Crystallization of Escherichia coli Catabolite Gene Activator Protein with its DNA Binding Site: The use of Modular DNA

To obtain crystals of the Escherichia coli catabolite gene activator protein (CAP) complexed with its DNA-binding site, we have searched for crystallization conditions with 26 different DNA segments ≥28 base-pairs in length that explore a variety of nucleotide sequences, lengths, and extended 5′ or 3′ termini. In addition to utilizing uninterrupted asymmetric lac site sequences, we devised a novel approach of synthesizing half-sites that allowed us to efficiently generate symmetric DNA segments with a wide variety of extended termini and lengths in the large size range (≥28 bp) required by this protein. We report three crystal forms that are suitable for X-ray analysis, one of which (crystal form III) gives measurable diffraction amplitudes to 3 Å resolution. Additives such as calcium, n-octyl-β-d-glucopyranoside and spermine produce modest improvements in the quality of diffraction from crystal form III. Adequate stabilization of crystal form III is unexpectedly complex, requiring a greater than tenfold reduction in the salt concentration followed by addition of 2-methyl-2,4-pentanediol and then an increase in the concentration of polyethylene glycol.

The Ciao clp(FD) library. A modular CLP extension for Prolog

We present a new free library for Constraint Logic Programming over Finite Domains, included with the Ciao Prolog system. The library is entirely written in Prolog, leveraging on Ciao's module system and code transformation capabilities in order to achieve a highly modular design without compromising performance. We describe the interface, implementation, and design rationale of each modular component. The library meets several design goals: a high level of modularity, allowing the individual components to be replaced by different versions; highefficiency, being competitive with other TT> implementations; a glass-box approach, so the user can specify new constraints at different levels; and a Prolog implementation, in order to ease the integration with Ciao's code analysis components. The core is built upon two small libraries which implement integer ranges and closures. On top of that, a finite domain variable datatype is defined, taking care of constraint reexecution depending on range changes. These three libraries form what we call the TT> kernel of the library. This TT> kernel is used in turn to implement several higher-level finite domain constraints, specified using indexicals. Together with a labeling module this layer forms what we name the TT> solver. A final level integrates the CLP (J7©) paradigm with our TT> solver. This is achieved using attributed variables and a compiler from the CLP (J7©) language to the set of constraints provided by the solver. It should be noted that the user of the library is encouraged to work in any of those levels as seen convenient: from writing a new range module to enriching the set of TT> constraints by writing new indexicals.

