Population parameters and reproductive biology of the Iberian hareLepus granatensis in southern Iberia

To analyse the population structure and reproductive biology of the Iberian hareLepus granatensis Rosenhauer, 1856, 498 hares (264 males and 234 females) were collected in monthly samples from October 1998 to September 1999. Females reached larger sizes than males, with approximately 400 g difference in body mass on reaching sexual maturity. The total sex ratio was 1∶1, with a bias in favour of males in winter. Sexually active males and females appeared in every month but August, when no sexually active female was found. Births occurred in every month and were more frequent between March and July. Seasonal variation in kidney fat index (KFI) followed a similar pattern in males and females, with an increase in mid-autumn and a decline at the end of winter. Reproductive activity appeared in every month, with a maximum from February to June. Reproductively hyperactive females (simultaneously pregnant and nursing) appeared in every month except in January, with a first peak in March and a second lower peak in May–June. Litter size fluctuated between 1 and 7 leverets. The most frequent gestations involved 1 or 2 foetuses. The mean annual litter size was 2.08 and the average number of litters per productive female per year was estimated to be 3.48. The maximum productivity was recorded between March and May. The total annual production of young per adult female was estimated to be 7.21. The minimum annual survival rate of young was 27.91%. On the basis of these results we propose to maintain the limit of the hunting period between October and December. We discuss the relationship between the low young/adult ratio obtained and the hunting method used.     

DNA methylation profile in chronic myelomonocytic leukemia associates with distinct clinical,biological and genetic features

Chromosomal abnormalities are detected in 20–30% of patients with chronic myelomonocytic leukemia (CMML) and correlate with prognosis. On the mutation level, disruptive alterations are particularly frequent in chromatin regulatory genes. However, little is known about the consequential alterations in the epigenetic marking of the genome. Here, we report the analysis of genomic DNA methylation patterns of 64 CMML patients and 10 healthy controls, using a DNA methylation microarray focused on promoter regions. Differential methylation analysis between patients and controls allowed us to identify abnormalities in DNA methylation, including hypermethylation of specific genes and large genome regions with aberrant DNA methylation. Unsupervised hierarchical cluster analysis identified two main clusters that associated with the clinical, biological, and genetic features of patients. Group 1 was enriched in patients with adverse clinical and biological characteristics and poorer overall and progression-free survival. In addition, significant differences in DNA methylation were observed between patients with low risk and intermediate/high risk karyotypes and between TET2 mutant and wild type patients. Taken together, our results demonstrate that altered DNA methylation patterns reflect the CMML disease state and allow to identify patient groups with distinct clinical features.     

What can conservation strategies learn from the ecosystem services approach? Insights from ecosystem assessments in two Spanish protected areas

Biodiversity conservation strategies that overlook the interests of local people are prone to create conflicts. The ecosystem service approach holds potential for more comprehensively integrating the social dimension into decision-making in protected areas, but its implementation in conservation policies is still in its infancy. This research assesses the extent to which ecosystem services have been implemented in conservation strategies in protected areas. The study was conducted in two outstanding Spanish protected areas, covering a wetland (Doñana Natural and National Parks) and a Mediterranean mountain system (Sierra Nevada Natural and National Parks). Data were collected from deliberative workshops with managers and researchers, face-to-face surveys with users and a review of management plans. We found that, beyond intrinsic values of ecosystems and biodiversity, these areas provide multiple ecosystem services that deserve further attention to ensure their sustained delivery. Our research shows that environmental managers and researchers have different perceptions and priorities regarding ecosystem services management compared with ecosystem service users. Environmental managers and researchers in both protected areas perceived that human-nature relationships and ecosystem services are already widely included in management plans, if often not explicitly. We found that different ecosystem service categories receive uneven attention in management plans. These contained measures to manage provisioning and cultural services whereas measures for managing regulating services were perceived to be largely absent. We conclude by summarizing insights on how the ecosystem service approach may enhance the consideration of social interests in the management of management protected areas.     

High‐Throughput DNA sequencing of ancient wood

Reconstructing the colonization and demographic dynamics that gave rise to extant forests is essential to forecasts of forest responses to environmental changes. Classical approaches to map how population of trees changed through space and time largely rely on pollen distribution patterns, with only a limited number of studies exploiting DNA molecules preserved in wooden tree archaeological and subfossil remains. Here, we advance such analyses by applying high‐throughput (HTS) DNA sequencing to wood archaeological and subfossil material for the first time, using a comprehensive sample of 167 European white oak waterlogged remains spanning a large temporal (from 550 to 9,800 years) and geographical range across Europe. The successful characterization of the endogenous DNA and exogenous microbial DNA of 140 (~83%) samples helped the identification of environmental conditions favouring long‐term DNA preservation in wood remains, and started to unveil the first trends in the DNA decay process in wood material. Additionally, the maternally inherited chloroplast haplotypes of 21 samples from three periods of forest human‐induced use (Neolithic, Bronze Age and Middle Ages) were found to be consistent with those of modern populations growing in the same geographic areas. Our work paves the way for further studies aiming at using ancient DNA preserved in wood to reconstruct the micro‐evolutionary response of trees to climate change and human forest management.     

