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Resumen Objetivos Determinar las propiedades metricas de la escala COMFORT Behavior Scale desarrollada por van Dijk et al. (2000) en el contexto del paciente critico pediatrico. Diseno Estudio observacional, psicometrico y prospectivo. Lugar de estudio Unidad de Cuidados Intensivos de un hospital pediatrico y universitario de tercer nivel asistencial. Pacientes Un total de 311 ninos con una mediana de edad de 5,07 anos (0,9-11,7). Intervenciones Ninguna. Principales variables y resultados La escala fue administrada de forma simultanea por 2 enfermeros a 311 pacientes ingresados en una unidad de criticos de 18 camas de un hospital pediatrico de tercer nivel asistencial. La version espanola de la COMFORT Behavior Scale obtuvo un alfa de Cronbach de 0,715 y esta compuesta por 3 factores, con 2 items cada uno: 1) alerta y movimiento fisico; 2) calma/agitacion y respuesta respiratoria/llanto, y 3) tono muscular y tension facial. Conclusiones La escala COMFORT B logro ser adaptada al idioma espanol y mostro ser valida para determinar y cuantificar el grado de confort en un grupo de ninos ingresados en una unidad de cuidados pediatricos espanola.

Objectives To determine the measurement properties of the Spanish version of the COMFORT Behavior Scale developed by van Dijk et al (2000) in paediatric critical care patients. Design Prospective observational and psychometric study. Setting Level III Intensive Care Area at a university’s children hospital. Patients A total of 311 children with median age of 5.07 years (IQR = 0.9-11.7). Interventions None. Principal variables and results To determine the measurement properties the Spanish version of the COMFORT Behavior Scale was simultaneous administered by two nurses to 311 patients admitted to an eighteen-bed critical care unit of a third level paediatric hospital. The Spanish version of the COMFORT Behavior Scale obtained a Cronbach alpha coefficient of 0.715 and it is a tool made up of three factors with two items in each domain: i) alertness and physical movement; ii) calmness/agitation and respiratory response/crying, and iii) muscle tone and facial tension. Conclusions The COMFORT B was adapted to Spanish and it has shown to be a valid an reliable tool to assess comfort in a group of children admitted to an Spanish Intensive Care Unit.

The detection of repetitive sequences with single-base resolution is becoming increasingly important aiming to understand the biological implications of genomic variation in these sequences. However, there is a lack of techniques to experimentally validate sequencing data from repetitive sequences obtained by Next-Generation Sequencing methods, especially in the case of Single-Nucleotide Variations (SNVs). That is one of the reasons why repetitive sequences have been poorly studied and excluded from most genomic studies. Therefore, in addition to sequencing data, there is an urgent need for efficient validation methods of genomic variation in these sequences. Herein we report the development of chemFISH, an alternative method for the detection of SNVs in repetitive sequences. ChemFISH is an innovative method based on dynamic chemistry labelling and abasic Peptide Nucleic Acid (PNA) probes to detect in situ the α-satellite DNA, organized in tandem repeats, with single-base resolution in a direct and rapid reaction. With this approach, we detected by microscopy the α-satellite DNA in a variety of human cell lines, we quantified the detection showing a low coefficient of variation among samples (13.16%-25.33%) and we detected single-base specificity with high sensitivity (82.41%-88.82%). These results indicate that chemFISH can serve as a rapid method to validate previously detected SNVs in sequencing data, as well as to find novel SNVs in repetitive sequences. Furthermore, the versatile chemistry behind chemFISH can lead to develop novel molecular assays for the in situ detection of nucleic acids.

miRNAs are well known for being implicated in a myriad of biological situations, including those related to serious diseases. Amongst miRNAs, miRNA-21 has the spotlight as it is reported to be up-regulated in multiple severe pathological conditions, being its quantification a key point in medicine. To date, most of the techniques for miRNA quantification have shown to be less effective than expected; thus, we herein present a novel, rapid, cost-effective, robust and PCR-free approach, based on dynamic chemistry, for the identification and quantification of miRNA directly from tumour cells using both FACS and a fluorescent microplate. This dynamic chemistry novel application involves bead based reagents and allows quantifying the number of miR-21 molecules presented in MDA-MB-468 and H1975 tumour cells.

