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Ante-mortem diagnosis of caprine tuberculosis, as in other ruminant species, is a complex process. Therefore, it is sometimes necessary to apply a combination of methods that increases the sensitivity of diagnosis while maintaining adequate specificity. In the present study, we analysed the use of ELISA based on a double-recognition methodology for the diagnosis of goat tuberculosis. Serum samples from animals with tuberculosis, confirmed by histopathological studies and presence of Ziehl-Neelsen-positive bacteria in the lesions, were used to assess sensitivity. Sera from flocks without tuberculosis were used to assess diagnostic specificity. The relationship between ELISA results and the type of tuberculous lesions, both macroscopic and microscopic, was analysed. The sensitivity and specificity of ELISA test were 69.9% and 97.9%, respectively. Sensitivity was higher in animals with macroscopic lesions, indicating more advanced forms of the disease. Similarly, a higher sensitivity was found in animals with microscopic lesions associated with active tuberculosis, such as cavitary lesions.

Geohazard prone areas require continuous monitoring to detect risks, understand the phenomena occurring in those regions and prevent disasters. Satellite interferometry (InSAR) has come to be a trustworthy technique for ground movement detection and monitoring in the last few years. InSAR based techniques allow to process large areas providing high number of displacement measurements at low cost. However, the results provided by such techniques are usually not easy to interpret by non-experienced users hampering its use for decision makers. This work presents a set of tools developed in the framework of different projects (Momit, Safety, U-Geohaz, Riskcoast) and an example of their use in the Granada Coastal area (Spain) is shown. The ADA (Active Displacement Areas) tool have been developed with the aim of easing the management, use and interpretation of InSAR based results. It provides a semi-automatic extraction of the most significant ADAs through the application ADAFinder tool. This tool aims to support the exploitation of the European Ground Motion Service (EU-GMS), which will provide consistent, regular and reliable information regarding natural and anthropogenic ground motion phenomena all over Europe .

Medicinal plants contain a large variety of chemical compounds in highly variable concentrations, so the quality control of these materials is especially complex. With this purpose, regulatory institutions have accepted chromatographic fingerprints as a valid tool to perform the analyses. In order to improve the results, separation conditions that maximise the number of detected peaks in these chromatograms are needed. This work reports the extension of a simulation strategy, based on global retention models previously developed for selected compounds, to all detected peaks in the full chromatogram. Global models contain characteristic parameters for each component in the sample, while other parameters are common to all components and describe the combined effects of column and solvent. The approach begins by detecting and measuring automatically the position of all peaks in a chromatogram, obtained preferably with the slowest gradient. Then, the retention time for each detected component is fitted to find the corresponding solute parameter in the global model, which leads to the best agreement with the measured experimental value. The process is completed by developing bandwidth models for the selected compounds used to build the global retention model based on gradient data, which are applied to all peaks in the chromatogram. The usefulness of the simulation approach is demonstrated by predicting chromatographic fingerprints for three medicinal plants with specific separation problems (green tea, lemon balm and linden), using several multi-linear gradients that lead to problematic predictions.

The advantages and disadvantages of the use of isocratic experimental designs including transient increments of organic solvent (i.e., pulses) in the mobile phase(s) of lowest elution strength are explored with modelling purposes. For retained solutes, this type of mixed design offers similar or better predictive capability than gradient designs, shorter measurement time than pure isocratic designs, and retention model parameters that agree with those derived from pure isocratic experiments, with similar uncertainties. The predicted retention times are comparable to those offered by models adjusted from pure isocratic designs, and the solvent waste is appreciably lower. Under a practical standpoint, mixed designs including pulse(s) can be easily constructed by replacing the slowest isocratic runs with runs containing a pulse of short duration at an intermediate time. This allows the elution of the fastest solutes with appreciable retention in the initial sector of the elution program, previous to the pulse, and the elution of the slow solutes after the pulse, also in acceptable times. The fitting of the retention data obtained with pulses is simpler compared to gradient elution, and involves solving the integral equation of gradient elution, simplified by the presence of isocratic sectors. Experiments involving pulses reveal the existence of discrepancies in the predictions for solutes eluting in the nearby of the pulse, offered by the fundamental equation of gradient elution when this is solved using numerical integration. The correction of such discrepancies implies the inclusion of intra-column delays, in the arrival of changes in the concentration of organic modifier in the gradient to the instantaneous position of the solute, along the whole migration.

The purpose of this work was to study the potential of recently developed ultra-short column hardware for ion exchange chromatography (IEX). Various prototype and commercial columns having lengths of 5, 10, 15, 20 and 50 mm and packed with non-porous 3 µm particles were systematically compared. Both pH and salt gradient modes of elution were evaluated. Similarly, what has been previously reported for reversed phase liquid chromatography (RPLC) mode, an “on-off” retention mechanism was observed in IEX for therapeutic proteins and their fragments (25–150 kDa range). Because of the non-porous nature of the IEX packing material, the column porosity was relatively low (e = 0.42) and therefore the volumes of ultra-short columns were very small. Based on this observation, it was important to reduce as much as possible all the sources of extra-column volumes (i.e. injection volume, extra-bed volume, detector cell volume and connector tubing volume), to limit peak broadening. With a fully optimized UHPLC system, very fast separations of intact and IdeS digested mAb products were successfully performed in about 1 min using an IEX column with dimensions of 15 × 2.1 mm. This column was selected for high-throughput separations, since it probably offers the best compromise between efficiency and analysis time. For such ultra-fast separations, PEEK tubing was applied to bypass the column oven (column directly connected) to the optical detector via a zero dead volume connection.

