Descripción: In 1833 during his journey across the Buenos Aires Pampas, Charles Darwin made observations that reflected his thoughts on two major landscape units, Pampa interserrana and Pampa deprimida, later identified by other authors. Darwin grouped the Pampean sediments into a single unit, the Pampean Formation, based upon the lithological homogeneity and the large extension of the deposits; the unit was thought to be of estuarine-marine origin and attributed to the Recent Epoch considering the paleontological content (vertebrates and mollusks). At present, the Pampean sedimentary succession, which accumulated approximately during the last 11-12 Ma, is interpreted as a pedosedimentary sequence due to the ubiquity of pedogenetic features throughout the deposits. Four main subcycles of sedimentation are identified related to reactivations of the Pampean landscape. At a regional scale, the outcrop distribution of Pampean sediments of different ages suggests the dominance of more stable conditions since the late Miocene-Pliocene in a vast area of Pampa interserrana, documented by the formation of calcretes. However, sedimentation during the late Pliocene-Pleistocene was active within the domain of the Salado tectonic basin and Sierras de Tandil. The regional disparity shown by the Pampean stratigraphic record reveals the major morphostructural differences of its basement.

Descripción: Addition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although tags can readily be included as fusions to head decoration proteins, many phages do not have decoration proteins as virion components. The addition of a small (10-amino-acid) Strep-tag II (STAG II) to the mycobacteriophage TM4 capsid subunit, gp9, was not tolerated as a genetically homogenous recombinant phage but could be incorporated into the head by growth of wild-type phage on a host expressing the capsid-STAG fusion. Particles with capsids composed of wild-type and STAG-tagged subunit mixtures could be grown to high titers, showed good infectivities, and could be used to isolate phage-bacterium complexes. Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence. © 2013, American Society for Microbiology.

Descripción: Bacteriophage Recombineering of Electroporated DNA (BRED) has been described for construction of gene deletion and point mutations in mycobacteriophages. Using BRED, we inserted a Phsp60-egfp cassette (1143 bp) into the mycobacteriophage D29 genome to construct a new reporter phage, which was used for detection of mycobacterial cells. The cassette was successfully inserted and recombinant mycobacteriophage purified. DNA sequencing of the cassette did not show any mutations even after several phage generations. Mycobacterium smegmatis mc2155 cells were infected with D29::Phsp60-egfp (MOI of 10) and evaluated for EGFP expression by microscopy. Fluorescence was observed at around 2 h after infection, but dissipated in later times because of cell lysis. We attempted to construct a lysis-defective mutant by deleting the lysA gene, although we were unable to purify the mutant to homogeneity even with complementation. These observations demonstrate the ability of BRED to insert c. 1 kbp-sized DNA segments into mycobacteriophage genomes as a strategy for constructing new diagnostic reporter phages. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd.

Descripción: A soluble Ca2+-dependent protein kinase (CDPK) was purified to homogeneity in potato (Solanum tuberosum L.) plants. Potato CDPK was strictly dependent on Ca2+ (one-half maximal activation 0.6 μM) and phosphorylated a wide diversity of substrates, in which Syntide 2 was the best phosphate acceptor (Michaelis constant = 30 μM). The kinase was inhibited by Ca2+- chelating agents, phenotiazine derivatives, and N-(6-aminohexyl)-5-chloro-1- naphthalenesulfonamide (one-half maximal inhibition = 0.25 mM). Polyclonal antibodies directed against the regulatory region of the soybean CDPK recognized a 53-kD polypeptide. In an autophosphorylation assay, this same band was strongly labeled with [γ-32P]ATP in the presence of Ca2+. CDPK activity was high in nontuberized plants, but increased 2.5-fold at the onset of tuber development and was reduced to one-half of its original activity when the tuber had completed formation. In the early stages of tuberization, Ca2+-dependent phosphorylation of endogenous targets (specific bands of 68, 51, and 46 kD) was observed. These polypeptides were not labeled in nontuberizing plants or in completely formed tubers, indicating that this phosphorylation is a stage-specific event. In addition, dephosphorylation of specific polypeptides was detected in tuberizing plants, suggesting the involvement of a phosphatase. Preincubation of crude extracts with phosphatase inhibitors rendered a 100% increase in CDPK activity.

Descripción: Nucleotide variation was studied in a 1.1 kb section of the coding region of an Esterase gene (Est-A) that maps in the center of the segments rearranged by polymorphic inversions in the cactophilic Drosophila buzzatii. We examine 30 homozygous second-chromosome lines differing in gene arrangement and three D. koepferae isofemale lines as outgroups. Our data show that Est-A is a highly polymorphic gene at both synonymous and replacement sites. Significant departures from homogeneity in the distribution of the ratio of silent polymorphism to divergence predicted by the neutral theory reveals a local excess of silent polymorphism. This is consistent with the presence of two apparent narrow peaks of elevated silent polymorphism surrounding nonconservative amino acid substitutions. These polymorphisms as well as others at synonymous and nonsynonymous sites are shared with D. koepferae. We suggest that the presence of shared nucleotide polymorphisms is probably due to interspecific gene flow and/or balancing selection acting on replacement variants and/or to a decreased probability of loss of ancestral polymorphisms caused by linkage to an adaptive inversion polymorphism. Recurrent mutation and persistence of neutral ancestral polymorphisms cannot, however, be ruled out. The analysis of the distribution of nucleotide variation among the three chromosomal arrangements sampled reveals that derived arrangements (J and JZ3) are less polymorphic than the ancestral ST, and that the widely distributed ST and J arrangements are genetically differentiated. However, a significant number of polymorphisms are shared between arrangements, suggesting frequent exchange either from gene conversion or from double crossovers in heterokaryotypes. Finally, our present results in combination with data of sequence variation at the breakpoints of inversion J suggest that this old gene arrangement has risen in frequency in relatively recent times.

Descripción: A database of daily extreme temperature was created for as many stations as possible for Uruguay, as far back as possible. This is the first attempt to gather all the different data sources together, perform a quality control and homogeneity assessment. We work with seven stations; it should be taken into account that Uruguay is a small country (around 177 000 km2) and this represents most of the available data. There are three old series with starting dates in 1930, and four that start around 1950. From this database, a set of four extreme temperature indices was constructed for the oldest five stations, warm days (TX90), cold days (TX10), warm nights (TN90) and cold nights (TN10). The index TN10 shows the largest significant negative trend for the period 1960-2002, while TN90 shows a positive but not significant trend for this period indicating a strong warming of nighttime temperature. A spectral analysis was performed using the multi taper methods (MTM) to the de-trended annual, summer Dec-Feb (DJF) and winter Jun-Aug (JJA) indices time-series. This analysis shows that on inter-annual timescales, the most significant range of frequencies is from 2 to 2.5 years and from 3 to 6 years. Low frequencies of variability were detected when the MTM was applied to de-trended smoothed annual time-series, around the range of frequencies of 15-25 years for almost all the indices analysed. Links with global sea surface temperature (SST) were studied for two stations (Paysandu and Rocha), and it was found that the indices showed largest correlations with SST anomalies in the Pacific Ocean. We detected changes in the response of the TN10 index for Rocha station when the series was split up into two different periods (1942-1976 and 1977-2005). Copyright © 2007 Royal Meteorological Society.