Nucleic acid-based molecular diagnosis has gained special importance for the detection and early diagnosis of genetic diseases as well as for the control of infectious disease outbreaks. The development of systems that allow for the detection and analysis of nucleic acids in a low-cost and easy-to-use way is of great importance. In this context, we present a combination of a nanotechnology-based approach with the already validated dynamic chemical labeling (DCL) technology, capable of reading nucleic acids with single-base resolution. This system allows for the detection of biotinylated molecular products followed by simple detection using a standard flow cytometer, a widely used platform in clinical and molecular laboratories, and therefore, is easy to implement. This proof-of-concept assay has been developed to detect mutations in KRAS codon 12, as these mutations are highly important in cancer development and cancer treatments., Graphical abstract Image 1, Highlights • A nanotechnology-based approach for capturing and detecting nucleic acids by flow cytometry has been developed. • A great advantage is that molecular reactions are taking place in solution while the reading is done by using flow cytometry. • Detection of KRAS codon 12 and miR-122 sequences using this approach has been successfully carried out. • This approach will have a clear impact in molecular diagnostic.

The life-long and life-threatening Chagas disease is one of the most neglected tropical diseases caused by the protozoan parasite Trypanosoma cruzi. It is a major public health problem in Latin America, as six to seven million people are infected, being the principal cause of mortality in many endemic regions. Moreover, Chagas disease has become widespread due to migrant populations. Additionally, there are no vaccines nor effective treatments to fight the disease because of its long-term nature and complex pathology. Therefore, these facts emphasize how crucial the international effort for the development of new treatments against Chagas disease is. Here, we present the in vitro and in vivo trypanocidal activity of some oxygenated abietane diterpenoids and related compounds. The 1,4-benzoquinone 15, not yet reported, was identified as a fast-acting trypanocidal drug with efficacy against different strains in vitro and higher activity and lower toxicity than benznidazole in both phases of murine Chagas disease. The mode of action was also evaluated, suggesting that quinone 15 kills T. cruzi by inducing mitochondrion-dependent necrosis through a bioenergetics collapse caused by a mitochondrial membrane depolarization and iron-containing superoxide dismutase inhibition. Therefore, the abietane 1,4-benzoquinone 15 can be considered as a new candidate molecule for the development of an appropriate and commercially accessible anti-Chagas drug.

The LHC collimation system must adopt materials with excellent thermal shock resistance, high electrical conductivity, geometrical stability, and radiation hardness. Two novel composites, Molybdenum–Carbide–Graphite and Copper–Diamond, are proposed for the LHC collimation upgrade. A postirradiation examination was performed to assess the status of the composites, tested under intense proton beam impacts at the CERN HiRadMat facility. Metrology measurements, computed tomography, and 3D topography allowed to evaluate the localized spallation induced by the beam. This article provides an overview of the thermophysical characterization of the two composites before irradiation and nondestructive postirradiation results.

The synthesis of podocarpanes, including 12,19-dihydroxy-13-acetyl-8,11,13-podocarpatriene (23), isolated from Gaultheria yunnanensis and not previously synthesized, and totarane-type terpenoids, starting from the natural labdane trans-communic acid (15), is described. Their antiproliferative activities against MCF-7, T-84 and A-549 human tumoural cell lines are studied. An antiproliferative effect was induced by compounds 23, 27 and 28, with IC50

A bioinspired synthesis of rearranged abietane diterpenes, related to pygmaeocins, is described. In this process, the key step is the 1,2-migration of the C-20 angular methyl to the C-5 position of the abietane skeleton, which occurs when a C6-C7 unsaturated dehydroabietane derivative is treated with SeO

A novel sensitive, specific and rapid method for the detection and quantification of microRNAs without requiring extraction from their biological sources is now available using a novel chemical based, PCR-free technology for nucleic acid testing. In this study, we both demonstrate how this method can be used to profile miR-451a, an important miRNA in erythropoiesis, and compare with the gold standard RT-qPCR.

