Clock Genes, Melanopsins, Melatonin, and Dopamine Key Enzymes and Their Modulation by Light and Glutamate in Chicken Embryonic Retinal Cells

The avian circadian system is composed of the retina, the mammalian homolog region of the suprachiasmatic nucleus (SNC), and the pineal gland. The retina, itself, displays many rhythmic physiological events, such as movements of photoreceptor cells, opsin expression, retinal reisomerization, and melatonin and dopamine production and secretion. Altogether, these rhythmic events are coordinated to predict environmental changes in light conditions during the day, optimizing retina function. The authors investigated the expression pattern of the melanopsin genes Opn4x and Opn4m, the clock genes Clock and Per2, and the genes for the key enzymes N-Acetyltransferase and Tyrosine Hidroxylase in chicken embryo dispersed retinal cells. Primary cultures of chicken retina from 8-day-old embryos were kept in constant dark (DD), in 12-h light/12-h dark (12L:12D), in 12L:12D followed by DD, or in DD in the absence or presence of 100 mu M glutamate for 12 h. Total RNA was extracted throughout a 24-h span, every 3 h starting at zeitgeber time 0 (ZT0) of the 6th day, and submitted to reverse transcriptase-polymerase chain reaction (RT-PCR) followed by quantitative PCR (qPCR) for mRNA quantification. The data showed no rhythmic pattern of transcription for any gene in cells kept in DD. However under a light-dark cycle, Clock, Per2, Opn4m, N-Acetyltransferase, and Tyrosine Hydroxylase exhibited rhythmic patterns of transcription. In DD, 100 mu M glutamate was able to induce rhythmic expression of Clock, strongly inhibited the expression of Tyrosine Hydroxylase, and, only at some ZTs, of Opn4x and Opn4m. The neurotransmitter had no effect on Per2 and N-Acetyltransferase transcription. The authors confirmed the expression of the protein OPN4x by immunocytochemistry. These results suggest that chicken embryonic retinal cells contain a functional circadian clock, whose synchronization requires light-dark cycle or glutamate stimuli. (Author correspondence: amdlcast@ib.usp.br).

A new genus of Ectinosomatidae (Copepoda, Harpacticoida) from sublittoral sediments in Ubatuba, Sao Paulo State (Brazil), an updated key to genera and notes on Noodtiella Wells, 1965

Both sexes of a new genus and species of Ectinosomatidae (Copepoda, Harpacticoida) from sublittoral sediments collected on the inner continental shelf in Ubatuba, Sao Paulo State (Brazil) are described in detail. Chaulionyx gen. n. (type species: C. paivacarvalhoi sp. n.) differs from all known genera in the presence of a conspicuous bifid spine on the prehensile P1 endopod. It can be differentiated from other genera with a prehensile endopod (Halophytophilus Brian, 1919; Bradyellopsis Brian, 1925; Klieosoma Hicks & Schriever, 1985) by the presence of distinctive subrectangular middorsal pores on the urosomites and the unarmed male sixth legs. The genus Lineosoma Wells, 1965 is recognized as a paraphyletic taxon and relegated to a junior subjective synonym of Noodtiella Wells, 1965. Arenosetella pectinata Chappuis, 1954a is removed from its floating position in Ectinosomoides Nicholls, 1945, transferred to the genus Noodtiella as N. pectinata comb. n. and considered the senior subjective synonym of N. toukae Mitwally & Montagna, 2001. Dichotomous keys are provided for the identification of the 18 valid species of Noodtiella and the 21 valid genera of the family Ectinosomatidae. Halophytophilus aberrans Wells & Rao, 1987 is placed species incertae sedis in the family.

New genera of philopotine spider flies (Diptera, Acroceridae) with a key to living and fossil genera

In this paper we describe two new genera of philopotine Acroceridae: Schlingeriella irwini gen. et sp. n. (New Caledonia) and Quasi fisheri gen. et sp. n. (Mexico). The Baltic amber species Eulonchiella eocenica Meunier, 1912 is rediagnosed and a neotype designated based on a newly discovered specimen. We also provide a dichotomous key to the world genera of Philopotinae, both living and fossil.