The basis of interpretive optimisation in liquid chromatography is the prediction of resolution, from appropriate solute retention models. The reliability of the process depends critically on the quality of the experimental design. This work develops, validates and applies a general methodology aimed to evaluate the quality of any training experimental design, which will be applied in Part II to design optimisation. The methodology is based on the systematic evaluation of the uncertainties associated to the prediction of retention times in comprehensive scans of both isocratic and gradient experimental conditions. It is able to evaluate comprehensively experimental designs of arbitrary complexity. Five common training experimental designs were used to model the retention, according to the Linear Solvent Strength (LSS) and the Neue-Kuss (NK) equations, using a set of 14 sulphonamides of different polarity. The results are presented in terms of relative uncertainties in predictions, which provide significant and interpretable results. The magnitude of such uncertainties, together with the systematic, coherent and logical changes observed at increasing solute hydrophobicity, give support to the results. The NK model gave smaller errors and unbiased predictions, whereas the LSS model gave rise to lack of fit. Isocratic training designs, which are widely accepted as the most informative, are confirmed as the best. As a general conclusion, gradients are predicted with intrinsically smaller uncertainties, independently of the training experimental design. In addition, gradients are more insensitive than isocratic predictions with regard to the type of training design used. Isocratic predictions deteriorate quickly with mobile phase composition. This explains the better performance of gradient predictions, even with biased models.

The search for best conditions in liquid chromatography is routinely carried out with information provided by chemical standards. However, sometimes there are samples with insufficient knowledge about their chemical composition. In other cases, identities of the components are known, but there are no standards available, and in other cases the identities of peaks in chromatograms taken under different conditions are ambiguous. Most resolution criteria used to measure separation performance cannot be applied to these samples. In this work, a global resolution function valid for all situations was developed based on automatic measurements of peak prominences (area fraction exceeding the line that joins the valleys delimiting each peak). Relative performance is evaluated against the peak purity criterion (which measures the area free of overlapping). Peak purity provides a truly comprehensive measurement of global resolution since the underlying signals for each compound are used. However, it is only accessible through in silico simulations. In contrast, peak prominences are not based on a comprehensive knowledge of the individual signals, and can be obtained from experimental chromatograms besides in silico simulations. Therefore, this criterion is suitable for direct evaluation of the resolution of chromatograms with high complexity. A comparison study was carried out based on the agreement of the gradients chosen as Pareto-optimal by both criteria, using information from standards of the 19 primary amino acids found in proteins. The developed global resolution function was applied with success to chromatographic fingerprints of medicinal herbs.

The development of a new comprehensive two-dimensional liquid chromatographic method is described, to obtain the profiles of polyphenolic compounds present in olive (Olea europaea L.) leaves and pulps from different genetic origin. Optimisation of the stationary phase nature, particle size, column length and internal diameter, as well as other separation conditions, was performed. Along the study, three stationary phases (C18, PFP and phenyl) in the first dimension (1D), and five (C18, amide, cyano, phenyl and PFP) in the second dimension (2D) were combined to obtain the maximal number of resolved peaks. The optimised method successfully characterised the presence of 26 and 29 common polyphenols in olive leaves and pulp extracts, respectively. Peak volume ratios were used to develop linear discriminant analysis models able to distinguish olive leaves and pulp extracts among seven cultivars from several Spanish regions. The results demonstrate that polyphenolic profiles were characteristic of each cultivar.

The development of methacrylate-based monolithic columns was studied for the separation of pharmaceutical hydrophilic compounds in nano-liquid chromatography. The selected polymerisation mixture consisted of 7.5% hexyl methacrylate, 4.5% methacrylic acid and 18.0% ethylene dimethacrylate (w/w), in a binary porogenic solvent (35:35 w/w 1-propanol/1,4-butanediol). The polymer synthesised with this mixture has a good permeability, not excessive back-pressure, and reasonable retention times for polar and non-polar solutes. Monolithic columns (12 cm total capillary length, 100 μm i.d.), prepared with this mixture, were able to produce hydrogen bonding and electrostatic interactions, giving rise to promising separations. To evaluate the chromatographic system, alkylbenzenes (neutral and hydrophobic compounds) and sulphonamides (hydrophilic drugs) were assayed. To optimise the chromatographic mobile phase in isocratic elution and characterise the retention mechanism for a mixture of eight sulphonamides, the performance of several mathematic models was checked in the description of retention. The behaviour of the monolithic capillary column was compared, in terms of selectivity and peak shape, to that obtained with a C18 column (9 cm × 4.6 mm i.d., 5 μm particle size) using a conventional HPLC equipment. The results revealed substantial differences in the interactions established for sulphonamides between the monolithic and C18 columns.

The resolution of samples containing unknown compounds of different nature, or without standards available, as is the case of chromatographic fingerprints, is still a challenge. Possibly, the most problematic aspect that prevents systematic method development is finding models that describe without bias the retention behaviour of the compounds in the samples. In this work, the use of global models (able to describe the whole sample) is proposed as an alternative to the use of individual models for each solute. Global models contain parameters that are specific for each solute, while other parameters ‒related to the column and solvent‒ are common for all solutes. A special regression procedure is presented for the construction of global models, which are applied to predict highly complex chromatograms, such as chromatographic fingerprints, for diverse experimental conditions in isocratic and gradient elution. Another interesting application is the prediction of molecular properties, such as log Po/w, from the specific solute parameters of the global models. The examined adapted models are based on the equations proposed by Snyder, Schoenmakers, Neue and Kuss, Jandera, and Bosch Roses to describe the retention. In all cases, the predictive capability was very satisfactory. Two cases of study were considered: chromatograms of camomile extracts analysed using acetonitrile gradients, and a set of 145 known compounds in a wide range of structures and functionalities, eluted isocratically with acetonitrile/water mobile phases.

