Multicelled Behaviour in E. coli: Design and experimental characterization of an engineered library of hybrid promoters.

Synthetic biology has recently had a great development, many papers have been published and many applications have been presented, spanning from the production of biopharmacheuticals to the synthesis of bioenergetic substrates or industrial catalysts. But, despite these advances, most of the applications are quite simple and don’t fully exploit the potential of this discipline. This limitation in complexity has many causes, like the incomplete characterization of some components, or the intrinsic variability of the biological systems, but one of the most important reasons is the incapability of the cell to sustain the additional metabolic burden introduced by a complex circuit. The objective of the project, of which this work is part, is trying to solve this problem through the engineering of a multicellular behaviour in prokaryotic cells. This system will introduce a cooperative behaviour that will allow to implement complex functionalities, that can’t be obtained with a single cell. In particular the goal is to implement the Leader Election, this procedure has been firstly devised in the field of distributed computing, to identify the process that allow to identify a single process as organizer and coordinator of a series of tasks assigned to the whole population. The election of the Leader greatly simplifies the computation providing a centralized control. Further- more this system may even be useful to evolutionary studies that aims to explain how complex organisms evolved from unicellular systems. The work presented here describes, in particular, the design and the experimental characterization of a component of the circuit that solves the Leader Election problem. This module, composed of an hybrid promoter and a gene, is activated in the non-leader cells after receiving the signal that a leader is present in the colony. The most important element, in this case, is the hybrid promoter, it has been realized in different versions, applying the heuristic rules stated in [22], and their activity has been experimentally tested. The objective of the experimental characterization was to test the response of the genetic circuit to the introduction, in the cellular environment, of particular molecules, inducers, that can be considered inputs of the system. The desired behaviour is similar to the one of a logic AND gate in which the exit, represented by the luminous signal produced by a fluorescent protein, is one only in presence of both inducers. The robustness and the stability of this behaviour have been tested by changing the concentration of the input signals and building dose response curves. From these data it is possible to conclude that the analysed constructs have an AND-like behaviour over a wide range of inducers’ concentrations, even if it is possible to identify many differences in the expression profiles of the different constructs. This variability accounts for the fact that the input and the output signals are continuous, and so their binary representation isn’t able to capture the complexity of the behaviour. The module of the circuit that has been considered in this analysis has a fundamental role in the realization of the intercellular communication system that is necessary for the cooperative behaviour to take place. For this reason, the second phase of the characterization has been focused on the analysis of the signal transmission. In particular, the interaction between this element and the one that is responsible for emitting the chemical signal has been tested. The desired behaviour is still similar to a logic AND, since, even in this case, the exit signal is determined by the hybrid promoter activity. The experimental results have demonstrated that the systems behave correctly, even if there is still a substantial variability between them. The dose response curves highlighted that stricter constrains on the inducers concentrations need to be imposed in order to obtain a clear separation between the two levels of expression. In the conclusive chapter the DNA sequences of the hybrid promoters are analysed, trying to identify the regulatory elements that are most important for the determination of the gene expression. Given the available data it wasn’t possible to draw definitive conclusions. In the end, few considerations on promoter engineering and complex circuits realization are presented. This section aims to briefly recall some of the problems outlined in the introduction and provide a few possible solutions.

Vascular hospitalization rates and costs in patients with peripheral artery disease in the United States

BACKGROUND: Peripheral artery disease (PAD) is common and imposes a high risk of major systemic and limb ischemic events. The REduction of Atherothrombosis for Continued Health (REACH) Registry is an international prospective registry of patients at risk of atherothrombosis caused by established arterial disease or the presence of 3 atherothrombotic risk factors. METHODS AND RESULTS: We compared the 2-year rates of vascular-related hospitalizations and associated costs in US patients with established PAD across patient subgroups. Symptomatic PAD at enrollment was identified on the basis of current intermittent claudication with an ankle-brachial index (ABI) <0.90 or a history of lower-limb revascularization or amputation. Asymptomatic PAD was diagnosed on the basis of an enrollment ABI <0.90 in the absence of symptoms. Overall, 25 763 of the total 68 236-patient REACH cohort were enrolled from US sites; 2396 (9.3%) had symptomatic and 213 (0.8%) had asymptomatic PAD at baseline. One- and cumulative 2-year follow-up data were available for 2137 (82%) and 1677 (64%) of US REACH patients with either symptomatic or asymptomatic PAD, respectively. At 2 years, mean cumulative hospitalization costs, per patient, were $7445, $7000, $10 430, and $11 693 for patients with asymptomatic PAD, a history of claudication, lower-limb amputation, and revascularization, respectively (P=0.007). A history of peripheral intervention (lower-limb revascularization or amputation) was associated with higher rates of subsequent procedures at both 1 and 2 years. CONCLUSIONS: The economic burden of PAD is high. Recurring hospitalizations and repeat revascularization procedures suggest that neither patients, physicians, nor healthcare systems should assume that a first admission for a lower-extremity PAD procedure serves as a permanent resolution of this costly and debilitating condition.