The Muscini flies of the world (Diptera, Muscidae): identification key and generic diagnoses

Flies of the tribe Muscini (Diptera, Muscidae) are worldwide in distribution and are represented by some 350 species in 18 genera. The present study provides an identification key and diagnoses for all the genera of world Muscini: Biopyrellia Townsend, Curranosia Paterson, Dasyphora Robineau-Desvoidy, Deltotus Seguy, Hennigmyia Peris, Mesembrina Meigen, Mitroplatia Enderlein, Morellia Robineau-Desvoidy, Musca Linnaeus, Myiophaea Enderlein, Neomyia Walker, Neorypellia Pont, Polietes Rondani, Polietina Schnabl & Dziedzicki, Pyrellia Robineau-Desvoidy, Pyrellina Malloch, Sarcopromusca Townsend, Ziminellia Nihei & de Carvalho. Most infrageneric taxa are also represented, namely, the sub-genera of Dasyphora and Morellia. Comments on phylogeny support (whenever pertinent) and the major references containing revisions and regional identification keys to species are provided for each genus and subgenus.

Mannose-binding lectin complement pathway plays a key role in complement activation by Paracoccidioides brasiliensis

Paracoccidioides brasiliensis (Pb) is a dimorphic fungal pathogen that causes paracoccidioidomycosis the most severe deep mycosis from South America Although cell mediated immunity is considered the most efficient protective mechanism against Pb infection mechanisms of innate immunity are poorly defined Herein we investigated the interaction of the complement system with high and low virulence isolates of Pb We demonstrated that Pb18 a high virulence Pb Isolate when incubated with normal human serum (NHS) induces consumption of hemolytic complement and when immobilized promotes binding of C4b C3b and C5b-C9 Both low virulence (Pb265) and high virulence (Pb18) isolates consumed C4 C3 and mannose-binding learn (MBL) of MBL-sufficient but not of MBL-deficient serum as revealed by deposition of residual C4 C3 and MBL on immune complexes and mannan However higher complement components consumption was observed with Pb265 as compared with Pb18 The suggested relationship between low virulence and significant complement activation properties of Pb isolates was confirmed by the demonstration that virulence attenuation of Pb 18 results in acquisition of the ability to activate complement Conversely reactivation of attenuated Pb18 results in loss of the ability to activate complement Our results demonstrate for the first time that Pb yeasts activate the complement system by the lectin pathway and there is an Inverse correlation between complement activating ability and Pb virulence These differences could exert an influence on Innate immunity and severity of the disease developed by infected hosts (C) 2010 Elsevier Ltd All rights reserved

The Key Role of the School Principal’s Leadership in the Schooling of Newly Arrived Immigrant Pupils in Sweden : Experiences from an Interactive Action Research Project

When newly immigrated children and young people begin school in Sweden, certain challengesarise. These may result from weak Swedish-language skills and different schooling backgrounds,as well as organizational and pedagogical limitations in the schools. This generates demands onschool leaders to lead and develop the organization and teachers competences to meet these pupils’needs. This situation was behind the initiation of the project “New Immigrants and Learning—Competence Development for Teachers and School Principals.” The project ran in schools infour Swedish municipalities, its aim was to develop leadership, organizational and pedagogicalskills that would facilitate the schooling and integration of newly arrived pupils. This article aimsto describe and discuss a Participant Action Research (PAR) based on a think tank and researchcircles, drawing special attention to the role of the school leaders. It will also examine whether theresearch circles and the project overall served to develop educational and intercultural leadership,organizational conditions, collegial learning, pedagogical methods and competence in terms ofschooling for this pupil group.

Winning the Accreditation Game: The Library as Key Player and Hidden Gold Mine

Libraries are a central hub of information resources supporting college and university curricula. Several library strategies, cross-campus collaborations, and philosophical considerations of electronic and print offerings led to successful accreditations in business and engineering programs.

National development strategy: the key growth institution

O Crescimento Econômico é Quase que Invariavelmente o Resultado de uma Estratégia Nacional De Desenvolvimento. o Efetivo Desenvolvimento Econômico Ocorre Historicamente Quando a Nação é Uma Nação Forte, e as Diferentes Classes Sociais são Capazes de Cooperar e Formular uma Efetiva Estratégia Para Promover o Crescimento e Enfrentar a Competição Internacional. uma Estratégia Nacional De Desenvolvimento é Essencialmente uma Instituição ou um Conjunto de Instituições que Estimulam A Acumulação de Capital e o Progresso Técnico. Segue uma Discussão das Principais Características de Tais Estratégias. o Artigo Finaliza com uma Análise dos Conflitos ou Tensões Envolvidos nas Estratégias Nacionais de Desenvolvimento.

