Methodology was developed to reveal in-depth perspectives regarding the work practices employed by 4 key informants as they combined intellectual and child-care responsibilities. Multiple data collection methods included journal reflections, day time observation sessions, a focus group, and a final evaluation questionnaire. Using research findings, together with information extrapolated from Three Models of the Family (Eichler, 1997), this study also took steps toward developing a Proposed "Three Models of the University," to offer explanation for the work practices of the key informants as academics/parents, and also for future consideration in university policy formation.

Kierkegaard : the temporality of becoming a self

Introduction In Difference and Repetition, Deleuze compares and contrasts Kierkegaard's and Nietzsche's ideas of repetition. He argues that neither of them really give a representation of repetition. Repetition for them is a sort of selective task: the way in which they determine what is ethical and eternal. With Nietzsche, it is a theater of un belie f. ..... Nietzsche's leading idea is to found the repetition in the etemal return at once on the death of God and the dissolution of the self But it is a quite different alliance in the theater of faith: Kierkegaard dreams of alliance between a God and a self rediscovered. I Repetition plays a theatrical role in their thinking. It allows them to dramatically stage the interplay of various personnae. Deleuze does give a positive account ofKierkegaard's "repetition"; however, he does not think that Kierkegaard works out a philosophical model, or a representation of what repetition is. It is true that in the book Repetition, Constantin Constantius does not clearly and fully work out the concept of repetition, but in Sickness Unto Death, Kierkegaard gives a full explanation of the self and its temporality which can be connected with repetition. When Sickness Unto Death is interpreted according to key passages from Repetition and The Concept of Anxiety, a clear philosophical concept of repetition can be established. In my opinion, Kierkegaard's philosophy is about the task of becoming a self, and I will be attempting to show that he does have a model of the temporality of self-becoming. In Sickness Unto Death, Kierkegaard explains his notions of despair with reference to sin, self, self-becoming, faith, and repetition. Despair is a sickness of the spirit, of the self, and accordingly can take three forms: in despair not to be conscious of having a self (not despair in the strict sense); in despair not to will to be oneself; in despair to will to be oneself2 In relation to this definition, he defines a self as "a relation that relates itself to itself and in relating itself to itself relates to another.''3 Thus, a person is a threefold relationship, and any break in that relationship is despair. Despair takes three forms corresponding to the three aspects of a self s relation to itself Kierkegaard says that a selfis like a house with a basement, a first floor, and a second floor.4 This model of the house, and the concept of the stages on life's way that it illustrates, is central to Kierkegaard's philosophy. This thesis will show how he unpacks this model in many of his writings with different concepts being developed in different texts. His method is to work with the same model in different ways throughout his authorship. He assigns many of the texts to different pseudonyms, but in this thesis we will treat the model and the related concepts as being Kierkegaard's and not only the pseudonyms. This is justified as our thesis will show this modelremains the same throughout Kierkegaard's work, though it is treated in different ways by different pseudonyms. According to Kierkegaard, many people live in only the basement for their entire lives, that is, as aesthetes ("in despair not to be conscious of having a self'). They live in despair of not being conscious of having a self They live in a merely horizontal relation. They want to get what they desire. When they go to the first floor, so to speak, they reflect on themselves and only then do they begin to get a self In this stage, one acquires an ideology of the required and overcomes the strict commands of the desired. The ethical is primarily an obedience to the required whereas the aesthetic is an obedience to desire. In his work Fear and Trembling (Copenhagen: 1843), Johannes de Silentio makes several observations concerning this point. In this book, the author several times allows the desired ideality of esthetics to be shipwrecked on the required ideality of ethics, in order through these collisions to bring to light the religious ideality as the ideality that precisely is the ideality of actuality, and therefore just as desirable as that of esthetics and not as impossible as the ideality of ethics. This is accomplished in such a way that the religious ideality breaks forth in the dialectical leap and in the positive mood - "Behold all things have become new" as well as in the negative mood that is the passion of the absurd to which the concept "repetition" corresponds.s Here one begins to become responsible because one seeks the required ideality; however, the required ideality and the desired ideality become inadequate to the ethical individual. Neither of them satisfy him ("in despair not to will to be oneself'). Then he moves up to the second floor: that is, the mystical region, or the sphere of religiousness (A) ("despair to will to be oneself). Kiericegaard's model of a house, which is connected with the above definition ofdespair, shows us how the self arises through these various stages, and shows the stages of despair as well. On the second floor, we become mystics, or Knights of Infinite Resignation. We are still in despair because we despair ofthe basement and the first floor, however, we can be fiill, free persons only ifwe live on all the floors at the same time. This is a sort of paradoxical fourth stage consisting of all three floors; this is the sphere of true religiousness (religiousness (B)). It is distinguished from religiousness (A) because we can go back and live on all the floors. It is not that there are four floors, but in the fourth stage, we live paradoxically on three at once. Kierkegaard uses this house analogy in order to explain how we become a self through these stages, and to show the various stages of despair. Consequently, I will be explaining self-becoming in relation to despair. It will also be necessary to explain it in relation to faith, for faith is precisely the overcoming of despair. After explaining the becoming of the self in relation to despair and faith, I will then explain its temporality and thereby its repetition. What Kierkegaard calls a formula, Deleuze calls a representation. Unfortunately, Deleuze does not acknowledge Kierkegaard's formula for repetition. As we shall see, Kierkegaard clearly gives a formula for despair, faith, and selfbecoming. When viewed properly, these formulae yield a formula for repetition because when one hasfaith, the basement, firstfloor, and secondfloor become new as one becomes oneself The self is not bound in the eternity ofthe first floor (ethical) or the temporality of the basement (aesthete). I shall now examine the two forms of conscious despair in such a way as to point out also a rise in the consciousness of the nature of despair and in the consciousness that one's state is despair, or, what amounts to the same thing and is the salient point, a rise in the consciousness of the self The opposite to being in despair is to have faith. Therefore, the formula set forth above, which describes a state in which there is not despair at all, is entirely correct, and this formula is also the formula for faMi in ^elating itself to itself and in willing to be itself, the self rests transparently in the power that established it.