The phosphodiesterase DibA interacts with the c-di-GMP receptor LapD and specifically regulates biofilm in Pseudomonas putida.

The ubiquitous bacterial second messenger c-di-GMP is synthesized by diguanylate cyclase and degraded by c-di-GMP-specific phosphodiesterase. The genome of Pseudomonas putida contains dozens of genes encoding diguanylate cyclase/phosphodiesterase, but the phenotypical-genotypical correlation and functional mechanism of these genes are largely unknown. Herein, we characterize the function and mechanism of a P. putida phosphodiesterase named DibA. DibA consists of a PAS domain, a GGDEF domain, and an EAL domain. The EAL domain is active and confers DibA phosphodiesterase activity. The GGDEF domain is inactive, but it promotes the phosphodiesterase activity of the EAL domain via binding GTP. Regarding phenotypic regulation, DibA modulates the cell surface adhesin LapA level in a c-di-GMP receptor LapD-dependent manner, thereby inhibiting biofilm formation. Moreover, DibA interacts and colocalizes with LapD in the cell membrane, and the interaction between DibA and LapD promotes the PDE activity of DibA. Besides, except for interacting with DibA and LapD itself, LapD is found to interact with 11 different potential diguanylate cyclases/phosphodiesterases in P. putida, including the conserved phosphodiesterase BifA. Overall, our findings demonstrate the functional mechanism by which DibA regulates biofilm formation and expand the understanding of the LapD-mediated c-di-GMP signaling network in P. putida.

Alkaline phosphatase LapA regulates quorum sensing-mediated virulence and biofilm formation in Pseudomonas aeruginosa PAO1 under phosphate depletion stress.

IMPORTANCE: Our previous study demonstrated that the expression of lapA was induced under phosphate depletion conditions, but its roles in virulence and biofilm formation by Pseudomonas aeruginosa remain largely unknown. This study presents a systematic investigation of the roles of lapA in virulence induction and biofilm formation by constructing a lapA-deficient strain with P. aeruginosa PAO1. The results showed that deletion of the lapA gene evidently reduced elastase activity, swimming motility, C4-HSL, and 3-oxo-C12-HSL production, and increased rhamnolipid production under phosphate depletion stress. Moreover, lapA gene deletion inhibited PAO1 biofilm formation in porcine skin explants by reducing the expression levels of las and rhl quorum sensing systems and extracellular polymeric substance synthesis. Finally, lapA gene deletion also reduced the virulence of PAO1 in Caenorhabditis elegans in fast-kill and slow-kill infection assays. This study provides insights into the roles of lapA in modulating P. aeruginosa virulence and biofilm formation under phosphate depletion stress.

Anafilaxia à lapa: um caso clínico raro / Limpet anaphylaxis: a rare case

A lapa (Patella vulgata) é um molusco frequentemente encontrado em regiões costeiras com clima quente. A alergia alimentar à lapa é muito rara, com poucos casos descritos na literatura. Os autores descrevem um caso de anafilaxia à lapa, com evidência de reação de hipersensibilidade do tipo I, através de IgE específica positiva à lapa, tanto com métodos in vivo, como in vitro.

Limpet (Patella vulgata) is a mollusk mainly found in warm coastal regions. Limpet allergy is considered rare, and few cases can be found in the literature. We describe a clinical case of limpet anaphylaxis, including in vitro and in vivo evidence of IgE mechanism involvement.

Anesthetic and surgical considerations for staged bilateral nephrectomies in a pediatric patient: A case report / Consideraciones anestésicas y quirúrgicas en nefrectomía bilateral en dos tiempos en un paciente pediátrico: Reporte de caso

Abstract We present a 9-year-old patient with end-stage renal disease, on peritoneal dialysis, who underwent a staged prone retroperitoneoscopic bilateral nephrectomy. Bilateral nephrectomy was indicated in preparation for renal transplant in the context of genetic predisposition malignancy when immunosuppressed. The two mirror-image surgeries enable the comparison of the anesthetic management and outcomes in a single patient. Features of interest to anesthesiologists include approach to a child with chronic kidney disease, different requirements for intraoperative antihypertensives; pain management strategies, including a comparison of erector spinae plane block with and without adjunct dexmedetomidine; anesthetic management of retroperitoneoscopic pediatric surgery and the first description of using a Foley bag attached to a peritoneal dialysis catheter to aid in diagnosis and repair of posterior peritoneal cavity entry.