Construcción modular ligera energéticamente eficiente

Esta tesis trata sobre la construcción modular ligera, dentro del contexto de la eficiencia energética y de cara a los conceptos de nZEB (near Zero Energy Building) y NZEB (Net Zero Energy Building) que se manejan en el ámbito europeo y específicamente dentro del marco regulador de la Directiva 2010/31 UE. En el contexto de la Unión Europea, el sector de la edificación representa el 40% del total del consumo energético del continente. Asumiendo la necesidad de reducir este consumo se han planteado, desde los organismos de dirección europeos, unos objetivos (objetivos 20-20-20) para hacer más eficiente el parque edificatorio. Estos objetivos, que son vinculantes en términos de legislación, comprometen a todos los estados miembros a conseguir la meta de reducción de consumo y emisiones de GEI (Gases de Efecto Invernadero) antes del año 2020. Estos conceptos de construcción modular ligera (CML) y eficiencia energética no suelen estar asociados por el hecho de que este tipo de construcción no suele estar destinada a un uso intensivo y no cuenta con unos cerramientos con niveles de aislamiento de acuerdo a las normativas locales o códigos de edificación de cada país. El objetivo de nZEB o NZEB, e incluso Energy Plus, según sea el caso, necesariamente (y así queda establecido en las normativas), dependerá no sólo de la mejora de los niveles de aislamiento de los edificios, sino también de la implementación de sistemas de generación renovables, independientemente del tipo de sistema constructivo con el que se trabaje e incluso de la tipología edificatoria. Si bien es cierto que los niveles de industrialización de la sociedad tecnológica actual han alcanzado varias de las fases del proceso constructivo - sobre todo en cuanto a elementos compositivos de los edificios- también lo es el hecho de que las cotas de desarrollo conseguidas en el ámbito de la construcción no llegan al nivel de evolución que se puede apreciar en otros campos de las ingenierías como la aeronáutica o la industria del automóvil. Aunque desde finales del siglo pasado existen modelos y proyectos testimoniales de construcción industrializada ligera (CIL) e incluso ya a principios del siglo XX, ejemplos de construcción modular ligera (CML), como la Casa Voisin, la industrialización de la construcción de edificios no ha sido una constante progresiva con un nivel de comercialización equiparable al de la construcción masiva y pesada. Los términos construcción industrializada, construcción prefabricada, construcción modular y construcción ligera, no siempre hacen referencia a lo mismo y no siempre son sinónimos entre sí. Un edificio puede ser prefabricado y no ser modular ni ligero y tal es el caso, por poner un ejemplo, de la construcción con paneles de hormigón prefabricado. Lo que sí es una constante es que en el caso de la construcción modular ligera, la prefabricación y la industrialización, casi siempre vienen implícitas en muchos ejemplos históricos y actuales. Con relación al concepto de eficiencia energética (nZEB o incluso NZEB), el mismo no suele estar ligado a la construcción modular ligera y/o ligera industrializada; más bien se le ve unido a la idea de cerramientos masivos con gran inercia térmica propios de estándares de diseño como el Passivhaus; y aunque comúnmente a la construcción ligera se le asocian otros conceptos que le restan valor (corta vida útil; función y formas limitadas, fuera de todo orden estético; limitación en los niveles de confort, etc.), los avances que se van alcanzando en materia de tecnologías para el aprovechamiento de la energía y sistemas de generación renovables, pueden conseguir revertir estas ideas y unificar el criterio de eficiencia + construcción modular ligera. Prototipos y proyectos académicos– como el concurso Solar Decathlon que se celebra desde el año 2002 promovido por el DOE (Departamento de Energía de los Estados Unidos), y que cuenta con ediciones europeas como las de los años 2010 y 2012, replantean la idea de la construcción industrializada, modular y ligera dentro del contexto de la eficiencia energética, con prototipos de viviendas de ± 60m2, propuestos por las universidades concursantes, y cuyo objetivo es alcanzar y/o desarrollar el concepto de NZEB (Net Zero Energy Building) o edificio de energía cero. Esta opción constructiva no sólo representa durabilidad, seguridad y estética, sino también, rapidez en la fabricación y montaje, además de altas prestaciones energéticas como se ha podido demostrar en las sucesivas ediciones del Solar Decathlon. Este tipo de iniciativas de desarrollo de tecnologías constructivas, no sólo apuntan a la eficiencia energética sino al concepto global de energía neta, Energía plus o cero emisiones de CO2. El nivel de emisiones por la fabricación y puesta en obra de los materiales de construcción depende, en muchos casos, no solo de la propia naturaleza del material, sino también de la cantidad de recursos utilizados para producir una unidad de medida determinada (kg, m3, m2, ml, etc). En este sentido podría utilizarse, en muchos casos, el argumento válido de que a menos peso, y a menos tamaño, menos emisiones globales de gases de efecto invernadero y menos contaminación. Para el trabajo de investigación de esta tesis se han tomado como referencias válidas para estudio, prototipos tanto de CML (Modular 3D) como de CIL (panelizado y elementos 2D), dado que para los fines de análisis de las prestaciones energéticas de los materiales de cerramiento, ambos sistemas son equiparables. Para poder llegar a la conclusión fundamental de este trabajo de tesis doctoral - que consiste en demostrar la viabilidad tecnológica/ industrial que supone la combinación de la eficiencia energética y la construcción modular ligera - se parte del estudio del estado de la técnica ( desde la selección de los materiales y los posibles procesos de industrialización en fábrica, hasta su puesta en obra, funcionamiento y uso, bajo los conceptos de consumo cero, cero emisiones de carbono y plus energético). Además -y con un estado de la técnica que identifica la situación actual- se llevan a cabo pruebas y ensayos con un prototipo a escala natural y células de ensayo, para comprobar el comportamiento de los elementos compositivos de los mismos, frente a unas condicionantes climáticas