Overexpression of csc1-1. A plausible strategy to obtain wine yeast strains undergoing accelerated autolysis

The potential of several alternative genetic engineering based strategies in order to accelerate Saccharomyces cerevisiae autolysis for wine production has been studied. Both constitutively autophagic and defective in autophagy strains have been studied. Although both alternatives lead to impaired survival under starvation conditions, only constitutively autophagic strains, carrying a multicopy plasmid with the csc1-1 allele under the control of the TDH3 promoter, undergo accelerated autolysis in the experimental conditions tested. Fermentation performance is impaired in the autolytic strains, but industrial strains carrying the above-mentioned construction are still able to complete second fermentation of a model base wine. We suggest the construction of industrial yeasts showing a constitutive autophagic phenotype as a way to obtain second fermentation yeast strains undergoing accelerated autolysis.     

A common haplotype of <Emphasis Type="Italic">DRD3</Emphasis> affected by recent positive selection is associated with protection from schizophrenia

The number and frequency of susceptibility alleles at loci associated to most psychiatric disorders is largely unknown, in spite of its relevance for the design of studies aiming to find these alleles. Both, common polymorphisms and rare mutations may contribute to the genetic susceptibility to complex psychiatric disorders, being the relative relevance of each type of variation currently under debate. Here, we confirmed the existence of a common protective haplotype against schizophrenia at the dopamine D₃ receptor (DRD3) gene, by replication and pooled analysis with previous data (Mantel–Haenszel χ² P value = 0.00227; OR = 0.79, 95% CI 0.68–0.92, based on 794 cases and 1,078 controls from three independent populations of European origin). This protective haplotype is at very low frequency in Sub-Saharan Africans (median 0.06) and at intermediate frequencies in other populations (median 0.25). We also revealed, by examining the patterns of linkage disequilibrium around this gene, that the protective haplotype has reached high frequency in non-African populations due to selection acting, most probably, on a linked functional polymorphism, the non-synonymous single nucleotide polymorphism Ser9Gly (rs6280), also at DRD3. Thus, this finding shows that the natural selection may play a role in the existence of common alleles conferring different susceptibility to schizophrenia. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Left-sided infective endocarditis caused by Pseudomonas aeruginosa treated medically

Infective endocarditis due to Pseudomonas aeruginosa is a rare clinical condition, difficult to diagnose and associated with high mortality. Herein we present a case of a 51 years old male without history of intravenous drug use or valvular disease, with past medical history of cholecystectomy in the previous month, who presented to the emergency department with fever, gastrointestinal symptoms, and subsequent signs of distant embolization, positive blood cultures for P. aeruginosa and development of multiple complications of the disease. The clinical presentation of infective endocarditis is nonspecific, leading to delayed diagnosis, and preventing early and effective treatment. Valvular replacement is indicated in fungal or P. aeruginosa endocarditis. This case is notable because of the resolution with amikacin combined with meropenem, in a patient with several complications that contraindicated surgery.     

RIP2 regulates growth and differentiation of normal myoblasts and of rhabdomyosarcoma cells

RIP2 is an important regulator of myoblast proliferation and differentiation. We have previously demonstrated that in the myoblast cell line C2C12 and in primary myoblasts, downregulation of rip2 gene expression is a prerequisite for differentiation. To further study the role of rip genes in myogenesis, we compared expression patterns of rip1–4 in two myoblast cell lines, C2C12 and C2F3, after the induction of differentiation. These two cell lines are derived from the same clonal origin, but differ with respect to their differentiation behaviour: specifically, the differentiation process is slower and more incomplete in C2F3 cells. When analyzing cells up to 4 days after the induction of differentiation, we found no downregulation of rip2 gene expression in C2F3 cells, which might be linked to the low differentiation potential of these cells. In addition, in contrast to C2C12 cells, the rip3 gene was not expressed in C2F3 cells. To further study the role of rip genes in the regulation of myoblast growth and differentiation, we analyzed expression patterns of rip1–4 in rhabdomyosarcoma cell lines. We found that in these cells, rip2 expression was not downregulated after the induction of differentiation. Furthermore, in contrast to normal myoblasts, they did not express the rip3 and rip4 genes. Thus, we focused on the functional role of RIP2 in rhabdomyosarcoma cells. Inhibition of rip2 gene expression in C2C12 and in rhabdomyosarcoma cells using specific siRNAs led to decreased proliferation and promoted the differentiation process of these cells. These data indicate that differential expression of rip genes can be associated with abnormal growth and differentiation behaviour of skeletal myoblasts.     

