BIOMATERIAIS: TIPOS, APLICAÇÕES E MERCADO

AbstractThe types of compounds used in the production of biomaterials, namely metals, ceramics, synthetic and natural polymers, as well as composite materials, are discussed in the present work, together with details of their application and evolution from biocompatible to bioactive, biodegradable, and biomimetic clinical products. The chemical structure, the three-dimensional structure, and the molecular organization of compounds frequently used in the manufacture of relevant classes of biomaterials are discussed, along with their advantages and some of their major limitations in specific clinical applications. The main chemical, physical, mechanical, and biological requirements of biomaterials categories are presented, as well as typical tissular responses to implanted biomaterials. Reasons for the recent economic growth of the biomaterials market segment are addressed, and the most successful biomaterial categories are discussed, emphasizing areas such as orthopedic and cardiovascular implants, regenerative medicine, tissue engineering, and controlled drug release devices. Finally, the need for the development of innovative and more accessible biomaterials, due to the expected increase in the number of elderly people and the growing trend of personalized medical procedures, is pointed out.

Structural studies on enzymes of biotechnological and biomedical interest

Structural studies of proteins aim at elucidating the atomic details of molecular interactions in biological processes of living organisms. These studies are particularly important in understanding structure, function and evolution of proteins and in defining their roles in complex biological settings. Furthermore, structural studies can be used for the development of novel properties in biomolecules of environmental, industrial and medical importance. X-ray crystallography is an invaluable tool to obtain accurate and precise information about the structure of proteins at the atomic level. Glutathione transferases (GSTs) are amongst the most versatile enzymes in nature. They are able to catalyze a wide variety of conjugation reactions between glutathione (GSH) and non-polar components containing an electrophilic carbon, nitrogen or sulphur atom. Plant GSTs from the Tau class (a poorly characterized class) play an important role in the detoxification of xenobiotics and stress tolerance. Structural studies were performed on a Tau class fluorodifen-inducible glutathione transferase from Glycine max (GmGSTU4-4) complexed with GSH (2.7 Å) and a product analogue Nb-GSH (1.7 Å). The three-dimensional structure of the GmGSTU4-4-GSH complex revealed that GSH binds in different conformations in the two subunits of the dimer: in an ionized form in one subunit and a non-ionized form in the second subunit. Only the ionized form of the substrate may lead to the formation of a catalytically competent complex. Structural comparison between the GSH and Nb-GSH bound complexes revealed significant differences with respect to the hydrogen-bonding, electrostatic interaction pattern, the upper part of -helix H4 and the C-terminus of the enzyme. These differences indicate an intrasubunit modulation between the G-and Hsites suggesting an induced-fit mechanism of xenobiotic substrate binding. A novel binding site on the surface of the enzyme was also revealed. Bacterial type-II L-asparaginases are used in the treatment of haematopoietic diseases such as acute lymphoblastic leukaemia (ALL) and lymphomas due to their ability to catalyze the conversion of L-asparagine to L-aspartate and ammonia. Escherichia coli and Erwinia chrysanthemi asparaginases are employed for the treatment of ALL for over 30 years. However, serious side-effects affecting the liver and pancreas have been observed due to the intrinsic glutaminase activity of the administered enzymes. Structural studies on Helicobacter pylori L-asparaginase (HpA) were carried out in an effort to discover novel L-asparaginases with potential chemotherapeutic utility in ALL treatment. Detailed analysis of the active site geometry revealed structurally significant differences between HpA and other Lasparaginases that may be important for the biological activities of the enzyme and could be further exploited in protein engineering efforts.