Resumen Se presenta un paciente de 9 años de edad con enfermedad renal terminal, en diálisis peritoneal, quien se sometió a nefrectomía bilateral retroperitoneoscópica estadificada en posición prona. Se indicó la nefrectomía bilateral en preparación para trasplante renal en el contexto de predisposición genética hacia desarrollar una patología maligna al estar inmunosuprimido. Las dos cirugías en espejo permiten hacer una comparación del manejo anestésico y de los desenlaces en un mismo paciente. Las características de interés para los anestesiólogos incluyen el abordaje de un niño con enfermedad renal crónica, con requisitos diferentes de antihipertensivos intraoperatorios; estrategias para el manejo del dolor, incluyendo una comparación de bloqueo del plano del erector espinal con y sin dexmedetomidina adyuvante; manejo anestésico de cirugía pediátrica retroperitoneoscópica y la primera descripción del uso de una bolsa Foley conectada a un catéter de diálisis peritoneal para ayudar en el diagnóstico y la reparación de la entrada de la cavidad peritoneal posterior.

Pseudomonas putida Biofilm Depends on the vWFa-Domain of LapA in Peptides-Containing Growth Medium.

The biofilm of Pseudomonas putida is complexly regulated by several intercellular and extracellular factors. The cell surface adhesin LapA of this bacterium is a central factor for the biofilm and, consequently, the regulation of lapA expression, for example, by Fis. It has been recently shown that peptides in growth media enhance the formation of P. putida biofilm, but not as a source of carbon and nitrogen. Moreover, the peptide-dependent biofilm appeared especially clearly in the fis-overexpression strain, which also has increased LapA. Therefore, we investigate here whether there is a relationship between LapA and peptide-dependent biofilm. The P. putida strains with inducible lapA expression and LapA without the vWFa domain, which is described as a domain similar to von Willebrand factor domain A, were constructed. Thereafter, the biofilm of these strains was assessed in growth media containing extracellular peptides in the shape of tryptone and without it. We show that the vWFa domain in LapA is necessary for biofilm enhancement by the extracellular peptides in the growth medium. The importance of vWFa in LapA was particularly evident for the fis-overexpression strain F15. The absence of the vWFa domain diminished the positive effect of Fis on the F15 biofilm.

Identification of c-di-GMP/FleQ-Regulated New Target Genes, Including cyaA, Encoding Adenylate Cyclase, in Pseudomonas putida.

The bacterial second messenger cyclic diguanylate (c-di-GMP) modulates plankton-to-biofilm lifestyle transition of Pseudomonas species through its transcriptional regulatory effector FleQ. FleQ regulates transcription of biofilm- and flagellum-related genes in response to c-di-GMP. Through transcriptomic analysis and FleQ-DNA binding assay, this study identified five new target genes of c-di-GMP/FleQ in P. putida, including PP_0681, PP_0788, PP_4519 (lapE), PP_5222 (cyaA), and PP_5586 Except lapE encoding an outer membrane pore protein and cyaA encoding an adenylate cyclase, the functions of the other three genes encoding hypothetical proteins remain unknown. FleQ and c-di-GMP coordinately inhibit transcription of PP_0788 and cyaA and promote transcription of PP_0681, lapE, and PP_5586 Both in vitro and in vivo assays show that FleQ binds directly to promoters of the five genes. Further analyses confirm that LapE plays a central role of in the secretion of adhesin LapA and that c-di-GMP/FleQ increases lapE transcription, thereby promoting adhesin secretion and biofilm formation. The adenylate cyclase CyaA is responsible for synthesis of another second messenger, cyclic AMP (cAMP). FleQ and c-di-GMP coordinate to decrease the content of cAMP, suggesting that c-di-GMP and FleQ coregulate cAMP by modulating cyaA expression. Overall, this study adds five new members to the c-di-GMP/FleQ-regulated gene family and reveals the role of c-di-GMP/FleQ in LapA secretion and cAMP synthesis regulation in P. putida IMPORTANCE c-di-GMP/FleQ promotes the plankton-to-biofilm lifestyle transition at the transcriptional level via FleQ in Pseudomonas species. Identification of new target genes directly regulated by c-di-GMP/FleQ helps to broaden the knowledge of c-di-GMP/FleQ-mediated transcriptional regulation. Regulation of lapE by c-di-GMP/FleQ guarantees highly efficient LapA secretion and biofilm formation. The mechanism of negative correlation between c-di-GMP and cAMP in both P. putida and P. aeruginosa remains unknown. Our result concerning transcriptional inhibition of cyaA by c-di-GMP/FleQ reveals the mechanism underlying the decrease of cAMP content by c-di-GMP in P. putida.

