Immediate and late analysis of dental pulp stem cells viability after indirect exposition to alternative in-office bleaching strategies

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Energia da fonti rinnovabili negli edifici esistenti. Integrazione architettonica di captatori solari ed eolici.

Questa tesi tratta dell'applicazione agli edifici esistenti di dispositivi per la produzione energetica da fonte rinnovabile: solare fotovoltaica, solare termica ed eolica. Dopo aver analizzato i motivi economici ed ambientali che oggi inducono ad affrontare il tema complesso dell'integrazione di queste tecnologie anche negli edifici esistenti, vengono documentate alcune decine di recenti istallazioni, reperite in ambito internazionale, costituendo così un piccolo repertorio di casi applicativi. Il ricorso alle fonti di energia rinnovabili in architettura ed il concetto di sostenibilità applicato alla progettazione sono temi di interesse mondiale che coinvolgono la riqualificazione energetica del parco edilizio esistente. Questo fenomeno sta generando una vera rivoluzione che, da una dimensione culturale promossa dalle politiche energetiche dei vari paesi, travolge svariati ambiti disciplinari, primo tra tutti l'architettura. L’elaborato è idealmente diviso in quattro parti, come di seguito specificato. Nel primo capitolo vengono trattate le problematiche che stanno portando alla diffusione delle energie rinnovabili nel mondo e descritte le varie forme e tecnologie con cui si utilizzano. Nel secondo e terzo capitolo sono trattati i temi dell’energia negli edifici e la sostenibilità nel progetto per la riqualificazione energetica in architettura. Negli ultimi tre capitoli si analizzano le caratteristiche, i componenti ed il funzionamento dei sistemi fotovoltaici, solari termici e micro/mini-eolici applicati ai manufatti edilizi. Si descrivono, inoltre, le tecnologie e le soluzioni per l’integrazione di detti impianti sugli edifici esistenti. L’ultima parte, costituita dall’allegato che contiene le schede di analisi di alcuni casi studio, descrive la realizzazione di progetti di riqualificazione energetica in cui sistemi fotovoltaici, solari termici ed eolici vengono applicati ad edifici di vario genere.

Valutazione della sostenibilità ambientale tramite metodologia LCA di sistemi per lo sfruttamento di fonti alternative di energia e materiali

La dissertazione ha riguardato l’analisi di sostenibilità di un sistema agronomico per la produzione di olio vegetale a fini energetici in terreni resi marginali dall’infestazione di nematodi. Il processo indagato ha previsto il sovescio di una coltura con proprietà biofumiganti (brassicacea) coltivata in precessione alla specie oleosa (soia e tabacco) al fine di contrastare il proliferare dell’infestazione nel terreno. Tale sistema agronomico è stato confrontato attraverso una analisi di ciclo di vita (LCA) ad uno scenario di coltivazione della stessa specie oleosa senza precessione di brassica ma con l’utilizzo di 1-3-dicloropropene come sistema di lotta ai nematodi. Allo scopo di completare l’analisi LCA con una valutazione dell’impatto sull’uso del suolo (Land use Impact) generato dai due scenari a confronto, sono stati costruiti due modelli nel software per il calcolo del Soil Conditioning Index (SCI), un indicatore quali-quantitativo della qualità del terreno definito dal Dipartimento per l’Agricoltura degli Stati Uniti d’America (USDA).

Current United States stem cell policy considerations and a recommendation for an alternative policy based on guidelines from existing international policies

Embryonic stem cell research is a widely debated topic in modern politics and religion. Differing views on the fetal rights conflict with the rights of an embryo. Those who believe an embryo has the same human qualities as a fetus accordingly believe embryonic stem cell research is unethical because it destroys a potential human life. However, scientists advocate the embryo does not have human qualities and should be used for valuable research in the stem cell field. Stem cell research may lead to vast developments in medical treatments, including cancer and brain conditions and injuries that are currently incurable. ^ The current stem cell policy introduced by President Bush in 2001 in an attempt to balance the moral issues with the need for scientific research has broad negative implications on the furthering of stem cell research. There is a limited diversity of available stem cell lines, there may be constitutional issues, there is an increasing disparity between the public and private research spheres, and the U.S. is struggling to maintain its scientific community. The U.S. must develop a new stem cell research policy that will balance the interest of science and public health with the moral issues that concern the public. ^ The United Kingdom allows researchers great liberty in conducting research, permitting the creation of embryos for the sole purpose of research, while Germany is equally conservative in their laws, as their policies support the philosophy that all embryos deserve the protection of full life. The United States should adopt a policy that takes the "middle ground" approach and permit research on excess embryos created for IVF purposes, rather than simply discarding those potentially valuable research tools. ^

