Co-expression of the Bcl-xL antiapoptotic protein enhances the induction of Th1-like immune responses in mice immunized with DNA vaccines encoding FMDV B and T cell epitopes

Foot-and-mouth disease (FMD) is one of the most devastating animal diseases, affecting all cloven-hoofed domestic and wild animal species. Previous studies from our group using DNA vaccines encoding FMD virus (FMDV) B and T cell epitopes targeted to antigen presenting cells, allowed demonstrating total protection from FMDV homologous challenge in those animals efficiently primed for both humoral and cellular specific responses (Borrego et al. Antivir Res 92:359-363, 2011). In this study, a new DNA vaccine prototype expected to induce stronger and cross-reactive immune responses against FMDV which was designed by making two main modifications: i) adding a new B-cell epitope from the O-serotype to the B and T-cell epitopes from the C-serotype and ii) using a dual promoter plasmid that allowed inserting a new cistron encoding the anti-apoptotic Bcl-xL gene under the control of the internal ribosomal entry site (IRES) of encephalomyocarditis virus aiming to increase and optimize the antigen presentation of the encoded FMDV epitopes after in vivo immunization. In vitro studies showed that Bcl-xL significantly prolonged the survival of DNA transfected cells (p?<?0.001). Accordingly, vaccination of Swiss out-bred mice with the dual promoter plasmid increased the total IgG responses induced against each of the FMDV epitopes however no significant differences observed between groups. The humoral immune response was polarized through IgG2a in all vaccination groups (p?<?0.05); except peptide T_3A; in correspondence with the Th1-like response observed, a clear bias towards the induction of specific IFN-γ secreting CD4⁺ and CD8⁺ T cell responses was also observed, being significantly higher (p?<?0.05) in the group of mice immunized with the plasmid co-expressing Bcl-xL and the FMDV B and T cell epitopes.     

Seasonal Evolution of Levels of Gaseous Pollutants in an Urban Area (Ciudad Real) in Central-Southern Spain: A Doas Study

Long term continuous monitoring measurements of urban atmospheric concentrations of O₃, NO₂, NO, and SO₂ were performed for the first time in Ciudad Real, a city in central-southern Spain. The measurements were carried out using the differential optical absorption spectroscopy (DOAS) technique, with a commercial system (OPSIS, Lund-Sweden), covering the summer and winter seasons (from 21st July 2000 to 23rd March 2001). Mean levels of O₃, NO₂ and SO₂ monitored during this period were: 27 μg m^?3, 50 μg m^?3 and 7 μg m^?3 respectively. The highest hourly averaged value of O₃ (160 μg m^?3) was measured during the summer period, while NO₂ was enhanced in wintertime (highest values 90 μg m^?3). In the coldest period, when central heating installations were operating, SO₂ showed maximum levels of 20 μg m^?3. The daily, weekly and seasonal analysis of the data shows that photochemical air pollution dominates in this urban atmosphere and is strongly influenced by levels of motor traffic and domestic heating system emissions. These measurements were compared with other studies in Spain and Europe. Also, the long path averaged DOAS measurements were compared with in situ observations made in Ciudad Real, from 23rd August 2000 to 25th September 2000, using a mobile air pollution control station. All gas concentrations reported in this paper are below the WHO guidelines and the different thresholds introduced by the European Environmental Legislation.     

High level of mislabeling in Spanish and Greek hake markets suggests the fraudulent introduction of African species

DNA analysis of hake products commercialized in southern European (Spanish and Greek) market chains have demonstrated more than 30% mislabeling, on the basis of species substitution. Tails and fillets were more mislabeled than other products, such as slices and whole pieces. African species were substitute species for products labeled as American and European species, and we suggest it is a case of deliberate economically profitable mislabeling because real market prices of European and American hake products are higher than those of African in Spanish market chains. The presented results suggest fraud detection that disadvantages African producers. Government-mandated genetic surveys of commercial hakes and the use of subsequent statements of fair trade on labels of seafood products could help to reduce fraud levels in a global market of increasingly conscious consumers sensitive to ethical issues.     

Connections between the internal and the external capsules and the globus pallidus in the sheep: A dichromate stain X-ray microtomographic study

Sheep are recognized as useful species for translational neurodegeneration research, in particular for the study of Huntington disease. There is a lack of information regarding the detailed anatomy and connections of the basal ganglia of sheep, in normal myeloarchitectonics and in tract-tracing studies. In this work, the organization of the corticostriatal projections at the level of the putamen and globus pallidus (GP) are explored. For the first time, the myeloarchitectonic pattern of connections between the internal (IC) and the external (EC) capsules with the GP have been investigated in the sheep. Formaldehyde-fixed blocks of the striatum were treated with a metallic stain containing potassium dichromate and visualized using micro-CT (µ-CT). The trivalent chromium (Cr3+), attached to myelin phospholipids, imparts a differential contrast to the grey and white matter compartments, which allows the visualization of myelinated fascicles in µ-CT images. The fascicles were classified according to their topographical location in dorsal supreme fascicles (X, Y, apex) arising from the IC and EC; pre-commissurally, basal fascicles connecting the ventral part of the EC with the lateral zone of the ventral pallidum (VP) and, post-commissurally, superior (Z₁), middle (Z₂) and lower (Z₃) fascicles, connecting at different levels the EC with the GP. The results suggest that the presumptive cortical efferent and afferent fibres to the pallidum could be organized according to a dorsal to ventrolateral topography in the sheep, similar to that seen in other mammals. The proposed methodology has the potential to delineate the myeloarchitectonic patterns of nervous systems and tracts.     

