Pulsatile Viscous Flows in Elliptical Vessels and Annuli: Solution to the Inverse Problem, with Application to Blood and Cerebrospinal Fluid Flow

We consider the fully-developed flow of an incompressible Newtonian fluid in a cylindrical vessel with elliptical cross-section and in an annulus between two confocal ellipses. Since flow rate is the main physical quantity which can be actually be derived from measurements, we address the extit {inverse problem} to compute the velocity field associated with a given, time-periodic flow rate. We propose a novel numerical strategy, which is nonetheless grounded on several analytical relations and which leads to the solution of systems of ordinary differential equations. Our method holds romise to be more amenable to implementation than previous ones based on challenging computation of Mathieu functions. We report numerical results based on measured data for human blood flow in the internal carotid artery, and cerebrospinal fluid flow in the upper cervical region of the human spine. Computational efficiency is shown, but the main goal of the present study is to provide an improved source of initial/boundary data, as well as a benchmark solution for pulsatile flows in elliptical sections and the proposed method has potential applications to bio-fluid dynamics investigations (e.g. to address aspects of relevant diseases), to biomedical applications (e.g. targeted drug delivery and energy harvesting for implantable devices), up to longer-term medical microrobotics applications

ISOLATION AND CHARACTERIZATION OF THE GM3 SYNTHASE cDNA FROM HUMAN PLACENTA

It is known that gangliosides have various important biological functions, and their functions as well as their biosynthesis are currently clarified (1, 2). In vertebrates, almost all the ganglio-series gangliosides are synthesized from a common precursor, ganglioside GM3, which has the simplest structure among the major gangliosides. GM3 itself is known to participate in induction of differentiation, modulation of proliferation, signal transduction and integrin-mediated cell adhesion. GM3 synthase (EC 2.4.99.9, ST3Gal V) is the enzyme involved in the last step of GM3 biosynthesis: it catalyses the transfer of a sialic acid moiety from CMP-sialic acid onto lactosylceramide, forming an a2-3 linkage. Whereas GM3 is ubiquitously distributed in the plasma membranes of all eukaryotic cells, GM3 synthase results expressed in a tissue specific manner, especially in brain, placenta, muscle and testis (3). Many important issues, such as human cDNA identification and characterization, genomic structure and regulation of gene expression, are still open. To isolate the coding sequence of the gene of GM3 synthase from human placenta we used the 5\u2019- and 3\u2019-Rapid Amplification of cDNA Ends technology (SMART RACE cDNA Amplification Kit, Clontech) using, as specific primers, oligonucleotides derived from the human GM3 synthase cDNA sequence from differentiated HL60 cells (3). The different PCR products were cloned into the pCR2.1 vector (TA Cloning Kit, InVitrogen) and the nucleotide sequence was determined. A cDNA, showing high sequence homology with that encoding the human GM3 synthase from TPA-differentiated HL60 cells (3), has been successfully isolated and cloned from human placenta. The major difference between these two cDNAs is in the 5\u2019-end, according to the existence of different promoter regions, responsible for tissue-specific expression of the gene. Furthermore, the cDNA from the human placenta contains, upstream and in frame with the ATG indicated as translation initiation site for the GM3 synthase of HL60 cells, another ATG codon inserted in a sequence compatible with Kozak\u2019s rule, suggesting that the protein of the human placenta has an additional portion in NH2-terminus. The complete coding region of the human placenta cDNA is going to be cloned in an expression vector, under the control of the CMV promoter, in order to evaluate its activity. On the other hand, in vitro translation experiments are going to be carried out to define the first start codon. 1) Hakomori S.I. (2000): Glycoconj. J. 17, 627-647 2) Kolter T. et al. (2002): J.Biol.Chem. 277, 25859-25862 3) Ishii A. et al. (1998): J.B.C. 273, 31652-3165