The High Luminosity upgrade of the Large Hadron Collider (HL-LHC) has prompted the investigation of novel materials for beam-intercepting devices, and in particular for the collimators responsible for protecting the machine from beam losses. The HL-LHC collimation system will inevitably experience increased levels of radiation damage and undergo changes in their crucial physio-mechanical properties. Graphite-matrix composite materials containing molybdenum carbide particles, along with small amounts of titanium carbide, were developed with the objective of enhanced in-beam performance and tested under proton irradiation. The physical degradation observed in early grades of molybdenum carbide compounds, even after modest proton fluences, has prompted the development of advanced compounds. In this work, we examine the effects of proton irradiation on the microstructural and thermophysical properties of new grades of Molybdenum-carbide-graphite compounds up to fluences of ~2 × 1020 p/cm2. We employ a combination of precision dilatometry and high-energy X-ray diffraction to quantify the dimensional stability and crystallographic phase evolution both pre- and post-irradiation. Our results reveal that these new compounds exhibit superior resilience to radiation damage than their predecessors.

The in vitro antiproliferative activities of some taiwaniaquinoids and related compounds with functionalized A, B, or C rings against human breast (MCF-7), colon (T-84), and lung (A-549) tumor cell lines were assayed. The most potent compounds, 16, 27, and 36, were more effective than the naturally occurring taiwaniaquinones A (4) and F (5) in all three cell lines. The structure-activity relationship study of these new taiwaniaquinoids highlighted the correlation between the bromo substituent and the antiproliferative activity, especially in MCF-7 cells. These findings indicate that some of the taiwaniaquinoids might be useful as cytostatic agents against breast, colon, and lung cancer cell lines.

Estudiamos las características clínicas y biológicas de 16 pacientes afectos de polimialgia arterítica. Doce de ellos fueron diagnosticados por biopsia de la arteria temporal y los 4 restantes por poseer criterios clínicos, analíticos y espectacular respuesta a los corticoides. No hemos hallado diferencias en cuanto a la presentación según el sexo. La mayoría (75 %) tenían síntomas previos entre 1 y 6 meses, siendo las cefaleas el más común (62 %). Por orden de frecuencia presentaron además síndrome febril (56 %) polimialgia (50 %), cuadro tóxico (37 %), claudicación intermitente, disminución de la agudeza visual, y artritis (12 %). Entre los hallazgos de laboratorio destacamos la elevación de la VSG en todos los casos, hemoglobina inferior a 8 g/100 mi. en 8 y elevación de alfa-2-globulina en otros 8 pacientes. La palpación de las arterias temporales fue anormal en 11 casos. Destacamos la positividad de la biopsia en 2 de los 5 pacientes cuya palpación arterial era normal. Todos los enfermos fueron tratados con 6-metilprednisolona, pudiendo valorar su evolución en 11, de los cuales 3 se hallan asintomáticos tras haber recibido tratamiento 3, 4 y 6 meses respectivamente. Los 8 restantes se hallan libres de síntomas recibiendo dosis mínimas.

Dynamic chemical labelling is a single-base specific method to enable detection and quantification of micro-Ribonucleic Acids in biological fluids without extraction and pre-amplification. In this study, dynamic chemical labelling was combined with the Luminex MAGPIX system to profile levels of microRNA-122 biomarker in serum from patients with Drug-Induced Liver Injury.

