Characterization of Microbial Mats from a Siliciclastic Tidal Flat (Bahía Blanca Estuary,Argentina)

Biofilms and microbial mats cover the tidal flats of the central zone of the Bahía Blanca estuary (Argentina), creating extensive layers. The objective of this study was to characterize the microphytobenthic communities in these biofilms and mats from sediment cores taken in March, June, September and December 2010. Microorganisms were identified and enumerated by microscopy, and their biomass (chlorophyll a, biovolume) quantified at two different stations in the lower supratidal zone, located ～210 m apart from each other (namely S1 and S2). Additionally, the colloidal carbohydrates produced by these microbial communities were quantified, together with physical parameters such as temperature, granulometry, moisture and organic matter content of the sediment layers that comprise a typical epibenthic mat. On the other hand, changes in biomass and colloidal carbohydrate content were studied through a half-tidal cycle (7 h). There were significant seasonal differences in microphytobenthic biovolume (P < 0.001) with a considerably lower biomass in summer, but no significant differences in microalgal biovolume between stations (P = 0.454). Cyanobacterial biomass (largely composed of the filamentous Microcoleus chthonoplastes) was dominant on all dates at both stations, followed by pennate diatoms. Chlorophyll a and colloidal carbohydrate contents in sediment presented a similar pattern to that of microalgal biovolume; with a 5-fold variation in chlorophyll a for S1 between consecutive sampling events on September and December. There were significant differences between sampling dates in colloidal carbohydrates (P < 0.001) with the lowest values recorded during fall and winter; conversely there were no significant differences between stations (P = 0.324). Silt was the dominant sediment fraction at S1 while sand dominated throughout the uppermost 20 mm at S2. Chlorophyll a contents did not show significant differences throughout a half-tidal cycle, likely the product of vertical migration along the section sampled. Conversely, the content of colloidal carbohydrates varied 5-fold, showing a significant (P < 0.001) and steady increase with time of exposure to air and pointing to the rapid metabolic rates of the community. In conclusion, the microphytobenthic community of the Bahía Blanca estuary presented marked seasonality in its biological parameters and overall physiognomy, also showing elevated metabolic rates when subject to tidal fluctuations.     

Mitochondrial c-Jun NH2-terminal kinase prevents the accumulation of reactive oxygen species and reduces necrotic damage in neural tumor cells that lack trophic support

In response to different stress signals, the c-Jun NH(2)-terminal kinase (JNK) can trigger cell death. However, JNK also facilitates the survival and cell cycle progression of tumor cells by mechanisms that are poorly defined. Here, we show that schwannoma RN22 cells can survive and proliferate under serum-free conditions although serum withdrawal rapidly induces mitochondrial fission and swelling. Although the morphologic changes observed in the mitochondria did not trigger cytochrome c release, they were accompanied by an increase in the mitochondrial membrane potential (DeltaPsi(M)) and of immunoreactivity for active JNK in these organelles. Pharmacologic inhibition of JNK provoked a further increase of the DeltaPsi(M), an increase in reactive oxygen species (ROS) production, and a sustained decrease in cell viability due to necrosis. This increase in necrosis was prevented by the presence of ROS scavengers. Immunoreactivity for active JNK was also observed in the mitochondria of neuroblastoma 1E-115 and neuroblastoma 2a neuroblastoma cell lines on serum withdrawal, whereas active JNK was barely detected in serum-deprived fibroblasts. Accordingly, the reduction in neural tumor cell viability induced by JNK inhibition was largely attenuated in serum-deprived fibroblasts. These data indicate that local activation of JNK in the mitochondria can protect against necrotic cell death associated with ROS production, facilitating the growth of neural tumor cells subjected to serum deprivation.     

Hydroxy-fatty acid production in a <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> 42A2 PHA synthase mutant generated by directed mutagenesis

Pseudomonas aeruginosa 42A2 growing on waste frying oils is capable to synthesize polyhydroxyalkanoic acids (PHAs) and hydroxy-fatty acids as a result of several enzymatic conversions. In order to study the physiological role of PHA biosynthesis in P. aeruginosa with respect to the synthesis of hydroxy-fatty acids, an unmarked deletion mutant deficient for PHA biosynthesis was generated in P. aeruginosa 42A2. A combination of the sacB-based negative selection system with a cre-lox antibiotic marker recycling method was used for mutant isolation. Electron microscopy, nuclear magnetic resonance analysis, and transmission electron microscopy confirmed that PHA accumulation was completely abolished in the mutant strain. Interestingly, the new mutant strain showed higher carbon and oxygen uptake rate than the wild-type strain and higher efficiency in the conversion of oleic acid into (E)-10-hydroxy-8-octadecenic acid-octadecenoic acid.     