Restoring Balance to the Outer Membrane: YejM's Role in LPS Regulation.

Gram-negative bacteria produce an asymmetric outer membrane (OM) that is particularly impermeant to many antibiotics and characterized by lipopolysaccharide (LPS) exclusively at the cell surface. LPS biogenesis remains an ideal target for therapeutic intervention, as disruption could kill bacteria or increase sensitivity to existing antibiotics. While it has been known that LPS synthesis is regulated by proteolytic control of LpxC, the enzyme that catalyzes the first committed step of LPS synthesis, it remains unknown which signals direct this regulation. New details have been revealed during study of a cryptic essential inner membrane protein, YejM. Multiple functions have been proposed over the years for YejM, including a controversial hypothesis that it transports cardiolipin from the inner membrane to the OM. Strong evidence now indicates that YejM senses LPS in the periplasm and directs proteolytic regulation. Here, we discuss the standing literature of YejM and highlight exciting new insights into cell envelope maintenance.

From Input to Output: The Lap/c-di-GMP Biofilm Regulatory Circuit.

Biofilms are the dominant bacterial lifestyle. The regulation of the formation and dispersal of bacterial biofilms has been the subject of study in many organisms. Over the last two decades, the mechanisms of Pseudomonas fluorescens biofilm formation and regulation have emerged as among the best understood of any bacterial biofilm system. Biofilm formation by P. fluorescens occurs through the localization of an adhesin, LapA, to the outer membrane via a variant of the classical type I secretion system. The decision between biofilm formation and dispersal is mediated by LapD, a c-di-GMP receptor, and LapG, a periplasmic protease, which together control whether LapA is retained or released from the cell surface. LapA localization is also controlled by a complex network of c-di-GMP-metabolizing enzymes. This review describes the current understanding of LapA-mediated biofilm formation by P. fluorescens and discusses several emerging models for the regulation and function of this adhesin.

Manejo laparoscópico de hernia diafragmática traumática: reporte de un caso / Laparoscopic management of traumatic diaphragmatic hernia: a case report

La hernia diafragmática traumática representa un reto al momento del diagnóstico. Una detección oportuna permite establecer un correcto tratamiento quirúrgico. Se presenta el caso de un paciente masculino de 37 años de edad con antecedente de trauma torácico por arma blanca 4 años antes; quien acude por cuadro de dolor abdominal intenso localizado en epigastrio que se irradia a hipocondrio izquierdo de aparición súbita 72 horas antes de su ingreso. Al examen físico murmullo vesicular disminuido en campo pulmonar izquierdo, y abdomen doloroso a la palpación a nivel de epigastrio e hipocondrio izquierdo. En exámenes de laboratorio no se evidencian alteraciones, mientras que la tomografía reporta hernia diafragmática izquierda; la misma que fue resuelta quirúrgicamente mediante técnica laparoscópica.

Traumatic diaphragmatic hernia represents a challenge at diagnosis. A timely detection allows to establish a correct surgical treatment. It is presented the case of a 37-year-old male patient with a history of stabbing chest trauma 4 years earlier; who comes for symptoms of intense abdominal pain located in the epigastrium that radiates to the left hypochondrium of sudden onset 72 hours before admission. On physical xamination, vesicular murmur decreased in the left lung field, and a painful abdomen on palpation at the level of the epigastrium and left hypochondrium were showed. Laboratory tests did not show any alterations, but tomography reports a left diaphragmatic hernia; this one was solved surgically by laparoscopic technique.