Micellar liquid chromatography (MLC) is a reversed-phase mode with mobile phases containing an organic solvent and a micellised surfactant. Most procedures developed in MLC are implemented in the isocratic mode, since the general elution problem in chromatography is less troublesome. However, gradient elution may be still useful in MLC to analyse mixtures of compounds within a wide range of polarities, in shorter times. MLC using gradients is attractive to determine by direct injection moderate to low polar compounds in physiological samples. In these analyses, the use of initial micellar conditions (isocratic or gradient) with a fixed amount of surfactant above the critical micellar concentration, keeping the organic solvent content low, will provide better protection of the column against the precipitation of the proteins in the physiological fluid. Once the proteins are swept away, the elution strength can be increased using a positive gradient of organic solvent to reduce the analysis time. This may give rise to the transition from the micellar to the submicellar mode, since micelles are destroyed at sufficiently high concentration of organic solvent. In this work, several retention models covering extended solvent domains in MLC are developed and tested, and applied to investigate the performance in isocratic, linear and multi-linear gradient separations. The study was applied to the screening of β-adrenoceptor antagonists in urine samples, using mobile phases prepared with sodium dodecyl sulphate and 1-propanol. Predicted chromatograms were highly accurate in all situations, although suffered of baseline problems and minor shifts for peaks eluting close to a steep gradient segment. Two columns (C18 and C8) were investigated, with the C8 column being preferable owing to the smaller amount of adsorbed surfactant.

Chlamydia abortus produces ovine enzootic abortion (OEA). Symptoms are not observed until the organism colonises the placenta, eventually causing abortion. Infected animals become carriers and will shed the organism in the following oestruses. This process suggests that sex hormones might play an important role in the physiopathology of OEA, affecting the success of chlamydial clearance and also jeopardising the effectiveness of vaccination. However, the mechanisms through which sex hormones are involved in chlamydial pathogenicity remain unclear. The aim of this study, therefore, was to determine the effect of progesterone on the immune response against C. abortus and on the protection conferred by an experimental inactivated vaccine in sheep. Eighteen sheep were ovariectomised and divided into four groups: vaccinated and progesterone-treated (V-PG), vaccinated and non-treated (V-NT), non-vaccinated and non-treated (NV-NT) and non-vaccinated and progesterone-treated sheep (NV-PG). Animals from both PG groups were treated with commercial medroxyprogesterone acetate impregnated intravaginal sponges before and during the vaccination (V-PG) or just before challenge (NV-PG). The animals from both V groups were subcutaneously immunised with an experimental inactivated vaccine, which was seen to confer high protection in previous studies. All sheep were challenged intratracheally with C. abortus strain AB7 and were sacrificed on day 8 post-infection. Morbidity was measured as the variation in rectal temperature and samples of sera were collected for antibody and cytokine (IFN-γ and IL-10) analysis by commercial ELISA. In addition, lung and lymph node samples were collected for chlamydial detection by qPCR and for histopathological and immunohistochemical analyses. Sheep from the V-PG group showed less severe or no lesions and lower morbidity than the other groups. They also had the highest abundance of regulatory T-cells. The sheep from V-NT also manifested high antibody levels against C. abortus and less severe lesions than those observed in non-vaccinated sheep, which showed high morbidity, low antibody levels and severe lesions, especially in NV-NT. These results confirm the effectiveness of the experimental vaccine employed and suggest that progesterone could enhance the effect.

Peak capacity (PC) is a key concept in chromatographic analysis, nowadays of great importance for characterising complex separations as a criterion to find the most promising conditions. A theoretical expression for PC estimation can be easily deduced in isocratic elution, provided that the column plate count is assumed constant for all analytes. In gradient elution, the complex dependence of peak width with the gradient program implies that an integral equation has to be solved, which is only possible in a limited number of situations. In 2005, Uwe Neue developed a comprehensive theory for the calculation of PC in gradient elution, which is only valid for certain situations: single linear gradients, absence of delays and extra-column effects, Gaussian peaks and constant plate count. Going beyond these limitations implies resolving algebraic expressions that unfortunately cannot be integrated. In this work, PC is predicted for multiple situations based on peak simulation. The approach is more general and can be applied for situations out of the scope of the Neue outline, such as complex multi-linear gradients, including asymmetrical peaks. The plots of PC versus retention time of the last eluted solute give rise to Pareto fronts, and can be useful for the probabilistic enhancement of peak resolution in situations where complex multi-analyte samples are processed.