determinadas. Este tipo de resultados se contrastan con los obtenidos mediante simulaciones informáticas basadas en los mismos parámetros y realizadas en su mayoría mediante métodos simplificados de cálculos, validados por los organismos competentes en materia de eficiencia energética en la edificación en España y de acuerdo a la normativa vigente. ABSTRACT This thesis discusses lightweight modular construction within the context of energy efficiency in nZEB (near Zero Energy Building) and NZEB (Net Zero Energy Building) both used in Europe and, specifically, within the limits of the regulatory framework of the EU Directive 2010/31. In the European Union the building sector represents 40% of the total energy consumption of the continent. Due to the need to reduce this consumption, European decision-making institutions have proposed aims (20-20-20 aims) to render building equipment more efficient. These aims are bound by law and oblige all member States to endeavour to reduce consumption and GEI emissions before the year 2020. Lightweight modular construction concepts and energy efficiency are not generally associated because this type of building is not normally meant for intensive use and does not have closures with insulation levels which fit the local regulations or building codes of each country. The objective of nZEB or NZEB and even Energy Plus, depending on each case, will necessarily be associated (as established in the guidelines) not only with the improvement of insulation levels in buildings, but also with the implementation of renewable systems of generation, independent of the type of building system used and of the building typology. Although it is true that the levels of industrialisation in the technological society today have reached several of the building process phases - particularly in the composite elements of buildings - it is also true that the quotas of development achieved in the area of construction have not reached the evolutionary levelfound in other fields of engineering, such as aeronautics or the automobile industry. Although there have been models and testimonial projects of lightweight industrialised building since the end of last century, even going back as far as the beginning of the XX century with examples of lightweight modular construction such as the Voisin House, industrialisation in the building industry has not been constant nor is its comercialisation comparable to massive and heavy construction. The terms industrialised building, prefabricated building, modular building and lightweight building, do not always refer to the same thing and they are not always synonymous. A building can be prefabricated yet not be modular or lightweight. To give an example, this is the case of building with prefabricated concrete panels. What is constant is that, in the case of lightweight modular construction, prefabrication and industrialisation are almost always implicit in many historical and contemporary examples. Energy efficiency (nZEB or even NZEB) is not normally linked to lightweight modular construction and/or industrialised lightweight; rather, it is united to the idea of massive closureswith high thermal inertia typical of design standards such as the Passive House; and although other concepts that subtract value from it are generally associated with lightweight building (short useful life, limited forms and function, inappropriate toany aesthetic pattern; limitation in comfort levels, etc.), the advances being achieved in technology for benefitting from energy and renewable systems of generation may well reverse these ideas and unify the criteria of efficiency + lightweight modular construction. Academic prototypes and projects - such as the Solar Decathlon competition organised by the US Department of Energy and celebrated since 2002, with its corresponding European events such as those held in 2010 and 2012, place a different slant on the idea of industrialised, modular and lightweight building within the context of energy efficiency, with prototypes of homes measuring approximately 60m2, proposed by university competitors, whose aim is to reach and/or develop the NZEB concept, or the zero energy building. This building option does not only signify durability, security and aesthetics, but also fast manufacture and assembly. It also has high energy benefits, as has been demonstrated in successive events of the Solar Decathlon. This type of initiative for the development of building technologies, does not only aim at energy efficiency, but also at the global concept of net energy, Energy Plus and zero CO2 emissions. The level of emissions in the manufacture and introduction of building materials in many cases depends not only on the inherent nature of the material, but also on the quantity of resources used to produce a specific unit of measurement (kg, m3, m2, ml, etc.). Thus in many cases itcould be validly arguedthat with less weight and smaller size, there will be fewer global emissions of greenhouse effect gases and less contamination. For the research carried out in this thesis prototypes such as the CML (3D Module) and CIL (panelled and elements) have been used as valid study references, becauseboth systems are comparablefor the purpose of analysing the energy benefits of closure materials. So as to reach a basic conclusion in this doctoral thesis - that sets out to demonstrate the technological/industrial viability of the combination of energy efficiency and lightweight modular construction - the departure point is the study of the state of the technique (from the selection of materials and the possible processes of industrialisation in manufacture, to their use on site, functioning and use, respecting the concepts of zero consumption, zero emissions of carbon and Energy Plus). Moreover, with the state of the technique identifying the current situation, tests and practices have been carried out with a natural scale prototype and test cells so as to verify the behaviour of the composite elements of these in certain climatic conditions. These types of result are contrasted with those obtained through computer simulation based on the same parameters and done, principally, using simplified methods of calculation, validated by institutions competent in energy efficiency in Spanish building and in line with the rules in force.