The introduction at CERN of new extremely energetic particle accelerators, such as the high-luminosity large hadron collider (HL-LHC) or the proposed future circular collider (FCC), will increase the energy stored in the circulating particle beams by almost a factor of two (from 360 to 680 MJ) and of more than 20 (up to 8500 MJ), respectively. In this scenario, it is paramount to assess the dynamic thermomechanical response of materials presently used, or being developed for future use, in beam intercepting devices (such as collimators, targets, dumps, absorbers, spoilers, windows, etc.) exposed to potentially destructive events caused by the impact of energetic particle beams. For this reason, a new HiRadMat experiment, named “MultiMat”, was carried out in October 2017, with the goal of assessing the behaviour of samples exposed to high-intensity, high-energy proton pulses, made of a broad range of materials relevant for collimators and beam intercepting devices, thin-film coatings and advanced equipment. This paper describes the experiment and its main results, collected online thanks to an extensive acquisition system and after the irradiation by non-destructive examination, as well as the numerical simulations performed to benchmark experimental data and extend materials constitutive models., peer-reviewed

Objetives: The aim of this study was to determine which of three methods for measuring BIC (bone-to-implant contact), using vestibular and lingual scanning electron microscopy (SEM) for different implant systems at 15, 30 and 90 days post-surgery was the most precise. An elemental analysis with SEM was used to evaluate neoformed bone composition for three implant systems at the same study times. Material and Methods: 36 implants were placed in eighteen Beagle dogs mandible about one year old and weighing approximately 12-13 kg in order to evaluate bone apposition to three different implant surfaces. It was used the third and fourth premolar and first molar distal sockets in both quadrants of the mandible (3P3, 4P4 and 1M1). Teeth were hemi-sected and the distal roots were removed. The specimens were prepared for histological examination and each section surface was stained using Masson’s trichrome and hematoxylin and eosin stains. BIC evaluations were performed by the three methods, BIC I (the quantity of mineralized bone in direct contact with the implant’s titanium surface across the entire threaded area); BIC II (along a line that passes from apex to apex of the implant threads); BIC III (both in areas around and above the threads and in between threads). Results: Both BIC and bone content were analyzed for all implants placed in P3, P4 y M1 alveoli on both, the buccal and palatine sides (elemental analysis quantified Ca, P, O and C). It was seen it was only at the ninety-day mark that high percentages of calcium were present. Conclusions: This study suggest that BIC III evaluation is the most certain method for establishing the quantity of bone formed as the BIC area. Key words:Bone-to-impant contact, dogs, extraction socket, implants.

The High-Luminosity LHC Project aims to increase the integrated luminosity that will be collected by the Large Hadron Collider for the needs of the high energy physics frontier by the end of its Run 3 by more than a factor ten. This will require doubling the beam intensity, and in order to ensure coherent stability until the brighter beams are put in collision, the transverse impedance of the machine has to be reduced. As the major portion of the ring impedance is generated by its collimation system, several low resistivity jaw materials have been considered to lower the collimator impedance and a special collimator has been built and installed in the machine to study their effect. In order to assess the performance of each material we performed a series of tune shift measurements with LHC beams. The results show a significant reduction of the resistive wall tune shift with novel materials, in good agreement with the impedance model and the bench impedance and resistivity measurements. The largest improvement is obtained with a molybdenum coating of a molybdenum-graphite jaw. This coating, applied to the most critical collimators, is estimated to lower the machine impedance by up to 30% and the stabilizing Landau octupole threshold by up to 240 A after accounting for uncertainties of the model and other destabilising effects. A half of the overall improvement can be obtained by coating the jaws of a subset of 4 out of 11 collimators identified as the highest contributors to machine impedance. This subset of low-impedance collimators is being installed during the Long Shutdown 2 in 2019-2020. The High-Luminosity upgrade of the Large Hadron Collider (HL-LHC) will double its beam intensity for the needs of High Energy Physics frontier. In order to ensure coherent stability until the beams are put in collision, the transverse impedance has to be reduced. As the major portion of the ring impedance is sup

Two new absorbing materials were developed as collimator inserts to fulfil the requirements of HL-LHC higher brightness beams: molybdenum-carbide graphite (MoGr) and copper-diamond (CuCD). These materials were tested under intense beam impacts at CERN HiRadMat facility in 2015, when full jaw prototypes were irradiated. Additional tests in HiRadMat were performed in 2017 on another series of material samples, including also improved grades of MoGr and CuCD, and different coating solutions. This paper summarizes the main results of the two experiments, with a main focus on the behaviour of the novel composite blocks, the metallic housing, as well as the cooling circuit. The experimental campaign confirmed the final choice for the materials and the design solutions for HL-LHC collimators, and constituted a unique chance of benchmarking numerical models. In particular, the tests validated the selection of MoGr for primary and secondary collimators, and CuCD as a valid solution for robust tertiary collimators., Proceedings of the 10th Int. Particle Accelerator Conf., IPAC2019, Melbourne, Australia