Pregnant ewes have been widely used to test vaccines against Chlamydia abortus . However, this model entails many disadvantages such as high economic costs and long periods of pregnancy. The murine model is very useful for specific studies but cannot replace the natural host for the later stages of vaccine evaluation. Therefore, a non-pregnant model of the natural host might be useful for a vaccine trial to select the best vaccine candidates prior to use of the pregnant model. With this aim, two routes of infection were assessed in young non-pregnant sheep, namely, intranasal (IN) and intratracheal (IT). In addition, groups of non-vaccinated sheep and sheep immunised with an inactivated vaccine were established to investigate the suitability of the model for testing vaccines. After the experimental infection, isolation of the microorganism in several organs, with pathological and immunohistochemical analyses, antibody production assessment and investigation by PCR of the presence of chlamydia in the vagina or rectum were carried out. Experimental IT inoculation of C. abortus induced pneumonia in sheep during the first few days post-infection, confirming the suitability of the IT route for testing vaccines in the natural host. The course of infection and the resulting pathological signs were less severe in vaccinated sheep compared with non-vaccinated animals, demonstrating the success of vaccination. IN infection did not produce evident lesions or demonstrate the presence of chlamydial antigen in the lungs and cannot be considered an appropriate model for testing vaccines.

The data processing step of complex signals in high-performance liquid chromatography may constitute a bottleneck to obtain significant information from chromatograms. Data pre-processing should be preferably done with little (or no) user supervision, for a maximal benefit and highest speed. In this work, a tool for the configuration of a state-of-the-art baseline subtraction algorithm, called BEADS (Baseline Estimation And Denoising using Sparsity) is developed and verified. A quality criterion based on the measurement of the autocorrelation level was designed to select the most suitable working parameters to obtain the best baseline. The use of a log transformation of the signal attenuated artifacts associated to a large disparity in signal size between sample constituents. Conventional BEADS makes use of trial-and-error strategies to set up the working parameters, which makes the process slow and inconsistent. This constitutes a major drawback in its successful application. In contrast, the assisted BEADS simplifies the setup, shortens the processing time and makes the baseline subtraction more reliable. The assisted algorithm was tested on several complex chromatograms corresponding to extracts of medicinal herbs analysed with acetonitrile-water gradients, and a mixture of sulphonamides eluted with acetonitrile gradients in the presence of the non-ionic surfactant Brij-35 under micellar conditions.

The maintenance of T-cell homeostasis must be tightly regulated. Here, we have identified a coordinated role of Poly(ADP-ribose) polymerase-1 (PARP-1) and PARP-2 in maintaining T-lymphocyte number and function. Mice bearing a T-cell specific deficiency of PARP-2 in a PARP-1-deficient background showed defective thymocyte maturation and diminished numbers of peripheral CD4 + and CD8 + T-cells. Meanwhile, peripheral T-cell number was not affected in single PARP-1 or PARP-2-deficient mice. T-cell lymphopenia was associated with dampened in vivo immune responses to synthetic T-dependent antigens and virus, increased DNA damage and T-cell death. Moreover, double-deficiency in PARP-1/PARP-2 in T-cells led to highly aggressive T-cell lymphomas with long latency. Our findings establish a coordinated role of PARP-1 and PARP-2 in T-cell homeostasis that might impact on the development of PARP-centred therapies., Spanish Ministerio de Economía y Competitividad (MINECO) (SAF2014-53467-R to JY; SAF2010-18917 and SAF2013-48754-C2-1-R to MDV; and BFU2015-68354 to THS), cofinanced by the European Regional Development Fund, Fundació La Marató de TV3 (20134130), and CIBERehd

Many solvents and additives are used to prepare mobile phases in liquid chromatography (LC). Also, mixtures of solvents at different ratios are used to modify the mobile-phase properties. This can make solvent selection for method development a puzzling task, unless suitable guidelines are followed. This chapter summarizes the most common strategies used by skilled chromatographers in reversed-phase, normal-phase, and hydrophilic interaction LC. These are based on considerations about the global polarity of solutes, stationary phase, and mobile phase, which determine the elution strength, and on the particular profile of the contributions of intermolecular interactions to the global polarity, which determines the selectivity. The optimization of selectivity using isoeluotropic mixtures on the rational basis provided by solvent-selectivity profiles and by systematic trial-and-error procedures in the isocratic and gradient elution modes is described. The modification of the selectivity by using combinations of solvents is discussed. Additional considerations for mobile-phase selection, including computer-assisted interpretive optimization, sustainability criteria, and use of high temperature, are also commented.

The addition of reagents to a reversed-phase liquid chromatography (RPLC) mobile phase enables the separation of ionizable compounds, inorganic anions, and metal ions, using conventional instrumentation, silica-based materials, and hydro-organic mixtures, thanks to a variety of secondary equilibria. This gives rise to several chromatographic modes, the main features of which are outlined in this chapter. The effect of the mobile phase pH on the retention of ionizable compounds is described, together with the recommended experimental practice. The mechanism of adsorption of amphiphilic anions or cations on the stationary phase to attract analytes with opposite charge, or suppress the silanol activity, is discussed. Different reagents such as alkylammonium salts, surfactants (below and above the critical micelle concentration or forming microemulsions), perfluorinated carboxylate anions, chaotropic ions, and ionic liquids are considered. The potential of metal chelation and redox reactions for the determination of metal ions and organic compounds are also summarized.

Intragastric infection mimics the natural route of infection of Chlamydia abortus (etiological agent of ovine enzootic abortion). In the mouse model, intragastric experimental infection induces very mild signs of infection followed by late term abortions, as it is shown by the natural ovine host. In order to evaluate the immune mechanisms associated to the dissemination of the pathogen from the gastrointestinal tract, we have administered an intragastric dose of C. abortus to pregnant mice. Systemic and local expression of cytokines, tissue colonization and excretion of bacteria after parturition were monitored during pregnancy. Susceptible CBA/J mice showed a higher bacterial colonization of the placenta and excretion of live bacteria after parturition that were related to a higher local IL-10 expression. By contrast, resistant C57BL/6 mouse strain had higher local IFN-γ mRNA expression in the placenta just before parturition and a transient bacterial colonization of the reproductive tract, with no excretion of C. abortus after parturition. In summary, intragastric infection not only mimics the natural route of infection of C. abortus, but can also be useful in order to understand the immunopathogenesis of chlamydial abortion in the mouse.