CLONING OF THE GM3 SYNTHASE cDNA FROM HUMAN PLACENTA AND GENOMIC ORGANISATION OF THE GENE

INTRODUCTION: Gangliosides are a large family of sialic acid-containing glycosphingolipids that play important roles in a large variety of biological processes. Both their functions and their biosynthetic pathways are currently clarified (1, 2). In vertebrates, almost all the ganglio-series gangliosides are synthesized from a common precursor, ganglioside GM3, which has the simplest structure among the major ganglioside. GM3 itself is known to participate in induction of differentiation, modulation of proliferation, signal transduction and integrin-mediated cell adhesion. GM3 synthase (EC 2.4.99.9, ST3Gal V) is the enzyme involved in the last step of GM3 biosynthesis: it catalyses the transfer of a sialic acid moiety from CMP-sialic acid onto lactosylceramide, forming an a2-3 linkage. Whereas GM3 is ubiquitously distributed in the plasma membranes of all eukaryotic cells, GM3 synthase results expressed in a tissue-specific manner, especially in brain, placenta, muscle and testis (3). Although its cDNA has been cloned from some mouse (4, 5) and human tissues (3, 6), studies on the genomic structure (7, 8) and on its transcriptional regulation (8, 9) provides contrasting results. MATERIALS AND METHODS: To isolate the complete coding sequence of the gene of GM3 synthase from human placenta we used the 5\u2019- and 3\u2019-Rapid Amplification of cDNA Ends technology (SMART RACE cDNA Amplification Kit, Clontech) using, as specific primers, oligonucleotides derived from the human GM3 synthase cDNA sequence from differentiated HL60 cells (3). The identity of some amplified DNA fragments was confirmed by Southern blot analysis (Gene ImagesTM AlkPhos DirectTM labelling and detection system, Amersham Pharmacia Biotech). The different PCR products were cloned into the pCR2.1 vector (TA Cloning Kit, InVitrogen) and the nucleotide sequence was determined (\u201cProgetto Camilla\u201d, M-Medical). The genomic structure of the human GM3 synthase gene has been determined through a human genome BLAST homology search of the public database (GenBank) using the GM3 synthase cDNA from human placenta as the query sequence. RESULTS: A cDNA, consisting of 2149 bp and showing high sequence homology with those encoding the human GM3 synthase from other human tissues (3, 6), has been successfully isolated and cloned from human placenta. Notwithstanding our approach, our cDNA has not the poli(A) tail. Between our cDNA and the other published ones, the major difference is in the 5\u2019-end, according to the existence of different promoter regions, responsible for tissue-specific expression of the gene. Furthermore, the cDNA from the human placenta contains, upstream and in frame with the ATG indicated as translation initiation site for the GM3 synthase of HL60 cells, another ATG codon inserted in a sequence compatible with Kozak\u2019s rule, suggesting that the protein of the human placenta could have an additional portion in NH2-terminus. The complete and partial coding regions of the human placenta cDNA are going to be cloned in an expression vector, under the control of the CMV promoter, in order to evaluate their GM3 synthase activity. The results of the human genome BLAST homology search of the public database using the GM3 synthase cDNA from human placenta as the query sequence showed that the gene consists of seven exons which span over 28.5 kb, with exons ranging in size up to 1242 bp. All exon-intron boundaries follows the GT-AG rule (10). 1) Hakomori S.I. (2000) Glycoconj. J. 17, 627-647 2) Kolter T. et al. (2002) J. Biol. Chem. 277, 25859-25862 3) Ishii A. et al. (1998) J. Biol. Chem. 273, 31652-31655 4) Kono M. et al. (1998) Biochem. Biophys. Res. Comm. 253, 170-175 5) Fukumoto S. et al. (1999) J. Biol. Chem. 274, 9271-9276 6) Kapitonov D. et al.(1999) Glycoconj J. 16, 337-350 7) Kim K.W. et al. (2001) Gene 273, 163-171 8) Kim S.W. et al. (2002) ) Biochim. Biophys. Acta 1578, 84-89 9) Zeng G. et al. (2003) Biochim. Biophys. Acta 1625, 30-35 10) Breathnach R. and Chambon P. (1981) Annu. Rev. Biochem. 50, 349-38