Analysis of Histone Deacetylase 7 (HDAC7) Alternative Splicing and Its Role in Embryonic Stem Cell Differentiation Toward Smooth Muscle Lineage

Histone deacetylases (HDACs) have a central role in the regulation of gene expression, which undergoes alternative splicing during embryonic stem cell (ES) cell differentiation. Alternative splicing gives rise to vast diversity over gene information, arousing public concerns in the last decade. In this chapter, we describe a strategy to detect HDAC7 alternative splicing and analyze its function on ES cell differentiation.

Autocrine fibroblast growth factor 2 increases the multipotentiality of human adipose-derived mesenchymal stem cells

Multipotent mesenchymal stem cells (MSCs), first identified in the bone marrow, have subsequently been found in many other tissues, including fat, cartilage, muscle, and bone. Adipose tissue has been identified as an alternative to bone marrow as a source for the isolation of MSCs, as it is neither limited in volume nor as invasive in the harvesting. This study compares the multipotentiality of bone marrow-derived mesenchymal stem cells (BMSCs) with that of adipose-derived mesenchymal stem cells (AMSCs) from 12 age- and sex-matched donors. Phenotypically, the cells are very similar, with only three surface markers, CD106, CD146, and HLA-ABC, differentially expressed in the BMSCs. Although colony-forming units-fibroblastic numbers in BMSCs were higher than in AMSCs, the expression of multiple stem cell-related genes, like that of fibroblast growth factor 2 (FGF2), the Wnt pathway effectors FRAT1 and frizzled 1, and other self-renewal markers, was greater in AMSCs. Furthermore, AMSCs displayed enhanced osteogenic and adipogenic potential, whereas BMSCs formed chondrocytes more readily than AMSCs. However, by removing the effects of proliferation from the experiment, AMSCs no longer out-performed BMSCs in their ability to undergo osteogenic and adipogenic differentiation. Inhibition of the FGF2/fibroblast growth factor receptor 1 signaling pathway demonstrated that FGF2 is required for the proliferation of both AMSCs and BMSCs, yet blocking FGF2 signaling had no direct effect on osteogenic differentiation. Disclosure of potential conflicts of interest is found at the end of this article.

Advanced Gen-1, 2 and 3 biofuels research in Australia - Fuelling advanced biofuels training for international scientists

Flinders University and Queensland University of Technology, biofuels research interests cover a broad range of activities. Both institutions are seeking to overcome the twin evils of "peak oil" (Hubbert 1949 & 1956) and "global warming" (IPPC 2007, Stern 2006, Alison 2010), through development of Generation 1, 2 and 3 (Gen-1, 2 & 3) biofuels (Clarke 2008, Clarke 2010). This includes development of parallel Chemical Biorefinery, value-added, co-product chemical technologies, which can underpin the commercial viability of the biofuel industry. Whilst there is a focused effort to develop Gen-2 & 3 biofuels, thus avoiding the socially unacceptable use of food based Gen-1 biofuels, it must also be recognized that as yet, no country in the world has produced sustainable Gen-2 & 3 biofuel on a commercial basis. For example, in 2008 the United States used 38 billion litres (3.5% of total fuel use) of Gen-1 biofuel; in 2009/2010 this will be 47.5 billion litres (4.5% of fuel use) and in 2018 this has been estimated to rise to 96 billion litres (9% of total US fuel use). Brazil in 2008 produced 24.5 billion litres of ethanol, representing 37.3% of the world’s ethanol use for fuel and Europe, in 2008, produced 11.7 billion litres of biofuel (primarily as biodiesel). Compare this to Australia’s miserly biofuel production in 2008/2009 of 180 million litres of ethanol and 75 million litres of biodiesel, which is 0.4% of our fuel consumption! (Clarke, Graiver and Habibie 2010) To assist in the development of better biofuels technologies in the Asian developing regions the Australian Government recently awarded the Materials & BioEnergy Group from Flinders University, in partnership with the Queensland University of Technology, an Australian Leadership Award (ALA) Biofuel Fellowship program to train scientists from Indonesia and India about all facets of advanced biofuel technology.

