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Tropical forests are known for their high diversity. Yet, forest patches do occur in the tropics where a single tree species is dominant. Such "monodominant" forests are known from all of the main tropical regions. For Amazonia, we sampled the occurrence of monodominance in a massive, basin-wide database of forest-inventory plots from the Amazon Tree Diversity Network (ATDN). Utilizing a simple defining metric of at least half of the trees ≥ 10 cm diameter belonging to one species, we found only a few occurrences of monodominance in Amazonia, and the phenomenon was not significantly linked to previously hypothesized life history traits such wood density, seed mass, ectomycorrhizal associations, or Rhizobium nodulation. In our analysis, coppicing (the formation of sprouts at the base of the tree or on roots) was the only trait significantly linked to monodominance. While at specific locales coppicing or ectomycorrhizal associations may confer a considerable advantage to a tree species and lead to its monodominance, very few species have these traits. Mining of the ATDN dataset suggests that monodominance is quite rare in Amazonia, and may be linked primarily to edaphic factors.

Acinetobacter baumannii ( Ab ) is one of the most treacherous pathogens among those causing hospital-acquired pneumonia (HAP). A. baumannii possesses an adaptable physiology, seen not only in its antibiotic resistance and virulence phenotypes but also in its metabolic versatility. In this study, we observed that A. baumannii undergoes global transcriptional changes in response to human pleural fluid (PF), a key host-derived environmental signal. Differential gene expression analyses combined with experimental approaches revealed changes in A. baumannii metabolism, affecting cytotoxicity, persistence, bacterial killing, and chemotaxis. Over 1,220 genes representing 55% of the differentially expressed transcriptomic data corresponded to metabolic processes, including the upregulation of glutamate, short chain fatty acid, and styrene metabolism. We observed an upregulation by 1.83- and 2.61-fold of the pyruvate dehydrogenase complex subunits E3 and E2, respectively. We also found that pyruvate (PYR), in conjunction with PF, triggers an A. baumannii pathogenic behavior that adversely impacts human epithelial cell viability. Interestingly, PF also amplified A. baumannii cytotoxicity against murine macrophages, suggesting an immune evasion strategy implemented by A. baumannii . Moreover, we uncovered opposing metabolic strategies dependent on the degree of pathogenicity of the strains, where less pathogenic strains demonstrated greater utilization of PYR to promote persister formation in the presence of PF. Additionally, our transcriptomic analysis and growth studies of A. baumannii suggest the existence of an alternative phenylalanine (PA) catabolic route independent of the phenylacetic acid pathway, which converts PA to phenylpyruvate (PP) and shuttles intermediates into styrene metabolism. This alternative route promoted a neutrophil-evasive state, as PF-induced degradation of PP significantly reduced overall human neutrophil chemotaxis in ex vivo chemotactic assays. Taken together, these data highlight A. baumannii pathoadaptabililty in response to host signals and provide further insight into the role of bacterial metabolism in virulence traits, antibiotic persistence strategies, and host innate immune evasion.

Background and Aims: Combining simethicone (SIM) with a colon preparation agent has been shown to improve mucosal visibility during screening colonoscopy, but its effect on the adenoma detection rate (ADR) remains unclear. SIM is commonly used through the endoscope to eliminate bubbles during endoscopy. However, this practice recently has been associated with endoscope-transmitted infections. Our aims were to determine the role of SIM added to a polyethylene glycol preparation on the ADR, procedure times, colon preparation, and intraprocedural use of SIM., Methods: This was a randomized, controlled, observer-blinded, clinical trial of patients undergoing screening colonoscopy. Patients with a high risk of colorectal cancer were excluded. Patients were randomly assigned to 2 different preparations: polyethylene glycol plus SIM or polyethylene glycol. Two endoscopists blinded to patient preparation regimens scored its quality by using the Boston Bowel Preparation scale (BBPS) and the bubble scale. Interobserver agreement was calculated. The polyp detection rate, ADR, intraprocedural use of SIM, cecal intubation time, and withdrawal time were recorded. For study purposes, cecal intubation time and withdrawal time were combined to determine the effective procedure time., Results: No significant difference between the polyethylene glycol plus SIM and polyethylene glycol arms was seen regarding the ADR (33.3% vs 38.8%; P = .881) and effective procedure time (759.3 ± 253.1 seconds vs 800.2 ± 459.6 seconds; P = .373), respectively. Intraprocedural use of SIM as well as the bubble scale score were significantly lower in the polyethylene glycol plus SIM arm (1.6% vs 48.9%; P ≤ .05) and (0.1 vs 2.1; P ≤ .05), respectively. Conversely, no difference was found in the BBPS scores. The interobserver agreement for both scores was strong (bubble scale score kappa = .537; P < .05; BBPS score kappa = .184; P <.05)., Conclusion: Adding SIM to a polyethylene glycol preparation did not improve the ADR or effective procedure time. Nevertheless, it resulted in lower bubble scale scores, and more importantly, in less intraprocedural use of SIM. This simple and inexpensive intervention may have the potential to reduce the risk of endoscope-transmitted infections. (Clinical trial registration number: NCT03119168.)., (Copyright © 2019 American Society for Gastrointestinal Endoscopy. Published by Elsevier Inc. All rights reserved.)