Green logistics: key elements for a well-functioning sustainable logistics system

A growing awareness of the modern society about the direct relationship between a growing global community with increasing total industrial activities on one hand and various environmental problems and a natural limitation of natural resources on the other hand set the base for sustainable or “green” approaches within the supply chain. This paper therefore will look at the issue of “Green Logistics” which seeks to reduce the environmental impact of logistics activities by taking into account functions such as recycling, waste and carbon emission reduction and the use of alternative sources of energy. In order to analyze how these approaches and ideas are being perceived by the system as a whole two models from the area of prospective and scenario planning are being used and described to identify the main drivers and tendencies within the system in order to create feasible hypothesis. Using the URCA/CHIVAS model allows us to identify the driver variables out of a high number of variables that best describe the system “Green Logistics”. Followed by the analysis of the actor’s strategies in the system with the Mactor model it is possible to reduce the complexity of a completely holistic system to a few key drivers that can be analyzed further on. Here the implications of URCA/CHIVAS and Mactor are being used to formulate hypotheses about the perception of Green Logistics and its successful implementation among logistics decision makers by an online survey. This research seeks to demonstrate the usefulness of scenario planning to a highly complex system observing it from all angles and extracting information about the relevant factors of it. The results of this demonstration indicate that there are drivers much beyond the factory walls that need to be considered when implementing successfully a system such as Green Logistics.

Judicial system performance and economic development

The role of judicial systems in determining economic perfonnance has gained increasingly attention in recent years. Nonetheless, the literature lacks a clearly articulated framework to examine how judicial systems influence the investment and production decisions of economic agents. This paper tries to till in this gap. It examines what constitutes a well-functioning judiciary, analyzes how dysfunctional judicial systems compromise economic growth, and reviews the relevant empirical literature. It concludes with some remarks about why, despite the widespread perception that well-functioning legal and judicial systems are key to the success of market-oriented reforms in developing and transition countries, judicial refonn has lagged so much behind other reforms.

Understanding e-government technology transfer: evidence from the transfer of a patent management system from EPO to INPI

Drawing upon Brazilian experience, this research explores some of the key issues to be addressed in using e-government technical cooperation designed to enhance service provision of Patent Offices in developing countries. While the development of software applications is often seen merely as a technical engineering exercise, localization and adaptation are context bounded matters that are characterized by many entanglements of human and non-humans. In this work, technical, legal and policy implications of technical cooperation are also discussed in a complex and dynamic implementation environment characterized by the influence of powerful hidden agendas associated with the arena of intellectual property (IP), which are shaped by recent technological, economic and social developments in our current knowledge-based economy. This research employs two different theoretical lenses to examine the same case, which consists of transfer of a Patent Management System (PMS) from the European Patent Office (EPO) to the Brazilian Patent Office that is locally named ‘Instituto Nacional da Propriedade Industrial’ (INPI). Fundamentally, we have opted for a multi-paper thesis comprising an introduction, three scientific articles and a concluding chapter that discusses and compares the insights obtained from each article. The first article is dedicated to present an extensive literature review on e-government and technology transfer. This review allowed the proposition on an integrative meta-model of e-government technology transfer, which is named E-government Transfer Model (ETM). Subsequently, in the second article, we present Actor-Network Theory (ANT) as a framework for understanding the processes of transferring e-government technologies from Patent Offices in developed countries to Patent Offices in developing countries. Overall, ANT is seen as having a potentially wide area of application and being a promising theoretical vehicle in IS research to carry out a social analysis of messy and heterogeneous processes that drive technical change. Drawing particularly on the works of Bruno Latour, Michel Callon and John Law, this work applies this theory to a longitudinal study of the management information systems supporting the Brazilian Patent Office restructuration plan that involved the implementation of a European Patent Management System in Brazil. Based upon the ANT elements, we follow the actors to identify and understand patterns of group formation associated with the technical cooperation between the Brazilian Patent Office (INPI) and the European Patent Office (EPO). Therefore, this research explores the intricate relationships and interactions between human and non-human actors in their attempts to construct various network alliances, thereby demonstrating that technologies embodies compromise. Finally, the third article applies ETM model as a heuristic frame to examine the same case previously studied from an ANT perspective. We have found evidence that ETM has strong heuristic qualities that can guide practitioners who are engaged in the transfer of e-government systems from developed to developing countries. The successful implementation of e-government projects in developing countries is important to stimulate economic growth and, as a result, we need to understand the processes through which such projects are being implemented and succeed. Here, we attempt to improve understanding on the development and stabilization of a complex social-technical system in the arena of intellectual property. Our preliminary findings suggest that e-government technology transfer is an inherently political process and that successful outcomes require continuous incremental actions and improvisations to address the ongoing issues as they emerge.