Mortality and morbidity in the city of Bern, Switzerland, 1805-1815 with special emphasis on infant, child and maternal deaths

This article contributes to the research on demographics and public health of urban populations of preindustrial Europe. The key source is a burial register that contains information on the deceased, such as age and sex, residence and cause of death. This register is one of the earliest compilations of data sets of individuals with this high degree of completeness and consistency. Critical assessment of the register's origin, formation and upkeep promises high validity and reliability. Between 1805 and 1815, 4,390 deceased inhabitants were registered. Information concerning these individuals provides the basis for this study. Life tables of Bern's population were created using different models. The causes of death were classified and their frequency calculated. Furthermore, the susceptibility of age groups to certain causes of death was established. Special attention was given to causes of death and mortality of newborns, infants and birth-giving women. In comparison to other cities and regions in Central Europe, Bern's mortality structure shows low rates for infants (q0=0.144) and children (q1-4=0.068). This could have simply indicated better living conditions. Life expectancy at birth was 43 years. Mortality was high in winter and spring, and decreased in summer to a low level with a short rise in August. The study of the causes of death was inhibited by difficulties in translating early 19th century nomenclature into the modern medical system. Nonetheless, death from metabolic disorders, illnesses of the respiratory system, and debilitation were the most prominent causes in Bern. Apparently, the worst killer of infants up to 12 months was the "gichteren", an obsolete German term for lethal spasmodic convulsions. The exact modern identification of this disease remains unclear. Possibilities such as infant tetanus or infant epilepsy are discussed. The maternal death rate of 0.72% is comparable with values calculated from contemporaneous sources. Relevance of childbed fever in the early 1800s was low. Bern's data indicate that the extent of deaths related to childbirth in this period is overrated. This research has an explicit interdisciplinary value for various fields including both the humanities and natural sciences, since information reported here represents the complete age and sex structure of a deceased population. Physical anthropologists can use these data as a true reference group for their palaeodemographic studies of preindustrial Central Europe of the late 18th and early 19th century. It is a call to both historians and anthropologists to use our resources to a better effect through combination of methods and exchange of knowledge.

'Initiators of Discursive Practices': Authorship, attribution, and intent in the debates between philosophes and anti-philosophes

This article focuses on the “social side” of pseudonymity—on how writers and readers compete to influence the critical destiny of a pseudonymous work. By analyzing pseudonymity and attribution in both the specific context of Voltaire’s 1760 staging of the play, Le café ou l’écossaise, and in larger debates in the emerging fields of anonymity, pseudonymity, and attribution studies, I hope to show how literary scholars at present can address the individuality of each pseudonymous case while not letting go of trans-historical, general problems of anonymous strategies. Voltaire’s use of multiple pseudonyms before and after releasing L’Ecossaise, a comédie sérieuse in which Voltaire attacks his enemy Elie-Cathérine Fréron, supports his philosophe friends at a crucial moment in history, and exemplifies his emerging taste for serious comedy and British drama calls into question traditional takes on pseudonymity, anonymity, and attribution by refusing to fit into the binary arguments of anonymous vs. attributed and authorial intent vs. the reader’s control.

