Extensive microRNA-mediated crosstalk between lncRNAs and mRNAs in mouse embryonic stem cells.

Recently, a handful of intergenic long noncoding RNAs (lncRNAs) have been shown to compete with mRNAs for binding to miRNAs and to contribute to development and disease. Beyond these reports, little is yet known of the extent and functional consequences of miRNA-mediated regulation of mRNA levels by lncRNAs. To gain further insight into lncRNA-mRNA miRNA-mediated crosstalk, we reanalyzed transcriptome-wide changes induced by the targeted knockdown of over 100 lncRNA transcripts in mouse embryonic stem cells (mESCs). We predicted that, on average, almost one-fifth of the transcript level changes induced by lncRNAs are dependent on miRNAs that are highly abundant in mESCs. We validated these findings experimentally by temporally profiling transcriptome-wide changes in gene expression following the loss of miRNA biogenesis in mESCs. Following the depletion of miRNAs, we found that >50% of lncRNAs and their miRNA-dependent mRNA targets were up-regulated coordinately, consistent with their interaction being miRNA-mediated. These lncRNAs are preferentially located in the cytoplasm, and the response elements for miRNAs they share with their targets have been preserved in mammals by purifying selection. Lastly, miRNA-dependent mRNA targets of each lncRNA tended to share common biological functions. Post-transcriptional miRNA-mediated crosstalk between lncRNAs and mRNA, in mESCs, is thus surprisingly prevalent, conserved in mammals, and likely to contribute to critical developmental processes.

The planarian flatworm: an in vivo model for stem cell biology and nervous system regeneration

Planarian flatworms are an exception among bilaterians in that they possess a large pool of adult stem cells that enables them to promptly regenerate any part of their body, including the brain. Although known for two centuries for their remarkable regenerative capabilities, planarians have only recently emerged as an attractive model for studying regeneration and stem cell biology. This revival is due in part to the availability of a sequenced genome and the development of new technologies, such as RNA interference and next-generation sequencing, which facilitate studies of planarian regeneration at the molecular level. Here, we highlight why planarians are an exciting tool in the study of regeneration and its underlying stem cell biology in vivo, and discuss the potential promises and current limitations of this model organism for stem cell research and regenerative medicine.

Self-Renewing Human Bone Marrow Mesenspheres Promote Hematopoietic Stem Cell Expansion.

Strategies for expanding hematopoietic stem cells (HSCs) include coculture with cells that recapitulate their natural microenvironment, such as bone marrow stromal stem/progenitor cells (BMSCs). Plastic-adherent BMSCs may be insufficient to preserve primitive HSCs. Here, we describe a method of isolating and culturing human BMSCs as nonadherent mesenchymal spheres. Human mesenspheres were derived from CD45- CD31- CD71- CD146+ CD105+ nestin+ cells but could also be simply grown from fetal and adult BM CD45--enriched cells. Human mesenspheres robustly differentiated into mesenchymal lineages. In culture conditions where they displayed a relatively undifferentiated phenotype, with decreased adherence to plastic and increased self-renewal, they promoted enhanced expansion of cord blood CD34+ cells through secreted soluble factors. Expanded HSCs were serially transplantable in immunodeficient mice and significantly increased long-term human hematopoietic engraftment. These results pave the way for culture techniques that preserve the self-renewal of human BMSCs and their ability to support functional HSCs.