Feline mammary carcinomas are highly aggressive neoplasms. Several mechanisms are thought to be involved in their progression, including the loss of epithelial adhesion molecules. The present study was carried out on 21 adenomas and 139 mammary carcinomas. Of the carcinomas, 66 were not reported to have metastasized, while the remaining 73 had evidence of regional lymph node metastasis at the moment of diagnosis. The relationship was examined between the expression of the E-cadherin–β-catenin complex and basal (CK5/6, CK14) and luminal (CK8/18) cytokeratin expression. In the medical literature, carcinomas expressing basal cytokeratins are reported as having a poor prognosis in human breast cancer. Results revealed that preservation of the expression of E-cadherin and β-catenin is a significant feature of carcinomas without metastasis, whereas carcinomas with metastasis reveal the loss of one or both adhesion molecules. Additionally, basal cytokeratin expression was statistically associated with the presence of regional metastasis. Furthermore, the expression of E-cadherin–β-catenin was significantly correlated with the high expression of CK18 and low expression of CK5/6.

Caprine tuberculosis is caused by bacteria of the Mycobacterium tuberculosis complex (Mycobacterium bovis and Mycobacterium caprae). Although typical tuberculoid granulomata are usually observed in the lungs and lymph nodes of infected goats, the presence of cavitary lesions with exuberant mycobacterial growth is also a common feature in this species. The aim of this study was to characterize the immunological mechanisms that lead to liquefaction and cavity formation by comparing granulomata and cavitary lesions. Samples from animals positive by skin testing were collected for microscopical and immunohistochemical examination. Samples were also collected for analysis of cytokine gene expression in the lesions by real time polymerase chain reaction. There were marked differences between granulomata and cavitary lesions. In cavitary lesions there was a substantial population of neutrophils and a significant decrease in the number of CD4(+) T cells, with concomitant increases in other T-cell populations (CD8(+) and cells expressing the γδ form of the T-cell receptor). The enzyme iNOS was strongly expressed by macrophages in the cavitary lesions. There was no difference in the balance of pro- and anti-inflammatory cytokine gene expression in the lesions. These findings suggest that cavitary lesions are reactivation sites, where conditions are optimal for Mycobacterium proliferation and that immunological mechanisms may underlie the severe destruction of lung tissue that characterizes the cavitary pathology.

Using thermoluminiscent dosimeters the absorbed dose in the bladder, rectum and thyroid have been evaluated when 200 cGy was applied to the prostate. The treatment was applied with a (60)Co unit. A water phantom was built and thermoluminiscent dosimeters were located in the position where the prostate, bladder, rectum and thyroid are located. The therapeutic beam was applied in 4 irradiations at 0, 90, 180 and 270° with the prostate at the isocenter. The TLDs readouts were used to evaluate the absorbed dose in each organ. The absorbed doses were used to estimate the effective doses and the probability of developing secondary malignacies in thyroid, rectum and bladder.

Chlamydophila (C.) abortus is an obligate intracellular bacterium able to colonize the placenta of several species of mammals, which may induce abortion in the last third of pregnancy. The infection affects mainly small ruminants resulting in major economic losses in farming industries worldwide. Furthermore, its zoonotic risk has been reported in pregnant farmers or abattoir workers. Mouse models have been widely used to study both the pathology of the disease and the role of immune cells in controlling infection. Moreover, this animal experimental model has been considered a useful tool to evaluate new vaccine candidates and adjuvants that could prevent abortion and reduce fetal death. Future studies using these models will provide and reveal information about the precise mechanisms in the immune response against C. abortus and will increase the knowledge about poorly understood issues such as chlamydial persistence.

Resumen Objetivo Valorar la eficacia de los bloqueos nerviosos como tecnica alternativa de analgesia postoperatoria en las protesis de rodilla, senalando la validez y ventajas del abordaje por via anterior del bloqueo ciatico. Material y metodo Estudiamos un grupo de pacientes consecutivos, desde abril de 2004 a marzo de 2006, intervenidos de artroplastia de rodilla, utilizando el bloqueo subaracnoideo como tecnica anestesica y los bloqueos perifericos combinados femoral y ciatico mediante su abordaje anterior como tecnica analgesica postoperatoria. Valoramos el tiempo medio libre de dolor, calidad de la analgesia y estancia hospitalaria. Resultados Incluimos 78 pacientes. El intervalo libre de dolor del conjunto de todos los pacientes fue de 42,1±3,9 h. Los pacientes que refirieron dolor leve, fue a las 34,8±4,1 h y dolor moderado-intenso a las 42,4±3,5 h. Alcanzaron 90° de flexion de la rodilla, al tercer dia, el 62,8% de los pacientes. No hubo complicacion derivada de la tecnica y el grado satisfactorio de los pacientes fue alto. Conclusiones El bloqueo combinado femoral y ciatico en cirugia protesica de la rodilla es eficaz, controla el dolor postoperatorio, permite una rehabilitacion precoz. El abordaje anterior del ciatico es relativamente sencillo de realizar sin retirar el vendaje compresivo del muslo tras la cirugia y ademas evita movilizar al paciente.

