Spatial patterns of white stork (Ciconia ciconia) migratory phenology in the Iberian Peninsula

In contrast to the attention attracted by temporal trends of phenology, the spatial patterns of arrivals, departures or stays of trans-Saharan birds are still nowadays largely unknown in most of their European breeding areas. In the case of the white stork (Ciconia ciconia), some studies have attempted to describe its migratory patterns throughout some European countries but, to our knowledge, no one has related these patterns to some kind of explanatory variable which offers an ecologically-based explanation for the heterogeneous phenology observable among populations. Here, arrivals, departures and stays of this species, recorded in hundreds of Spanish localities, were related to a set of environmental, geographical, biological and spatial predictors, and modeled by multiple regression. The best model for arrival dates accounted for up to 34% of variability of data and pointed towards an earlier arrival in those populations located in south-western Iberia and with higher population densities. This last relationship is probably due to the competition for nest-site fidelity maintenance. However, no variable was able to explain properly the blurred spatial pattern recorded for departure dates. Departure decisions are strongly influenced by social behavior in this species and depend on collective decisions influenced by peculiar local environmental conditions of each year rather than macrogeographical gradients. Environmental, geographical or spatial variables also did not capture much of the observed variability in the length of the stays among populations. However, this variable was strongly related to the arrival and departure dates of populations. White storks stay longer in localities with earlier arrivals and, especially, later departures.     

Expression and crystallization of a Cry3Aa–Cry1Ac chimerical proteinof Bacillus thuringiensis

The insecticidal Cry1 proteins of Bacillus thuringiensis form a typical bipyramidal parasporal crystal and their protoxins contain a highly conserved C-terminal region. A chimerical gene was constructed with the coding regions of the Cry3Aa protein's toxic domain, and of the Cry1Ac protoxin's C-terminal fragment. This chimerical construction expressed a truncated (70kDa) protein in the acrystalliferous strain 4Q7 of B. thuringiensis, assembled in spherical to amorphous parasporal crystals. This protein was recognized only by antibodies raised against the Cry3Aa protein. When the protease-deficient mutant BL21 of Escherichia coli was transformed with the same chimerical construction, a complete (140kDa) chimerical protein was expressed. However, the formation of a crystalline inclusion was unclear. This protein was recognized by antibodies raised against the proteins Cry1Ac and Cry3Aa. Both chimerical proteins showed toxicity against larvae of Leptinotarsa texana, being much more active when expressed truncated in B. thuringiensis. These results suggest that the formation of bipyramidal crystals requires more than just the presence of the C-terminal region of Cryl protoxins. They also suggest that proteolysis plays an important role during the post-translational processing of Cry proteins.     

Permanent Stained Preparations of Synovial Fluid for Detection of Calcium Compounds Using Alizarin Red S

Permanent preparations of air dried synovial fluids were prepared by staining calcium compounds with alizarin red S stain; each slide was coverslipped with Permount. Variables studied were: (a) concentration of the solution of alizarin red S, (b) pH of staining solution, (c) time of incubation in staining solution and aqueous and ethanolic content of staining solution. The staining effect of each solution was tested on calcium pyrophosphate dihydrate, calcium oxalate, apatite and monosodium urate (MSU). Of all the solutions, best results were obtained with 0.25% alizarin red S in 50% ethanol at pH 7.0 for 30 min. With this solution, the calcium-containing compounds were well stained. MSU did not stain and still preserved negative birefringence on polarizaton. Fixation of smears with ethanol served a double purpose: It fixed the slides without dissolving or removing MSU or the calcium compounds, yet it did dissolve five corticosteroids commonly used for intra-articular injection which may interfere with interpretation of compensated polarized light microscopy of synovial fluids.     