Sla1p Is a Functionally Modular Component of the Yeast Cortical Actin Cytoskeleton Required for Correct Localization of Both Rho1p-GTPase and Sla2p, a Protein with Talin Homology

SLA1 was identified previously in budding yeast in a genetic screen for mutations that caused a requirement for the actin-binding protein Abp1p and was shown to be required for normal cortical actin patch structure and organization. Here, we show that Sla1p, like Abp1p, localizes to cortical actin patches. Furthermore, Sla1p is required for the correct localization of Sla2p, an actin-binding protein with homology to talin implicated in endocytosis, and the Rho1p-GTPase, which is associated with the cell wall biosynthesis enzyme β-1,3-glucan synthase. Mislocalization of Rho1p in sla1 null cells is consistent with our observation that these cells possess aberrantly thick cell walls.  Expression of mutant forms of Sla1p in which specific domains were deleted showed that the phenotypes associated with the full deletion are functionally separable. In particular, a region of Sla1p encompassing the third SH3 domain is important for growth at high temperatures, for the organization of cortical actin patches, and for nucleated actin assembly in a permeabilized yeast cell assay. The apparent redundancy between Sla1p and Abp1p resides in the C-terminal repeat region of Sla1p. A homologue of SLA1 was identified in Schizosaccharomyces pombe. Despite relatively low overall sequence homology, this gene was able to rescue the temperature sensitivity associated with a deletion of SLA1 in Saccharomyces cerevisiae.

Modular construction for function of a ribonucleoprotein enzyme: the catalytic domain of Bacillus subtilis RNase P complexed with B.subtilis RNase P protein

The bacterial RNase P holoenzyme catalyzes the formation of the mature 5′-end of tRNAs and is composed of an RNA and a protein subunit. Among the two folding domains of the RNase P RNA, the catalytic domain (C-domain) contains the active site of this ribozyme. We investigated specific binding of the Bacillus subtilis C-domain with the B.subtilis RNase P protein and examined the catalytic activity of this C-domain–P protein complex. The C-domain forms a specific complex with the P protein with a binding constant of ∼0.1 µM. The C-domain–P protein complex and the holoenzyme are equally efficient in cleaving single-stranded RNA (∼0.9 min–1 at pH 7.8) and substrates with a hairpin–loop 3′ to the cleavage site (∼40 min–1). The holoenzyme reaction is much more efficient with a pre-tRNA substrate, binding at least 100-fold better and cleaving 10–500 times more efficiently. These results demonstrate that the RNase P holoenzyme is functionally constructed in three parts. The catalytic domain alone contains the active site, but has little specificity and affinity for most substrates. The specificity and affinity for the substrate is generated by either the specificity domain of RNase P RNA binding to a T stem–loop-like hairpin or RNase P protein binding to a single-stranded RNA. This modular construction may be exploited to obtain RNase P-based ribonucleoprotein complexes with altered substrate specificity.

A modular holistic approach to prosody modelling for Standard Yorùbá speech synthesis

This paper presents a novel prosody model in the context of computer text-to-speech synthesis applications for tone languages. We have demonstrated its applicability using the Standard Yorùbá (SY) language. Our approach is motivated by the theory that abstract and realised forms of various prosody dimensions should be modelled within a modular and unified framework [Coleman, J.S., 1994. Polysyllabic words in the YorkTalk synthesis system. In: Keating, P.A. (Ed.), Phonological Structure and Forms: Papers in Laboratory Phonology III, Cambridge University Press, Cambridge, pp. 293–324]. We have implemented this framework using the Relational Tree (R-Tree) technique. R-Tree is a sophisticated data structure for representing a multi-dimensional waveform in the form of a tree. The underlying assumption of this research is that it is possible to develop a practical prosody model by using appropriate computational tools and techniques which combine acoustic data with an encoding of the phonological and phonetic knowledge provided by experts. To implement the intonation dimension, fuzzy logic based rules were developed using speech data from native speakers of Yorùbá. The Fuzzy Decision Tree (FDT) and the Classification and Regression Tree (CART) techniques were tested in modelling the duration dimension. For practical reasons, we have selected the FDT for implementing the duration dimension of our prosody model. To establish the effectiveness of our prosody model, we have also developed a Stem-ML prosody model for SY. We have performed both quantitative and qualitative evaluations on our implemented prosody models. The results suggest that, although the R-Tree model does not predict the numerical speech prosody data as accurately as the Stem-ML model, it produces synthetic speech prosody with better intelligibility and naturalness. The R-Tree model is particularly suitable for speech prosody modelling for languages with limited language resources and expertise, e.g. African languages. Furthermore, the R-Tree model is easy to implement, interpret and analyse.

Chronic Telogen Effluvium And Female Pattern Hair Loss Are Separate And Distinct Forms Of Alopecia: A Histomorphometric And Immunohistochemical Analysis.