Chagas disease in Virgem da Lapa, Minas Gerais, Brazil: left ventricle aneurysm and the risk of death in the 24-year interval

BACKGROUND Left ventricular aneurysm (LVA) is indicator of high morbidity in Chagas' disease. A cross-sectional study performed identified LVA in 18.8% of the chronic chagasic patients (CCP). OBJECTIVE Determine the risk of death of patients with chronic chagasic cardiopathy (CCC) and LVA in 24-year interval. MATERIAL AND METHODS In 1995 a cohort of 298 CCP was evaluated by anamnesis, physical examination, EKG and ECHO and classified in groups: G0 = 86 without cardiopathy; G1 = 156 with cardiopathy without LVA and G2 = 56 with cardiopathy and LVA. 38 patients of G0 and G1 used benznidazole. Information about the deaths was obtained in the notary, death certificates, hospital records and family members. FINDINGS Were registered 113 deaths (37.9%): 107 (35.9%) attributed to cardiopathy and 6 (2.0%) to other causes (p < 0.05). Amongst these 107 deaths, 10 (11.6%) occurred in G0; 49 (31.4%) occurred in G1 and 48 (85.7%) occurred in G2 (p < 0.05). The risk of death was 2.7 and 7.4 times significantly higher in G2, than in G1 and G0, respectively. CONCLUSION Chronic chagasic patients with LVA and ejection fraction < 45% have a higher risk of death than those without.

Digestive tract reconstruction with tubular stapler after 4K laparoscopic total gastrectomy / 中华消化外科杂志

Digestive tract reconstruction after laparoscopic total gastrectomy is one of the difficult problems faced by surgeons. The classical reconstruction method is Roux-en-Y esophagojejunostomy, which is mainly completed by two instruments: linear stapler and tubular stapler. Due to the advantages of wide application range and safety of anastomosis, the tubular stapler is most commonly used in clinical practice. The traditional method of esophagojejunostomy with tubular stapler is end-to-side esophagojejunostomy. Because this method is directly transplanted from open surgery, there are many difficulties in the actual operation. Semi end to end esophagojejunostomy is an improved reconstruction method with tubular stapler. It is easy to operate and has less complications. It is the optimal choice for digestive tract reconstruction after laparoscopic total gastrectomy. With the help of 4K laparoscopic system, the operation is more accurate. The authors summarize key points of digestive tract reconstruction with tubular stapler after 4K laparoscopic total gastrectomy.

Modified strategy of anesthesia for laparoscopic radical resection of rectal carcinoma: ultrasound-guided anterior quadratus lumborum block combined with general anesthesia / 中华麻醉学杂志

Objective To evaluate the efficacy of ultrasound-guided anterior quadratus lumborum block combined with general anesthesia for laparoscopic radical resection of rectal carcinoma. Methods A total of 80 patients of both sexes, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 40-64 yr, scheduled for elective laparoscopic radical resection of rectal carcinoma, were divided into 2 groups ( n=40 each) using a random number table method: anterior quadratus lumborum block combined with general anesthesia group ( group QG) and general anesthesia group ( group G) . In group QG, anteri-or quadratus lumborum block was performed with 0. 33％ ropivacaine 25 ml and dexamethasone 5 mg under ultrasound guidance before operation, and the same procedure was performed on the other side. Combined intravenous-inhalational anesthesia was applied, propofol 3-5μg∕ml and remifentanil 3-5 ng∕ml were given by target-controlled infusion, and cisatracurium was intermittently injected in two groups. Patient-controlled intravenous analgesia with sufentanil 2μg∕kg was used for postoperative analgesia. The analgesic pump was set up to deliver a 2 ml bolus dose with a 15-min lockout interval. Bruggrmann comfort scale ( BCS) scores were recorded at 1, 6, 12, 24 and 48 h after operation ( T1-5 ) . Tramadol was used for rescue analgesic after operation. The consumption of remifentanil and sufentanil, requirement for tramadol, occurrence of adverse reactions and patients' satisfaction with postoperative analgesia were recorded. The emergence time, first ambulation time, time to first flatus∕poo and length of hospital stay were also recorded. The develop-ment of anterior quadratus lumborum block-related complications was recorded. Results Compared with group G, BCS scores were significantly increased at T4,5 , the consumption of remifentanil, requirement for tramadol and incidence of nausea and vomiting were decreased, patients' satisfaction with postoperative an-algesia was increased, and the emergence time, first ambulation time, time to first flatus∕poo and length of hospital stay were shortened in group QG (P<0. 05). Conclusion Ultrasound-guided anterior quadratus lumborum block combined with general anesthesia can reduce the consumption of opioids in the perioperative period and is helpful in improving outcomes when used for laparoscopic radical resection of rectal carcinoma.