The complex geological history of Central America has been useful for understanding the processes influencing the distribution and diversity of multiple groups of organisms. Anurans are an excellent choice for such studies because they typically exhibit site fidelity and reduced movement. The objective of this work was to identify the impact of recognized geographic barriers on the genetic structure, phylogeographic patterns and divergence times of a wide-ranging amphibian species, Hyalinobatrachium fleischmanni . We amplified three mitochondrial regions, two coding (COI and ND1) and one ribosomal (16S), in samples collected from the coasts of Veracruz and Guerrero in Mexico to the humid forests of Chocó in Ecuador. We examined the biogeographic history of the species through spatial clustering analyses (Geneland and sPCA), Bayesian and maximum likelihood reconstructions, and spatiotemporal diffusion analysis. Our data suggest a Central American origin of H. fleischmanni and two posterior independent dispersals towards North and South American regions. The first clade comprises individuals from Colombia, Ecuador, Panama and the sister species Hyalinobatrachium tatayoi ; this clade shows little structure, despite the presence of the Andes mountain range and the long distances between sampling sites. The second clade consists of individuals from Costa Rica, Nicaragua, and eastern Honduras with no apparent structure. The third clade includes individuals from western Honduras, Guatemala, and Mexico and displays deep population structure. Herein, we synthesize the impact of known geographic areas that act as barriers to glassfrog dispersal and demonstrated their effect of differentiating H. fleischmanni into three markedly isolated clades. The observed genetic structure is associated with an initial dispersal event from Central America followed by vicariance that likely occurred during the Pliocene. The southern samples are characterized by a very recent population expansion, likely related to sea-level and climatic oscillations during the Pleistocene, whereas the structure of the northern clade has probably been driven by dispersal through the Isthmus of Tehuantepec and isolation by the Motagua-Polochic-Jocotán fault system and the Mexican highlands., Competing Interests: Gabriela Parra Olea is an Academic Editor for PeerJ.

Introduction: the preliminary nutritional screening tool CIPA (food intake, proteins, anthropometry) is positive when it fulfills one of the following: control food intake (CI) 48-72 h < 50%, albumin < 3 g/dl, total protein < 5 g/dl, body mass index (BMI) < 18,5 kg/m2 or mid-upper arm circumference (MUAC) ≤ 22,5 cm. The use of two protein parameters increases costs and difficulty; one of them can be suppressed without affecting validity., Objectives: to evaluate the effectiveness of screening CIPA after exclusion of total protein., Method: prospective study of hospitalized patients; prevalence or risk of malnutrition was evaluated through CIPA and Subjective Global Assessment (SGA). Hospital malnutrition according to complete CIPA screening (with total proteins and albumin, [CIPAc] and without total proteins [CIPAw/p]) and concordance between both methods were analyzed, as well as the association of the positive screening result with clinical outcomes., Results: three hundred and forty-three patients were analyzed. The prevalence or risk of malnutrition identified by complete CIPA (c) was 38.19% (33.02-43.36); by CIPA without protein (w/p), 37.32% (32.17-42.46); and SGA was 29.15% (24.32-33.99). Kappa index: 0.981 between both CIPAs, p < 0.001. Both CIPA and SGA detect patients with higher mortality in hospital and one month after discharge. Early readmission was higher in positive CIPA, statistical significantly in CIPAw/p (screening with positive results 21.88% vs screening with negative results 13.49%, p = 0.044), SGA 20.01% vs 15.23%, p = 0.28. Length of stay was higher in patients with positive screening in CIPAc, CIPAw/p and SGA., Conclusions: CIPAw/p is equally or more effective than the previously validated full CIPA; therefore, it could replace the latter saving costs. The prevalence of malnutrition is high in both variants and they are able to predict which patient has worse clinical prognosis.

Species distribution models (SDMs) are widely used in ecology and conservation. Presence-only SDMs such as MaxEnt frequently use natural history collections (NHCs) as occurrence data, given their huge numbers and accessibility. NHCs are often spatially biased which may generate inaccuracies in SDMs. Here, we test how the distribution of NHCs and MaxEnt predictions relates to a spatial abundance model, based on a large plot dataset for Amazonian tree species, using inverse distance weighting (IDW). We also propose a new pipeline to deal with inconsistencies in NHCs and to limit the area of occupancy of the species. We found a significant but weak positive relationship between the distribution of NHCs and IDW for 66% of the species. The relationship between SDMs and IDW was also significant but weakly positive for 95% of the species, and sensitivity for both analyses was high. Furthermore, the pipeline removed half of the NHCs records. Presence-only SDM applications should consider this limitation, especially for large biodiversity assessments projects, when they are automatically generated without subsequent checking. Our pipeline provides a conservative estimate of a species' area of occupancy, within an area slightly larger than its extent of occurrence, compatible to e.g. IUCN red list assessments.

Background: The aim of this study was to report the clinical outcome and toxicity of radiochemotherapy in locally advanced gastric cancer (LAGC) patients treated according to the Intergroup 116 trial protocol in our institution., Methods: We retrospectively reviewed 105 patients with LAGC treated with radical surgery and adjuvant radiochemotherapy. We analyzed overall survival (OS), disease-free survival (DFS), locoregional failure-free survival (LFS), prognostic factors and toxicity., Results: The mean follow-up was 96.48 months. The majority of tumors were T3-T4 (75%) and 86.6% had nodal metastases. The OS, DFS and LFS rates to 3 years were 53.48%, 52.75% and 81.65%, respectively and to 5 years 40%, 46.73% and 76.77% respectively. The univariate analysis showed that N stage < N2, TN stage < IIIA, R0 resection and N‑ratio < 3 were statistically significant prognostic factors for OS and DFS, T stage < T4 for OS and N‑ratio < 3 for LFS. The group with D2 lymphadenectomy had worse LFS than the D1 group (65.2% vs 88.1%, respectively, p = 0.039) probably due to a significant difference in the proportion node positive patients in the D2 group (94% vs. 78%; p = 0.027). In the multivariate analysis, only R0 resection was statistically significant factor for improved OS (p = 0.018). Acute grade III-IV gastrointestinal and hematologic toxicity rates were 8.5% and 15.2%, respectively and 89.5% completed treatment as planned., Conclusion: Our results are consistent with those of the Intergroup-0116 trial for LAGC in terms of survival. This regimen is well tolerated and with acceptable toxicity. An R0 resection was an independent prognostic factor for improved OS.