A family of novel graphite-based composites reinforced with a dispersion of molybdenum carbide particles, with very high thermal and electrical properties, has been recently developed at CERN, in collaboration with Brevetti Bizz (IT), for applications in particle accelerators. These materials, produced by spark plasma sintering, assisted by liquid phase, have been extensively investigated and characterized. The influence of the initial constituents, together with the sintering parameters, have been studied in view of optimizing the thermo-physical and mechanical properties as well as ultra-high vacuum compatibility. The carbide phases and the microstructures have been analysed. Their excellent thermal conductivity (approaching 800 W m − 1 K − 1 ), low thermal expansion and low density make these materials appealing for high-end thermal management applications.

The CERN Large Hadron Collider is currently being upgraded to operate at a stored beam energy of 680 MJ through the High Luminosity upgrade. The LHC performance is dependent on the functionality of beam collimation systems, essential for safe beam cleaning and machine protection. A dedicated beam experiment at the CERN High Radiation to Materials facility is created under the HRMT-23 experimental campaign. This experiment investigates the behavior of three collimation jaws having novel composite absorbers made of copper diamond, molybdenum carbide graphite, and carbon fiber carbon, experiencing accidental scenarios involving the direct beam impact on the material. Material characterization is imperative for the design, execution, and analysis of such experiments. This paper presents new data and analysis of the thermostructural characteristics of some of the absorber materials commissioned within CERN facilities. In turn, characterized elastic properties are optimized through the development and implementation of a mixed numerical-experimental optimization technique., peer-reviewed

The mechanism and kinetics of pterostilbene reactions with OH and OOH radicals were studied in gas and aqueous phases through Density Functional Theory (DFT). For the pterostilbene + OH reaction in gas phase, the addition of OH to unsaturated carbons is kinetically favored with respect to the direct hydrogen-abstraction from the hydroxyl group. In aqueous solution, OH should react at diffusion controlled rate with pterostilbene via hydrogen abstraction or adduct formation. For pterostilbene + OOH reaction in gas phase, the highest rate constant was obtained for hydrogen abstraction pathway in the hydroxyl group. In solution, only the hydrogen-abstraction mechanism is thermodynamically favored.

Chagas disease, caused by the protozoa parasite Trypanosoma cruzi, is an example of extended parasitaemia with unmet medical needs. Current treatments based on old-featured benznidazole (Bz) and nifurtimox are expensive and do not fulfil the criteria of effectiveness, and a lack of toxicity devoid to modern drugs. In this work, a group of abietic acid derivatives that are chemically stable and well characterised were introduced as candidates for the treatment of Chagas disease. In vitro and in vivo assays were performed in order to test the effectiveness of these compounds. Finally, those which showed the best activity underwent additional studies in order to elucidate the possible mechanism of action. In vitro results indicated that some compounds have low toxicity (i.e. >150 μM, against Vero cell) combined with high efficacy (i.e.