Exploring transparent approaches to the authentication of signatures on artwork

This thesis is composed of three main parts. The first consists of a state of the art of the different notions that are significant to understand the elements surrounding art authentication in general, and of signatures in particular, and that the author deemed them necessary to fully grasp the microcosm that makes up this particular market. Individuals with a solid knowledge of the art and expertise area, and that are particularly interested in the present study are advised to advance directly to the fourth Chapter. The expertise of the signature, it's reliability, and the factors impacting the expert's conclusions are brought forward. The final aim of the state of the art is to offer a general list of recommendations based on an exhaustive review of the current literature and given in light of all of the exposed issues. These guidelines are specifically formulated for the expertise of signatures on paintings, but can also be applied to wider themes in the area of signature examination. The second part of this thesis covers the experimental stages of the research. It consists of the method developed to authenticate painted signatures on works of art. This method is articulated around several main objectives: defining measurable features on painted signatures and defining their relevance in order to establish the separation capacities between groups of authentic and simulated signatures. For the first time, numerical analyses of painted signatures have been obtained and are used to attribute their authorship to given artists. An in-depth discussion of the developed method constitutes the third and final part of this study. It evaluates the opportunities and constraints when applied by signature and handwriting experts in forensic science. A brief summary covering each chapter allows a rapid overview of the study and summarizes the aims and main themes of each chapter. These outlines presented below summarize the aims and main themes addressed in each chapter. Part I - Theory Chapter 1 exposes legal aspects surrounding the authentication of works of art by art experts. The definition of what is legally authentic, the quality and types of the experts that can express an opinion concerning the authorship of a specific painting, and standard deontological rules are addressed. The practices applied in Switzerland will be specifically dealt with. Chapter 2 presents an overview of the different scientific analyses that can be carried out on paintings (from the canvas to the top coat). Scientific examinations of works of art have become more common, as more and more museums equip themselves with laboratories, thus an understanding of their role in the art authentication process is vital. The added value that a signature expertise can have in comparison to other scientific techniques is also addressed. Chapter 3 provides a historical overview of the signature on paintings throughout the ages, in order to offer the reader an understanding of the origin of the signature on works of art and its evolution through time. An explanation is given on the transitions that the signature went through from the 15th century on and how it progressively took on its widely known modern form. Both this chapter and chapter 2 are presented to show the reader the rich sources of information that can be provided to describe a painting, and how the signature is one of these sources. Chapter 4 focuses on the different hypotheses the FHE must keep in mind when examining a painted signature, since a number of scenarios can be encountered when dealing with signatures on works of art. The different forms of signatures, as well as the variables that may have an influence on the painted signatures, are also presented. Finally, the current state of knowledge of the examination procedure of signatures in forensic science in general, and in particular for painted signatures, is exposed. The state of the art of the assessment of the authorship of signatures on paintings is established and discussed in light of the theoretical facets mentioned previously. Chapter 5 considers key elements that can have an impact on the FHE during his or her2 examinations. This includes a discussion on elements such as the skill, confidence and competence of an expert, as well as the potential bias effects he might encounter. A better understanding of elements surrounding handwriting examinations, to, in turn, better communicate results and conclusions to an audience, is also undertaken. Chapter 6 reviews the judicial acceptance of signature analysis in Courts and closes the state of the art section of this thesis. This chapter brings forward the current issues pertaining to the appreciation of this expertise by the non- forensic community, and will discuss the increasing number of claims of the unscientific nature of signature authentication. The necessity to aim for more scientific, comprehensive and transparent authentication methods will be discussed. The theoretical part of this thesis is concluded by a series of general recommendations for forensic handwriting examiners in forensic science, specifically for the expertise of signatures on paintings. These recommendations stem from the exhaustive review of the literature and the issues exposed from this review and can also be applied to the traditional examination of signatures (on paper). Part II - Experimental part Chapter 7 describes and defines the sampling, extraction and analysis phases of the research. The sampling stage of artists' signatures and their respective simulations are presented, followed by the steps that were undertaken to extract and determine sets of characteristics, specific to each artist, that describe their signatures. The method is based on a study of five artists and a group of individuals acting as forgers for the sake of this study. Finally, the analysis procedure of these characteristics to assess of the strength of evidence, and based on a Bayesian reasoning process, is presented. Chapter 8 outlines the results concerning both the artist and simulation corpuses after their optical observation, followed by the results of the analysis phase of the research. The feature selection process and the likelihood ratio evaluation are the main themes that are addressed. The discrimination power between both corpuses is illustrated through multivariate analysis. Part III - Discussion Chapter 9 discusses the materials, the methods, and the obtained results of the research. The opportunities, but also constraints and limits, of the developed method are exposed. Future works that can be carried out subsequent to the results of the study are also presented. Chapter 10, the last chapter of this thesis, proposes a strategy to incorporate the model developed in the last chapters into the traditional signature expertise procedures. Thus, the strength of this expertise is discussed in conjunction with the traditional conclusions reached by forensic handwriting examiners in forensic science. Finally, this chapter summarizes and advocates a list of formal recommendations for good practices for handwriting examiners. In conclusion, the research highlights the interdisciplinary aspect of signature examination of signatures on paintings. The current state of knowledge of the judicial quality of art experts, along with the scientific and historical analysis of paintings and signatures, are overviewed to give the reader a feel of the different factors that have an impact on this particular subject. The temperamental acceptance of forensic signature analysis in court, also presented in the state of the art, explicitly demonstrates the necessity of a better recognition of signature expertise by courts of law. This general acceptance, however, can only be achieved by producing high quality results through a well-defined examination process. This research offers an original approach to attribute a painted signature to a certain artist: for the first time, a probabilistic model used to measure the discriminative potential between authentic and simulated painted signatures is studied. The opportunities and limits that lie within this method of scientifically establishing the authorship of signatures on works of art are thus presented. In addition, the second key contribution of this work proposes a procedure to combine the developed method into that used traditionally signature experts in forensic science. Such an implementation into the holistic traditional signature examination casework is a large step providing the forensic, judicial and art communities with a solid-based reasoning framework for the examination of signatures on paintings. The framework and preliminary results associated with this research have been published (Montani, 2009a) and presented at international forensic science conferences (Montani, 2009b; Montani, 2012).

