Neanderthal and carnivore activities at Llonin Cave,Asturias, northern Iberian Peninsula: Faunal study of Mousterian levels (MIS 3)

This paper presents a study of the macromammalian fauna recovered from Mousterian levels of Llonin Cave. The sample is highly heterogeneous and comprises six species of ungulates, including Rupicapra pyrenaica, Capra pyrenaica, and Cervus elaphus, and seven species of carnivores, predominantly Ursus spelaeus, Crocuta spelaea, Canis/Cuon and Panthera pardus. The archaeozoological and taphonomic study of the remains shows preferential use of basal levels of the cave as a den for hyenas and leopards. Neanderthals were also present during this phase and they would have acted mainly on deer and some caprines, while the action of hyenas would mainly have been linked to scavenging of elements left by humans and the introduction of bear remains. Leopards would have transported caprines in order to consume them. The study of several coprolites confirms that hyenas and leopards were the main occupants of the cave. The information from the animals processed by humans together with other archaeological evidence and the intervention of various carnivores in these basal levels enables us to characterise a palimpsest of occupations that would have been short and sporadic in the case of humans.     

A new approach to drug discovery: high-throughput screening of microbial natural extracts against Aspergillus fumigatus using resazurin

Natural products are an inexhaustible source for drug discovery. However, the validation and selection of primary screening assays are vital to guarantee a selection of extracts or molecules with relevant pharmacological action and worthy of following up. The assay must be rapid, simple, easy to implement, and produce quick results and preferably at a low cost. In this work, we developed and validated a colorimetric microtiter assay using the resazurin viability dye. The parameters of the resazurin method for high-throughput screening (HTS) using natural extracts against Aspergillus fumigatus were optimized and set up. The extracts plus RPMI-1640 modified medium containing the spores and 0.002% resazurin were added per well. The fluorescence was read after 24 to 30 h of incubation. The resazurin proved to be as suitable as Alamar Blue for determining the minimal inhibitory concentration of different antifungals against A. fumigatus and effective to analyze fungicidal and fungistatic compounds. An HTS of 12 000 microbial extracts was carried out against two A. fumigatus strains, and 2.7% of the extracts displayed antifungal activity. Our group has been the first to use this methodology for screening a collection of natural extracts to identify compounds with antifungal activity against the medically important human pathogen A. fumigatus.     

Variations in airborne pollen in central and south-western Spain in relation to the distribution of potential sources

The present study seeks to compare daily and hourly airborne pollen concentrations at eight different stations in Castilla-La Mancha (central Spain) and Extremadura (south-western Spain) and assess pollen distribution sources. Sampling stations were located 69–440 km apart in a straight line in Albacete, Toledo, Talavera de la Reina and Ciudad Real in Castilla-La Mancha, and in Badajoz, Plasencia, Santa Amalia and Zafra in Extremadura. Airborne pollen was collected using Hirst-type volumetric spore traps. Quercus was the most abundant pollen type at all stations except for Ciudad Real, where Olea pollen predominated. Comparisons of daily data between pairs of stations revealed statistically significant positive correlations in all cases for Poaceae and Olea. Comparisons of hourly data between stations indicated greater differences than daily data. Analysis of correlation coefficients and straight-line distances between stations revealed a strong negative correlation. Analysis of total airborne pollen data for the eight sampling stations suggests that airborne pollen concentrations decrease from west to east and from south to north, partly reflecting dominant wind patterns. A clear correlation was observed between airborne pollen concentrations and the surface area covered by olive crops in a 50 km radius around the sampling stations.     

Binding of the natural killer cell inhibitory receptor Ly49A to its major histocompatibility complex class I ligand. Crucial contacts include both H-2Dd AND beta 2-microglobulin.

Ly49A, an inhibitory C-type lectin-like mouse natural killer cell receptor, functions through interaction with the major histocompatibility complex class I molecule, H-2D(d). The x-ray crystal structure of the Ly49A.H-2D(d) complex revealed that homodimeric Ly49A interacts at two distinct sites of H-2D(d): Site 1, spanning one side of the alpha1 and alpha2 helices, and Site 2, involving the alpha1, alpha2, alpha3, and beta(2)m domains. Mutants of Ly49A, H-2D(d), and beta(2)-microglobulin at intermolecular contacts and the Ly49A dimer interface were examined for binding affinity and kinetics. Although mutations at Site 1 had little affect, several at Site 2 and at the dimer interface hampered the Ly49A.H-2D(d) interaction, with no effect on gross structure or T cell receptor interaction. The region surrounding the most critical residues (in H-2D(d), Asp(122); in Ly49A, Asp(229), Ser(236), Thr(238), Arg(239), and Asp(241); and in beta(2)-microglobulin, Gln(29) and Lys(58)) of the Ly49A.H-2D(d) interface at Site 2 includes a network of water molecules, suggesting a molecular basis for allelic specificity in natural killer cell recognition.     

Environmental Factors Affecting Airborne Pollen Concentration in Anemophilous Species of Plantago

Pollination of Plantago species in the Extremadura Region (SWof the Iberian Peninsula) was studied using three volumetrictraps located in the cities of Badajoz, Mérida and Cáceresbetween 1994 and 1999. Variations in atmospheric concentrationof Plantago pollen were analysed between locations, and annual,daily, and hourly variations recorded for each location. Thehighest concentrations of pollen were recorded at Cáceres,while the Mérida and Badajoz values were similar. Thisis explained by the nature of the surroundings of each city.Interannual variations in pollination levels were significantlycorrelated with autumn rains, which determine the extent ofdevelopment of the populations. The hourly patterns of pollencapture were well-defined and similar for all three study sites.Maximum levels were reached between 1000 and 1200 h. Nocturnalconcentrations were very low. Furthermore, this pattern wasmaintained throughout the flowering period, impling a very closelink with the patterns of anthesis of the species. The threestations showed similar patterns of daily variation, which weresignificantly correlated with certain meteorological parameters.Pollen concentrations were affected positively by temperature,but negatively by relative humidity. The influence of wind directionalso seemed to be explicable in these terms since the easterlywinds, which are dry and hot in this region, had a positiveinfluence, and the westerly winds which are moist and cool hada negative effect. The most relevant factor influencing levelsofPlantago pollen in the atmosphere was wind speed, which wasnegatively correlated with pollen levels. Copyright 2001 Annalsof Botany Company Plantago, aerobiology, reproduction, anemophily, pollen     

surA, an Escherichia coli gene essential for survival in stationary phase.