Chronic telogen effluvium (CTE), a poorly understood condition, can be confused with or may be a prodrome to female pattern hair loss (FPHL). The pathogenesis of both is related to follicle cycle shortening and possibly to blood supply changes. To analyze a number of histomorphometric and immunohistochemical findings through vascular endothelial growth factor (VEGF), Ki-67, and CD31 immunostaining in scalp biopsies of 20 patients with CTE, 17 patients with mild FPHL and 9 controls. Ki-67 index and VEGF optical density were analyzed at the follicular outer sheath using ImageJ software. CD31 microvessel density was assessed by a Chalkley grid. Significant follicle miniaturization and higher density of nonanagen follicles were found in FPHL, compared with patients with CTE and controls. Ki-67+ index correlated positively with FPHL histological features. The FPHL group showed the highest VEGF optical density, followed by the CTE and control groups. No differences were found in CD31 microvessel density between the three groups. Histomorphometric results establish CTE as a distinct disorder, separate from FPHL from its outset. Its pathogenic mechanisms are also distinct. These findings support the proposed mechanism of 'immediate telogen release' for CTE, leading to cycle synchronization. For FPHL, accelerated anagen follicular mitotic rates and, thus, higher Ki-67 and VEGF values, would leave less time for differentiation, resulting in hair miniaturization.

Use Of Experimental Design To Optimize A Triple-potential Waveform To Develop A Method For The Determination Of Streptomycin And Dihydrostreptomycin In Pharmaceutical Veterinary Dosage Forms By Hplc-pad.

An HPLC-PAD method using a gold working electrode and a triple-potential waveform was developed for the simultaneous determination of streptomycin and dihydrostreptomycin in veterinary drugs. Glucose was used as the internal standard, and the triple-potential waveform was optimized using a factorial and a central composite design. The optimum potentials were as follows: amperometric detection, E1=-0.15V; cleaning potential, E2=+0.85V; and reactivation of the electrode surface, E3=-0.65V. For the separation of the aminoglycosides and the internal standard of glucose, a CarboPac™ PA1 anion exchange column was used together with a mobile phase consisting of a 0.070 mol L(-1) sodium hydroxide solution in the isocratic elution mode with a flow rate of 0.8 mL min(-1). The method was validated and applied to the determination of streptomycin and dihydrostreptomycin in veterinary formulations (injection, suspension and ointment) without any previous sample pretreatment, except for the ointments, for which a liquid-liquid extraction was required before HPLC-PAD analysis. The method showed adequate selectivity, with an accuracy of 98-107% and a precision of less than 3.9%.

Pode a atividade física materna modular a programação fetal induzida pela nutrição?

Existe considerável evidência para a indução de diferentes fenótipos em reposta às variações no ambiente fetal e neonatal. O aporte inadequado de nutrientes no período crítico do desenvolvimento está associado ao risco alto de doenças metabólicas na vida adulta, este fenômeno biológico é chamado de programação. A atividade física durante a gestação resulta em adaptações fisiológicas da mãe e no aumento da disponibilidade de nutrientes e oxigênio no espaço feto-placentário. Este trabalho tem como objetivo discutir os mecanismos da indução de programação fetal pela nutrição e o provável efeito modulador da atividade física durante a gestação. Foram utilizadas as bases de dados do Medline Pubmed, Lilacs e Bireme, com publicações entre 1990 até 2008. Os termos de indexação utilizados foram: nutrition, fetal programming, gestation, physical activity, physical exercise, metabolism. Em conclusão, o aporte inadequado de nutrientes programa o aparecimento de doenças metabólicas na vida adulta, enquanto que a atividade física durante a gestação aumenta a disponibilidade de nutrientes e oxigênio, repercutindo positivamente no crescimento fetal e no peso ao nascer.

Interleukin-22 forms dimers that are recognized by two interleukin-22R1 receptor chains

