Kainate receptor-mediated depression of glutamatergic transmission involving protein kinase A in the lateral amygdala

Kainate receptors (KARs) have been described as modulators of synaptic transmission at different synapses. However, this role of KARs has not been well characterized in the amygdala. We have explored the effect of kainate receptor activation at the synapse established between fibers originating at medial geniculate nucleus and the principal cells in the lateral amygdala. We have observed an inhibition of evoked excitatory postsynaptic currents (eEPSCs) amplitude after a brief application of KARs agonists KA and ATPA. Paired-pulse recordings showed a clear pair pulse facilitation that was enhanced after KA or ATPA application. When postsynaptic cells were loaded with BAPTA, the depression of eEPSC amplitude observed after the perfusion of KAR agonists was not prevented. We have also observed that the inhibition of the eEPSCs by KARs agonists was prevented by protein kinase A but not by protein kinase C inhibitors. Taken together our results indicate that KARs present at this synapse are pre-synaptic and their activation mediate the inhibition of glutamate release through a mechanism that involves the activation of protein kinase A.     

The Epstein-Barr virus (EBV)-induced tumor suppressor microRNA MiR-34a is growth promoting in EBV-infected B cells

Epstein-Barr virus (EBV) infection of primary human B cells drives their indefinite proliferation into lymphoblastoid cell lines (LCLs). B cell immortalization depends on expression of viral latency genes, as well as the regulation of host genes. Given the important role of microRNAs (miRNAs) in regulating fundamental cellular processes, in this study, we assayed changes in host miRNA expression during primary B cell infection by EBV. We observed and validated dynamic changes in several miRNAs from early proliferation through immortalization; oncogenic miRNAs were induced, and tumor suppressor miRNAs were largely repressed. However, one miRNA described as a p53-targeted tumor suppressor, miR-34a, was strongly induced by EBV infection and expressed in many EBV and Kaposi's sarcoma-associated herpesvirus (KSHV)-infected lymphoma cell lines. EBV latent membrane protein 1 (LMP1) was sufficient to induce miR-34a requiring downstream NF-κB activation but independent of functional p53. Furthermore, overexpression of miR-34a was not toxic in several B lymphoma cell lines, and inhibition of miR-34a impaired the growth of EBV-transformed cells. This study identifies a progrowth role for a tumor-suppressive miRNA in oncogenic-virus-mediated transformation, highlighting the importance of studying miRNA function in different cellular contexts.     

Grain Yield and Charcoal Rot Resistance Stability in Common Beans under Terminal Drought Conditions

Charcoal rot (Macrophomina phaseolina) is a major disease of beans (Phaseolus vulgaris L.) in Mexico. The use of germplasm combining high‐yield stability with resistance to drought and charcoal rot could reduce damage from this disease. In this study, we compared the Eberhart and Russell method and the Additive Main Effect and Multiplicative Interaction (AMMI) model plus biplot analysis for measuring grain yield (GY) and charcoal rot resistance (CHRR) stabilities in 98 F_{8 : 10} recombinant inbred lines (RILs) derived from a cross between bean adapted to the tropics (BAT) 477 (resistant) × Pinto UI‐114 (susceptible). Experiments were conducted from 2007 to 2009 in Isla, Cotaxtla, Río Bravo and Díaz Ordaz, México, under irrigated or terminal drought conditions. anova detected significant differences (P ≤ 0.05) in GY and CHRR among environments, genotypes and genotype × environment interactions (GEI). Most RILs showed good responses to unfavourable environments based on GY (48) and CHRR (40). AMMI anova s for both traits showed that all sources of variation in the model accounted for approximately 49% of the total squared sum. For the first principal component (PC1), we found 13 RILs that were stable for GY, and for the second (PC2), we found 9 that were stable for GI. For CHRR, we detected 14 stable RILs (PC1) and eight (PC2). Biplot analysis showed the largest vectors for Díaz Ordaz (irrigated and drought, 2008), where the highest and most variable GYs were detected. The shortest vectors were found in Isla (drought, 2007) and Río Bravo (irrigated and drought, 2008), where the lowest and least variable GY were found. We found differential responses of RILs to locations, years and soil humidity conditions as well as significant GEI based on GY and CHRR. The two methods were complementary, and both gave us information to select stable, high‐yield germplasm associated with resistance to charcoal rot disease.     

Integrated archaeobotanical research into vegetation management and land use in El Llano de la Horca (Santorcaz, Madrid, central Spain)

This paper presents an integrated archaeobotanical analysis carried out at the site known as “El Llano de la Horca”, a Bronze and Late Iron Age (3rd–1st century b.c.) Carpetanian settlement in central Spain (3600?±?80 b.p.). Pollen and spore analysis indicates an open landscape dominated by the herb taxa Asteraceae, Chenopodiaceae and Poaceae with an important representation of Glomus. The combination of pollen and charcoal studies also reveals the presence of Pinus sylvestris-nigra, P. pinaster-pinea, Juniperus thurifera, Quercus faginea, Q. ilex and Q. suber as the main trees used. Occurrences of Alnus, Corylus, Fraxinus and Juglans suggest the existence of water courses nearby. The archaeobotanical data is correlated with the geographical setting and archaeological context. The evolution of settlement patterns in central Spain, as well as the economy of other Carpetanian sites, is also considered to evaluate how this territory was managed for its vegetational resources since the Bronze Age (BA). Changes are expected as a consequence of different socio-economic factors at this site, especially when the settlement became an important Carpetanian oppidum. The combined results demonstrate a subsistence system based on cereal agriculture, in which naked wheat was dominant, together with some hulled barley and some erratic occurrences of hulled wheat. Furthermore, livestock breeding and metalwork activities seem to have resulted in a significant loss of woodland in order to clear land for crop fields and cattle grazing to cope with the needs of the growing Carpetanian community.     