Mesenchymal stem cells

Cell based therapies as they apply to tissue engineering and regenerative medicine, require cells capable of self renewal and differentiation, and a prerequisite is to be able to prepare an effective dose of ex vivo expanded cells for autologous transplants. The in vivo identification of a source of physiologically relevant cell types suitable for cell therapies therefore figures as an integral part of tissue engineering. Stem cells serve as a reserve for biological repair, having the potential to differentiate into a number of specialised cell types within the body; they therefore represent the most useful candidates for cell based therapies. The primary goal of stem cell research is to produce cells that are both patient specific, as well as having properties suitable for the specific conditions for which they are intended to remedy. From a purely scientific perspective, stem cells allow scientists to gain a deeper understanding of developmental biology and regenerative therapies. Stem cells have acquired a number of uses for applications in regenerative medicine, immunotherapy, gene therapy, but it is in the area of tissue engineering that they generate most excitement, primarily as a result of their capacity for self-renewal and pluripotency. A unique feature of stem cells is their ability to maintain an uncommitted quiescent state in vivo and then, once triggered by conditions such as disease, injury or natural wear or tear, serve as a reservoir and natural support system to replenish lost cells. Although these cells retain the plasticity to differentiate into various tissues, being able to control this differentiation process is still one of the biggest challenges facing stem cell research. In an effort to harness the potential of these cells a number of studies have been conducted using both embryonic/foetal and adult stem cells. The use of embryonic stem cells (ESC) have been hampered by strong ethical and political concerns, this despite their perceived versatility due to their pluripotency. Ethical issues aside, other concerns raised with ESCs relates to the possibility of tumorigenesis, immune rejection and complications with immunosuppressive therapies, all of which adds layers of complications to the application ESC in research and which has led to the search for alternative sources for stem cells. The adult tissues in higher organisms harbours cells, termed adult stem cells, and these cells are reminiscent of unprogrammed stem cells. A number of sources of adult stem cells have been described. Bone marrow is by far the most accessible source of two potent populations of adult stem cells, namely haematopoietic stem cells (HSCs) and bone marrow mesenchymal stem cells (BMSCs). Autologously harvested adult stem cells can, in contrast to embryonic stem cells, readily be used in autografts, since immune rejection is not an issue; and their use in scientific research has not attracted the ethical concerns which have been the case with embryonic stem cells. The major limitation to their use, however, is the fact that adult stem cells are exceedingly rare in most tissues. This fact makes identifying and isolating these cells problematic; bone marrow being perhaps the only notable exception. Unlike the case of HSCs, there are as yet no rigorous criteria for characterizing MSCs. Changing acuity about the pluripotency of MSCs in recent studies has expanded their potential application; however, the underlying molecular pathways which impart the features distinctive to MSCs remain elusive. Furthermore, the sparse in vivo distribution of these cells imposes a clear limitation to their study in vitro. Also, when MSCs are cultured in vitro, there is a loss of the in vivo microenvironment, resulting in a progressive decline in proliferation potential and multipotentiality. This is further exacerbated with increased passage numbers in culture, characterized by the onset of senescence related changes. As a consequence, it is necessary to establish protocols for generating large numbers of MSCs but without affecting their differentiation potential. MSCs are capable of differentiating into mesenchymal tissue lineages, including bone, cartilage, fat, tendon, muscle, and marrow stroma. Recent findings indicate that adult bone marrow may also contain cells that can differentiate into the mature, nonhematopoietic cells of a number of tissues, including cells of the liver, kidney, lung, skin, gastrointestinal tract, and myocytes of heart and skeletal muscle. MSCs can readily be expanded in vitro and can be genetically modified by viral vectors and be induced to differentiate into specific cell lineages by changing the microenvironment–properties which makes these cells ideal vehicles for cellular gene therapy. MSCs can also exert profound immunosuppressive effects via modulation of both cellular and innate immune pathways, and this property allows them to overcome the issue of immune rejection. Despite the many attractive features associated with MSCs, there are still many hurdles to overcome before these cells are readily available for use in clinical applications. The main concern relates to in vivo characterization and identification of MSCs. The lack of a universal biomarker, sparse in vivo distribution, and a steady age related decline in their numbers, makes it an obvious need to decipher the reprogramming pathways and critical molecular players which govern the characteristics unique to MSCs. This book presents a comprehensive insight into the biology of adult stem cells and their utility in current regeneration therapies. The adult stem cell populations reviewed in this book include bone marrow derived MSCs, adipose derived stem cells (ASCs), umbilical cord blood stem cells, and placental stem cells. The features such as MSC circulation and trafficking, neuroprotective properties, and the nurturing roles and differentiation potential of multiple lineages have been discussed in details. In terms of therapeutic applications, the strengths of MSCs have been presented and their roles in disease treatments such as osteoarthritis, Huntington’s disease, periodontal regeneration, and pancreatic islet transplantation have been discussed. An analysis comparing osteoblast differentiation of umbilical cord blood stem cells and MSCs has been reviewed, as has a comparison of human placental stem cells and ASCs, in terms of isolation, identification and therapeutic applications of ASC in bone, cartilage regeneration, as well as myocardial regeneration. It is my sincere hope that this book will update the reader as to the research progress of MSC biology and potential use of these cells in clinical applications. It will be the best reward to all contributors of this book, if their efforts herein may in some way help the readers in any part of their study, research, and career development.