Eros and Transgression in an Age of Immanence: Georges Bataille’s (Religious) Critique of Kinsey

This paper explores the religious implications of eroticism in Western culture since the Sexual Revolution, a period at once applauded for its open and immanent view of sexuality and denounced for its shamelessness and promiscuity. After discussing the work and effects of Alfred C. Kinsey, the father of the Sexual Revolution, I focus on a critical appraisal of Kinsey written by French theorist Georges Bataille (“Kinsey, the Underworld and Work,” in L’Erotisme, 1957). Bataille situates contemporary Western sexuality within a larger historical movement towards the “desacralization” of all aspects of human life: sex, under the scientific gaze of the Kinsey team, became simply another “object” to be analyzed and classified, and “good” sex defined solely in terms of frequency and explosiveness of orgasm. For many, including Hugh Hefner, this approach to sex occasioned a refreshing awakening from the long dark night of Victorian sexual repression. However, as Bataille’s protégé Foucault has shown, the scientific approach to sexuality often masks a desire to control and delimit sexual behaviour, not “liberate” it. Moreover, Bataille makes the point that the desacralization of sexuality denudes sex of a vital component—eroticism—which is necessary for real pleasure and ecstasy. Beyond the “moral” critiques one often hears leveled against Kinsey and his work, Bataille provides a “religious” critique, one that stands, perhaps surprisingly, on the “near side” of sexuality.

Parameters of Reform and Unification in Modern Japanese Buddhist Thought: Murakami Senshō and Critical Buddhism

Reform is a word that, one might easily say, characterizes more than any other the history and development of Buddhism. Yet, it must also be said that reform movements in East Asian Buddhism have often taken on another goal—harmony or unification; that is, a desire not only to reconstruct a more worthy form of Buddhism, but to simultaneously bring together all existing forms under a single banner, in theory if not in practice. This paper explores some of the tensions between the desire for reform and the quest for harmony in modern Japanese Buddhism thought, by comparing two developments: the late 19th century movement towards ‘New Buddhism’ (shin Bukkyō) as exemplified by Murakami Senshō 村上専精 (1851–1929), and the late 20th century movement known as ‘Critical Buddhism’ (hihan Bukkyō), as found in the works of Matsumoto Shirō 松本史朗 and Hakamaya Noriaki 袴谷憲昭. In all that has been written about Critical Buddhism, in both Japanese and English, very little attention has been paid to the place of the movement within the larger traditions of Japanese Buddhist reform. Here I reconsider Critical Buddhism in relation to the concerns of the previous, much larger trends towards Buddhist reform that emerged almost exactly 100 years previous—the so-called shin Bukkyō or New Buddhism of the late-Meiji era. Shin Bukkyō is a catch-all term that includes the various writings and activities of Inoue Enryō, Shaku Sōen, and Kiyozawa Manshi, as well as the so-called Daijō-hibussetsuron, a broad term used (often critically) to describe Buddhist writers who suggested that Mahāyāna Buddhism is not, in fact, the Buddhism taught by the ‘historical’ Buddha Śākyamuni. Of these, I will make a few general remarks about Daijō-hibusseturon, before turning attention more specifically to the work of Murakami Senshō, in order to flesh out some of the similarities and differences between his attempt to construct a ‘unified Buddhism’ and the work of his late-20th century avatars, the Critical Buddhists. Though a number of their aims and ideas overlap, I argue that there remain fundamental differences with respect to the ultimate purposes of Buddhist reform. This issue hinges on the implications of key terms such as ‘unity’ and ‘harmony’ as well as the way doctrinal history is categorized and understood, but it also relates to issues of ideology and the use and abuse of Buddhist doctrines in 20th-century politics.

The Pursuit of Tangible Happiness: Religion and Politics in a Japanese New, New Religion

The decline of traditional religions in Japan in the past century, and especially since the end of World War Two, has led to an explosion of so-called “new religions” (shin shūkyō 新宗教), many of which have made forays into the political realm. The best known—and most controversial—example of a “political” new religion is Sōka Gakkai 創価学会, a lay Buddhist movement originally associated with the Nichiren sect that in the 1960s gave birth to a new political party, Komeitō 公明党 (lit., Clean Government Party), which in the past several decades has emerged as the third most popular party in Japan (as New Komeitō). Since the 1980s, Japan has also seen the emergence of so-called “new, new religions” (shin shin shūkyō 新新宗教), which tend to be more technologically savvy and less socially concerned (and, in the eyes of critics, more akin to “cults” than the earlier new religions). One new, new religion known as Kōfuku-no-Kagaku 幸福の科学 (lit., Institute for Research in Human Happiness or simply Happy Science), founded in 1986 by Ōkawa Ryūho 大川隆法, has very recently developed its own political party, Kōfuku Jitsugentō 幸福実現党 (The Realization of Happiness Party). This article will analyse the political ideals of Kōfuku Jitsugentō in relation to its religious teachings, in an attempt to situate the movement within the broader tradition of religio-political syncretism in Japan. In particular, it will examine the recent “manifesto” of Kōfuku Jitsugentō in relation to those of New Komeitō and “secular” political parties such as the Liberal Democratic Party (Jimintō 自民党) and the Democratic Party (Minshutō 民主党).