Injection of xenogeneic cells into teleost fish elicits systemic and local cellular innate immune responses

The early innate immune response of the teleost gilthead seabream (Sparus aurata L.) against xenogeneic cells was studied. Fish received a single intraperitoneal injection of xenogeneic cells (tumour cell line), following which leucocyte mobilization, degranulation, peroxidase content, respiratory burst and phagocytic and cytotoxic activities were determined in both peritoneal exudate leucocytes (PELs) and head-kidney leucocytes (HKLs). The total number of PELs increased from 4 h post-injection until the end of the experiment (3 days). Interestingly, flow cytometric analysis of PEL and HKL suspensions revealed variations in the proportion of cell types. The percentage of HK acidophilic granulocytes significantly increased after 72 h, whereas PE acidophils increased after 4 h. Moreover, numbers of PE lymphocytes and monocyte-macrophages significantly increased during the experiment. The peroxidase content of the leucocytes was unaffected, although PEL degranulation was largely enhanced. This liberation of peroxidases correlated well with the enhancement of the oxidative respiratory burst activity in PELs, reflecting leucocyte activation. However, phagocytosis only increased in PELs 4 h after intraperitoneal injection, whereas the cytotoxic activity of HKLs increased 1 and 2 days post-injection but, in general, decreased in the PELs. Our data thus demonstrate that the appearance of xenogeneic cells involves leucocyte mobilization and innate immune-response activation at the site of invasion and in the head-kidney. Involvement of the various leucocyte types and potential modes of activation are discussed.This work was partially funded by the European Commission (QLRT-2001-00722). A. Cuesta and I. Salinas are fellows of Fundación CajaMurcia and Fundación Séneca, respectively.     

Genetic structure of two Thalassia testudinum populations from the south Texas Gulf coast

The south Texas Gulf coast is a unique ecosystem that contains a number of different bay systems. We used random amplification of polymorphic DNA (RAPD) markers to assess genetic diversity, differentiation and genetic distance between populations from two different bays that differed significantly in terms of flowering rate and disturbance. We found that while each bay contained a number of unique RAPD profiles, the average genetic diversity in each population was low. Genetic distance between the two populations was also low (F_st = 0.084) and the majority (92%) of the genetic variation was attributed to differences between individuals within populations. The population from the Laguna Madre location, however, was polymorphic for a larger number of markers, had a higher average genetic diversity and a larger number of unique RAPD profiles. The higher level of flowering at this location most likely accounts for the higher diversity.     

Cyclo-oxygenase 2 expression impairs serum-withdrawal-induced apoptosis in liver cells

We have investigated the mechanism of COX-2 (cyclo-oxygenase 2)-dependent inhibition of apoptosis in liver, a key pathway underlying proliferative actions of COX-2 in liver cancers, cirrhosis, chronic hepatitis C infection and regeneration after partial hepatectomy. Stable expression of COX-2 in CHL (Chang liver) cells induced proliferation, with an increase in the proportion of cells in S-phase, but no other significant changes in cell-cycle distribution. This was associated with a marked inhibition of the apoptotic response to serum deprivation, an effect mimicked by treating empty-vector-transfected control cells (CHL-V cells) with prostaglandin E2 and prevented in COX-2-expressing cells (CHL-C cells) treated with selective inhibitors of COX-2. Serum-deprived CHL-V cells displayed several indicators of activation of intrinsic apoptosis: caspases 9 and 3 activated within 6 h and caspase 8 within 18 h, Bax expression was induced, cytochrome c was released to the cytosol, and PARP-1 [poly(ADP-ribose) polymerase 1] cleavage was evident in nuclei. COX-2 expression blocked these events, concomitant with reduced expression of p53 and promotion of Akt phosphorylation, the latter indicating activation of survival pathways. CHL cells were resistant to stimulation of the extrinsic pathway with anti-Fas antibody. Moreover, in vivo expression of GFP (green fluorescent protein)-labelled COX-2 in mice by hydrodynamics-based transient transfection conferred resistance to caspase 3 activation and apoptosis induced by stimulation of Fas.     

Aggression, dominance, and affiliation: Their relationships with androgen levels and intelligence in 5-year-old children

This study explores the potential relationship between social behavior (aggression, dominance, and affiliation) and testosterone, androstenedione, and DHEA measurements in 5-year-old children while also analyzing the moderating effect of IQ on the hormone-behavior relationship. 129 healthy normal Iberian children (60 boys and 69 girls) were videotaped in free play interactions in the school playground. Their behavior was then evaluated with particular emphasis on aggression, government, and affiliation. Testosterone, androstenedione, and DHEA levels were measured using an enzyme immunoassay technique in saliva samples. A test (K-BIT) which provides an IQ measurement for children was also administered to subjects. The correlation analysis revealed a positive relationship between the behavioral factor of Provocation and androstenedione in boys, and a regression analysis indicated that this relationship was moderated in a positive direction by the subject's intelligence. In girls, we observed a positive relationship between testosterone and Affectivity, with this relationship being moderated in a negative direction by intelligence.