Model-based analysis and optimization of bioreactor for hematopoietic stem cell cultivation

One of the problems to be solved in attaining the full potentials of hematopoietic stem cell (HSC) applications is the limited availability of the cells. Growing HSCs in a bioreactor offers an alternative solution to this problem. Besides, it also offers the advantages of eliminating labour intensive process as well as the possible contamination involved in the periodic nutrient replenishments in the traditional T-flask stem cell cultivation. In spite of this, the optimization of HSC cultivation in a bioreactor has been barely explored. This manuscript discusses the development of a mathematical model to describe the dynamics in nutrient distribution and cell concentration of an ex vivo HSC cultivation in a microchannel perfusion bioreactor. The model was further used to optimize the cultivation by proposing three alternative feeding strategies in order to prevent the occurrence of nutrient limitation in the bioreactor. The evaluation of these strategies, the periodic step change increase in the inlet oxygen concentration, the periodic step change increase in the media inflow, and the feedback control of media inflow, shows that these strategies can successfully improve the cell yield of the bioreactor. In general, the developed model is useful for the design and optimization of bioreactor operation.

Integrated disease management of stem end rot of mango in the Southern Philippines

This research aimed to develop and evaluate pre- and postharvest management strategies to reduce stem end rot (SER) incidence and extend saleable life of 'Carabao' mango fruits in Southern Philippines. Preharvest management focused on the development and improvement of fungicide spray program, while postharvest management aimed to develop alternative interventions aside from hot water treatment (HWT). Field evaluation of systemic fungicides, namely azoxystrobin ( Amistar 25SC), tebuconazole ( Folicur 25WP), carbendazim ( Goldazim 500SC), difenoconazole ( Score 250SC) and azoxystrobin+difenoconazole ( Amistar Top), reduced blossom blight severity and improved fruit setting and retention, resulting in higher fruit yield but failed to sufficiently suppress SER incidence. Based on these findings, an improved fungicide spray program was developed taking into account the infection process of SER pathogens and fungicide resistance. Timely application of protectant (mancozeb) and systemic fungicides (azoxystrobin, carbendazim and difenoconazole) during the most critical stages of mango flower and fruit development ensured higher harvestable fruit yield and minimally lowered SER incidence. Control of SER was also achieved by employing postharvest treatment such as HWT (52-55°C for 10 min), which significantly prolonged the saleable life of mango fruits. However, extended hot water treatment (EHWT; 46°C pulp temperature for 15 min), rapid heat treatment (RHT; 59°C for 30-60 sec), fungicide dip and promising biological control agents failed to satisfactorily reduce SER and prolong saleable life. In contrast, the integration of the improved spray program as preharvest management practice, and postharvest treatments such as HWT and fungicide dips (azoxystrobin, 150-175 ppm; carbendazim, 312.5 ppm; and tebuconazole, 125-156 ppm), significantly reduced disease and extended marketable life for utmost 8 days.

