MPA network design based on graph network theory and emergent properties of larval dispersal

Despite the recognised effectiveness of networks of Marine Protected Areas (MPAs) as a biodiversity conservation instrument, nowadays MPA network design frequently disregards the importance of connectivity patterns. In the case of sedentary marine populations, connectivity stems not only from the stochastic nature of the physical environment that affects early-life stages dispersal, but also from the spawning stock attributes that affect the reproductive output (e.g., passive eggs and larvae) and its survivorship. Early-life stages are virtually impossible to track in the ocean. Therefore, numerical ocean current simulations coupled to egg and larval Lagrangian transport models remain the most common approach for the assessment of marine larval connectivity. Inferred larval connectivity may be different depending on the type of connectivity considered; consequently, the prioritisation of sites for marine populations' conservation might also differ. Here, we introduce a framework for evaluating and designing MPA networks based on the identification of connectivity hotspots using graph theoretic analysis. We use as a case of study a network of open-access areas and MPAs, off Mallorca Island (Spain), and test its effectiveness for the protection of the painted comber Serranus scriba. Outputs from network analysis are used to: (1) identify critical areas for improving overall larval connectivity; (2) assess the impact of species' biological parameters in network connectivity; and (3) explore alternative MPA configurations to improve average network connectivity. Results demonstrate the potential of graph theory to identify non-trivial egg/larval dispersal patterns and emerging collective properties of the MPA network which are relevant for increasing protection efficiency.Comment: 8 figures, 3 tables, 1 Supplementary material (including 4 table; 3 figures and supplementary methods

Validation of the Spanish Acute Cystitis Symptoms Score (ACSS) in native Spanish‐speaking women of Europe and Latin America

Introduction and Objectives The Acute Cystitis Symptom Score (ACSS) is a patient self-reporting questionnaire for clinical diagnostics and patient-reported outcome (PRO), which may assess the symptoms and the effect on the quality of life in women with acute cystitis (AC). The current study aimed to create a validated Spanish version of the ACSS questionnaire. Material and Methods The process of linguistic validation of the Spanish version of the ACSS consisted of the independent forward and backward translations, revision and reconciliation, and cognitive assessment. Clinical evaluation of the study version of the ACSS was carried out in clinics in Spain and Latin America. Statistical tests included the calculation of Cronbach's α, split-half reliability, specificity, sensitivity, diagnostic odds ratio, positive and negative likelihood ratio, and area under the receiver-operating characteristic curve (AUC). Results The study was performed on 132 patients [age (mean;SD) 45.0;17.8 years] with AC and 55 controls (44.5;12.2 years). Cronbach's α of the ACSS was 0.86, and the split-half reliability was 0.82. The summary scores of the ACSS domains were significantly higher in patients than in controls, 16.0 and 2.0 (p < 0.001), respectively. The predefined cut-off point of ≥6 for a summary score of the “Typical” domain resulted in a specificity of 83.6% and a sensitivity of 99.2% for the Spanish version of the ACSS. AUC was 0.91 [0.85; 0.97]. Conclusions The validated Spanish ACSS questionnaire evaluates the symptoms and clinical outcomes of patients with AC. It can be used as a patient's self-diagnosis of AC, as a PRO measure tool, and help to rule out other pathologies in patients with voiding syndrome

La lesion en el derecho civil

La lesión en el Código Civil Colombiano: A imitación del Derecho Romano, del español antiguo y del francés, nuestro Código Civil ha consagrado un sistema injusto, fragmentario y empírico en materia de la lesión en los actos jurídicos..

Changes of operative performance of pulse pressure variation as a predictor of fluid responsiveness in endotoxin shock.

Several limitations regarding pulse pressure variation (PPV) use have been reported. Our aim was to describe changes in the PPV operative performance as a predictor of fluid responsiveness during the development of a swine endotoxin shock model and to assess hemodynamic variables associated with PPV changes. A swine porcine endotoxin shock model was established (Escherichia Coli 055:B5 endotoxin) in 7 pigs, and 3 pigs were included in the control group. The endotoxin was infused until the mean arterial pressure (MAP) dropped below 50 mmHg (TH0); then, the model animal was reanimated with fluids and vasopressors. We performed fluid challenges every hour for 6 h. ROC curve analysis and a linear mixed model were performed. The area under the curve of PPV decreased from 0.95 (0.81-1.00) to 0.60 (0.17-1.00) at TH0. Its cutoff increased from 10.5 to 22.00% at TH0. PPV showed an inverse relationship with stroke volume, mean systemic filling pressure, MAP, and systemic vascular resistance (SVR) (p

The 25 kDa Subunit of Cleavage Factor Im Is a RNA-Binding Protein That Interacts with the Poly(A) Polymerase in <i>Entamoeba histolytica</i>