El castillo-palacio de La Calahorra es un edificio histórico (s. XVI) con una doble funcionalidad. El exterior es un castillo-fortaleza de estilo tardo-medieval, construido con mampuestos y sillarejos de una caliza cristalina de gran dureza y resistencia. El interior es un palacio renacentista, hecho con sillares de caliza micrítica y arenisca dolomítica, ambos materiales pétreos son porosos, blandos y poco resistentes. Todo el edificio tiene una tonalidad anaranjada debido a la pátina de hierro que tiñe sus fábricas y que es consecuencia de la acción del viento. Existe un deterioro diferencial entre sus fábricas externas e internas, según el tipo de piedra empleado, la acción del agua de lluvia y el viento, y los defectos constructivos detectados. Mientras que la piedra exterior está en buen estado de conservación, las interiores sufren importantes procesos de deterioro, principalmente la caliza micrítica presente en las zonas ornamentales y decorativas del patio central., The castle-palace of La Calahorra is a historical building (s. XVI) with a dual-function. The exterior is a castlefortress of late-medieval style, built with masonry and ashlar of a crystalline limestone of great hardness and strength. The interior is a Renaissance palace, made with ashlar limestone micritic and dolomitic sandstone, both stone materials are porous, soft and little resistant. The entire building has an orange hue due to the patina of iron that stained their factories and that is a consequence of the action of the wind. There is a differential deterioration among its external and internal, depending on the type of stone used factories, the action of rain and wind and the construction defect detected. While the exterior stone is in good state of conservation, the interiors are important processes of deterioration, mainly the limestone micritic present in the ornamental and decorative areas of the central courtyard., Este trabajo ha sido cofinanciado por la Comunidad de Madrid a través del programa Geomateriales (S2009/MAT- 1629) y por el Ministerio de Economía y Competitividad (MINECO) a través del programa Consolider-Ingenio 2007 (CSD2007-0058). Además forma parte de las actividades del grupo de investigación de la UCM “Alteración y Conservación de los Materiales Pétreos del Patrimonio” (ref. 921349).

A new strategy for synthesizing taiwaniaquinoids, a group of terpenoids with an unusual rearranged 5(6→7) or 6-nor-5(6→7)abeo-abietane skeleton, which exhibit promising biological activities, is reported. The procedure, based on the cleavage of the C7-C8 double bond of abietane diterpenes, is the only one yet reported for synthesizing C(20) taiwaniaquinoids bearing a carbon function on the cyclopentane B ring; it is also applicable to the synthesis of the wide variety of existing taiwaniaquinoids. Utilizing this, (-)-taiwaniaquinone A, F, G, and H, (-)-taiwaniaquinol B, and (-)-dichroanone have been synthesized from (+)-abietic acid. The versatility of this strategy allows us to propose the abietane C7-C8 cleavage as a possible biosynthetic pathway to this type of rearranged diterpenes; this proposal seems to be supported by phytochemical evidence.

A series of 12-hydroxy-substituted abietane diterpenes, functionalized on C19 or C 18, have been synthesized starting from 18-hydroxyferruginol. The first synthesis of antibacterial hanagokenol A and fortunins E, G, and H, and a new procedure for preparing sugikurojin A and the immunosuppressive 19-hydroxyferruginol are reported.

Resumen Presentamos el caso de un varon de 85 anos que presentaba un dolor lumbar de larga y torpida evolucion, que no mejoraba con los tratamientos convencionales. Aunque el paciente lo relacionaba de forma clara con un esfuerzo realizado, el resultado final fue que la fractura aplastamiento se origino por una tuberculosis vertebral o mal de Pott. El objetivo de este trabajo es la revision clinica y actualizacion de esta patologia realmente poco frecuente.

Innovative collimators are being investigated to handle the high energy particle beams foreseen in future upgrades of CERN Large Hadron Collider (LHC). This calls for the development of novel advanced materials, as no existing metal- or carbon-based material possesses the combination of physical, thermal, electrical and mechanical properties, imposed by extreme collimator's working conditions. An ambitious research program has been undertaken at CERN and collaborating partners to investigate, process and characterize novel composite materials. A promising new family of materials has been identified in metal/ceramiccarbon composites: these are intended to combine optimum mechanical, thermal and electrical properties, such as mechanical strength, melting temperature, thermal shock resistance, electrical conductivity, and energy absorption. Besides High Energy Physics equipment, these materials are of particular interest for thermal management applications such as high power density electronic packaging, aerospace, automotive, nuclear fusion and solar energy. With that in mind, this paper aims at discussing the properties of investigated materials with respect to their relevance for the various application domains.