Immunomodulatory effect of swine CCL20 chemokine in DNA vaccination against CSFV

The objective of this work was to explore whether a plasmid expressing CCL20 chemokine could improve the immune response against CSFV in co-administration with a DNA vaccine expressing the E2 protein. The immunization of pigs with the DNA vaccine formulation, that contains swine CCL20 chemokine, resulted in the homogenous induction of detectable levels of CSFV antibodies at 36 days after the first injection. Remarkably, immunized animals with E2 DNA vaccine in co-administration with the plasmid containing swine CCL20 developed high titers of neutralizing antibodies against homologous and heterologous CSFV strains and were totally protected upon a lethal viral challenge (sterilizing protection). Our results confirm the role of CCL20 to increase antibody-mediated responses. At the same time suggest the ability of CCL20 to enhance the T helper cell response associated with the induction of neutralizing antibodies against CSFV in pigs previously reported. Systemic replication of virulent CSFV in vivo during the acute phase of infection induces type I IFN. Lower average values of IFN alpha were detected in the serum of pigs immunized with pE2 and pCCL20 at 3 days after challenge. The levels of IFN-alpha detected in pigs immunized with pE2 and principally in non-vaccinated challenged animals can be related to viral load in serum at 3 and 7 days post infection and the clinical signs observed. Our results emphasized the capacity of swine CCL20 chemokine to enhance cellular, humoral and anti viral response with an adjuvant effect in the immune response elicited by E2-DNA vaccination against CSFV. To our knowledge, this is the first report demonstrating the adjuvant effect of swine CCL20 to effectively enhance the potential of DNA vaccine in the immune induction and protection against virus challenge in swine infection model.     

Comparison Between Industrial�CUrban and Rural Particle Stations in a Ceramic Cluster (NE, Spain)

Since the beginning of the twentieth century, the province of Castellón, in eastern Spain, has been one of the most important areas of floor and wall tile production on both a national and worldwide scale. As a result of this sector??s productive rate of development (in the 1970?C2005 interval), a series of pollution-producing chemical substances has been introduced into the atmosphere. The objective of this study is the comparison between industrial?Curban and rural zones in this province, with the goal of establishing the causes of the contamination in order to apply corrective measures upon the different emission sources. Sustainable development is sought after to guarantee that, in the future, the environmental quality parameters fall within legal limits, ensuring the population??s well-being as well as conserving natural ecosystems and material assets. Total suspended particles and PM10 are the parameters studied. The reason for choosing this pollutant type is because particulate matter may present a much higher potential risk despite its low representativeness as compared to the gas pollutant??s group. A positive correlation between high particle concentrations and deterioration in public health has been shown in recent studies. The elements As, Cd, Ni, and Pb in PM10 were also analyzed to determine the toxicity of these particles. This study has demonstrated the different behaviors of the parameters studied at different types of stations (industrial?Curban and rural), leading to the conclusion that anthropogenic factors are very important in the area studied and that they determine the area??s air quality to a great extent.     

Die Pflanzenläuse (Aphidina) der Nadelhölzer (Coniferae) in Spanien

A view is given of the species of Aphidina (families Adelgidae, Mindaridae and Lachnidae) of conifers in Spain. Together with the list of species informations are given on the knowledges made hitherto especially on distribution, outbreaks and damages of the most important species.     

Production of pantothenic acid and thiamine by Azotobacter vinelandii in a chemically defined medium and a dialysed soil medium

The production of pantothenic acid and thiamine by Azotobacter vinelandii was studied in a chemically defined medium and a dialysed soil medium amended with different amounts of carbohydrate and NH _inf4 ^sup+ . Pantothenic acid and thiamine were produced in large amounts in the culture medium containing 2.0–5.0% glucose and 0.1–0.3% NH₄Cl. The results showed that the liberation of these vitamins was increased under adiazotrophic conditions and with an excess of C source.Group of Environmental Microbiology, Department of Microbiology, Faculty of Pharmacy and Institute of Water Research, University of Granada, 18071 Granada, Spain.     

Abrupt temperature changes in the Western Mediterranean over the past 250,000 years

A continuous high-resolution Western Mediterranean sea surface temperature (SST) alkenone record spanning the past 250,000 years shows that abrupt changes were more common at warming than at cooling. During marine isotope stage (MIS) 6, SST oscillated following a stadial-interstadial pattern but at lower intensities and rates of change than in the Dansgaard/Oeschger events of MIS 3. Some of the most prominent events occurred over MISs 5 and 7, after prolonged warm periods of high stability. Climate during the whole period was predominantly maintained in interglacial-interstadial conditions, whereas the duration of stadials was much shorter.