N-terminal amino acids of bovine alpha interferons are relevant for the neutralization of their antiviral activity

The structure-function relationship of interferons (IFNs) has been studied by epitope mapping. Epitopes of bovine IFNs, however, are practically unknown, despite their importance in virus infections and in the maternal recognition of pregnancy. It has been shown that recombinant bovine (rBo)IFN-alphaC and rBoIFN-alpha1 differ only in 12 amino acids and that the F12 monoclonal antibody (mAb) binds to a linear sequence of residues 10 to 34. We show here that the antiviral activities of these two IFNs were neutralized by the F12 mAb to different extents using two tests. In residual activity tests the antiviral activity dropped by more than 99% with rBoIFN-alphaC and by 84% with rBoIFN-alpha1. In checkerboard antibody titrations, the F12 mAb titer was 12,000 with rBoIFN-alphaC and only 600 with rBoIFN-alpha1. Since these IFNs differ in their amino acid sequence at positions 11, 16 and 19 of the amino terminus, only these amino acids could account for the different neutralization titers, and they should participate in antibody binding. According to the three-dimensional structure described for human and murine IFNs, these amino acids are located in the alpha helix A; amino acids 16 and 19 of the bovine IFNs would be expected to be exposed and could bind to the antibody directly. The amino acid at position 11 forms a hydrogen bond in human IFNs-alpha and it is possible that, in bovine IFNs-alpha, the F12 mAb, binding near position 11, would disturb this hydrogen bond, resulting in the difference in the extent of neutralization observed.

Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

Enveloped viruses always gain entry into the cytoplasm by fusion of their lipid envelope with a cell membrane. Some enveloped viruses fuse directly with the host cell plasma membrane after virus binding to the cell receptor. Other enveloped viruses enter the cells by the endocytic pathway, and fusion depends on the acidification of the endosomal compartment. In both cases, virus-induced membrane fusion is triggered by conformational changes in viral envelope glycoproteins. Two different classes of viral fusion proteins have been described on the basis of their molecular architecture. Several structural data permitted the elucidation of the mechanisms of membrane fusion mediated by class I and class II fusion proteins. In this article, we review a number of results obtained by our laboratory and by others that suggest that the mechanisms involved in rhabdovirus fusion are different from those used by the two well-studied classes of viral glycoproteins. We focus our discussion on the electrostatic nature of virus binding and interaction with membranes, especially through phosphatidylserine, and on the reversibility of the conformational changes of the rhabdovirus glycoprotein involved in fusion. Taken together, these data suggest the existence of a third class of fusion proteins and support the idea that new insights should emerge from studies of membrane fusion mediated by the G protein of rhabdoviruses. In particular, the elucidation of the three-dimensional structure of the G protein or even of the fusion peptide at different pH's might provide valuable information for understanding the fusion mechanism of this new class of fusion proteins.

The P48T germline mutation and polymorphism in the CDKN2A gene of patients with melanoma

CDKN2A has been implicated as a melanoma susceptibility gene in some kindreds with a family history of this disease. Mutations in CDKN2A may produce an imbalance between functional p16ink4a and cyclin D causing abnormal cell growth. We searched for germline mutations in this gene in 22 patients with clinical criteria of hereditary cancer (early onset, presence of multiple primary melanoma or 1 or more first- or second-degree relatives affected) by secondary structural content prediction, a mutation scanning method that relies on the propensity for single-strand DNA to take on a three-dimensional structure that is highly sequence dependent, and sequencing the samples with alterations in the electrophoretic mobility. The prevalence of CDKN2A mutation in our study was 4.5% (1/22) and there was a correlation between family history and probability of mutation detection. We found the P48T mutation in 1 patient with 2 melanoma-affected relatives. The patient descends from Italian families and this mutation has been reported previously only in Italian families in two independent studies. This leads us to suggest the presence of a mutational "hotspot" within this gene or a founder mutation. We also detected a high prevalence (59.1%) of polymorphisms, mainly alleles 500 C/G (7/31.8%) or 540 C/T (6/27.3%), in the 3' untranslated region of exon 3. This result reinforces the idea that these rare polymorphic alleles have been significantly associated with the risk of developing melanoma.