Caída de bloques de hielo en los frentes del glaciar de casquete de la Isla Livingston (Archipiélago de la Shetland del Sur). Detección por métodos sísmicos. Influencia de las variables ambientales.

Se han utilizado los datos procedentes de una red (array) sísmica situada en la isla Livingston para estudiar la evolución de las caídas de bloques de hielo en los frentes de los glaciares (calving). Pa rtiendo de que la causa fundamental de las caídas de bloques es el flujo del hielo, se ha estudiado la influencia de la temperatura ambiental, la variación del nivel de la marea, la humedad y la velocidad del viento sobre este fenómeno. Como principal conclusión se deduce que la temperatura ambiental es el factor que ejerce una mayor y más directa influencia sobre las caídas de los bloques de hielo.

Obstructive apneas induce early activation of mesenchymal stem cells and enhancement of endothelial wound healing

Background: The aim was to test the hypothesis that the blood serum of rats subjected to recurrent airway obstructions mimicking obstructive sleep apnea (OSA) induces early activation of bone marrow-derived mesenchymal stem cells (MSC) and enhancement of endothelial wound healing. Methods: We studied 30 control rats and 30 rats subjected to recurrent obstructive apneas (60 per hour, lasting 15 s each, for 5 h). The migration induced in MSC by apneic serum was measured by transwell assays. MSC-endothelial adhesion induced by apneic serum was assessed by incubating fluorescent-labelled MSC on monolayers of cultured endothelial cells from rat aorta. A wound healing assay was used to investigate the effect of apneic serum on endothelial repair. Results: Apneic serum showed significant increase in chemotaxis in MSC when compared with control serum: the normalized chemotaxis indices were 2.20 +- 0.58 (m +- SE) and 1.00 +- 0.26, respectively (p < 0.05). MSC adhesion to endothelial cells was greater (1.75 +- 0.14 -fold; p < 0.01) in apneic serum than in control serum. When compared with control serum, apneic serum significantly increased endothelial wound healing (2.01 +- 0.24 -fold; p < 0.05). Conclusions: The early increases induced by recurrent obstructive apneas in MSC migration, adhesion and endothelial repair suggest that these mechanisms play a role in the physiological response to the challenges associated to OSA.

Synergies between optical and physical variables in intercepting parabolic targets.

Interception requires precise estimation of time-to-contact (TTC) information. A long-standing view posits that all relevant information for extracting TTC is available in the angular variables, which result from the projection of distal objecs on to the retina. The diferent timing models rooted in this tradition have consequently relied on combining visual angle and its rate of expansion in diferent ways with tau being the most well-known solution for TTC...

Immunomodulatory effect of mesenchymal stem cells on B cells

The research on T cell immunosuppression therapies has attracted most of the attention in clinical transplantation. However, B cells and humoral immune responses are increasingly acknowledged as crucial mediators of chronic allograft rejection. Indeed, humoral immune responses can lead to renal allograft rejection even in patients whose cell-mediated immune responses are well controlled. On the other hand, newly studied B cell subsets with regulatory effects have been linked to tolerance achievement in transplantation. Better understanding of the regulatory and effector B cell responses may therefore lead to new therapeutic approaches. Mesenchymal stem cells (MSC) are arising as a potent therapeutic tool in transplantation due to their regenerative and immunomodulatory properties.The research on MSCs has mainly focused on their effects onT cells and although data regarding the modulatory effects of MSCs on alloantigen-specific humoral response in humans is scarce, it has been demonstrated that MSCs significantly affect B cell functioning. In the present review we will analyze and discuss the results in this field.

