SP-Sephadex equilibrium chromatography of bradykinin and related peptides: Application to trypsin-treated human plasma

An analytical method is deseribed for the separation of bradykinin, Lys-bradykinin, and Met-Lys-bradykinin by equilibrium chromatography on SP-Sephadex C-25 eluted in 0.02 Tris-HCl buffer, pH 8.10, 0.12 NaCl. A second elution buffer, 0.02 Tris-HCl buffer, pH 7.70, 0.06 NaCl, serves as a second parameter for the identification of bradykinin and also separates the hormone from plasma bradykinin-potentiating peptides. Ten to one-hundred nanomoles of each peptide can be recovered in high yields, identified by elution position, and measured by bioassay with the isolated guinea pig ileum. The identification of bradykinin as the peptide released by trypsin acting on acid-denatured plasma is documented as an illustration of the method

Use of resistivity measurements to detect urban caves in Mexico City and to assess the related hazard

International audienceIn the XIX century when Mexico City was much smaller than at present, there was non-regulated mining of building materials in a region of tuffs northwest of the city in an inhabited countryside. With the growth of the city during the XX century, this region was increasingly populated and in the 1970's many two-level bricks houses were built, without regard for underground caves created by the earlier extractions. Some ground sinkings in adjacent areas alarmed the residents who now are worried about this permanent hazard. An association of residents contracted a private company for a geophysical study in order to know the distribution of the caves. Resistivity measurements were taken in the area to detect the caves in order to alert city authorities. Resistivity data along most of the streets were collected with the array pole-dipole that consisted of three grounded electrodes. We performed 2-D dimensional inversions to the data in order to get a 2-D resistivity image of every street. This is similar to a resistivity cross-section of the ground but obtained from the inversion of pole-dipole and Schlumberger resistivity data simultaneously. Using the information of previous drills we modified our programming code in order to perform constrained inversion and to get more accurate resistivity models in agreement with the drills. From the resistivity models obtained for every street it was possible to produce a map which shows the horizontal distribution of the resistive bodies at a depth of 12m. These resistive bodies show coherent alignments that seem to correspond with a distributions of interconnected caves or tunnels used for extracting the sandy-tuffs. From these kind of interpretation method it was intended to get a more accurate horizontal distribution of the excavated areas in order to better know the urbanized area affected and lead the authorities to remedy the area with refill material

De Novo Transcriptome Assembly And Analysis To Identify Potential Gene Targets For Rnai-mediated Control Of The Tomato Leafminer (tuta Absoluta)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Background: Providing double-stranded RNA (dsRNA) to insects has been proven to silence target genes, and this approach has emerged as a potential method to control agricultural pests by engineering plants to express insect dsRNAs. A critical step of this technology is the screening of effective target genes essential for insect development and/or survival. The tomato leafminer (Tuta absoluta Meyrick) is a major Solanum lycopersicum (tomato) pest that causes significant yield losses and has recently invaded Europe, from where it is spreading at an alarming rate. To explore RNA interference (RNAi) against T. absoluta, sequence information on potential target genes is necessary, but only a few sequences are available in public databases. Results: We sequenced six libraries from RNA samples from eggs, adults, and larvae at four stages, obtaining an overall total of around 245 million reads. The assembled T. absoluta transcriptome contained 93,477 contigs with an average size of 1,574 bp, 59.8 % of which presented positive Blast hits, with 19,995 (21.4 %) annotated by gene ontology. From the transcriptome, most of the core genes of the RNAi mechanism of Lepidoptera were identified indicating the potential suitability of T. absoluta for gene silencing. No contigs displayed significant similarity with a RNA-dependent RNA Polymerase. Genes from the juvenile hormone and ecdysteroid biosynthetic pathways were identified, representing potential target genes for systemic silencing. Comparisons of transcript profiles among stages revealed 1,577 genes differentially expressed at earlier larval stages, from which potential gene targets were identified. Five of these genes were evaluated using in vitro transcribed dsRNA absorbed by tomato leaflets, which were fed to 1st instar T. absoluta larvae, resulting in significant reduction of larval body weight while exhibiting significant knockdown for three of the genes. Conclusions: The transcriptome we generated represents a valuable genomic resource for screening potential gene targets that affect the development or survival of T. absoluta larvae. Five novel genes that showed greater expression at the 1st larval stage were demonstrated to be effective potential RNAi targets by reducing larval weight and can be considered good candidates for use in RNAi-mediated crop protection.16Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP [2011/12869-6, 2012/24990-8, 2010/11313-1