Community-acquired methicillin resistant Staphylococcus aureus emerged as a worldwide health problem in the last few years. In Argentina, it is found in 70% of skin and skin structure infections in previously healthy adult patients and causes severe invasive diseases. The ST30-SCCmecIVc-spat019 clone is predominant in adult infections and has displaced the previously prevalent ST5-SCCmecIVa-spat311 clone in community settings. In the present work we compared the virulence of both clones in order to explain the displacement, and found that ST30-IVc is associated with invasive infections in adult patients from Argentina and possesses a different virulence-associated genes profile compared to ST5-IVa. A representative strain of ST30 lineage has a more aggressive behavior in animal models of infection and expresses higher level of Fibronectin binding protein A coding gene, which could enhance the bacterial invasion capacity., (Copyright © 2017. Published by Elsevier GmbH.)

The extent to which pre-Columbian societies altered Amazonian landscapes is hotly debated. We performed a basin-wide analysis of pre-Columbian impacts on Amazonian forests by overlaying known archaeological sites in Amazonia with the distributions and abundances of 85 woody species domesticated by pre-Columbian peoples. Domesticated species are five times more likely than nondomesticated species to be hyperdominant. Across the basin, the relative abundance and richness of domesticated species increase in forests on and around archaeological sites. In southwestern and eastern Amazonia, distance to archaeological sites strongly influences the relative abundance and richness of domesticated species. Our analyses indicate that modern tree communities in Amazonia are structured to an important extent by a long history of plant domestication by Amazonian peoples., (Copyright © 2017, American Association for the Advancement of Science.)

Detailed understanding and control of the intermolecular forces that govern molecular assembly are necessary to engineer structure and function at the nanoscale. Liquid crystal (LC) assembly is exceptionally sensitive to surface properties, capable of transducing nanoscale intermolecular interactions into a macroscopic optical readout. Self-assembled monolayers (SAMs) modify surface interactions and are known to influence LC alignment. Here, we exploit the different dipole magnitudes and orientations of carboranethiol and -dithiol positional isomers to deconvolve the influence of SAM-LC dipolar coupling from variations in molecular geometry, tilt, and order. Director orientations and anchoring energies are measured for LC cells employing various carboranethiol and -dithiol isomer alignment layers. The normal component of the molecular dipole in the SAM, toward or away from the underlying substrate, was found to determine the in-plane LC director orientation relative to the anisotropy axis of the surface. By using LC alignment as a probe of interaction strength, we elucidate the role of dipolar coupling of molecular monolayers to their environment in determining molecular orientations. We apply this understanding to advance the engineering of molecular interactions at the nanoscale.

Estimates of extinction risk for Amazonian plant and animal species are rare and not often incorporated into land-use policy and conservation planning. We overlay spatial distribution models with historical and projected deforestation to show that at least 36% and up to 57% of all Amazonian tree species are likely to qualify as globally threatened under International Union for Conservation of Nature (IUCN) Red List criteria. If confirmed, these results would increase the number of threatened plant species on Earth by 22%. We show that the trends observed in Amazonia apply to trees throughout the tropics, and we predict that most of the world's >40,000 tropical tree species now qualify as globally threatened. A gap analysis suggests that existing Amazonian protected areas and indigenous territories will protect viable populations of most threatened species if these areas suffer no further degradation, highlighting the key roles that protected areas, indigenous peoples, and improved governance can play in preventing large-scale extinctions in the tropics in this century.

Objective: Post-operative meningitis, caused mainly by Staphylococcus aureus and Gram-negative rods, is a life-threatening complication after neurosurgery, and its pathogenesis is far from clear. The purpose of this work was to study the experimental infection of human dura-mater fibroblasts and whole human dura by S. aureus., Methods: In vitro cultures of human dura-mater fibroblasts and organotypic cultures of small pieces of human dura mater were inoculated with a human-derived S. aureus strain. The pattern of bacterial infection as well as cytokines secretion by the infected fibroblasts was studied., Results: Our results suggest that colonisation of human dura-mater fibroblasts in culture and whole dura-mater tissue by S. aureus includes bacterial growth on the cell surface, fibroblast intracellular invasion by bacteria and a significant synthesis of interleukin 1beta (IL-1beta) by the infected cells., Conclusion: This is the first report of human dura-mater fibroblast infection by S. aureus. Hopefully, these results can lead to a better understanding of the pathogenesis of meningitis caused by this bacterial species and to a more rational therapeutic approach.