Mitigating high ‘equity capital’ risk exposure to ‘small cap’ sector in India: analysing ‘key factors of success’ for ‘Institutional Investors’ whilst Investing in small cap sector in India

This paper deals with the subject of mitigating high ‘Equity Capital’ Risk Exposure to ‘Small Cap’ Sector in India. Institutional investors in India are prone to be risk averse when it comes to investing in the small cap sector in India as they find the companies risky and volatile. This paper will help analyse ‘Key Factors of success’ for ‘Institutional Investors’ whilst investing in Small Cap sector in India as some of these Indian small cap stocks offer handsome returns despite economic downturn. This paper has been harnessed carefully under the influence of expert investors, which includes Benjamin Graham (Security Analysis); Warren Buffet; Philip Fisher (Common Stocks and Uncommon Profits); and Aswath Damodaran.

The Limits of political compromise: debt ceilings and political competition

We study the desirability of limits on the public debt and of political competition in an economy where political parties alternate in office. Due to rent-seeking motives, incumbents have an incentive to set public expenditures above the socially optimal level. Parties cannot commit to future policies, but they can forge a political compromise where each party curbs excessive spending when in office if it expects future governments to do the same. In contrast to the received literature, we find that strict limits on government borrowing can exacerbate political-economy distortions by rendering a political compromise unsustainable. This tends to happen when political competition is limited. Conversely, a tight limit on the public debt fosters a compromise that yields the efficient outcome when political competition is vigorous, saving the economy from immiseration. Our analysis thus suggests a legislative tradeoff between restricting political competition and constraining the ability of governments to issue debt.

Functionalization of dendrimers for improved gene delivery to mesenchymal stem cell