Proteomic analysis of phytopathogenic fungus <Emphasis Type="Italic">Botrytis cinerea</Emphasis> as a potential tool for identifying pathogenicity factors,therapeutic targets and for basic research

Botrytis cinerea is a phytopathogenic fungus causing disease in a substantial number of economically important crops. In an attempt to identify putative fungal virulence factors, the two-dimensional gel electrophoresis (2-DE) protein profile from two B. cinerea strains differing in virulence and toxin production were compared. Protein extracts from fungal mycelium obtained by tissue homogenization were analyzed. The mycelial 2-DE protein profile revealed the existence of qualitative and quantitative differences between the analyzed strains. The lack of genomic data from B. cinerea required the use of peptide fragmentation data from MALDI-TOF/TOF and ESI ion trap for protein identification, resulting in the identification of 27 protein spots. A significant number of spots were identified as malate dehydrogenase (MDH) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The different expression patterns revealed by some of the identified proteins could be ascribed to differences in virulence between strains. Our results indicate that proteomic analysis are becoming an important tool to be used as a starting point for identifying new pathogenicity factors, therapeutic targets and for basic research on this plant pathogen in the postgenomic era.     

Hospital costs associated with delirium in older medical patients

Introduction

Delirium is a common and serious complication in older patients, associated with increased, potentially preventable, morbidity and mortality. The aim of this study was to evaluate the associated costs of delirium during hospitalization in a university affiliated hospital in Chile.

Materials and methods

Prospective cohort study of consecutive patients 65 years and older, admitted to a medical ward. A psychogeriatric team assessed patients during the first and every 48 h until discharge using the Confusion Assessment Method (CAM-S), length of hospital stay, pharmacy and total hospitalization costs were analyzed. Statistical analysis was performed using bivariate and multivariate analysis according to delirium diagnosis.

Results

Data from 454 patients was analyzed, 160 of them in a delirium cohort (35.2%) and 294 in a non-delirium cohort (64.8%). The delirium cohort had a longer hospital stay (DATA) and higher mortality (7.0% versus 1.7%). The median of total costs of delirium during hospital stay was 38.7% higher than the non-delirium cohort (P < .001). Total costs were significantly higher in the delirium cohort after adjustment of covariables (P = .01).

Conclusions

This study confirms that delirium is associated with significantly greater costs. Considering that effective delirium prevention is possible, the knowledge of associated costs can help health care providers to justify prevention strategies and finally give better care for older patients.     

Structural basis of the lisinopril-binding specificity in N- and C-domains of human somatic ACE

Angiotensin I-converting enzyme (ACE) is a dipeptidyl carboxypeptidase which converts angiotensin I into the vasopressor peptide angiotensin II and also inactivates the hypotensive peptide bradykinin, playing an important role in blood pressure regulation. The present work describes the molecular modeling of the N-terminal human somatic ACE in complex with the inhibitor lisinopril, identifying the residues involved in the inhibitor-binding pocket. The obtained results identify differences in the lisinopril lysine moiety-binding residues for N- and C-terminals of sACE domains and an important carboxy-terminal proline hydrophobic accommodations mediated by the aromatic ring of Tyr532 and Tyr1128 residues, respectively. The present model will be useful for the development of a new inhibitor family based on the natural BPP peptides and derivatives, or even to improve the binding capacities and the domain specificity of the already known inhibitors.     

A new methodology combining PCR, cloning, and sequencing of clones discriminated by RFLP for the study of microbial populations: application to an UASB reactor sample

This work describes a methodology combining DNA extraction, polymerase chain reaction amplification with primers targeting 16S ribosomal RNA genes, cloning, and sequencing of clones previously analyzed by restriction fragment length polymorphism (RFLP), which can be applied to study the microbial diversity in a given habitat. The methodology allows the minimization of the sequencing effort, which is particularly relevant when analyzing large numbers of clones. The methodology does not require particularly skilled personnel and can easily be adaptable to the molecular characterization of virtually any particular microbial population, provided that both adequate primers and suitable restriction enzymes for RFLP analysis of the clone library have been chosen. An example of application is presented, in which a sample taken from a continuously operating upflow anaerobic sludge blanket reactor was analyzed. RFLP analysis of the initial 162 clones with HaeIII allowed the identification of only 28 distinct profiles. As expected, identical RFLP profiles corresponded to identical nucleotide sequences.