The electroencephalogram in the parkinsonian syndrome

The electroencephalograms of 50 parkinsonian patients with no convulsive antecedents were studied. EEG abnormalities were found in 20 patients (40% of the cases). The incidence of abnormal records was larger in the bilateral parkinsonian syndrome than in the unilateral one (52% and 25%, respectively). Paroxysmal abnormalities characterized mainly by bursts of theta waves were the most frequent ones (65% of the cases). Sharp activity found in the EEG patterns of three patients is emphasized. Alterations of the base rhythm were recorded in 30% of the abnormal EEG cases. The patient's age doesn't seem to have an important role in the genesis of the electroencephalographic abnormalities.Foram estudados os eletrencefalogramas de 50 pacientes parkinsonianos livres de qualquer antecedente convulsivo, sendo registrados traçados anormais em 40% dos casos. A incidência de traçados anormais foi maior na síndrome parkinsoniana bilateral do que na unilateral (52% e 25%, respectivamente). Entre as anormalidades assinaladas foram as paroxísticas as mais freqüentes (65% dos casos), caracterizando-se predominantemente por surtos de ondas teta. O registro de anormalidades paroxísticas do tipo irritativo encontrado em três casos é salientado. Alterações da atividade de base foram assinaladas em 35% dos pacientes. A idade dos pacientes não parece ter tido importante papel na gênese das anormalidades eletrencefalográficas.Escola Paulista de Medicina Departamento de Neurologia e NeurocirurgiaUNIFESP, EPM, Depto. de Neurologia e NeurocirurgiaSciEL

Estados fallidos: ¿amenaza global o regional?

Failed States have been understood as a global threat by virtue of the terrorist groups that emerge and consolidate in their interior, where they are supposed to plan international actions. This document, however, delimits the impact of the state failure to regional confines, inasmuch as the neighboring countries of a failed State are the ones that, through their borders, see their internal dynamics modified due to two main factors: the illegal traffic caused by the absence of effective controls on the part of the State, and the large quantities of refugees that cross the borders closest to their failed place of origin and settle in nearby countries. Therefore, it is refuted in good measure the idea that repercussions of the failure of some States, bring the invariable consequence of a terrorist attack of global dimensions; instead, it is a matter of recognizing that their regional neighbors are the ones that are impacted by the anomalies of the Failed States.Los Estados fallidos han sido entendidos como una amenaza global en virtud de los grupos terroristas que emergen y se consolidan en su interior, desde donde se supone planean acciones internacionales. El presente documento, sin embargo, deslinda el impacto de la falla estatal al ámbito regional, como quiera que son los países vecinos de un Estado fallido los que, a través de sus fronteras, ven modificadas sus dinámicas internas debido a dos factores principales: los tráficos ilegales causados por la ausencia de controles efectivos por parte del Estado, y las grandes cantidades de refugiados que cruzan las fronteras más inmediatas de su fallido lugar de origen y se asientan en los países más cercanos. Por tanto, se desmiente en buena medida la idea según la cual la repercusión del fracaso de algunos Estados trae consigo la invariable consecuencia de un ataque terrorista de dimensiones globales, más bien se trata de reconocer que son sus vecinos regionales los que resultan impactados por sus anomalías

Towards a precise test for malaria diagnosis in the Brazilian Amazon: comparison among field microscopy, a rapid diagnostic test, nested PCR, and a computational expert system based on artificial neural networks