Anomalous origin of the left pulmonary artery from the ascending aorta is a rare cardiac malformation more commonly described with tetralogy of Fallot (TOF). The association of absent right carotid artery with TOF and anomalous left pulmonary artery from the ascending aorta has been rarely reported in the past. This case highlights the need for careful evaluation of the aortic arch and head vessels anatomy in patients with TOF because of the potential neurologic complications after surgery., (© The Author(s) 2014.)

Background: Proton pump inhibitors (PPIs) are commonly overused in hospitalized patients. The objectives of this study were to determine the extent of their inappropriate initiation in patients with low risk for gastrointestinal hemorrhage, factors associated with their continuation on discharge and potential cost of this trend., Methods: Retrospective examination of patients with low risk for gastrointestinal hemorrhage admitted to a tertiary-care teaching hospital over a 3-month period who received esomeprazole. The following information was collected: age, gender, PPI status (de novo or continued) and admitting diagnoses. Additional information collected from the de novo subgroup included indication for PPI, number of days on PPI and continuation of the drug on discharge. The cost of the medication was obtained from pharmacy records., Results: Four hundred nine patients were admitted during the study period and 204 (49.9%) received PPI de novo. Among these, 155 patients (76%) had an inappropriate indication for PPI. Of these, 62 (40%) patients were continued on PPI on discharge. Older age was a significant predictor of continuation of PPI at discharge. The estimated cost of the inpatient and outpatient inappropriate use of PPI was $12,272 and $59,272, respectively., Conclusions: PPIs are overused in the majority of hospitalized patients with low risk for gastrointestinal bleeding and this practice gets perpetuated at discharge, especially in older patients. The cost of this phenomenon is alarming.

Forest inventory studies in the Amazon indicate a large terrestrial carbon sink. However, field plots may fail to represent forest mortality processes at landscape-scales of tropical forests. Here we characterize the frequency distribution of disturbance events in natural forests from 0.01 ha to 2,651 ha size throughout Amazonia using a novel combination of forest inventory, airborne lidar and satellite remote sensing data. We find that small-scale mortality events are responsible for aboveground biomass losses of ~1.7 Pg C y(-1) over the entire Amazon region. We also find that intermediate-scale disturbances account for losses of ~0.2 Pg C y(-1), and that the largest-scale disturbances as a result of blow-downs only account for losses of ~0.004 Pg C y(-1). Simulation of growth and mortality indicates that even when all carbon losses from intermediate and large-scale disturbances are considered, these are outweighed by the net biomass accumulation by tree growth, supporting the inference of an Amazon carbon sink.

Nutcracker esophagus is an esophageal motility disorder characterized by the presence of hypertensive contraction waves. These waves can have very high amplitudes, but they maintain a peristaltic pattern and therefore, bolus passage is minimally affected. Esophageal food impactions are rare in nutcracker esophagus. Our patient was a previously asymptomatic man who presented with an esophageal meat impaction due to nutcracker esophagus in which high-resolution manometry played a key role in the diagnosis. Although a rare etiology, nutcracker esophagus can result in esophageal food impaction. High-resolution manometry plays a critical role in the diagnosis of specific motility disorders, even in the setting of minimal symptoms.

Bacteria of the genus Azospirillum colonize roots of important cereals and grasses, and promote plant growth by several mechanisms, notably phytohormone synthesis. The genomes of several Azospirillum strains belonging to different species, isolated from various host plants and locations, were recently sequenced and published. In this study, an additional genome of an A. brasilense strain, isolated from maize grown on an alkaline soil in the northeast of Mexico, strain CBG497, was obtained. Comparative genomic analyses were performed on this new genome and three other genomes (A. brasilense Sp245, A. lipoferum 4B and Azospirillum sp. B510). The Azospirillum core genome was established and consists of 2,328 proteins, representing between 30% to 38% of the total encoded proteins within a genome. It is mainly chromosomally-encoded and contains 74% of genes of ancestral origin shared with some aquatic relatives. The non-ancestral part of the core genome is enriched in genes involved in signal transduction, in transport and in metabolism of carbohydrates and amino-acids, and in surface properties features linked to adaptation in fluctuating environments, such as soil and rhizosphere. Many genes involved in colonization of plant roots, plant-growth promotion (such as those involved in phytohormone biosynthesis), and properties involved in rhizosphere adaptation (such as catabolism of phenolic compounds, uptake of iron) are restricted to a particular strain and/or species, strongly suggesting niche-specific adaptation.

In molecular biology studies of Anura, nondestructive methods to obtain genetic material are needed as alternatives to toe clipping. This work evaluates a nondestructive method for sampling DNA from blood puncture, comparing the performance of three different extraction protocols (Qiagen Kit, Salting-out and Chelex). We collected 134 individuals of Eleutherodactylus johnstonei, extracting blood via puncture of the medial vein using commercial-grade glucometer lancets. We extracted 100-1880 ng DNA, finding no differences between the extraction protocols. We compared the quality of the resulting DNA through amplification and sequencing of the 16S mitochondrial gene. Amplification was successful for the three extraction protocols, although Chelex showed better performance, making it the most recommendable protocol for extraction of DNA from blood. The resulting sequences corresponded to those registered in the GenBank for this species. Additionally, we found no significant differences in survival or weight change between the individuals that were manipulated and a control group (mean survival 66.7% treated, 62.9% untreated). Data reveal that blood samples obtained by puncture are a convenient alternative to other tissues (phalange, buccal swab, liver) that have traditionally been used as DNA sources for anurans. The technique is applicable to small and large species, covering most anuran diversity, provides enough DNA for many genetic applications and produces no noticeable effect on the survival or performance, given that it does not affect the motor parts or the dexterity of the animals., (© 2012 Blackwell Publishing Ltd.)