Mouse models have been widely used to test candidate vaccines against Chlamydophila abortus infection in mice. Although the induction of a systemic infection by endogenous or intraperitoneal inoculation is a useful tool for understanding the immune mechanism involved in the protection conferred by the vaccination, a different approach is necessary to understand other factors of the infection, such as mucosal immunity or the colonization of target organs. To test whether C. abortus intranasal model of infection in mice is a useful tool for testing vaccines in a first group of experiments mice, were infected intranasally with C. abortus to characterize the model of infection. When this model was used to test vaccines, two inactivated experimental vaccines, one of them adjuvated with QS-21 and another with aluminium hydroxide, and a live attenuated vaccine (strain 1B) were used. Non-vaccinated control mice died within the first 8 days, after displaying substantial loss of weight. Histologically, the mice showed lobar fibrinopurulent bronchointerstitial pneumonia. Prior immunization with QS-21 adjuvated vaccine or 1B vaccine presented mortality and the recipients showed a greater number of T cells in the lesions, especially CD8+ T cells, than the control mice and mice immunized with vaccine adjuvated with aluminium hydroxide. The results confirm that the C. abortus intranasal model of infection in mice is a useful tool for testing vaccines

Monoclonal antibodies (mAbs) have become a successful therapeutic approach in cancer. However, some patients do not achieve long-term clinical benefit and most mAbs only exert modest effects as monotherapies. Therefore, combinations with chemotherapy are currently being investigated. Emerging studies have shown a synergistic therapeutic effect of PARP inhibitors and mAbs in cancer. PARP enzymes catalytically cleave β-NAD+ and transfer the ADP-ribose moiety to acceptor proteins, modifying their function. In here, we update recent data about the therapeutic effect of the combination of PARP inhibitors with mAbs in cancer treatment and discuss the molecular mechanisms involved in this synergy.

Presentamos dos casos de afectación pulmonar por BCG diagnosticados recientemente en nuestro servicio. Un primer paciente varón de 81 años de edad que desarrolló un cuadro de neumonitis intersticial granulomatosa severa en el contexto de un tratamiento mediante instilación endovesical de BCG por tumor vesical de alto grado. Este paciente fue tratado mediante corticoterapia y triple asociación de antituberculosos con una respuesta rápida y completa. El segundo caso fue un hallazgo radiológico en un paciente de 56 años que presentaba fiebre persistente como único síntoma y que mejoró tras iniciar doble asociación de tuberculostáticos.

The increased susceptibility during pregnancy to certain pathogens that cause abortions may be related to changes in the distribution and phenotype of lymphocyte subpopulations in the uterus. Histological, electron microscopic and immunocytochemical techniques were used in this study to examine whether such variations occur in different stages of the reproductive cycle of goats. The study of non-pregnant goats showed that most uterine lymphocytes were T cells and displayed both an intraepithelial and stromal distribution. CD8 + T lymphocytes were more numerous than CD4 + T lymphocytes. In the endometrial epithelium two lymphocyte subpopulations were observed: non-granulated CD2 + CD8 + T lymphocytes and granulated CD2 + CD8 ) T lymphocytes. During gestation, no lymphocytes were observed in the placentomal area, while a decreased number of T lymphocyte subpopulations were found in the inter-placentomal area. In the inter-caruncular epithelium, non-granulated CD2 + CD8 + T lymphocytes disappeared, whereas the granulated CD2 + CD8 ) T lymphocyte subpopulations increased their number and changed their morphology.

This work presents a novel system for sensing electric signals using an electrooptic modulator and optical coherence modulation. The system consists of an integrated optics coherence modulator acting as electric signal sensor in the frequency range up to 900 MHz. The main features of coherence-modulated transmissions include the need of using broad-band optical sources and lithium niobate electrooptic retarders. When low-coherence optical sources are used, the retarders introduce optical delays which are modulated by detected electric signals. The optical delays carry electric signals and are transmitted in the optical channel to a receiver. In this case, a scanning Michelson interferometer is used as an optical receiver, which measures the autocorrelation of light. The sensed electric signal is obtained when the interferometer is matched with optical delays on the detecting devices. Coherence-modulated systems have been studied for the last few years as new potential high-speed optical links useful for local area networks, point-to-point, and bidirectional transmissions.

Resumen Es dificil diferenciar entre tumor de celulas germinales retroperitoneal primario y enfermedad metastasica de un tumor gonadal indetectado o que ha regresado espontaneamente. Presentamos un caso clinico de “burn out” o tumor quemado, es decir un tumor retroperitoneal seminomatoso, metastasico con importantes alteraciones testiculares bilaterales tanto ecograficas como anatomopatologicas, pero sin evidenciar tumor. Pese a las controversias que detallamos, nuestra opcion terapeutica fue la extirpacion de la masa residual tras la quimioterapia asociada a orquiectomia bilateral en el mismo acto. Concluimos que ante un tumor retroperitoneal, la existencia de alteraciones ecograficas en los testiculos, pese a una palpacion normal, debe hacer pensar en un “Sindrome de burn out”, siendo obligatoria la biopsia testicular bilateral. La exploracion quirurgica de los mismos e incluso la orquiectomia son opciones a valorar en funcion de cada paciente.