A new route to 15-hydroxydehydroabietic acid derivatives from abietic acid ( 11 ), via abieta-8,13(15)-dien-18-oic acid ( 12 ), is reported. Utilizing this, the first synthesis of bioactive terpenes 3 , 6 , 9 and 10 , as well as natural diol 4 and lactones 7 – 8 was achieved.

Concepto. La hepatitis virica se define como la inflamacion difusa del parenquima hepatico secundaria a la infeccion por un virus hepatotropo. Clasificacion. Existen cinco virus hepatotropos denominados por las letras A-E, de los que los VHA y VHE se transmiten por via oro-fecal, y los VHB, VHC y VHD por via parenteral y/o sexual y con frecuencia evolucionan a la cronicidad. Clinica. El sindrome general de la hepatitis aguda virica es comun a los distintos tipos con algunas manifestaciones diferenciales * La hepatitis por VHA puede ser autolimitada, aunque a veces se comporta de forma colestasica y recurrente * La hepatitis por VHB puede debutar con manifestaciones diversas extrahepaticas * La infeccion por VHD puede presentarse en el curso de una infeccion aguda o cronica por VHB * La hepatitis aguda por VHC es asintomatica en la mayoria de los casos. Evolucion. Las hepatitis por VHA y VHE nunca cronifican. La hepatitis aguda por VHB pasa a la cronicidad en un porcentaje del 1%-5%. Tanto la hepatitis cronica por VHB como por VHC se han relacionado con el desarrollo de hepatocarcinoma. Tratamiento. La mayoria de las hepatitis agudas pueden ser vigiladas en domicilio. En la hepatitis aguda por VHB con ictericia importante, lamivudina podria disminuir la mortalidad. En la hepatitis por VHA y VHB existen diversas pautas de profilaxis pasiva y activa segun condiciones clinicas.

Epidemiological, viral and host factors are associated with the outcome of hepatitis C virus (HCV) infection, and strong host immune responses against HCV favour viral clearance. Recently, genome-wide association studies have shown a strong correlation between single-nucleotide polymorphisms (SNPs) near the interleukin-28B (IL28B) gene and spontaneous or treatment-induced HCV clearance. We have investigated whether protective IL28B genetic variants are associated with HCV-specific T-cell responses among Spanish blood donors. The rs12979860 IL28B haplotype was determined in 69 anti-HCV-positive blood donors (21 HCV RNA negative and 48 HCV RNA positive) and 30 seronegative donors. In all cases, HCV-specific CD4(+) T-cell responses to HCV recombinant proteins (core, NS3 and NS3 helicase) were assessed by ex vivo interferon-γ ELISpot assay. The rs12979860-CC genotype was highly overrepresented in donors with spontaneous HCV clearance when compared to those with chronic infection (76.2%vs 29.2%, P 0.001; odds ratio, 7.77; 95% confidence interval, 2.4-25.3, P 0.001). HCV-specific CD4(+) T-cell responses were detected in 16 (76.2%) spontaneous resolvers especially towards nonstructural proteins, but with no correlation with IL28B genotype. Chronic individuals had a significantly lower overall T-cell response again irrespective of IL28B genotype. When spontaneous resolvers and chronic individuals were stratified according to their IL28B genotype, significantly stronger T-cell responses were only observed among those with non-CC haplotypes. Although the protective rs12979860 IL28B CC genotype is associated with spontaneous HCV clearance, stronger CD4(+) T-cell responses towards NS3 were only evident among those with non-CC haplotypes.