Hematopoietic Stem Cell Development and Transcriptional Regulation

The continuous production of blood cells, a process termed hematopoiesis, is sustained throughout the lifetime of an individual by a relatively small population of cells known as hematopoietic stem cells (HSCs). HSCs are unique cells characterized by their ability to self-renew and give rise to all types of mature blood cells. Given their high proliferative potential, HSCs need to be tightly regulated on the cellular and molecular levels or could otherwise turn malignant. On the other hand, the tight regulatory control of HSC function also translates into difficulties in culturing and expanding HSCs in vitro. In fact, it is currently not possible to maintain or expand HSCs ex vivo without rapid loss of self-renewal. Increased knowledge of the unique features of important HSC niches and of key transcriptional regulatory programs that govern HSC behavior is thus needed. Additional insight in the mechanisms of stem cell formation could enable us to recapitulate the processes of HSC formation and self-renewal/expansion ex vivo with the ultimate goal of creating an unlimited supply of HSCs from e.g. human embryonic stem cells (hESCs) or induced pluripotent stem cells (iPS) to be used in therapy. We thus asked: How are hematopoietic stem cells formed and in what cellular niches does this happen (Papers I, II)? What are the molecular mechanisms that govern hematopoietic stem cell development and differentiation (Papers III, IV)? Importantly, we could show that placenta is a major fetal hematopoietic niche that harbors a large number of HSCs during midgestation (Paper I)(Gekas et al., 2005). In order to address whether the HSCs found in placenta were formed there we utilized the Runx1-LacZ knock-in and Ncx1 knockout mouse models (Paper II). Importantly, we could show that HSCs emerge de novo in the placental vasculature in the absence of circulation (Rhodes et al., 2008). Furthermore, we could identify defined microenvironmental niches within the placenta with distinct roles in hematopoiesis: the large vessels of the chorioallantoic mesenchyme serve as sites of HSC generation whereas the placental labyrinth is a niche supporting HSC expansion (Rhodes et al., 2008). Overall, these studies illustrate the importance of distinct milieus in the emergence and subsequent maturation of HSCs. To ensure proper function of HSCs several regulatory mechanisms are in place. The microenvironment in which HSCs reside provides soluble factors and cell-cell interactions. In the cell-nucleus, these cell-extrinsic cues are interpreted in the context of cell-intrinsic developmental programs which are governed by transcription factors. An essential transcription factor for initiation of hematopoiesis is Scl/Tal1 (stem cell leukemia gene/T-cell acute leukemia gene 1). Loss of Scl results in early embryonic death and total lack of all blood cells, yet deactivation of Scl in the adult does not affect HSC function (Mikkola et al., 2003b. In order to define the temporal window of Scl requirement during fetal hematopoietic development, we deactivated Scl in all hematopoietic lineages shortly after hematopoietic specification in the embryo . Interestingly, maturation, expansion and function of fetal HSCs was unaffected, and, as in the adult, red blood cell and platelet differentiation was impaired (Paper III)(Schlaeger et al., 2005). These findings highlight that, once specified, the hematopoietic fate is stable even in the absence of Scl and is maintained through mechanisms that are distinct from those required for the initial fate choice. As the critical downstream targets of Scl remain unknown, we sought to identify and characterize target genes of Scl (Paper IV). We could identify transcription factor Mef2C (myocyte enhancer factor 2 C) as a novel direct target gene of Scl specifically in the megakaryocyte lineage which largely explains the megakaryocyte defect observed in Scl deficient mice. In addition, we observed an Scl-independent requirement of Mef2C in the B-cell compartment, as loss of Mef2C leads to accelerated B-cell aging (Gekas et al. Submitted). Taken together, these studies identify key extracellular microenvironments and intracellular transcriptional regulators that dictate different stages of HSC development, from emergence to lineage choice to aging.