Disease, injury, and age problems compromise human quality of life and continuously motivate the search for new and more efficacious therapeutic approaches. The field of Tissue Regeneration and Engineering has greatly evolved over the last years, mainly due to the combination of the important advances verified in Biomaterials Science and Engineering with those of Cell and Molecular Biology. In particular, a new and promising area arose – Nanomedicine – that takes advantage of the extremely small size and especial chemical and physical properties of Nanomaterials, offering powerful tools for health improvement. Research on Stem Cells, the self-renewing progenitors of body tissues, is also challenging to the medical and scientific communities, being expectable the appearance of new and exciting stem cell-based therapies in the next years. The control of cell behavior (namely, of cell proliferation and differentiation) is of key importance in devising strategies for Tissue Regeneration and Engineering. Cytokines, growth factors, transcription factors and other signaling molecules, most of them proteins, have been identified and found to regulate and support tissue development and regeneration. However, the application of these molecules in long-term regenerative processes requires their continuous presence at high concentrations as they usually present short half-lives at physiological conditions and may be rapidly cleared from the body. Alternatively, genes encoding such proteins can be introduced inside cells and be expressed using cell’s machinery, allowing an extended and more sustained production of the protein of interest (gene therapy). Genetic engineering of stem cells is particularly attractive because of their self-renewal capability and differentiation potential. For Tissue Regeneration and Engineering purposes, the patient’s own stem cells can be genetically engineered in vitro and, after, introduced in the body (with or without a scaffold) where they will not only modulate the behavior of native cells (stem cell-mediated gene therapy), but also directly participate in tissue repair. Cells can be genetically engineered using viral and non-viral systems. Viruses, as a result of millions of years of evolution, are very effective for the delivery of genes in several types of cells, including cells from primary sources. However, the risks associated with their use (like infection and immunogenic reactions) are driving the search for non-viral systems that will efficiently deliver genetic material into cells. Among them, chemical methods that are promising and being investigated use cationic molecules as carriers for DNA. In this case, gene delivery and gene expression level remain relatively low when primary cells are used. The main goal of this thesis was to develop and assess the in vitro potential of polyamidoamine (PAMAM) dendrimers based carriers to deliver genes to mesenchymal stem cells (MSCs). PAMAM dendrimers are monodispersive, hyperbranched and nanospherical molecules presenting unique characteristics that make them very attractive vehicles for both drug and gene delivery. Although they have been explored for gene delivery in a wide range of cell lines, the interaction and the usefulness of these molecules in the delivery of genes to MSCs remains a field to be explored. Adult MSCs were chosen for the studies due to their potential biomedical applications (they are considered multipotent cells) and because they present several advantages over embryonic stem cells, such as easy accessibility and the inexistence of ethical restrictions to their use. This thesis is divided in 5 interconnected chapters. Chapter I provides an overview of the current literature concerning the various non-viral systems investigated for gene delivery in MSCs. Attention is devoted to physical methods, as well as to chemical methods that make use of polymers (natural and synthetic), liposomes, and inorganic nanoparticles as gene delivery vectors. Also, it summarizes the current applications of genetically engineered mesenchymal stem cells using non-viral systems in regenerative medicine, with special focus on bone tissue regeneration. In Chapter II, the potential of native PAMAM dendrimers with amine termini to transfect MSCs is evaluated. The level of transfection achieved with the dendrimers is, in a first step, studied using a plasmid DNA (pDNA) encoding for the β-galactosidase reporter gene. The effect of dendrimer’s generation, cell passage number, and N:P ratio (where N= number of primary amines in the dendrimer; P= number of phosphate groups in the pDNA backbone) on the level of transfection is evaluated, being the values always very low. In a second step, a pDNA encoding for bone morphogenetic protein-2, a protein that is known for its role in MSCs proliferation and differentiation, is used. The BMP-2 content produced by transfected cells is evaluated by an ELISA assay and its effect on the osteogenic markers is analyzed through several classical assays including alkaline phosphatase activity (an early marker of osteogenesis), osteocalcin production, calcium deposition and mineralized nodules formation (late osteogenesis markers). Results show that a low transfection level is enough to induce in vitro osteogenic differentiation in MSCs. Next, from Chapter III to Chapter V, studies are shown where several strategies are adopted to change the interaction of PAMAM dendrimers with MSCs cell membrane and, as a consequence, to enhance the levels of gene delivery. In Chapter III, generations 5 and 6 of PAMAM dendrimers are surface functionalized with arginine-glycine-aspartic acid (RGD) containing peptides – experiments with dendrimers conjugated to 4, 8 and 16 RGD units were performed. The underlying concept is that by including the RGD integrin-binding motif in the design of the vectors and by forming RGD clusters, the level of transfection will increase as MSCs highly express integrins at their surface. Results show that cellular uptake of functionalized dendrimers and gene expression is enhanced in comparison with the native dendrimers. Furthermore, gene expression is dependent on both the electrostatic interaction established between the dendrimer moiety and the cell surface and the nanocluster RGD density. In Chapter IV, a new family of gene delivery vectors is synthesized consisting of a PAMAM dendrimer (generation 5) core randomly linked at the periphery to alkyl hydrophobic chains that vary in length and number. Herein, the idea is to take advantage of both the cationic nature of the dendrimer and the capacity of lipids to interact with biological membranes. These new vectors show a remarkable capacity for internalizing pDNA, being this effect positively correlated with the –CH2– content present in the hydrophobic corona. Gene expression is also greatly enhanced using the new vectors but, in this case, the higher efficiency is shown by the vectors containing the smallest hydrophobic chains. Finally, chapter V reports the synthesis, characterization and evaluation of novel gene delivery vectors based on PAMAM dendrimers (generation 5) conjugated to peptides with high affinity for MSCs membrane binding - for comparison, experiments are also done with a peptide with low affinity binding properties. These systems present low cytotoxicity and transfection efficiencies superior to those of native dendrimers and partially degraded dendrimers (Superfect®, a commercial product). Furthermore, with this biomimetic approach, the process of gene delivery is shown to be cell surface receptor-mediated. Overall, results show the potential of PAMAM dendrimers to be used, as such or modified, in Tissue Regeneration and Engineering. To our knowledge, this is the first time that PAMAM dendrimers are studied as gene delivery vehicles in this context and using, as target, a cell type with clinical relevancy. It is shown that the cationic nature of PAMAM dendrimers with amine termini can be synergistically combined with surface engineering approaches, which will ultimately result in suitable interactions with the cytoplasmic membrane and enhanced pDNA cellular entry and gene expression. Nevertheless, the quantity of pDNA detected inside cell nucleus is always very small when compared with the bigger amount reaching cytoplasm (accumulation of pDNA is evident in the perinuclear region), suggesting that the main barrier to transfection is the nuclear membrane. Future work can then be envisaged based on the versatility of these systems as biomedical molecular materials, such as the conjugation of PAMAM dendrimers to molecules able to bind nuclear membrane receptors and to promote nuclear translocation.