A Chlamydophila abortus-induced abortion model was carried out on the basis of the experimental infection of ewes at day 75 of gestation. The infection induced abortions and the birth of weak lambs during the last 3 weeks of pregnancy. To study the kinetics of the infection in the placenta and in other organs, infected ewes were killed at 105, 120, and 130 days of gestation and also several days after abortion or parturition. Infected ewes developed a systemic infection that caused a mild and transient pneumonia and focal hepatitis. Pathologic changes were observed in placentas at 120 day of gestation, although the lesions varied between animals and even between placentomes of the same placenta. The first placental area infected was the maternal stroma and epithelium next to the intercaruncular areas, where neutrophilic response seemed to control the infection. A substantial degree of multiplication of C. abortus was then observed in the trophoblast cells of the placentome, periplacentomal choriallantoic membranes, and hilius, with an inflammatory exudate composed mainly of neutrophils, some macrophages, and very scarce lymphocytes. After abortion, the lesions affected the intercotyledonary areas of the aborted placentas, whereas in the uterus significant lymphocyte infiltration was observed, together with a rapid decrease of the C. abortus antigen in the degenerated caruncular tissues.

RESUMEN El Registro Nominal de Vacunaciones de la Region de Murcia esta implantado desde 1991. Utiliza como base de datos poblacional a las personas residentes en nuestra Comunidad nacidas hasta 1938 (> 65 anos) y desde 1991 hasta la fecha. Cada dosis administrada se registra con los siguientes datos: fecha de administracion, puesto de vacunacion, fabricante, lote, indicacion y reaccion adversa si se produce. La calidad de los datos registrados es fundamental en un registro nominal, mas aun en lo concerniente a la seguridad vacunal e identificacion exacta de los preparados y lotes administrados. La base de datos incluye a un total de 449.918 personas y 2.429.861 dosis de vacunas registradas como administradas. Se analizaron las dosis administradas entre el 1 de enero de 1991 y el 31 de agosto de 2003. Se realizo un estudio descriptivo de los 2 campos estudiados (fabricante y lote), en funcion del ano de administracion (quinquenios) y del centro gestor de vacunas (hay 4 en la region). El porcentaje de registros con el campo fabricante "vacio" es del 2,85% y "desconocido", del 2,26% para toda la region. El porcentaje de registros con lote en "blanco" es de un 39% y el porcentaje de registros con lote "desconocido" alcanza un 2,4%. Urge un aumento de la motivacion del personal implicado y una serie de mejoras en el proceso de introduccion de datos. El empleo de etiquetas con codigos de barras en las vacunas para su mecanizacion mediante lectores opticos podria ser una herramienta para mejorar la calidad del proceso.

Objetivo: La disponibilidad de una vacuna neumocócica conjugada plantea la necesidad de conocer la incidencia y características de la enfermedad neumocócica invasiva en niños de la Región de Murcia, con la finalidad de obtener información que pueda ser de utilidad para establecer adecuadamente las indicaciones de vacunación. Métodos: Se ha realizado una búsqueda retrospectiva de casos de enfermedad invasiva por Streptococcus pneumoniae en menores de 15 años de edad atendidos en hospitales de la Región de Murcia durante el período 1991-2000. Las fuentes de datos utilizadas han sido las bases de datos de los Servicios de Microbiología, el Conjunto Mínimo de Datos Básicos, registro de altas pediátricas y Registro EDO. Resultados: La tasa de incidencia para el período 1996-2000 fue, para los menores de 1 año, de 18,25 por 10(5) personas-año en el caso de enfermedad invasiva (10,6 para meningitis); para los menores de 2 años, de 13,6 para enfermedad invasiva (6 para meningitis); para menores de 5 años, de 8,9 (1,35 para meningitis), y para los menores de 15 años, de 3,7 (1,3 para meningitis). El 28% de los casos presentaba factores de riesgo. Las complicaciones alcanzaron el 35,2% y las secuelas el 5%. La letalidad fue del 11,8%. Los serogrupos prevalentes fueron el 19, el 6, el 18, el 5, el 14 y el 23. Conclusiones: El alto porcentaje de casos con factores de riesgo de enfermedad neumocócica invasiva aconseja la implantación de programas de vacunación dirigidos a todos los niños con factores de riesgo. La incidencia de enfermedad neumocócica invasiva encontrada en la Región de Murcia difiere de la de otras zonas geográficas; sin embargo, la incidencia de meningitis es similar a la de otros estudios. La gravedad de la enfermedad justifica la realización de estudios coste-efectividad para valorar la posible incorporación de la vacuna en el calendario vacunal.

We have analyzed a set of 27 patients diagnosed of penile cancer who have been treated in our Hospital during the last 17 years (since january 1984 to december 2000). The annual incidence was set in 1.7 patients/100,000 men/year. The average age has been 64 years (range 42-85). Patients delayed medical consulting of their lesions for 15 months (2-120). Histologic analysis found an epidermoid carcinoma in 19 patients, and verrucous carcinoma in 8. The average follow-up has been 48 months (range 2-120). As a conclusion we don't belive necessary prophylactic inguinal lymphadenectomy. Correct followup, for early detection of lymph nodes does not worsen survival and it avoids unnecessary operations with a high rate of morbidity. The prognostic factors in our patients have been the presence of lymph nodes and the degree of local extension. We lack of experience with radiotherapy and chemotherapy.