Type 1 Does the Two-Step: Type 1 Secretion Substrates with a Functional Periplasmic Intermediate.

Bacteria have evolved several secretion strategies for polling and responding to environmental flux and insult. Of these, the type 1 secretion system (T1SS) is known to secrete an array of biologically diverse proteins-from small, <10-kDa bacteriocins to gigantic adhesins with a mass >1 MDa. For the last several decades, T1SSs have been characterized as a one-step translocation strategy whereby the secreted substrate is transported directly into the extracellular environment from the cytoplasm with no periplasmic intermediate. Recent phylogenetic, biochemical, and genetic evidences point to a distinct subgroup of T1SS machinery linked with a bacterial transglutaminase-like cysteine proteinase (BTLCP), which uses a two-step secretion mechanism. BTLCP-linked T1SSs transport a class of repeats-in-toxin (RTX) adhesins that are critical for biofilm formation. The prototype of this RTX adhesin group, LapA of Pseudomonas fluorescens Pf0-1, uses a novel N-terminal retention module to anchor the adhesin at the cell surface as a secretion intermediate threaded through the outer membrane-localized TolC-like protein LapE. This secretion intermediate is posttranslationally cleaved by the BTLCP family LapG protein to release LapA from its cognate T1SS pore. Thus, the secretion of LapA and related RTX adhesins into the extracellular environment appears to be a T1SS-mediated two-step process that involves a periplasmic intermediate. In this review, we contrast the T1SS machinery and substrates of the BLTCP-linked two-step secretion process with those of the classical one-step T1SS to better understand the newly recognized and expanded role of this secretion machinery.

To reduce routine computed tomographic angiography for thoracic aortic injury assessment in level II blunt trauma patients using three mediastinal signs on the initial chest radiograph: a preliminary report.

PURPOSE: CTA is routinely ordered on level II blunt thoraco-abdominally injured patients for assessment of injury to the thoracic aorta. The vast majority of such assessments are negative. The question being asked is, Does the accurate interpretation of the three mediastinal signs permit reliable determination of which patients need CTA for aortic assessment? The purpose of this investigation was to evaluate the role of three specifically selected mediastinal anatomic signs on the initial supine chest radiograph (CXR) of adult level II blunt thoraco-abdominally injured patients for the presence or absence of a mediastinal hematoma. The presence of a mediastinal hematoma is typically used as an indicator for computed tomographic angiography (CTA). The three mediastinal signs are the right para-tracheal stripe (RPTS), left para-spinal line (LPSL), and the left apical extra-pleural area (LAPA). MATERIALS AND METHODS: The patient triage designation (level II trauma) was made by the attending physician at the time of admission. The initial CXR image and the CTA report of the 197 adult blunt level II thoraco-abdominally injured patients obtained on the day of admission were compared. The CXR of each of the 197 patients was independently assessed by each of four observers specifically for the status of the three mediastinal signs. Each observer was blinded to the CTA report until after the status of the three mediastinal sign evaluation had been determined. Two or three of the mediastinal signs being positive were required to determine that the CXR was positive for a mediastinal hematoma. RESULTS: Two or three of the selected mediastinal signs were normal in 192 (97.5%) patients. None of these patients had either a mediastinal hematoma or a major aortic injury on CTA. In each of the remaining five (2.5%) patients, two or three of the mediastinal signs were abnormal. Each of these patients had a mediastinal hematoma and a major thoracic aortic injury on CTA. CONCLUSIONS: This preliminary study suggests that the accurate interpretation of the three specifically selected mediastinal signs on the initial supine CXR of adult level II blunt thoraco-abdominally injured patients could reduce the need for routine CTA for thoracic aortic injury assessment, and requires verification by an additional study.