<div><p>In eukaryotes, polyadenylation of pre-mRNA 3´ end is essential for mRNA export, stability and translation. Taking advantage of the knowledge of genomic sequences of <i>Entamoeba histolytica</i>, the protozoan responsible for human amoebiasis, we previously reported the putative polyadenylation machinery of this parasite. Here, we focused on the predicted protein that has the molecular features of the 25 kDa subunit of the Cleavage Factor Im (CFIm25) from other organisms, including the Nudix (nucleoside diphosphate linked to another moiety <u>X</u>) domain, as well as the RNA binding domain and the PAP/PAB interacting region. The recombinant EhCFIm25 protein (rEhCFIm25) was expressed in bacteria and used to generate specific antibodies in rabbit. Subcellular localization assays showed the presence of the endogenous protein in nuclear and cytoplasmic fractions. In RNA electrophoretic mobility shift assays, rEhCFIm25 was able to form specific RNA-protein complexes with the <i>EhPgp5</i> mRNA 3´ UTR used as probe. In addition, Pull-Down and LC/ESI-MS/MS tandem mass spectrometry assays evidenced that the putative EhCFIm25 was able to interact with the poly(A) polymerase (EhPAP) that is responsible for the synthesis of the poly(A) tail in other eukaryotic cells. By Far-Western experiments, we confirmed the interaction between the putative EhCFIm25 and EhPAP in <i>E. histolytica</i>. Taken altogether, our results showed that the putative EhCFIm25 is a conserved RNA binding protein that interacts with the poly(A) polymerase, another member of the pre-mRNA 3´ end processing machinery in this protozoan parasite.</p> </div

Molecular characteristics of predicted EhCFIm25.

<p>A) Comparative molecular organization of CFIm25 proteins from <i>E. histolytica</i> and human. Upper panel, Schematic representation. The scale at the top indicates the size in aa. Numbers at the right are relative to the initial methionine in each protein. Lower panel, ClustalW sequence alignment of Nudix box. Black box, identical aa; grey box, similar aa. X, any residue; U, hydrophobic residue. D) Three dimensional organization of CFIm25 proteins from <i>E. histolytica</i> (left) and human (right). 3D modeling of EhCFIm25 was obtained using crystal data from human CFIm25 as template. The Nudix box is in black color. The UniProt KnowledgeBase database accession number for each predicted protein is indicated.</p

Expression of the putative EhCFIm25 in <i>E. histolytica</i> trophozoites.

<p>A and B) Western blot assays. A) Cytoplasmic (lanes 1, 3, 5 and 7) and nuclear (lanes 2, 4, 6 and 8) extracts of <i>E. histolytica</i> trophozoites were separated through 10% SDS-PAGE, electrotransferred to a nitrocellulose membrane and incubated with antibodies. Lanes 1 and 2, anti-EhCFIm25 antibodies; lanes 3 and 4, pre-immune serum; lanes 5 and 6, anti-EhPAP antibodies; lanes 7 and 8, anti-EhPC4 antibodies. B) Cytoplasmic (lanes 1, 3, 5 and 7) and nuclear (lanes 2, 4, 6 and 8) extracts were treated with 5% β-mercaptoethanol (lanes 1 to 4) or 8% β-mercaptoethanol (lanes 5 to 8) and separated through 10% SDS-PAGE in the presence of 8 M urea, and electrotransferred to a nitrocellulose membrane before being incubated with antibodies. Lanes 1, 2, 5 and 6: anti-EhCFIm25 antibodies; lanes 3, 4, 7 and 8: pre-immune serum. Arrowhead, endogenous EhCFIm25. C) Primers to evidence mRNA expression of genes at locus EHI_077110 (up) and EHI_077000 bottom) in RT-PCR assays. D) RT-PCR assays. <i>EhCFIm25</i> transcript was PCR amplified using cDNA synthesized from total RNA and products were analyzed through ethidium bromide stained polyacrylamide gels. Lane 1, molecular size markers; lane 2, cDNA; lane 3, control using genomic DNA from <i>E. histolytica</i>; lane 4, control using oligonucleotides for <i>actin</i> gene; lane 5, control using pRSET<i>-EhCFIm25</i>; lane 6, control without cDNA.</p

Expression of the recombinant EhCFIm25.

<p>A) Expression of the 6x-His-labeled EhCFIm25 protein. Bacteria <i>E. coli</i> were transformed with pRSET<i>-EhCFIm25</i> plasmid and protein expression was induced by the addition of 1 mM IPTG for 3 h. Proteins extracts (30 µg) were separated through 10% SDS-PAGE and gels were stained with Coomassie blue. Lane 1, molecular weight; lane 2, non-induced bacterial extract; lane 3, IPTG-induced bacterial extract. B) Immunodetection of rEhCFIm25 polypeptide by Western blot assays using anti-6x His tag antibodies. Lane 1, non-induced bacterial extract (30 μg); lane 2, IPTG-induced bacterial extract (30 μg). C) Purification of the 6x-His-labeled EhCFIm25 protein through affinity chromatography using a Ni-NTA column. Lane 1, molecular weight; lane 2, IPTG-induced bacterial extract; lane 3, unbound proteins; lane 4, wash using 150 mM imidazole; lanes 5-8, elution with 250 mM imidazole. D) Immunodetection of rEhCFIm25 polypeptide by Western blot assays using specific rabbit antibodies anti-EhCFm25Im. Lane 1, non-induced bacterial extract; lane 2, IPTG-induced bacterial extract; lane 3, IPTG-induced bacterial extract and anti-6x His tag antibodies used as control. E) Immunodetection of purified rEhCFIm25 polypeptide by Western blot assays. Lane 1, anti-6x His tag antibodies; lane 2, specific rabbit antibodies anti-EhCFIm25; lane 3, pre-immune serum; lane 4, control without primary antibody. Arrowhead, EhCFIm25.</p