The Andes of South America hosts perhaps the highest amphibian species diversity in the world, and a sizable component of that diversity is comprised of direct-developing frogs of the genus Pristimantis (Anura: Craugastoridae). In order to better understand the initial stages of species formation in these frogs, this study quantified local-scale spatial genetic structuring in three species of Pristimantis. DNA sequences of two mitochondrial gene fragments (16S and COI) were obtained from P. brevifrons, P. palmeri and P. jubatus at different locations in the Cordillera Occidental. We found high levels of genetic diversity in the three species, with highly structured populations (as measured by F(ST)) in P. brevifrons and P. palmeri while P. jubatus showed panmixia. Large effective population sizes, inferred from the high levels of genetic diversity, were found in the three species and two highly divergent lineages were detected within P. jubatus and P. palmeri. Estimated divergence times among populations within P. brevifrons and P. palmeri coincide with the Pleistocene, perhaps due to similar responses to climatic cycling or recent geological history. Such insights have important implications for linking alpha and beta diversity, suggesting regional scale patterns may be associated with local scale processes in promoting differentiation among populations in the Andes.

Experimental research has produced evidence in recent years underlying the beneficial effects that exercise can have in preventing and deceleration of the development of Parkinson disease. These beneficial effects are exerted through various mechanisms such as neuroprotection, neurotransmission, plasticity, neurogenesis, homeostasis, and neurotrophic factors. Studies on clinical application at an early stage are still scarce, although some results are encouraging. There are still many questions to determine the most suitable type of exercise (forced/voluntary), the time of its implementation, the duration, and the combination of strategies. Nonconventional therapies can play an important role in addition to exercise, and are so numerous that they could be adapted to the circumstances of patients, although there is no evidence to date that they could have a neuroprotective effect.

In order to evaluate the protection conferred by an experimental inactivated vaccine against infectious coryza, three challenge trials were undertaken using 112 1-day-old broilers. The vaccine "Hepa Inmuno NC" included bacterial antigens of Avibacterium paragallinarum (serogroups A, B, variant B, and C) as well as antigens of Newcastle virus and hepatitis virus. Fifty-six broiler chicks were vaccinated at the first day of life at the hatchery while another 56 chicks were left unvaccinated. Three infection trials were conducted simultaneously using each of the three serogroups A, B, or C of Av. paragallinarum. In each trial, 17 vaccinated and 17 unvaccinated broilers were used. Challenge was performed at day 31 of life by injection, into the left infraorbital sinus, of approximately 1 x 10(5) colony forming units of the corresponding Av. paragallinarum strain. Clinical signs were recorded on day 2 postchallenge. All broilers were euthanatized and both infraorbital sinuses were bacteriologically examined for the presence of Av. paragallinarum on day 5 postchallenge. In comparison with the unvaccinated broilers, the vaccine significantly reduced the number of broilers with clinical signs after challenge with serogroup B, and significantly fewer vaccinated broilers were positive for the presence of Av. paragallinarum after challenge with serogroup C. On the other hand, no significant protection was observed when broilers were challenged with Av. paragallinarum from serogroup A. Despite the high infection rates in vaccinated chicks after an experimental infection with Av. paragallinarum, it was possible to reduce colonization of Av. paragallinarum (serogroup B) and clinical signs (serogroup C) in broiler chicks by vaccination at the first day of life. Further cross-protection trials should be done, including other Av. paragallinarum strains in the vaccine, especially those from serogroup A.

Purpose: To evaluate the evolution of apical formation on 28 necrotic immature permanent teeth treated with calcium hydroxide at different stages of root development., Methods: Apical formation in 28 necrotic incisors was carried out (27 upper and one lower incisors), in children between the ages of 6 and 13 years old (11 males, 10 females). Following anesthesia and rubber dam isolation, the chamber was opened and the coronal and root pulp tissue was removed. Next, after measuring the canal, the root canal was irrigated with 5% sodium hypochlorite. Once the canal was dried, it was filled with the CaOH2 powder mixed with physiologic saline solution to a dense consistency but malleable. This paste was compressed into the canal using a cotton pellet. The canal was completely filled up to the apex. Finally, the cavity opening was sealed with zinc oxide-eugenol (IRM) and glass-ionomer (Vitrebond). Follow-up appointments were made every 3 months in order to evaluate the evolution of the periapical radiolucency and the formation of the apical barrier., Results: The duration of the apical induction was 8.6 +/- 5.36 months. Increase or lack of growth in the length of the root canal and the type of root end closing was as follows: cementoid tissue (85.72%); osseous tissue (14.28%), with a 100% overall success rate. The evolution of these teeth was monitored over a 2-year period with re-infections occurring in 7.1 percent of the cases.

Purpose: To study the evolution of 308 necrotic primary teeth after pulpectomy technique with resorbable paste (Kri-1 + calcium hydroxide + metacresol-formaldehyde) after 30 months., Methods: 134 children between 2-9 years were included, among which they had 348 pulpectomized teeth. Out of these 348 teeth, 308 had necrotic pulp and were included in the study. The filling material consisted of a paste based on Walkhoff's master formula and consisted of the combination of Kri-1a (80.8% iodoform, 2.025% parachlorophenol, 1.215% menthol, 4.68% camphor and excipient 15 mg), pure calcium hydroxide and metacresol formaldehyde (metacresol 20 mg, formaldehyde 20 mg, eugenol 20 mg, eucalyptus 6 mg and excipient 40 mg). Root canal filling was performed in one session by one operator., Results: Pain, swelling, and fistula resolution was observed in most cases at the first recall. Progressive remission of radiolucent areas was also observed. A relationship between fistula and radiolucency in pre-treated furcation area was observed, and, in some cases, premature eruption of permanent teeth was recorded. Only two cases out of 308 treated teeth required extraction.