Resumen Las migraciones suponen hoy dia uno de los mayores retos que tienen que afrontar los paises que gozan de una estabilidad politica y de una situacion de medio-alto desarrollo economico y social. Es primordial no solamente preservar la salud de los ciudadanos de los paises receptores sino tambien, e igual de importante, proporcionar a los inmigrantes los medios que les ayuden a alcanzar un optimo estado de salud. En este sentido, la prevencion de las enfermedades infecciosas mediante la vacunacion adquiere un protagonismo especial. En este articulo se aborda la situacion reciente de las enfermedades prevenibles por vacunacion, en regiones o zonas geograficas de emigracion preferente a Espana, las pautas de actuacion que se pueden adoptar y, para finalizar, cuales son las estrategias de comunidad con altas tasas de inmigracion.

Resumen Objetivo Estudio de coste-efectividad acerca de la introduccion de la vacuna conjugada frente al meningococo C de acuerdo con varias estrategias, todas ellas valoradas en el seno del Consejo Interterritorial del Sistema Nacional de Salud, en el«Acuerdo de Intenciones de la Comision de Salud Publica sobre la utilizacion de la vacuna conjugada contra la Meningitis meningococica C». Sujetos y metodos Se ha seleccionado la incidencia de Navarra por ser la comunidad con serie mas larga de diagnostico por serogrupo que conocemos en el pais (temporadas 1991-1992 a 1998-1999) y donde no se ha modificado la incidencia reciente por la accion de la vacuna. Resultados La mejor relacion coste-efectividad de la vacunacion antimeningococica frente al serogrupo C se observa situando como poblacion diana a ninos comprendidos entre 1 y 5 anos de edad. Asi, para evitar un caso de enfermedad meningococica en un nino de ese grupo de edad serian necesarias 850 dosis de vacuna. En el caso de los recien nacidos, necesitariamos 2.114 dosis, mientras que vacunando a ninos y jovenes de 6 a 15 anos unicamente precisariamos administrar 1.917 dosis. Este numero de dosis por caso evitado se traduciria en un coste de 3.196.000 ptas. (19.210 A), 7.948.000 ptas. (47.776 A) y 7.207.920 ptas. (43.324 A), respectivamente. Conclusiones La mejor relacion coste-efectividad de la vacunacion antimeningococica frente al serogrupo C se origina vacunando a ninos de 1 a 5 anos de edad, grupo de edad incluido en la campana de vacunacion. De las estrategias estudiadas, la que ocupa el segundo lugar en terminos de coste-efectividad es la vacunacion a ninos y jovenes de 6 a 15 anos de edad. Sin embargo, las campanas disenadas hasta ahora por las diferentes comunidades autonomas no incluyen a estos ninos y adolescentes. Asi, la vacunacion de recien nacidos es un 10% menos coste-efectiva que la anterior, ocupa el tercer lugar en terminos de coste-efectividad y esta incluida en todos los nuevos calendarios vacunales.Como alternativa a una campana de vacunacion activa, en el grupo de edad de 6 a 15 anos, se plantearia, estando la vacuna disponible en farmacias, la vacunacion individualizada mediante receta y visado posterior, con todos los inconvenientes de falta de control y equidad que se derivan de ello.

The androgen insensitivity syndrome is the most frequent form of masculine psedohermafroditism. The affected patients present female phenotype without sexual ambiguity but with karyotype 46 XY. In this syndrome the frequency of malignizacion of the testicles increases progressively with the age, because of this, the importance of an earlier diagnosis. We present a case of later diagnosis late of the androgen insensitivity syndrome, that debut with a great inguinal mass.

Resumen Objetivos Aunque las coberturas de vacunacion en nuestra comunidad son muy elevadas, es necesario seguir contando con un alto grado de participacion de la poblacion, para lo que es preciso conocer su nivel de conocimientos, creencias y fuentes de informacion que tienen sobre las vacunas, asi como el grado de satisfaccion que les reporta el Programa Regional de Vacunaciones. Metodos Estudio descriptivo transversal, mediante dos encuestas postales dirigidas a dos muestras representativas de padres de ninos recien nacidos y de ninos que habian cumplido 18 meses, de la region de Murcia. Resultados Casi el 99% de ambos grupos consideran que las vacunas siguen siendo necesarias. Practicamente el 100% de los en-cuestados vacunaron a sus hijos dentro del Sistema Publico. Los conocimientos generales sobre las enfermedades vacunables y vacunas son, en general, suficientes, aunque un 57,6% en el primer grupo y un 41,8% del segundo los consideran insuficientes. El pediatra fue la fuente informante mas destacada; por el contrario, otro personal sanitario y los medios de comunicacion audiovisual obtuvieron el porcentaje mas bajo. Aunque el 87% de los padres reconocen que las reacciones a las vacunas son leves, solo alrededor del 40% manifiestan estar tranquilos y sin miedo al ir a vacunar a sus hijos por primera vez. Una gran mayoria vacunarian a sus hijos en verano; pero no si estan resfriados y/o con fiebre. Los aspectos mejor valorados del programa de vacunaciones fueron la gestion y el trato dispensado en los centros de salud. Conclusiones Aunque el nivel de conocimiento sobre las vacunas en nuestra poblacion es elevado y las actitudes son positivas, debemos insistir mas en la transmision de informacion a los padres.

Cases of uterine perforation through to the bladder during the placement of an intrauterine device are rare. There have been only 25 previous reported cases. This type of perforation is no different to any other by an intravesical foreign body. A detailed anamnesis and sonographic examination is usually sufficient to provide a diagnosis. Complications can be diagnosed by an uretrocytoscopy which is also therapeutic in non-complicated cases. If there are associated complications surgery can be necessary. In conclusion, quick detection of any complication, by sonographic detection after placement, assists in the extraction of the foreign body and avoids the appearance of associated complications such as lithiasis.