The novel p.E89K mutation in the SRY gene inhibits DNA binding and causes the 46,XY disorder of sex development

Male sex determination in humans is controlled by the SRY gene, which encodes a transcriptional regulator containing a conserved high mobility group box domain (HMG-box) required for DNA binding. Mutations in the SRY HMG-box affect protein function, causing sex reversal phenotypes. In the present study, we describe a 19-year-old female presenting 46,XY karyotype with hypogonadism and primary amenorrhea that led to the diagnosis of 46,XY complete gonadal dysgenesis. The novel p.E89K missense mutation in the SRY HMG-box was identified as a de novo mutation. Electrophoretic mobility shift assays showed that p.E89K almost completely abolished SRY DNA-binding activity, suggesting that it is the cause of SRY function impairment. In addition, we report the occurrence of the p.G95R mutation in a 46,XY female with complete gonadal dysgenesis. According to the three-dimensional structure of the human SRY HMG-box, the substitution of the conserved glutamic acid residue by the basic lysine at position 89 introduces an extra positive charge adjacent to and between the positively charged residues R86 and K92, important for stabilizing the HMG-box helix 2 with DNA. Thus, we propose that an electrostatic repulsion caused by the proximity of these positive charges could destabilize the tip of helix 2, abrogating DNA interaction.

Études Structurales par Résonance Magnétique Nucléaire (RMN) du Site Actif du Ribozyme VS de Neurospora.

Nous étudions le ribozyme VS de Neurospora, en tant que système modèle, pour augmenter nos connaissances sur la relation entre la structure et la fonction chez les ARNs, ainsi que pour mieux comprendre le mécanisme de clivage de ce ribozyme. Il a été proposé précédemment que la boucle interne A730 dans la tige-boucle VI (SLVI) contient le site actif du ribozyme et lie un ou plusieurs ions métalliques qui pourraient participer au mécanisme réactionnel. Nous avons déterminé par spectroscopie RMN la structure de la tige-boucle SLVI contenant la boucle A730 afin d’éclaircir ce mécanisme. La structure obtenue est en accord avec les études biochimiques antérieures et présente un ou plusieurs sites de liaison au magnésium associé à la boucle interne. Suite à des études de cinétique et de mutagenèse, il a été proposé qu’une adénine localisée dans le site actif, A756, participe à la catalyse par acide/base générale. Des études de pH effectuées précédemment ont identifié un pKa catalytique (5.2-5.8) qui correspond probablement à l’équilibre de protonation du A756. À l’aide de méthodes utilisant le carbone-13, nous avons identifié un pKa modifié appartenant au A756, ce qui supporte le rôle de ce résidu dans la catalyse par acide/base générale. Les études structurales présentées ici aident donc à augmenter notre compréhension du mécanisme de clivage chez le ribozyme VS.

El paradigma del Desarrollo Urbano Sostenible -DUS- como referente del proceso de toma de decisiones legislativas en Colombia. Análisis de la agenda legislativa del Congreso de la República entre 1991 y 2006

Este trabajo contribuye al análisis de la incidencia del paradigma del Desarrollo Urbano Sostenible en el proceso de toma de decisiones legislativas en Colombia, concentrándose en la discusión y definición de la agenda legislativa sobre asuntos urbanos, durante el periodo 1991-2006.

Diseño y Pedagogía en el ciclo de BUP.

Hacer un estudio profundo acerca del Diseño y todo su entorno y posteriormente marcar un programa, una metodología sobre el Diseño. Didáctica del Diseño. Qué se entiende por Diseño. Cómo y cuándo nace el Diseño. Diseño y comunicación visual. Diseño: forma-función. Retroguardia, vanguardia, investigación. Metodología del Diseño. Necesidad de nuevos métodos. Adaptar el programa a los individuos y no a la inversa. Crítica a los métodos y temario en las asignaturas de Diseño. Índices de las asignaturas Diseño 1 y 2. Diseño bi-dimensional: nuevo esquema planteado por el autor. Diseño tri-dimensional: nuevo esquema planteado por el autor. Bibliografía. Análisis teórico. El objetivo principal del Diseño se refiere a la forma de las cosas producidas pero no hay que olvidar que lo que se percibe de modo inmediato está determinado por la disposición y estructura interna, por el material utilizado y por el procedimiento de fabricación, todos ellos aspectos del Diseño. El Diseño nace con el interés de muchos países industrializados durante los años 1959-60. Los nuevos tipos de complejidad están fuera del alcance del proceso tradicional del Diseño: son necesarios nuevos métodos. Un programa de enseñanza tiene que adaptarse a los individuos y no a la inversa: el programa de base se prepara teniendo en cuenta los elementos principales y la finalidad del curso; el que enseña ha de tener la elasticidad y rapidez necesarias para preparar las lecciones de acuerdo con las necesidades que se presenten en cada caso. Elabora el autor dos esquemas de diseño bidimensional y tridimensional plantados por él, los cuales no se pueden reseñar. Fecha finalización tomada del código del documento.