The LIFEBRIDGE B2T is a new portable cardiopulmonary bypass (CPB) system designed for temporary circulatory support. The LIFEBRIDGE B2T consists of a disposable patient unit with a CPB circuit, a control, and a base unit. The system weighs 20 kg. We used the LIFEBRIDGE B2T in four patients for circulatory support in beating heart coronary artery bypass graft for complete revascularization. The LIFEBRIDGE B2T was connected via femoral cannulation. Concentrations of free hemoglobin (fHb), interleukin (IL)-6, and -8 were measured. For venous blood drainage, 22-24 Fr cannulae and for arterial cannulation, 16-20 Fr cannulae were used. Average extracorporeal circulation (ECC) time was 61 +/- 18 minutes. During circulatory support, the system delivered an arterial blood flow between 3.1 and 4.1 L/min. The negative pressure at the venous drainage was between -79 and -45 mm Hg. During circulatory support, fHb concentration increased from 5.8 +/- 1.7 mg/dL to a maximum of 10.2 +/- 6.2 mg/dL. Also, IL-6 and -8 increased from 2.1 +/- 0.06 to 503.3 +/- 400.7 U/L and 5.9 +/- 0.9 to 66.5 +/- 46.8 U/L, respectively. The LIFEBRIDGE B2T is a new portable and safe circulatory support system. Connected via femoral cannulation, the system provides adequate arterial blood flow and an acceptable negative pressure at the venous cannula. The fHb concentration showed only a moderate increase during ECC.

Cytomegalovirus infection is usually asymptomatic or resembles infectious mononucleosis with fever, pharyngitis, arthralgias, lymphadenopathy, and atypical lymphocytosis. Even though primary CMV infection is usually self-limited in healthy individuals, significant complications can develop in immunocompromised patients. Venous or arterial thromboembolic phenomena are uncommon, yet very serious complications of CMV infection. Most published reports describe immunosupressed patients after organ transplantation or in the presence of HIV co-infection. However, thrombotic events in CMV infected immunocompetent individuals may occur. We describe the case of an immunocompetent adolescent with acute cytomegalovirus hepatitis that was complicated with pulmonary embolism and portal vein thrombosis. To our knowledge, this is the first reported case in which these two thrombotic phenomena occurred simultaneously in an adolescent with no obvious predisposing factors for thrombosis in the setting of an acute CMV infection.

Malaria is a serious public health problem in over 90 countries worldwide. In Brazil the disease is prevalent in the Amazon and the control rationale is based on early diagnosis and treatment. Quality pharmaceutical services are considered a key element for control. A proposal for evaluating pharmaceutical services for malaria is presented here. A theoretical outline composed of a logical model and an indicator framework is discussed and strives to establish a basis for assessment and judgment of the way in which these services are actually delivered. The aim is to contribute to the understanding of pharmaceutical services for malaria and other endemic diseases, complying with the directives of the Brazilian National Medicines Policy.

The Menidia humboldtiana silverside is considered one of the most important species in fisheries in some states of Mexico. Knowing the choice that this species exercises over its food gives us an understanding of the behavior of zooplankton in presence of zooplanktonic fish, and tells us if the silverside choose amongst bigger prey fish, and if there is competition for food between the native and introduced fish in the reservoir. The purpose of this study was to determine if M. humboldtiana selects its food, and in which genera, and to establish whether there was an overlap in the diet of the different sizes of silverside. The fish were catched with a 88 mm mesh net, in six sampling points during one year, from Danxh6 (19 degrees 5'17" - 53'46" N, 99 degrees 32'42" - 35'40" W) reservoir. Samples of filtered zooplankton were also taken with a 125 micron net. The analysis of stomach contents was carried out in fish grouped by regular size intervals during each season of the year, using the volumetric method and Chesson's selectivity coefficient. To evaluate the trophic overlap between the different size groups, the Morisita index, modified by Horn, was used. The zooplankton community was represented by twelve genera: Mastigodiaptomus and Cyclops (Copepoda); Bosmina, Diaphanosoma, Daphnia, Ceriodaphnia, Moina, Alonopsis and Camptocercus, (Cladocera); Asplanchna, Conochillus and Filinia (Rotifera). Mastigodiaptomus was the most abundant throughout the year. The silverside consumed only four genera: Bosmina, Mastigodiaptomus, Daphnia and Ceriodaphnia; the group measuring 5 to 8.9 cm consumed a high percentage of Bosmina, Mastigodiaptomus and Daphnia, and a lesser percentage of Ceriodaphnia. The larger fish (9 to 10.9 cm) consumed only Mastigodiaptomus and Daphnia. In accordance with the selectivity values, the small and medium size silverside selected their prey, with more intensity on the genera Bosmina and Daphnia, while the bigger fish concentrated on Daphnia and Mastigodiaptomus. During spring, the silverside between 3 to 8.9 cm showed overlap in diet; in summer the bigger size group (7-10.9 cm) showed the overlap. In autumm the overlap ocurred in all sizes, and in winter it was shown only in the medium size fish (5-8.9 cm).