Narrating Boundaries: Framing and Contesting Suffering, Community, and Belonging in Enclaves along the India-Bangladesh Border

This paper explores the politics of community making at the India-Bangladesh border by examining the public and private narratives of history and belonging in a Bangladeshi enclave-a sovereign piece of Bangladesh completely territorially surrounded by India. Drawing on framings of political society, this paper argues that understanding populations at the margins of South Asia and beyond requires attention to two processes: first, to the ways that para-legal activities are part and parcel of daily life; and second, to the strategies through which these groups construct themselves as moral communities deserving of inclusion within the state. Border communities often articulate narratives of dispossession, exceptionality, and marginalization to researchers and other visitors-narratives that are often unproblematically reproduced in academic treatments of the border. However, such articulations mask both the complicated histories and quotidian realities of border life. This paper views these articulations as political projects in and of themselves. By reading the more hidden histories of life in this border enclave, this article reconstructs the notion of borders as experienced by enclave residents themselves. It shows the ways that the politics of the India-Bangladesh border are constitutive of (and constituted by) a range of fractures and internal boundaries within the enclave. These boundaries are as central to forging community-to articulating who belongs and why-as are more public narratives that frame enclave residents as victims of confused territorial configurations. (C) 2012 Elsevier Ltd. All rights reserved.

[Juvenile sterile granulomatous dermatitis and lymphadenitis in the dog]

Juvenile sterile granulomatous dermatitis and lymphadenitis is a rare immune-mediated skin disease in young dogs. History, signalment, diagnostics, treatment, and outcome in 10 dogs are described. The age ranged from 8 - 36 weeks. The lymph nodes were enlarged in all dogs, especially the mandibular and prescapular lymph nodes. Systemic signs including fever were present in 8 dogs. Seven dogs suffered from blepharitis and painful edema of the muzzle with hemorrhagic discharge, pustules and papules. Cytology of pustules and lymph node aspirates revealed a pyogranulomatous inflammation. In 7 cases the diagnosis of juvenile sterile granulomatous dermatitis and lymphadenitis was confirmed by histology. Nine dogs were treated with prednisolone (0.5 - 1.25 mg/kg BID), H2-receptor antagonists and analgetics; all dogs were treated with antibiotics. Four dogs were treated with eye ointment containing antibiotics and glucocorticoids. The prednisolone dosage was tapered over 3 - 8 weeks. One dog had a relapse.

Falls and fractures in the elderly with sinus node disease: the impact of pacemaker implantation

Background. Falls and fractures in the elderly are among the leading causes of disability. We investigated whether pacemaker implantation prevents falls in patients with SND in a large cohort of patients. Methods. Patient demographics and medical history were collected prospectively. Fall history was retrospectively reconstituted from available medical records. The 10-year probability for major osteoporotic fractures was calculated retrospectively from available medical records using the Swiss fracture risk assessment tool FRAX-Switzerland. Results. During a mean observation period of 2.3 years after implantation, the rates of fallers and injured fallers with fracture were reduced to 15% and 6%, respectively. This corresponds to a relative reduction in the number of fallers of 75% (P < 0.001) and of injured fallers of 63% (P = 0.014) after pacemaker implantation. Similarly, the number of falls was reduced from 60 (48%) before pacemaker implantation to 22 (18%) thereafter (relative reduction 63%, P = 0.035) and the number of falls with injury from 22 (18%) to 7 (6%), which corresponds to a relative reduction of 67%, P = 0.013. Conclusion. In patients with SND, pacemaker implantation significantly reduces the number of patients experiencing falls, the total number of falls, and the risk for osteoporotic fractures.

Tuberculosis and the risk of opportunistic infections and cancers in HIV-infected patients starting ART in Southern Africa

To investigate the incidence of selected opportunistic infections (OIs) and cancers and the role of a history of tuberculosis (TB) as a risk factor for developing these conditions in HIV-infected patients starting antiretroviral treatment (ART) in Southern Africa.