Ewing sarcoma cancer stem cells : from mechanisms to therapeutic targeting

Le sarcome d'Ewing (SE) est la 2ème tumeur des os la plus fréquente chez les enfants, et le pronostic est sombre au stade métastatique. La pathogenèse du SE repose sur une translocation, provocant la fusion du domaine activateur du facteur de transcription EWS, avec la partie liant l'ADN de la protéine FLI-1. Les cellules souches cancéreuses (CSC) sont supposées être les moteurs de la croissance tumorale, et représente de ce fait des cibles thérapeutiques préférentielles. Dans ce travail nous nous sommes efforcés de comprendre, ainsi que de cibler les mécanismes liés à l'émergence des CSC dans le sarcome d'Ewing. La formation des CSC du ES est liée à un défaut de maturation des miRNAs provoqué par une sous-expression d'un gène, TARBP2, dans les CSC. Ce défaut de maturation peut être corrigé par un traitement des cellules avec de l'enoxacine, une fluoroquinolone utilisée pour traiter les infections urinaires. L'enoxacine seule n'étant pas suffisante pour éradiquer les tumeurs in vivo, nous avons testé la combinaison d'une thérapie ciblée sur les CSC avec une chimiothérapie classique, la doxorubicine, ciblant les cellules différentiées. In vitro l'enoxacine induit l'apoptose dans les CCS sans affecter les cellules différentiées, alors que à l'inverse, la doxorubicine n'affecte que les cellules de la « masse » tumorale. In vivo la combinaison de ces deux drogues inhibe la croissance de tumeurs provenant de cellules primaires xenotranplantées et éradique les CSCs. Nos résultats mettent en lumière une nouvelle approche thérapeutique directement applicable pour le sarcome d'Ewing, et pourraient ainsi rapidement déboucher sur des essais cliniques. Dans la deuxième partie de ce travail nous avons essayé de comprendre comment EWS-FLI1, la protéine de fusion issue de la translocation chromosomique du sarcome d'Ewing conduit à la génération des CSC. Pour cela nous avons effectué des ChIPseq (immunoprecipitation de la chromatine suivi de séquençage) pour EWS-FLI1 ainsi que pour certaines modifications histoniques. -- Ewing sarcoma family tumors (ESFT) are the second most frequent bone tumors in children and have a high rate of recurrence when metastatic at presentation. The pathogenesis of Ewing sarcoma is underlayed by a translocation, leading to the fusion of the trans-activating domain of EWS with the FLU DNA binding domain. Cancer stem cells (CSCs) are thought to be the driving force of tumor growth. In this work we focused on understanding the mechanisms underlying ESFT CSC emergence as well as defining targeted therapeutic strategies. Emergence of CSCs in ESFT has been shown to arise from a defect in TARBP2-dependent microRNA maturation, which can be corrected by exposure to the fluoroquinolone enoxacin. As enoxacin alone is not sufficient to reverse tumor growth in vivo, we assessed the effect of combining a drug that abrogates CSC properties with doxorubicin, a standard-of-care therapy in ESFT. Primary ESFT CSCs and bulk tumor cells were treated with different concentration of drugs and displayed divergent responses to doxorubicin and enoxacin. Doxorubicin, which targets the tumor bulk, displayed toxicity toward primary adherent ESFT cells in culture but not to CSC-enriched ESFT spheres. Conversely, enoxacin induced apoptosis but only in ESFT spheres and specifically on the CD133+ population. In combination, the two drugs markedly depleted CSC and strongly reduced primary growth in xenograft assays of two primary ESFT. Our results identify a potentially attractive therapeutic strategy for ESFT that combines mechanism-based targeting of CSC using a low toxicity antibiotic with a standard-of-care cytotoxic drug, offering immediate applications for clinical evaluation. In the second part of this work we performed chromatin immunopercipitation on CSCs and bulk cells for EWS-FLI1 binding as well as some chromatin modifications, and concluded that EWS-FLI1 shows cell context dependent binding.

Production and quality of Anacardium othonianum Rizz. seedlings grown in different substrates