Characterization of cytotoxic ribonucleases: from the internalization pathway to the importance of dimeric structures

En aquesta tesi s'ha caracteritzat la ruta d'internalització de l'onconasa, una RNasa citotòxica. Els resultats indiquen que l'onconasa entra a les cèl·lules per la via dependent de clatrina i del complex AP-2. Seguidament es dirigeix als endosomes de reciclatge i es a través d'aquesta ruta que la proteïna exerceix la citotoxicitat. Per altra banda, els resultats d'aquest treball demostren que PE5, una variant citotòxica de la ribonucleasa pancreàtica humana (HP-RNasa), interacciona amb la importina  mitjançant diferents residus que tot i que no són seqüencials, es troben propers en l'estructura tridimensional d'aquesta proteïna. PM8 és una HP-RNasa amb estructura cristal·logràfica dimèrica constituïda per intercanvi de dominis N-terminals. En aquesta tesi s'han establert les condicions per estabilitzar aquest dimer en solució i també es proposa un mecanisme per la dimerització.

How may tropical cyclones change in a warmer climate?

Tropical Cyclones (TC) under different climate conditions in the Northern Hemisphere have been investigated with the Max Planck Institute (MPI) coupled (ECHAM5/MPIOM) and atmosphere (ECHAM5) climate models. The intensity and size of the TC depend crucially on resolution with higher wind speed and smaller scales at the higher resolutions. The typical size of the TC is reduced by a factor of 2.3 from T63 to T319 using the distance of the maximum wind speed from the centre of the storm as a measure. The full three dimensional structure of the storms becomes increasingly more realistic as the resolution is increased. For the T63 resolution, three ensemble runs are explored for the period 1860 until 2100 using the IPCC SRES scenario A1B and evaluated for three 30 year periods at the end of the 19th, 20th and 21st century, respectively. While there is no significant change between the 19th and the 20th century, there is a considerable reduction in the number of the TC by some 20% in the 21st century, but no change in the number of the more intense storms. Reduction in the number of storms occurs in all regions. A single additional experiment at T213 resolution was run for the two latter 30-year periods. The T213 is an atmospheric only experiment using the transient Sea Surface Temperatures (SST) of the T63 resolution experiment. Also in this case, there is a reduction by some 10% in the number of simulated TC in the 21st century compared to the 20th century but a marked increase in the number of intense storms. The number of storms with maximum wind speeds greater than 50ms-1 increases by a third. Most of the intensification takes place in 2 the Eastern Pacific and in the Atlantic where also the number of storms more or less stays the same. We identify two competing processes effecting TC in a warmer climate. First, the increase in the static stability and the reduced vertical circulation is suggested to contribute to the reduction in the number of storms. Second, the increase in temperature and water vapor provide more energy for the storms so that when favorable conditions occur, the higher SST and higher specific humidity will contribute to more intense storms. As the maximum intensity depends crucially on resolution, this will require higher resolution to have its full effect. The distribution of storms between different regions does not, at first approximation, depend on the temperature itself but on the distribution of the SST anomalies and their influence on the atmospheric circulation. Two additional transient experiments at T319 resolution where run for 20 years at the end of the 20th and 21st century, respectively using the same conditions as in the T213 experiments. The results are consistent with the T213 study. The total number of tropical cyclones were similar to the T213 experiment but were generally more intense. The change from the 20th to the 21st century was also similar with fewer TC in total but with more intense cyclones.