GM3 IN THE REGULATION OF THE EXPRESSION AND ACTIVATION OF ErbB-2/EGFR HETERODIMERS

Gangliosides are a large and heterogeneous family of sialic acid containing glycosphingolipids, ubiquitous components of all eukaryotic cell membranes. They can partially segregate, together with cholesterol and specific signaling transduction proteins, such as receptor tyrosine-kinases, into unique more or less stable clusters or microdomains, indicated as \u201cglycolipid-enriched domains\u201d, \u201clipid rafts\u201d, \u201ccaveolae\u201d, contributing to the membrane structure, organization and function (1). Gangliosides are well-characterized modulators of receptor tyrosine-kinase (RTKs) phophorylation and dimerization (2). Our interest is particularly directed to study the relationship between gangliosides and two members of the tyrosine kinase ErbB receptor family: the epidermal growth factor receptor, EGFR or ErbB-1, and the structurally related protein ErbB-2. Unlike EGFR, ErbB-2 is a ligandless receptor: it can be activated by heterodimerization and cross-phosphorylation by other ligand-activated ErbB receptors (3,4). Our previous experimental evidence supports the functional relationship between ErbB-2 and gangliosides, in particular GM3 (5,6). In the present study, using the HC11 mouse mammary epithelial cell line, we investigated the ErbB-2 activation state and its tendency to form stable molecular complexes with EGFR and with ganglioside GM3, before and after EGF stimulation, by co-immunoprecipitation experiments with anti-ErbB-2 antibody and Western blot analyses. As HC11 cells express different ganglioside species, the exclusive association of GM3 with ErbB-2 and EGFR was ascertained by high performance-thin layer chromatography (HP-TLC) and TLC-immunostaining analyses of gangliosides extracted from the immunoprecipitates. Results display that in EGF-stimulated HC11 cells stable and tyrosine-phosphorylated ErbB2/EGFR dimers are formed and that GM3 is the unique ganglioside tightly associated to ErbB-2/EGFR dimers and to EGFR monomers, but not to ErbB2 monomers. In cells not stimulated with EGF a spontaneous but unproductive dimerization of ErbB2 and EGFR takes place and no ganglioside is found associated to the receptors. These observations indicate that the modulation of ErbB2 activation by GM3 may be mediated by EGFR, but that EGF stimulation is indispensable. After ganglioside depletion by [D]-PDMP, phosphorylated EGFR monomers are observed both before and after EGF stimulation, whereas ErbB-2 is present in the monomeric and unphosphorylated state even after EGF stimulation, suggesting that GM3 might have a bivalent key role in modulating the activation of ErbB-2 and EGFR. References 1. Fivaz, M., Abrami, L., Van Der Goot. F.G. Trends Cell Biol. 9(6), 212-213 (1999); 2. Bremer, E.G., Current topics in membranes 40, 387-411(1999); 3. Qian, X., LeVea, C.M., Freeman, J.K., Dougall, W.C., Greene, M.I., Proc. Natl. Acad. Sci. USA 91, 1500-1504 (1994); 4. Gulliford, T.J., Huang, G.C., Ouyang, X., Epstein, R.J., Oncogene 15, 2219-2223 (1997); 5. Sottocornola E., Berra B., and Colombo I., Biochim. Biophys. Acta-Molecolar and cell Biology of Lipids, 1635, 55-66 (2003); 6. Sottocornola E., Misasi R., Mattei V., Ciarlo L., Gradini R., Garofalo T., Berra B., Colombo I., and Sorice M., FEBS J. 273, 1821-1830 (2006)

Descrição de sistemas de criação tradicionais de ovinos da Nhecolândia, Pantanal, MS.

Este trabalho tem como objetivo principal resgatar o conhecimento empírico sobre o sistema de criação tradicional de ovinos no Pantanal, caracterizando-o e descrevendo-o a partir de oito fazendas da sub-região da Nhecolândia, Pantanal, MS. Devido a crescente demanda do mercado por carne ovina, os produtores do Pantanal podem diversificar a produção animal. A região apresenta potencial para a produção de ovelhas pantaneiras para cruzamentos no planalto, produção de cordeiros (orgânicos), produção de subsistência da fazenda, entre outros. Para tanto, há a necessidade de desenvolver práticas sustentáveis de manejo da ovinocultura no Pantanalbitstream/item/80589/1/CT94.pd

Identificação preliminar de grupos funcionais em pastagens nativas no Pantanal.