<p>Abstract</p> <p>Background</p> <p>Accurate malaria diagnosis is mandatory for the treatment and management of severe cases. Moreover, individuals with asymptomatic malaria are not usually screened by health care facilities, which further complicates disease control efforts. The present study compared the performances of a malaria rapid diagnosis test (RDT), the thick blood smear method and nested PCR for the diagnosis of symptomatic malaria in the Brazilian Amazon. In addition, an innovative computational approach was tested for the diagnosis of asymptomatic malaria.</p> <p>Methods</p> <p>The study was divided in two parts. For the first part, passive case detection was performed in 311 individuals with malaria-related symptoms from a recently urbanized community in the Brazilian Amazon. A cross-sectional investigation compared the diagnostic performance of the RDT Optimal-IT, nested PCR and light microscopy. The second part of the study involved active case detection of asymptomatic malaria in 380 individuals from riverine communities in Rondônia, Brazil. The performances of microscopy, nested PCR and an expert computational system based on artificial neural networks (MalDANN) using epidemiological data were compared.</p> <p>Results</p> <p>Nested PCR was shown to be the gold standard for diagnosis of both symptomatic and asymptomatic malaria because it detected the major number of cases and presented the maximum specificity. Surprisingly, the RDT was superior to microscopy in the diagnosis of cases with low parasitaemia. Nevertheless, RDT could not discriminate the <it>Plasmodium </it>species in 12 cases of mixed infections (<it>Plasmodium vivax </it>+ <it>Plasmodium falciparum</it>). Moreover, the microscopy presented low performance in the detection of asymptomatic cases (61.25% of correct diagnoses). The MalDANN system using epidemiological data was worse that the light microscopy (56% of correct diagnoses). However, when information regarding plasma levels of interleukin-10 and interferon-gamma were inputted, the MalDANN performance sensibly increased (80% correct diagnoses).</p> <p>Conclusions</p> <p>An RDT for malaria diagnosis may find a promising use in the Brazilian Amazon integrating a rational diagnostic approach. Despite the low performance of the MalDANN test using solely epidemiological data, an approach based on neural networks may be feasible in cases where simpler methods for discriminating individuals below and above threshold cytokine levels are available.</p

Severe Plasmodium vivax malaria exhibits marked inflammatory imbalance

<p>Abstract</p> <p>Background</p> <p>Despite clinical descriptions of severe vivax malaria cases having been reported, data regarding immunological and inflammatory patterns are scarce. In this report, the inflammatory and immunological status of both mild and severe vivax malaria cases are compared in order to explore immunopathological events in this disease.</p> <p>Methods and Results</p> <p>Active and passive malaria case detections were performed during 2007 in Buritis, Rondônia, in the Brazilian Amazon. A total of 219 participants enrolled the study. Study individuals were classified according to the presence of <it>Plasmodium vivax </it>infection within four groups: non-infected (n = 90), asymptomatic (n = 60), mild (n = 50) and severe vivax infection (n = 19). A diagnosis of malaria was made by microscopy and molecular assays. Since at present no clear criteria define severe vivax malaria, this study adapted the consensual criteria from falciparum malaria. Patients with severe <it>P. vivax </it>infection were younger, had lived for shorter time in the endemic area, and recalled having experienced less previous malaria episodes than individuals with no malaria infection and with mild or asymptomatic infection. Strong linear trends were identified regarding increasing plasma levels of C reactive protein (CRP), serum creatinine, bilirubins and the graduation of disease severity. Plasma levels of tumour necrosis factor (TNF), interferon-gamma(IFN-gamma) and also IFN-gamma/interleukin-10 ratios were increased and exhibited a linear trend with gradual augmentation of disease severity. Both laboratory parameters of organ dysfunction and inflammatory cytokines were reduced during anti-parasite therapy in those patients with severe disease.</p> <p>Conclusion</p> <p>Different clinical presentations of vivax malaria infection present strong association with activation of pro-inflammatory responses and cytokine imbalance. These findings are of utmost importance to improve current knowledge about physiopathological concepts of this serious widespread disease.</p