Mutations in genes not required for exponential growth but essential for survival in stationary phase were isolated in an effort to understand the ability of wild-type Escherichia coli cells to remain viable during prolonged periods of nutritional deprivation. The phenotype of these mutations is referred to as Sur- (survival) and the genes are designated sur. The detailed analysis of one of these mutations is presented here. The mutation (surA1) caused by insertion of a mini-Tn10 element defined a new gene located near 1 min on the E. coli chromosome. It was located directly upstream of pdxA and formed part of a complex operon. Evidence is presented supporting the interpretation that cells harboring the surA1 mutation die during stationary phase while similar insertion mutations in other genes of the operon do not lead to a Sur- phenotype. Strains harboring surA1 had a normal doubling time in both rich and minimal medium, but cultures lost viability after several days in stationary phase. Analysis of revertants and suppressors of surA1, which arose after prolonged incubation in stationary phase, indicates that DNA rearrangements (excisions and duplications) occurred in cultures of this strain even when the viable-cell counts were below 10(2) cells per ml. Cells containing suppressing mutations then grew in the same culture to 10(8) cells per ml, taking over the population. The implications of these observations to our understanding of stationary-phase mutagenesis are discussed.     

Staphylococcus aureus pathogenicity island DNA is packaged in particles composed of phage proteins

Staphylococcus aureus pathogenicity islands (SaPIs) have an intimate relationship with temperate staphylococcal phages. During phage growth, SaPIs are induced to replicate and are efficiently encapsidated into special small phage heads commensurate with their size. We have analyzed by amino acid sequencing and mass spectrometry the protein composition of the specific SaPI particles. This has enabled identification of major capsid and tail proteins and a putative portal protein. As expected, all these proteins were phage encoded. Additionally, these analyses suggested the existence of a protein required for the formation of functional phage but not SaPI particles. Mutational analysis demonstrated that the phage proteins identified were involved only in the formation and possibly the function of SaPI or phage particles, having no role in other SaPI or phage functions.     

Reliability of bulky DNA adducts measurement by the nuclease P1 32P-post-labelling technique.

The aim was to assess the reliability of bulky DNA adducts measurement by means of the 32P-post-labelling assay. The research design consisted of an intramethod reliability study. Buffy coats from 41 subjects were used to obtain two aliquots of 1-5 microg DNA for each subject; bulky DNA adducts were measured using the nuclease P1 32P-post-labelling technique. The reliability of the measurement was assessed by means of the intraclass correlation coefficient (ICC), the distribution of the differences between the two measurements and the limits of agreement. The estimated ICC was 0.977, with a 95% confidence interval between 0.921 and 0.977. The limits of agreement were +/- 0.44 (DNA adducts per 10(8) nucleotides). Only three subjects had differences lying out of such limits. Bulky DNA adduct levels measured by the 32P-post-labelling technique showed good reliability. Only one measurement is needed to use DNA adducts as a biomarker of exposure and, possibly, cancer risk. Besides, as a validation analysis, 32P-post-labelling measurements can be repeated in only 20-30% of samples.     

Crystallization and preliminary x-ray diffraction studies of a monoclonal antibody fab fragment against foot-and-mouth disease virus and of its complex with the main antigenic site peptide

The Fab fragment of the neutralizing monoclonal antibody SD6 elicited against foot-and-mouth disease virus (FMDV) C-SBcl and its complex with a peptide, corresponding to the major antigenic site of FMDV (VPl residues 136–150, YTASARGDLAHLTTT), have been crystallized using the hanging drop vapor diffusion techniques. For the isolated Fab, crystals diffracting to 2.5 Å resolution were obtained at room temperature using ammonium sulfate as precipitant. These crystals are monoclinic, space group C2, and unit cell parameters a = 109.53 Å, b = 89.12 Å, c = 64.04 Å, and β = 112.9° and contain one Fab molecule per asymmetric unit. Crystals from the complex diffract, at least, to 2.8 Å resolution and were obtained, at room temperature, using PEG as precipitant. These crystals are monoclinic, space group P2, and unit cell parameters a = 56.11 Å, b = 60.67 Å, c = 143.45 Å, and β = 95.4°, Density packing considerations indicate that there are two Fab molecules in the asymmetric unit. © 1994 John Wiley & Sons, Inc.     

Virulence factors in clinical and food isolates ofAeromonas species

Virulence factors were compared in 15Aeromonas spp. isolated from faeces of patients withAeromonas-associated gastroenteritis and in 81 strains isolated from food. Strains from food did not show differences in the distribution of virulence factors when compared with strains isolated from faeces. However, 88.8% ofAeromonas strains isolated from food were capable of producing possible virulence factors. Characterization of 28 autoagglutinating (AA⁺)Aeromonas spp. indicated that the human strains differed from the food strains in hemagglutinating and hemolytic capacities. These results suggest that auto-agglutination associated with hemagglutinating and hemolytic capacities in food strains may be a helpful indicator of potential pathogenicity.