Dynamical evolution of the 2003 southern hemisphere stratospheric winter using Envisat trace-gas observations

Data from the MIPAS instrument on Envisat, supplemented by meteorological analyses from ECMWF and the Met Office, are used to study the meteorological and trace-gas evolution of the stratosphere in the southern hemisphere during winter and spring 2003. A pole-centred approach is used to interpret the data in the physically meaningful context of the evolving stratospheric polar vortex. The following salient dynamical and transport features are documented and analysed: the merger of anticyclones in the stratosphere; the development of an intense, quasi-stationary anticyclone in spring; the associated top-down breakdown of the polar vortex; the systematic descent of air into the polar vortex; and the formation of a three-dimensional structure of a tracer filament on a planetary scale. The paper confirms and extends existing paradigms of the southern hemisphere vortex evolution. The quality of the MIPAS observations is seen to be generally good. though the water vapour retrievals are unrealistic above 10 hPa in the high-latitude winter.

Atmospheric factors governing banded orographic convection

Atmospheric factors Governing Banded Orographic Convection The three-dimensional structure of shallow orographic convection is investigated through simulations performed with a cloud-resolving numerical model. In moist flows that overcome a given topographic barrier to form statically unstable cap clouds, the organization of the convection depends on both the atmospheric structure and the mechanism by which the convection is initiated. Convection initiated by background thermal fluctuations embedded in the flow over a smooth mountain (without any small-scale topographic features) tends to be cellular and disorganized except that shear-parallel bands may form in flows with strong unidirectional vertical shear. The development of well-organized bands is favored when there is weak static instability inside the cloud and when the dry air surrounding the cloud is strongly stable. These bands move with the flow and distribute their cumulative precipitation evenly over the mountain upslope. Similar shear-parallel bands also develop in flows where convection is initiated by small-scale topographic noise superimposed onto the main mountain profile, but in this case stronger circulations are also triggered that create stationary rainbands parallel to the low-level flow. This second dominant mode, which is less sensitive to the atmospheric structure and the strength of forcing, is triggered by lee waves that form over small-scale topographic bumps near the upstream edge of the main orographic cloud. Due to their stationarity, these flow-parallel bands can produce locally heavy precipitation amounts.

The synthesis and characterisation of four new antimony sulphides incorporating transition-metal complexes

Four new antimony sulphides, [T(dien)(2)]Sb6S10 center dot xH(2)O [T = Ni (1), Co (2) x approximate to 0.45], [Co(en)(3)]SbsSI(3) (3) and [Ni(en)(3)]Sb12S19 (4), have been synthesised under solvothermal conditions. In compounds (1) - (3), Sb12S228- secondary building units are connected to form layered structures. In (1) and (2), Sb-6 S-2- layers containing Sb16S16 heterorings are separated by [T(dien]2](2+) cations, whilst in (3), Sb8 S2- layers 10 13 contain [Co(en)3]2+ cations within large Sb22S22 pores. Compound (4) adopts a three-dimensional structure in which [Ni(en)3 12 cations lie within ca. 5 A wide channels. (c) 2007 Elsevier Ltd. All rights reserved.

Structural studies on acridine derivatives binding to telomeric DNA

Acridine derivatives can inhibit a variety of nuclear enzymes by binding or intercalating to DNA. This class of compounds is of great interest in the development of novel anticancer agents. Despite the availability of crystallographic data for some of the compounds complexed with DNA, uncertainties remain about the mechanisms of action, binding preferences and biological targets. To investigate the intercalation of several acridine derivatives, a variety of techniques are being employed. Single-crystal X-ray diffraction is being used to determine the high resolution three-dimensional structure of short sequences of quadruplex telomeric DNA with bound drug. This will be compared to the effect of drug binding to long segments of double-stranded DNA using fibre diffraction, with neutron diffraction studies planned to analyse the hydrogen bonding patterns of the DNA-drug complexes. Small-angle neutron scattering (SANS) will also be applied to study drug binding to both short and long sequences of quadruplex and double-stranded DNA in solution. Initial SANS measurements of the telomeric repeat d(TGGGGT) imply that this hexamer is present as a quadruplex. (c) 2006 Elsevier B.V. All rights reserved.