Aging Modulates the Hemispheric Specialization during Word Production.

Although older adults exhibit normal accuracy in performing word retrieval and generation (lexical production; e.g., object naming), they are generally slower in responding than younger adults. To maintain accuracy, older adults recruit compensatory mechanisms and strategies. We focused on two such possible compensatory mechanisms, one semantic and one executive. These mechanisms are reflected at inter- and intra-hemispheric levels by various patterns of reorganization of lexical production cerebral networks. Hemispheric reorganization (HR) changes were also evaluated in relation to increase naming latencies. Using functional magnetic resonance imaging (fMRI), we examined 27 healthy participants (from 30 years to 85 years) during an object naming task, exploring and identifying task-related patterns of cerebral reorganization. We report two main results. First, we observed a left intra-hemispheric pattern of reorganization, the left anterior-posterior aging (LAPA) effect, consisting of supplementary activation of left posterior (temporo-parietal) regions in older adults and asymmetric activation along the left fronto-temporal axis. This pattern suggests that older adults recruit posterior semantic regions to perform object naming. The second finding consisted of bilateral recruitment of frontal regions to maintain appropriate response times, especially in older adults who were faster performers. This pattern is discussed in terms of compensatory mechanism. We suggest that aging is associated with multiple, co-existing compensation and reorganization mechanisms and patterns associated with lexical production.

Pedicle omentum flap harvested by laparoscopy in breast oncoplastic sugery / 中国癌症杂志

Surgery is one of the most important treatments for breast cancer. A part of the early breast cancer patients demand further oncoplastic breast surgery to reconstruct or restore the breast cosmosis after conventional breast conserving surgery, by oncoplastic breast surgery, which is a modern conception and technique including volume dis-placement and volume replacement. Oncoplastic breast surgery using pedicled omental flap is a new approach among volume replacement techniques. It has made some progress in recently years. A systematic review was therefore con-ducted to analyze and illuminate the present status.

New records of two endemic troglobitic and threatened arachnids (Amblypygi and Opiliones) from limestone caves of Minas Gerais state, southeast Brazil.

BACKGROUND: The endemic and threatened troglobites (organisms restricted to caves) Charinus eleonorae (Amblypygi) and Iandumoema uai (Opiliones), both from Olhos d'Água cave, located at Peruaçu Caves National Park (southeast Brazil), have their distribution expanded for a new locality inside of the National Park (Lapa do Cipó cave), extending their distribution at least in 9.5km(2). NEW INFORMATION: This new data suggest that these arachnids can be in a differentiation process and/or there are several possibilities of dispersion in the karst of Peruaçu. Indeed, a revision of their categorical status at IUCN Red List is necessary. We herein report a new distribution range (Lapa do Cipó cave) of the troglobitic species I. uai and C. eleonorae, which are, to date, known to occur in the Olhos d'Água cave, located at the Peruaçu Caves National Park (PCNP).

Beach almond (Terminalia catappa, Combretaceae) seed production and predation by scarlet macaws (Ara macao) and variegated squirrels (Sciurus variegatoides) / Producción de semillas y depredación del almendro de playa (Terminalia catappa, Combretaceae) por lapas rojas (Ara macao) y ardillas chizas (Sciurus variegatoides)

Knowledge of ecological impacts of exotic beach almond (Terminalia catappa) in the central Pacific of Costa Rica are little known, but studies have found this species to be a potentially important food source for endangered scarlet macaws (Ara macao). In this study, reproductive phenology and seed predation by variegated squirrels (Sciurus variegatoides) and scarlet macaws were measured during March and April 2011 on beaches of central Pacific coastal Costa Rica. Seed productivity and predation levels were quantified on a weekly basis for 111 beach almond trees to assess the importance of beach almond as a food source for scarlet macaws and the extent of resource partitioning between seed predators. Seed production of the trees was great (about 194 272 seeds) and approximately 67% of seeds were predated by seed predators. Macaws consumed an estimated 49% of seeds while squirrels consumed 18%. Additionally, evidence of resource partitioning between squirrels and macaws was found. Scarlet macaws preferred to feed on the northern side and edge of the canopy while squirrels preferred to feed on the southern and inside parts of the canopy. Both species ate most seeds on the ocean side of the tree. Despite the status of this tree as an exotic species, the beach almond appears to be an important resource for scarlet macaw population recovery. The resource produced by this tree should be taken into account as reforestation efforts continue in Costa Rica. Rev. Biol. Trop. 62 (3): 929-938. Epub 2014 September 01.