As pastagens nativas do Pantanal são dinâmicas, principalmente devido às inundações periódicas. O conhecimento sobre os grupos funcionais dos estados de transição dessas comunidades é de extrema importância no manejo para resiliência do ecossistema. Este artigo objetivou identificar grupos funcionais de plantas em uma borda de baía, influenciada por inundação e superpastejo, de bovinos ao longo do tempo. O trabalho foi desenvolvido na borda de uma lagoa superpastejada na fazenda Nhumirim, sub-região da Nhecolândia, Pantanal, no período de setembro 2007 a março 2010

Therapeutic Peptides and Proteins: Stabilization Challenges and Biomedical Applications by Means of Nanodelivery Systems

The delivery of peptides and proteins usually faces formulation development challenges attributed to the difficulties encountered in their stabilization. Nanoparticles offer an alternative to improve the physicochemical stability of such biomacromolecules, while increasing their bioavailability by overcoming biological absorption barriers. With this review, we aim to discuss the stability problems of proteins and peptides that have driven the scientific community to find in nanotechnology a valid alternative for oral administration of biomolecules. In addition, we describe the most commonly used nanoparticles for this purpose (e.g., polymers such as polylactic acid, poly(lactic-co-glycolic acid), polycaprolactone, modified chitosan, and lipids such as oil-in-water nanoemulsions, self-emulsified drug delivery systems, solid lipid nanoparticles, nanostructured lipid carriers, liposomes, as well as hybrid systems like micelles), and we show some of the most important recent applications of these nanoparticles for the delivery of proteins and peptides, including for the treatment of diabetes, viruses (such as HIV), cancer, as well as in the development of vaccines

When Does Eddy Viscosity Damp Subfilter Scales Sufficiently?

Large eddy simulation (LES) seeks to predict the dynamics of spatially filtered turbulent flows. The very essence is that the LES-solution contains only scales of size ≥Δ, where Δ denotes some user-chosen length scale. This property enables us to perform a LES when it is not feasible to compute the full, turbulent solution of the Navier-Stokes equations. Therefore, in case the large eddy simulation is based on an eddy viscosity model we determine the eddy viscosity such that any scales of size <Δ are dynamically insignificant. In this paper, we address the following two questions: how much eddy diffusion is needed to (a) balance the production of scales of size smaller than Δ; and (b) damp any disturbances having a scale of size smaller than Δ initially. From this we deduce that the eddy viscosity νe has to depend on the invariants q = ½tr(S^2) and r =−⅓tr(S^3) of the (filtered) strain rate tensor S. The simplest model is then given by νe = 3/2(Δ/π)^2|r|/q. This model is successfully tested for a turbulent channel flow (Reτ = 590).

Managing Pantanal rangelands for optimizing carbon flow: effects of growing season and pasture type on dry mass accumulation.

This work aimed to assess the forage mass accumulation in the main grazing sites in two pastures systems to identify resting decisions to maximize the carbon flow in the system.(Embrapa Gado de Corte. Documentos, 216). Coordenador Roberto Giolo de Almeida. II SIGEE. Disponível em: . Acesso em: 30 nov. 2016

Estilosantes Campo Grande como alternativa para recuperação e enriquecimento de pastagens nativas degradadas do Pantanal.

Este estudo objetivou avaliar a consorciação da gramínea nativa Mesosetum chaseae com estilosantes Campo Grande (ECG) para recuperação de pastagens degradadas no Pantanal. O experimento foi conduzido na sub-região da Nhecolândia, Pantanal, MS, em área de campo com sinais evidentes de degradação. O delineamento foi inteiramente casualizado com três repetições. O plantio de M. chaseae foi feito por mudas e ECG por sementes, no início das chuvas, em novembro de 2014. No final do período chuvoso foram feitas avaliações do valor nutritivo das pastagens e constatado o enriquecimento com a Introdução do ECG. Os teores médios de proteína bruta (%) e NDT (%) para M. chaseae e ECG foram de 10,9 e 57,2 e 5,3 e 52,3, respectivamente. A cobertura vegetal evoluiu de 75% para quase 100% no segundo ano, destacando algumas espécies forrageiras do banco natural de sementes e problemas no estabelecimento de ECG, provavelmente devido a inundação na área, reconhecendo a importância do manejo adaptativo.Edição dos anais do VI Congresso Latino-Americano de Agroecologia (CLAA), X Congresso Brasileiro de Agroecologia (CBA), V Seminário de Agroecologia do Distrito Federal e Entorno (SEMDF), set. 2017, Brasília, DF