Anthropogenic edge effects can compromise the conservation value of mature tropical forests. To date most edge-effect research in Amazonia has concentrated on forests in relatively seasonal locations or with poor soils in the east of the basin. We present the first evaluation from the relatively richer soils of far western Amazonia on the extent to which mature forest biomass, diversity, and composition are affected by edges. In a southwestern Amazonian landscape we surveyed woody plant diversity, species composition, and biomass in 88x0.1 ha samples of unflooded forest that spanned a wide range in soil properties and included samples as close as 50 m and as distant as >10 km from anthropogenic edges. We applied Mantel tests, multiple regression on distance matrices, and other multivariate techniques to identify anthropogenic effects before and after accounting for soil factors and spatial autocorrelation. The distance to the nearest edge, access point, and the geographical center of the nearest community ("anthropogenic-distance effects") all had no detectable effect on tree biomass or species diversity. Anthropogenic-distance effects on tree species composition were also below the limits of detection and were negligible in comparison with natural environmental and spatial factors. Analysis of the data set's capacity to detect anthropogenic effects confirmed that the forests were not severely affected by edges, although because our study had few plots within 100 m of forest edges, our confidence in patterns in the immediate vicinity of edges is limited. It therefore appears that the conservation value of most "edge" forests in this region has not yet been compromised substantially. We caution that because this is one case study it should not be overinterpreted, but one explanation for our findings may be that western Amazonian tree species are naturally faster growing and more disturbance adapted than those farther east.

An important strategy for "shotgun proteomics" profiling involves solution proteolysis of proteins, followed by peptide separation using multidimensional liquid chromatography and automated sequencing by mass spectrometry (LC-MS/MS). Several protocols for extracting and handling membrane proteins for shotgun proteomics experiments have been reported, but few direct comparisons of different protocols have been reported. We compare four methods for preparing membrane proteins from human cells, using acid labile surfactants (ALS), urea, and mixed organic-aqueous solvents. These methods were compared with respect to their efficiency of protein solubilization and proteolysis, peptide and protein recovery, membrane protein enrichment, and peptide coverage of transmembrane proteins. Overall, approximately 50-60% of proteins recovered were membrane-associated, identified from Gene Ontology annotations and transmembrane prediction software. Samples extracted with ALS, extracted with urea followed by dilution, or extracted with urea followed by desalting yielded comparable peptide recoveries and sequence coverage of transmembrane proteins. In contrast, suboptimal proteolysis was observed with organic solvent. Urea extraction followed by desalting may be a particularly useful approach, as it is less costly than ALS and yields satisfactory protein denaturation and proteolysis under conditions that minimize reactivity with urea-derived cyanate. Spectral counting was used to compare datasets of proteins from membrane samples with those of soluble proteins from K562 cells, and to estimate fold differences in protein abundances. Proteins most highly abundant in the membrane samples showed enrichment of integral membrane protein identifications, consistent with their isolation by differential centrifugation.

Correct identification of a peptide sequence from MS/MS data is still a challenging research problem, particularly in proteomic analyses of higher eukaryotes where protein databases are large. The scoring methods of search programs often generate cases where incorrect peptide sequences score higher than correct peptide sequences (referred to as distraction). Because smaller databases yield less distraction and better discrimination between correct and incorrect assignments, we developed a method for editing a peptide-centric database (PC-DB) to remove unlikely sequences and strategies for enabling search programs to utilize this peptide database. Rules for unlikely missed cleavage and nontryptic proteolysis products were identified by data mining 11 849 high-confidence peptide assignments. We also evaluated ion exchange chromatographic behavior as an editing criterion to generate subset databases. When used to search a well-annotated test data set of MS/MS spectra, we found no loss of critical information using PC-DBs, validating the methods for generating and searching against the databases. On the other hand, improved confidence in peptide assignments was achieved for tryptic peptides, measured by changes in DeltaCN and RSP. Decreased distraction was also achieved, consistent with the 3-9-fold decrease in database size. Data mining identified a major class of common nonspecific proteolytic products corresponding to leucine aminopeptidase (LAP) cleavages. Large improvements in identifying LAP products were achieved using the PC-DB approach when compared with conventional searches against protein databases. These results demonstrate that peptide properties can be used to reduce database size, yielding improved accuracy and information capture due to reduced distraction, but with little loss of information compared to conventional protein database searches.

Identifying proteins in cell extracts by shotgun proteomics involves digesting the proteins, sequencing the resulting peptides by data-dependent mass spectrometry (MS/MS), and searching protein databases to identify the proteins from which the peptides are derived. Manual analysis and direct spectral comparison reveal that scores from two commonly used search programs (Sequest and Mascot) validate less than half of potentially identifiable MS/MS spectra (class positive) from shotgun analyses of the human erythroleukemia K562 cell line. Here we demonstrate increased sensitivity and accuracy using a focused search strategy along with a peptide sequence validation script that does not rely exclusively on XCorr or Mowse scores generated by Sequest or Mascot, but uses consensus between the search programs, along with chemical properties and scores describing the nature of the fragmentation spectrum (ion score and RSP). The approach yielded 4.2% false positive and 8% false negative frequencies in peptide assignments. The protein profile is then assembled from peptide assignments using a novel peptide-centric protein nomenclature that more accurately reports protein variants that contain identical peptide sequences. An Isoform Resolver algorithm ensures that the protein count is not inflated by variants in the protein database, eliminating approximately 25% of redundant proteins. Analysis of soluble proteins from a human K562 cells identified 5130 unique proteins, with approximately 100 false positive protein assignments.