El conocimiento de los impactos ecológicos del almendro de playa exótico (Terminalia catappa) en el Pacífico Central de Costa Rica son poco conocidos, pero los estudios han encontrado que esta especie es una fuente de alimento potencialmente importante para la lapa roja (Ara macao), en peligro de extinción. En este estudio, se midieron la fenología reproductiva y la depredación de semillas por las ardillas (Sciurus variegatoides) y lapas rojas durante marzo y abril 2011 en las playas de la costa Pacífica Central de Costa Rica. Los niveles de productividad y depredación de semillas se cuantificaron semanalmente para 111 almendros de playa, para evaluar la importancia del almendro de playa como fuente de alimento para la lapa roja y el grado de repartición de recursos entre los depredadores de semillas. La producción de semillas de los árboles fue alta (cerca de 194 272 semillas) y aproximadamente el 67% de las semillas fueron comidas por los depredadores, las lapas rojas consumen un estimado de 49% de las semillas, mientras que las ardillas consumen el 18%. Adicionalmente, se encontró evidencia de la repartición de recursos entre las ardillas y las lapas. Las lapas rojas prefieren alimentarse en el lado norte y el borde de la copa, mientras que las ardillas prefieren las regiones del sur y el interior de la copa. Ambas especies se comieron la mayoría de las semillas en la parte del árbol con lado al mar. A pesar de la situación de este árbol como una especie exótica, la playa de almendras parece ser un recurso importante para la recuperación de la población de lapas rojas. El recurso que produce este árbol debe tomarse en cuenta para continuar con los esfuerzos de reforestación en Costa Rica.

Structure of tomato wound-induced leucine aminopeptidase sheds light on substrate specificity.

The acidic leucine aminopeptidase (LAP-A) from tomato is induced in response to wounding and insect feeding. Although LAP-A shows in vitro peptidase activity towards peptides and peptide analogs, it is not clear what kind of substrates LAP-A hydrolyzes in vivo. In the current study, the crystal structure of LAP-A was determined to 2.20 Šresolution. Like other LAPs in the M17 peptidase family, LAP-A is a dimer of trimers containing six monomers of bilobal structure. Each monomer contains two metal ions bridged by a water or a hydroxyl ion at the active site. Modeling of different peptides or peptide analogs in the active site of LAP-A reveals a spacious substrate-binding channel that can bind peptides of five or fewer residues with few geometric restrictions. The sequence specificity of the bound peptide is likely to be selected by the structural and chemical restrictions on the amino acid at the P1 and P1' positions because these two amino acids have to bind perfectly at the active site for hydrolysis of the first peptide bond to occur. The hexameric assembly results in the merger of the open ends of the six substrate-binding channels from the LAP-A monomers to form a spacious central cavity allowing the hexameric LAP-A enzyme to simultaneously hydrolyze six peptides containing up to six amino acids each. The hexameric LAP-A enzyme may also hydrolyze long peptides or proteins if only one such substrate is bound to the hexamer because the substrate can extend through the central cavity and the two major solvent channels between the two LAP-A trimers.

Crystallization and preliminary X-ray diffraction analysis of a high-affinity phosphate-binding protein endowed with phosphatase activity from Pseudomonas aeruginosa PAO1.

In prokaryotes, phosphate starvation induces the expression of numerous phosphate-responsive genes, such as the pst operon including the high-affinity phosphate-binding protein (PBP or pstS) and alkaline phosphatases such as PhoA. This response increases the cellular inorganic phosphate import efficiency. Notably, some Pseudomonas species secrete, via a type-2 secretion system, a phosphate-binding protein dubbed LapA endowed with phosphatase activity. Here, the expression, purification, crystallization and X-ray data collection at 0.87 Šresolution of LapA are described. Combined with biochemical and enzymatic characterization, the structure of this intriguing phosphate-binding protein will help to elucidate the molecular origin of its phosphatase activity and to decipher its putative role in phosphate uptake.