Interleukin-22 (IL-22) is a class 2 cytokine whose primary structure is similar to that of interleukin 10 (IL-10) and interferon-gamma (IFN-gamma). IL-22 induction during acute phase immune response indicates its involvement in mechanisms of inflammation. Structurally different from IL-10 and a number of other members of IL-10 family, which form intertwined inseparable V-shaped dimers of two identical polypeptide chains, a single polypeptide chain of IL-22 folds on itself in a relatively globular structure. Here we present evidence, based on native gel electrophoresis, glutaraldehyde cross-linking, dynamic light scattering, and small angle x-ray scattering experiments, that human IL-22 forms dimers and tetramers in solution under protein concentrations assessable by these experiments. Unexpectedly, low-resolution molecular shape of IL-22 dimers is strikingly similar to that of IL-10 and other intertwined cytokine dimeric forms. Furthermore, we determine an ab initio molecular shape of the IL-22/IL-22R1 complex which reveals the V-shaped IL-22 dimer interacting with two cognate IL-22R1 molecules. Based on this collective evidence, we argue that dimerization might be a common mechanism of all class 2 cytokines for the molecular recognition with their respective membrane receptor. We also speculate that the IL-22 tetramer formation could represent a way to store the cytokine in nonactive form at high concentrations that could be readily converted into functionally active monomers and dimers upon interaction with the cognate cellular receptors.

Test of a Modular Fault Current Limiter for 220 V Line Using YBCO Coated Conductor Tapes With Shunt Protection

A modular superconducting fault current limiter (SFCL) consisting of 16 elements was constructed and tested in a 220 V line for a fault current between 1 kA to 7.4 kA. The elements are made up of second generation (2G) YBCO-coated conductor tapes with stainless steel reinforcement. For each element four tapes were electrically connected in parallel with effective length of 0.4 m per element, totaling 16 elements connected in series. The evaluation of SFCL performance was carried out under DC and AC tests. The DC test was performed through pulsed current tests and its recovery characteristics under load current were analysed by changing the shunt resistor value. The AC test performed using a 3 MVA/220 V/60 Hz transformer has shown the current limiting ratio achieved a factor higher than 10 during fault of up to five cycles without conductor degradation. The measurement of the voltage for each element during the AC test showed that in this modular SFCL the quench is homogeneous and the transition occurs similarly in all the elements.

Towards modular and coordinated manufacturing systems oriented to services

Nowadays, there is a trend for industry reorganization in geographically dispersed systems, carried out of their activities with autonomy. These systems must maintain coordinated relationship among themselves in order to assure an expected performance of the overall system. Thus, a manufacturing system is proposed, based on ""web services"" to assure an effective orchestration of services in order to produce final products. In addition, it considers special functions, such as teleoperation and remote monitoring, users` online request, among others. Considering the proposed system as discrete event system (DES), techniques derived from Petri nets (PN), including the Production Flow Schema (PFS), can be used in a PFS/PN approach for modeling. The system is approached in different levels of abstraction: a conceptual model which is obtained by applying the PFS technique and a functional model which is obtained by applying PN. Finally, a particular example of the proposed system is presented.

Synthesis of Cyclic Guanidine Intermediates of Anatoxin-a(s) in Both Racemic and Enantiomerically Pure Forms

The alkyl chain of anatoxin-a(s) (cyclic guanidines), which can be used as an intermediate in the total synthesis of anatoxin-a(s), was synthesized in both racemic and enantiomerically pure forms. These enantiomerically pure cyclic compounds can be used as chiral inductors in some reactions. The two racemic routes disclosed herein have the advantages of high overall yield and mild reaction conditions. Both routes proceed through an intermediate 2,3-diaminoacid - an important synthetic scaffold - with good yields. Furthermore, the N,N-dimethyl-2(tosylimino)imidazolidine-4-carboxamide might be obtained from 2-(tosylimino)imidazolidine-4-carboxylic acid followed by selective reduction of the carbonyl functionality. All synthesized compounds were analyzed by mass spectrometry and (1)H NMR and (13)C NMR spectroscopy.

Monocyte-derived dendritic cells from patients with severe forms of chromoblastomycosis induce CD4(+) T cell activation in vitro

Dendritic cells (DCs) have been described as initiators and modulators of the immune response. Recently we have shown a predominant production of interleukin-10 cytokine, low levels of interferon-gamma and inefficient T cell proliferation in patients with severe forms of chromoblastomycosis. Chromoblastomycosis starts with subcutaneous inoculation of Fonsecaea pedrosoi into tissue where DCs are the first line of defence against this microorganism. In the present study, the interaction of F. pedrosoi and DCs obtained from patients with chromoblastomycosis was investigated. Our results showed that DCs from patients exhibited an increased expression of human leucocyte antigen D-related (HLA-DR) and co-stimulatory molecules. In the presence of conidia, the expression of HLA-DR and CD86 was up-regulated by DCs from patients and controls. Finally, we demonstrate the reversal of antigen-specific anergy and a T helper type 1 response mediated by DCs incubated with F. pedrosoi conidea.