This study aimed to evaluate the effect of substrate on growth, emergence, nutrition, and quality of Anacardium othonianum Rizz. (cerrado cashew tree) seedlings. The experiment was conducted in a greenhouse at the Plant Tissue Culture Laboratory on the Rio Verde campus. The following substrates were used: 1) Bioplant®, 2) Mecplant® (MP) + carbonized rice husk (CRH) (7:3), 3) fine-grained vermiculite (FGV), 4) FGV+CRH (3:1), 5) FGV+CRH (1:1), 6) FGV+CRH (1:3), and 7) sugarcane bagasse (SB) + sugarcane mill filter cake (FC) (3:2). Emerged seedlings were counted at 2-day intervals for 38 days following emergence of the first seedling. At 39, 64, and 89 days after seeding (DAS), the following variables were measured: stem length (SL), stem diameter (SD), and number of leaves (NL). Accumulated dry weight, quality indices, and leaf macro- and micronutrient levels were determined at 89 DAS. Plants grown in the FGV and FGV+CFH (1:3) substrates had shorter stem lengths than the plants grown in other substrates. Increases in seedling growth were smaller between 64 and 89 DAS compared to the initial period of the experiment. The highest leaf N concentrations were found in the SB+FC substrate treatment group; P and K concentrations were higher for the MP+CRH (7:3), SB+FC, and Bioplant® treatments; and Ca levels were higher for the SB+FC and MP+CRH (7:3) substrate treatments. The MP+CRH (7:3) substrate treatment group had the highest leaf B and Mn micronutrient concentrations, and plants from the Bioplant® substrate group had the highest leaf B micronutrient content. Mg, S, Cu, Zn, and Fe concentrations did not differ among the different substrates. The plant traits that differed most among the treatments included stem length for the FGV and FGV+CRH (1:3) substrate groups and leaf nutrient concentrations, which were higher for the SB+FC group followed by the MP+CRH and Bioplant® treatments.

Growth and production of irrigated vitória pineapple grown in semi-arid conditions

This study aimed to evaluate the growth characteristics of irrigated Vitória pineapple plants grown in semi-arid conditions and determine its developmental stages based on those characteristics. It was used a randomized block design with four replicates. The experimental treatments were: plant harvest at 270, 330, 390, 450, 510, 570, 690, 750, and 810 days after planting (DAP). The following variables were determined: plant height, stem diameter, D-leaf length, D-leaf fresh and dry mass, biomass production of plants and plant parts (organs), and vegetative biomass. Five phenological stages are proposed based on vegetative biomass production: < 20% biomass production (V1); 21-40% (V2); 41-60% (V3); 61-80% (V4); and > 80% (V5). The maximum growth rate for plant height, D-leaf length, and stem diameter was observed at the end of the phenological stage V1 (390-411 DAP), and at the end of stage V5 these plant traits had average values of 106, 82, and 7 cm, respectively. The maximum biomass accumulation rates were observed at stages V4 and V5, resulting in a final fruit yield and total fresh biomass of 72 t ha-1 and 326 t ha-1, respectively. Finally, we estimated that 80% of the accumulated biomass may remain in the field after fruit and slip harvest, and could be incorporated as plant residue into the soil.

Assessing positive mental health in people with chronic physical health problems: correlations with socio-demographic variables and physical health status

Background: A holistic perspective on health implies giving careful consideration to the relationship between physical and mental health. In this regard the present study sought to determine the level of Positive Mental Health (PMH) among people with chronic physical health problems, and to examine the relationship between the observed levels of PMH and both physical health status and socio-demographic variables. Methods: The study was based on the Multifactor Model of Positive Mental Health (Lluch, 1999), which comprises six factors: Personal Satisfaction (F1), Prosocial Attitude (F2), Self-control (F3), Autonomy (F4), Problem-solving and Self-actualization (F5), and Interpersonal Relationship Skills (F6). The sample comprised 259 adults with chronic physical health problems who were recruited through a primary care center in the province of Barcelona (Spain). Positive mental health was assessed by means of the Positive Mental Health Questionnaire (Lluch, 1999). Results: Levels of PMH differed, either on the global scale or on specific factors, in relation to the following variables: age: global PMH scores decreased with age (r=-0.129; p=0.038); b) gender: men scored higher on F1 (t=2.203; p=0.028) and F4 (t=3.182; p=0.002), while women scored higher on F2 (t -3.086; p=0.002) and F6 (t=-2.744; p=0.007); c) number of health conditions: the fewer the number of health problems the higher the PMH score on F5 (r=-0.146; p=0.019); d) daily medication: polymedication patients had lower PMH scores, both globally and on various factors; e) use of analgesics: occasional use of painkillers was associated with higher PMH scores on F1 (t=-2.811; p=0.006). There were no significant differences in global PMH scores according to the type of chronic health condition. The only significant difference in the analysis by factors was that patients with hypertension obtained lower PMH scores on the factor Autonomy (t=2.165; p=0.032). Conclusions: Most people with chronic physical health problems have medium or high levels of PMH. The variables that adversely affect PMH are old age, polypharmacy and frequent consumption of analgesics. The type of health problem does not influence the levels of PMH. Much more extensive studies with samples without chronic pathology are now required in order to be able to draw more robust conclusions.