Introduction: Ocular hypotony is an indirect open-globe injury sign that can be evaluated during primary care. A study was performed to ascertain the chances of a patient with ocular trauma and hypotony during initial evaluation of having open-globe injury compared with another patient without hypotony, and to estimate the value of hypotony as a predictor of open-globe injury., Material and Methods: Patients with ocular trauma evaluated between 1995 and 2003 with intraocular pressure records were included; patients with other causes of hypotony were excluded. Patients with hypotony were assigned to group 1 and those without hypotony to group 2. Rate of open-globe injury was compared between groups with chi2, odds ratio (OR), and 95% confidence intervals (95% CI)., Results: Three hundred sixty one patients (aged 1-90 years average age 26.3 years) were evaluated; 329 patients had closed-globe (91.1%) and 32, open-globe injury (8.9%). Thirty six were assigned to group 1 (10%) and 325 (90%) to group 2. Twenty three patients in group 1 had open-globe injury (63.9%) while nine in group 2 had open-globe injury (2.8%, p <0.001, OR 62.12, 95% CI 21.81-183.6). There was hypotony in 71.9% of eyes with open-globe injury and in 4% of eyes with closed-globe injury., Discussion: Although hypotony was not present in every open-globe injury, the chance of finding the latter when there was hypotony was 62 times greater than without it. Absence of hypotony might be a useful tool to rule out open-globe injury. It would be convenient to validate ocular hypotony as a diagnostic test of open-globe injury by means of a prospective study.

In an acute assay, rats fasted for 24 h were sounded with 2 ml of fresh yogurt, sterilised yogurt or distilled water, and sacrificed at 2, 4, 8 and 24 h. They were compared with non-sounded rats. The survival of the lactic bacteria of the yogurt administered in the animals' stomach and intestines was determined, and the bacteria isolated were to the Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus profiles belonging to the strains isolated originally from the yogurt. Counts of the total number of microorganisms that were grown in lactobacilli and lactic streptococci media were also made. Analyses of the different types of resulting colonies in the lactobacilli and lactic streptococci media was performed. No L. delbrueckii subsp. bulgaricus or S. thermophilus were characterized in this acute assay. The statistical analysis of the results did not show significant differences in the log UFC. g-1, of the intestinal microbiota microorganisms of the three groups of animals in any of the times. In a chronic assay, three groups of animals, after 30 days of nourishment ad libitum with either a semisynthetic diet with 10% of fresh yogurt, or with 10% of sterilised yogurt or controls, housed in individual metabolism cells, were sacrificed. The results did not show meaningful differences in the long UFC. g-1 of the intestinal microbiota microorganisms from the three groups of animals in any of the times. In turn, in the study of the different types of colonies resulting in the lactobacilli and lactic streptococci media, no L. delbrueckii subsp. bulgaricus or S. thermophilus were characterised in this chronic assay. The results did not show meaningful differences in the log CFU. g-1 of the intestinal flora microorganisms from the three groups of animals in any of the times. In turn, no L. delbrueckii subsp. bulgaricus or S. thermophilus were characterised in this chronic assay.

Toxicity of cyanide is related to its inhibitory action on cytochrome c oxidase (COx). The alpha-keto acids pyruvate and alpha-ketoglutarate are known to exert in vivo certain protective effect against CN- toxicity if present when the poison is administered. We characterized in vitro their protection of oxidative phosphorylation and of the activity of COx in rat testis, heart, and liver mitochondria, as well as that of the beef heart enzyme in the pure state. In all cases the keto acids proved to have a protective action, even when KCN was previously added to the incubation mixtures and inhibition had already been established. However, the extent of the protection seemed to depend upon the degree of aeration. In the presence of the alpha-keto acids, O2 succeeded with 77% efficiency in reversing CN- inhibition of [gamma-32P]ATP synthesis under high aeration, whereas only 15% was achieved if aeration was poor. In poisoned heart mitochondria (1 mM KCN) simultaneous estimations of ATP synthesis and COx activity in a closed oxygraph chamber showed a recovery of only 6% of both activities upon the addition of 12 mM pyruvate. Our results suggest that O2 displaced cyanide from the enzyme and the poison was then trapped by the keto acids to form the respective nontoxic cyanohydrin. The combination of both high oxygen concentration and the presence of either pyruvate or alpha-ketoglutarate was necessary to effectively protect COx against cyanide poisoning.

Two patients experienced sensorineural hearing losses for different reasons following stapedectomy. One had an immediate severe sensorineural hearing loss following a fat-wire implant. Postmortem histologic study revealed severe cochlear degeneration, presumably caused by necrosis of the fat graft. The other patient experienced progressive fluctuating sensorineural hearing loss and vertigo several years after the introduction of the Teflon-wire prosthesis. Postmortem histologic study of this ear showed that the prosthesis had impaled the saccule, causing collapse of its wall with subsequent cochlear endolymphatic hydrops. These findings underscore the importance of atraumatic surgical technique in the performance of successful stapedectomy.

Presentation is made of a case in a premature newborn, the offspring of a mother with bilateral pulmonary tuberculosis, of placental lesions suggestive of tuberculosis and acute miliary congenital tuberculosis with lesions in liver, spleen, lymph nodes and lungs due to a possible blood stream dissemination starting from the placental infection.