Spatio-Temporal Distribution and Ecological Preferences of Coexisting Hydropsychid Species (Trichoptera) in Two Mediterranean River Basins (S Spain)

The hydropsychids of two Mediterranean river basins in southern Spain (Guadaira and Guadalete) were investigated from January 1988 to January 1989. The distribution of hydropsychids along an environmental gradient related to altitude varied depending on species. Two distinct species assemblages were observed. one located in the lower altitude sites (the “downstream assemblage” comprising Hydropsyche pictetorum, Hydropsyche exocellata and Cheumatopsyche lepida) and the other in the upper-most areas (the “upstream assemblage” comprising Hydropsyche punica, Hydropsyche infernalis, Hydropsyche instabilis), which displayed a statistically significant preference for this type of habitat. Finally, H. pictetorum, selected the lowest sites. From a temporal point of view most species displayed a high variability in density dynamics between sites, suggesting a strong plasticity in life-cycle phenology.     

Glacial refugia of temperate, Mediterranean and Ibero-North African flora in south-eastern Spain: new evidence from cave pollen at two Neanderthal man sites

Aim To locate glacial refugia of thermophilous plant species in Spain. Location Two south‐eastern Spanish Neanderthal man sites in Murcia; namely, the inland Cueva Negra del Estrecho del Río Quípar and the coastal Sima de las Palomas del Cabezo Gordo. Methods We use pollen found in cave sediments as a source of palaeobotanical and palaeoecological information. The findings are discussed with regard both to animal remains from both sites, and also to other refugia in south‐eastern Spain and elsewhere in the Iberian Peninsula. Results Both sequences show persistence of abundant mesothermophilous trees during the last glacial stage, suggesting both localities were reservoirs of phytodiversity and woodland species. At both sites, deciduous and evergreen oaks are the most abundant components, followed by a wide variety of deciduous trees and sclerophyllous shrubs, including Ibero‐North African xerothermic scrub near the coast. Conclusions Incomplete information underlies a common misapprehension that Iberian glacial refugia were confined to southernmost parts of the peninsula. A rather different picture of Quaternary refugia emerges from consideration of pollen sequences from caves (and other inputs such as macroscopic charcoal, spatial genetic structure of present‐day populations, faunal remains, and present‐day distribution of thermophilous species). This picture offers a view of numerous viable areas for woodland species in southern Spain, in addition to others in the mountain ranges, both in continental central Spain and those of northern Spain: these stretch from the Mediterranean coast of Catalonia to the westernmost extent of the Bay of Biscay.     

The phosphoinositide 3-kinase/AKT1 pathway involvement in drug and all-trans-retinoic acid resistance of leukemia cells

Disruption of the apoptotic pathways may account for resistance to chemotherapy and treatment failures in human neoplastic disease. To further evaluate this issue, we isolated a HL-60 cell clone highly resistant to several drugs inducing apoptosis and to the differentiating chemical all-trans-retinoic acid (ATRA). The resistant clone displayed an activated phosphoinositide 3-kinase (PI3K)/AKT1 pathway, with levels of phosphatidylinositol (3,4,5) trisphosphate higher than the parental cells and increased levels of both Thr 308 and Ser 473 phosphorylated AKT1. In vitro AKT1 activity was elevated in resistant cells, whereas treatment of the resistant cell clone with two inhibitors of PI3K, wortmannin or Ly294002, strongly reduced phosphatidylinositol (3,4,5) trisphosphate levels and AKT1 activity. The inhibitors reversed resistance to drugs. Resistant cells overexpressing either dominant negative PI3K or dominant negative AKT1 became sensitive to drugs and ATRA. Conversely, if parental HL-60 cells were forced to overexpress an activated AKT1, they became resistant to apoptotic inducers and ATRA. There was a tight relationship between the activation of the PI3K/AKT1 axis and the expression of c-IAP1 and c-IAP2 proteins. Activation of the PI3K/AKT1 axis in resistant cells was dependent on enhanced tyrosine phosphorylation of the p85 regulatory subunit of PI3K, conceivably due to an autocrine insulin-like growth factor-I production. Our findings suggest that an up-regulation of the PI3K/AKT1 pathway might be one of the survival mechanisms responsible for the onset of resistance to chemotherapeutic and differentiating therapy in patients with acute leukemia.