Solid-phase chemical amination of a lipase from Bacillus thermocatenulatus to improve its stabilization via covalent immobilization on highly activated glyoxyl-agarose

In this paper, the stabilization of a lipase from Bacillus thermocatenulatus (BTL2) by a new strategy is described. First, the lipase is selectively adsorbed on hydrophobic supports. Second, the carboxylic residues of the enzyme are modified with ethylenediamine, generating a new enzyme having 4-fold more amino groups than the native enzyme. The chemical amination did not present a significant effect on the enzyme activity and only reduced the enzyme half-life by a 3-4-fold factor in inactivations promoted by heat or organic solvents. Next, the aminated and purified enzyme is desorbed from the support using 0.2% Triton X-100. Then, the aminated enzyme was immobilized on glyoxyl-agarose by multipoint covalent attachment. The immobilized enzyme retained 65% of the starting activity. Because of the lower p K of the new amino groups in the enzyme surface, the immobilization could be performed at pH 9 (while the native enzyme was only immobilized at pH over 10). In fact, the immobilization rate was higher at this pH value for the aminated enzyme than that of the native enzyme at pH 10. The optimal stabilization protocol was the immobilization of aminated BTL2 at pH 9 and the further incubation for 24 h at 25 degrees C and pH 10. This preparation was 5-fold more stable than the optimal BTL2 immobilized on glyoxyl agarose and around 1200-fold more stable than the enzyme immobilized on CNBr and further aminated. The catalytic properties of BTL2 could be greatly modulated by the immobilization protocol. For example, from (R/S)-2- O-butyryl-2-phenylacetic acid, one preparation of BTL2 could be used to produce the S-isomer, while other preparation produced the R-isomer.     

Heavy Metal Distribution in Open Canals and Drains in the Upper Rio Grande Basin

The inclusion of reclaimed effluents for irrigated agriculture may allow communities along the U.S.-Mexico border to use water resources more efficiently and reduce urban costs of wastewater treatment. However, metal inflow from industry and urbanization may threaten food safety. This study examines metal concentrations in sediments from open canal systems charged with flow from the Rio Grande and from effluents discharged from border communities. At the surface of canal beds, sediments were collected from six canal segments that began at the fringe of the El Paso/Juarez metroplex to rural areas downstream, and ranged from 9 to 24?km in length. Sediments were analyzed for Cd, Co, Cr, Cu, Ni, Pb, and Zn. These metals rarely exceeded 20?mg kg^?1. Drainage and effluent conveyance increased the variability of metal concentrations in sediments. Geostatistical models did not significantly account for spatial variability of metals, except in Mexico. This may implicate multiple rather than single inflow sources of metals. Peaks in metal concentrations often coincided with growing rural communities. However, most metal concentrations were within conventional global ranges and were not at levels high enough to threaten food safety. Future sampling strategies will require reduced sampling intervals from 1 to <0.13?km.     

Enhancement of Novozym-435 catalytic properties by physical or chemical modification

Physical (ionic exchange of ionic polymers) or chemical (aminoethylamidation, succinylation, hydroxyethylamidation) modifications of Novozym 435 have been performed and the resulting biocatalysts have been assayed in diverse reactions. The coating of the immobilized enzyme with dextran-sulphate via ionic exchange permitted to increase the asymmetric factor of the biocatalyst from A = 13 (ee = 83%) to 24 (ee > 90%) in the hydrolysis of 3-phenylglutaric acid dimethyl diester, producing the (R)-monomethyl ester. The chemical succinylation of Novozym 435 permitted to enhance the biocatalyst enantiospecificity from E = 1 to 13 in the hydrolysis of (±)-mandelic acid methyl ester. In the hydrolysis of (±)-2-O-butyryl-2-phenylacetic acid, the enantiospecificity of Novozym 435 was very high towards the S-enantiomer (E > 100) but it was inverted after the chemical hydroxyethylamidation of the immobilized enzyme (E = 6.6 towards R-enantiomer).Thus, these simple protocols seem to be a very powerful tool to generate a library of biocatalysts from Novozym 435 with very different catalytic properties.     

Stable low molecular weight RNA analyzed by staircase electrophoresis, a molecular signature for both prokaryotic and eukaryotic microorganisms

Low-molecular weight RNA (LMW RNA) analysis using staircase electrophoresis was performed for several species of eukaryotic and prokaryotic microorganisms. According to our results, the LMW RNA profiles of archaea and bacteria contain three zones: 5S RNA, class 1 tRNA and class 2 tRNA. In fungi an additional band is included in the LMW RNA profiles, which correspond to the 5.8S RNA. In archaea and bacteria we found that the 5S rRNA zone is characteristic for each genus and the tRNA profile is characteristic for each species. In eukaryotes the combined 5.8S and 5S rRNA zones are characteristic for each genus and, as in prokaryotes, tRNA profiles are characteristic for each species. Therefore, stable low molecular weight RNA, separated by staircase electrophoresis, can be considered a molecular signature for both prokaryotic and eukaryotic microorganisms. Analysis of the data obtained and construction of the corresponding dendrograms afforded relationships between genera and species; these were essentially the same as those obtained with 16S rRNA sequencing (in prokaryotes) and 18S rRNA sequencing (in eukaryotes).