Novel Alternative Splice Variants of Mouse Cdk5rap2

Autosomal recessive primary microcephaly (MCPH) is a rare neurodevelopmental disorder characterized by a pronounced reduction of brain volume and intellectual disability. A current model for the microcephaly phenotype invokes a stem cell proliferation and differentiation defect, which has moved the disease into the spotlight of stem cell biology and neurodevelopmental science. Homozygous mutations of the Cyclin-dependent kinase-5 regulatory subunit-associated protein 2 gene CDK5RAP2 are one genetic cause of MCPH. To further characterize the pathomechanism underlying MCPH, we generated a conditional Cdk5rap2 LoxP/hCMV Cre mutant mouse. Further analysis, initiated on account of a lack of a microcephaly phenotype in these mutant mice, revealed the presence of previously unknown splice variants of the Cdk5rap2 gene that are at least in part accountable for the lack of microcephaly in the mice.

Análise de viabilidade técnica e econômica da geração de energia através do biogás de lixo em aterros sanitários.

A geração de energia a partir do biogás do lixo em aterros sanitários é uma maneira de produzir energia elétrica renovável e limpa, reduzindo os impactos globais provocados pela queima dos resíduos sólidos urbanos. A contribuição ambiental mais relevante é a redução de emissões dos gases de efeito estufa (GEE), por meio da conversão do metano em dióxido de carbono, visto que o metano possui um potencial de aquecimento global cerca de 21 vezes maior, quando comparado ao dióxido de carbono (através da combustão do mesmo). De acordo com o Mecanismo de Desenvolvimento Limpo (MDL), os países ricos podem comprar créditos de carbono (CERs) dos países em desenvolvimento (que possuam projetos sustentáveis) para cumprir suas metas ambientais. O objetivo é transformar um passivo ambiental (destinação final dos resíduos sólidos urbanos) em um recurso energético, além do estudo da alternativa de obtenção de recursos financeiros através dos CERs. São analisadas as tecnologias de conversão energética (tecnologia de gás de lixo, incineração, entre outras), com a seleção da melhor alternativa para a geração de energia através do biogás de lixo em aterros sanitários. A metodologia utilizada é a recomendada pela Agência de Proteção Ambiental dos Estados Unidos - USEPA (2005). Serão apresentadas outras duas metodologias de cálculo da geração de metano: a do Banco Mundial e a do IPCC (Painel Intergovernamental sobre Mudanças Climáticas). São apresentados estudos comparativos demonstrando quando as turbinas a gás, motores de combustão interna (ciclos Otto ou Diesel) ou outras tecnologias de conversão energética serão viáveis na área técnica e econômica para implantação de Unidades Termoelétricas a biogás. No caso do Aterro de Gramacho, o projeto é viável com a utilização de motores a combustão interna e a obtenção de receitas com a venda da produção de energia e créditos de carbono. Por fim, será apresentada a alternativa do uso do biogás como substituto do gás natural para fins energéticos ou outros fins industriais.

Potencial de energia solar e de geração com sistemas fotovoltaicos conectados à rede em Moçambique.

A energia solar fotovoltaica é uma alternativa de energia limpa que está se tornando uma importante opção para se utilizar eletricidade em regiões desprovidas de energia elétrica. Um estudo das condições energéticas da região através de consulta às concessionárias de energia elétrica (Eletricidade de Moçambique - EDM no caso de Moçambique) e do Ministério de Energia do Governo, são fontes importantes de dados para se conhecer o cenário energético de cada região. Outra fonte de dados importante para se estimar o potencial de geração de energia solar fotovoltaico sãoos indices de irradiação da região. Estes podemser obtidos através do órgão meteorológico da região (INAM no caso de Moçambique) que forneceesses dados para as principais cidades através de suas estações meteorológicas espalhadas na região. Laboratórios de pesquisa internacionais sãooutra importante fonte de dados solarimétricos, pois utilizam satélites capazes de cobrir todo globo terrestre e os dados obtidos são publicados e permitem, a partir das coordenadas do local, identificar a irradiação solar em diversos locais do mundo. Os dados desses laboratórios podem ser comparados com dados de órgãos oficiais da região. Utilizando o banco de dados solarimétricospode-se calcular o potencial fotovoltaico de geração de energia de cada localização e também definir qual região seria mais propícia a utilização de energiasolar fotovoltaica conectada à rede.