Hepatitis C virus (HCV)-specific T cell responses are essential for HCV control, and chronic infection is characterized by functionally altered antigen-specific T cells. It has been proposed that the early inactivation of specific CD4(+) T cell responses may be involved in establishment of HCV persistence. We have investigated whether HCV-specific CD4(+) T cells dysfunction can be reversed in vitro. Nonstructural protein 3 (NS3) and core-specific CD4(+) T cells from eight chronically infected and eight spontaneously resolved HCV individuals were selected through transient CD154 (CD40 ligand) expression, and their functional profile (IFN-γ, IL-2, TNF-α, IL-10 and IL-4 production by enzyme-linked immunospot assay, cytometric bead array and intracellular cytokine staining, and proliferation by carboxy-fluorescein diacetate succinimidyl ester dilution assay) was determined both ex vivo and after in vitro expansion of sorted CD154-expressing cells in the absence of specific antigen in IL-7/IL-15-supplemented medium. Ex vivo bulk CD4(+) T cells from chronic patients expressed CD154 in most cases, albeit at lower frequencies than those of resolved patients (0.11%vs 0.41%; P = 0.01), when stimulated with NS3, but not core, although they had a markedly impaired capacity to produce IL-2 and IFN-γ. Antigen-free in vitro expansion of NS3-specific CD154(+) cells from chronic patients restored IFN-γ and IL-2 production and proliferation to levels similar to those of patients with spontaneously resolved infection. Hence, NS3-specific CD4(+) T cell response can be rescued in most chronic HCV patients by in vitro expansion in the absence of HCV-specific antigen. These results might provide a rationale for adoptive immunotherapy.

The in vitro leishmanicidal ( Leishmania infantum and Leishmania braziliensis ) and trypanocidal ( Trypanosoma cruzi ) activities of different compounds were evaluated. These compounds, of vegetal origin but synthesised in our laboratory, included five taiwaniaquinoid derivatives ( S-567 ; S-569 ; S-589; S-602 and A-246) and one abietane quinone ( P-1 ). The in vitro activity of the compounds on extracellular and intracellular forms of the two Leishmania species and T. cruzi was assayed. Infectivity and cytotoxicity tests for the Leishmania species were conducted on J774.2 macrophage cells using Glucantime as the reference drug. From all the compounds assayed, the derivatives P-1 > S-567 were more active and less toxic than Glucantime. Infection rates and amastigote means indicated that these two compounds were the most active in both Leishmania species . In the case of T. cruzi , the best derivatives were P-1 and S-567 , at the same levels as for the Leishmania species. These compounds exhibited the most potent anti-proliferative activity against the extracellular vector form (the epimastigote), the extracellular host form (the trypomastigote), and the intracellular host form (the amastigote), with lower toxicity than that of the reference drug Benznidazole. Metabolite excretion studies showed that alterations mainly at the level of the mitochondria may explain observed metabolic changes in succinate and acetate production, perhaps due to the disturbance of enzymes involved in sugar metabolism within the mitochondrion. The in vivo studies for T. cruzi provided results consistent with those found in vitro. No signs of toxicity were detected in mice treated with the compounds tested, and the parasitic charge was slightly lower than in the control. The effects of these two compounds were also demonstrated with the change in the anti- T. cruzi antibody levels during the chronic stage.

Mammalian cells are widely used in the production of viral vaccines and therapeutic proteins. Most of those processes employ cells growing in suspension; however, some, especially those in vaccine production, use adherent cells. Stirred-tank bioreactors are most widely used for mammalian cell culture processes. When a stirred-tank bioreactor is used for growing adherent cells, a supporting surface for cell attachment is provided by the use of microcarriers or macroporous microcarriers. Alternatively, adherent cells are grown as aggregates in suspension. Fed-batch culture is commonly practised since it reaches high cell and product concentrations. When a stirred-tank bioreactor is operated in a continuous mode, it is a common practice to employ a cell retention device for perfusion in order to increase cell and product concentrations. As mammalian cell culture processes are increasingly being used in the manufacturing, one also witnesses an increasing adoption of disposable (or single-use) cell culture devices. Traditional roller bottles are still often seen. Additionally, multiple flat plates or parallel trays, along with bag type cell growth chambers and even plastic stirred tanks, are used in moderate-scale production and in the seed culture propagation for large-scale bioreactors. Those disposable devices offer simplicity in operations and ease process validation in the production of biopharmaceuticals. Recently, some innovative bioreactors developed nearly two decades ago are finding applications in tissue engineering. Furthermore, the demand for high throughput operations in cell culture processing will continue to drive the innovations in bioreactor technology. Keywords: cell culture; bioreactor; single-use bioreactor; scale-down