The multiple personalities of the chemokine receptor CCR7 in dendritic cells

CCR7 was described initially as a potent leukocyte chemotactic receptor that was later shown to be responsible of directing the migration of dendritic cells (DCs) to the lymph nodes where these cells play an important role in the initiation of the immune response. Recently, a variety of reports have indicated that, apart from chemotaxis, CCR7 controls the cytoarchitecture, the rate of endocytosis, the survival, the migratory speed, and the maturation of the DCs. Some of these functions of CCR7 and additional ones also have been described in other cell types. Herein we discuss how this receptor may contribute to modulate the immune response by regulating different functions in DCs. Finally, we also suggest a possible mechanism whereby CCR7 may control its multiple tasks in these cells.     

Genomic selection of purebreds for crossbred performance

Background

One of the main limitations of many livestock breeding programs is that selection is in pure breeds housed in high-health environments but the aim is to improve crossbred performance under field conditions. Genomic selection (GS) using high-density genotyping could be used to address this. However in crossbred populations, 1) effects of SNPs may be breed specific, and 2) linkage disequilibrium may not be restricted to markers that are tightly linked to the QTL. In this study we apply GS to select for commercial crossbred performance and compare a model with breed-specific effects of SNP alleles (BSAM) to a model where SNP effects are assumed the same across breeds (ASGM). The impact of breed relatedness (generations since separation), size of the population used for training, and marker density were evaluated. Trait phenotype was controlled by 30 QTL and had a heritability of 0.30 for crossbred individuals. A Bayesian method (Bayes-B) was used to estimate the SNP effects in the crossbred training population and the accuracy of resulting GS breeding values for commercial crossbred performance was validated in the purebred population.

Results

Results demonstrate that crossbred data can be used to evaluate purebreds for commercial crossbred performance. Accuracies based on crossbred data were generally not much lower than accuracies based on pure breed data and almost identical when the breeds crossed were closely related breeds. The accuracy of both models (ASGM and BSAM) increased with marker density and size of the training data. Accuracies of both models also tended to decrease with increasing distance between breeds. However the effect of marker density, training data size and distance between breeds differed between the two models. BSAM only performed better than AGSM when the number of markers was small (500), the number of records used for training was large (4000), and when breeds were distantly related or unrelated.

Conclusion

In conclusion, GS can be conducted in crossbred population and models that fit breed-specific effects of SNP alleles may not be necessary, especially with high marker density. This opens great opportunities for genetic improvement of purebreds for performance of their crossbred descendents in the field, without the need to track pedigrees through the system.     

The Depletion of Nuclear Glutathione Impairs Cell Proliferation in 3t3 Fibroblasts

Background

Glutathione is considered essential for survival in mammalian cells and yeast but not in prokaryotic cells. The presence of a nuclear pool of glutathione has been demonstrated but its role in cellular proliferation and differentiation is still a matter of debate.

Principal Findings

We have studied proliferation of 3T3 fibroblasts for a period of 5 days. Cells were treated with two well known depleting agents, diethyl maleate (DEM) and buthionine sulfoximine (BSO), and the cellular and nuclear glutathione levels were assessed by analytical and confocal microscopic techniques, respectively. Both agents decreased total cellular glutathione although depletion by BSO was more sustained. However, the nuclear glutathione pool resisted depletion by BSO but not with DEM. Interestingly, cell proliferation was impaired by DEM, but not by BSO. Treating the cells simultaneously with DEM and with glutathione ethyl ester to restore intracellular GSH levels completely prevented the effects of DEM on cell proliferation.

Conclusions

Our results demonstrate the importance of nuclear glutathione in the control of cell proliferation in 3T3 fibroblasts and suggest that a reduced nuclear environment is necessary for cells to progress in the cell cycle.