The transcriptional repressor protein NsrR senses nitric oxide directly via a [2Fe-2S] cluster

The regulatory protein NsrR, a member of the Rrf2 family of transcription repressors, is specifically dedicated to sensing nitric oxide (NO) in a variety of pathogenic and non-pathogenic bacteria. It has been proposed that NO directly modulates NsrR activity by interacting with a predicted [Fe-S] cluster in the NsrR protein, but no experimental evidence has been published to support this hypothesis. Here we report the purification of NsrR from the obligate aerobe Streptomyces coelicolor. We demonstrate using UV-visible, near UV CD and EPR spectroscopy that the protein contains an NO-sensitive [2Fe-2S] cluster when purified from E. coli. Upon exposure of NsrR to NO, the cluster is nitrosylated, which results in the loss of DNA binding activity as detected by bandshift assays. Removal of the [2Fe-2S] cluster to generate apo-NsrR also resulted in loss of DNA binding activity. This is the first demonstration that NsrR contains an NO-sensitive [2Fe-2S] cluster that is required for DNA binding activity

A novel malaria vaccine candidate antigen expressed in Tetrahymena thermophila

Development of effective malaria vaccines is hampered by the problem of producing correctly folded Plasmodium proteins for use as vaccine components. We have investigated the use of a novel ciliate expression system, Tetrahymena thermophila, as a P. falciparum vaccine antigen platform. A synthetic vaccine antigen composed of N-terminal and C-terminal regions of merozoite surface protein-1 (MSP-1) was expressed in Tetrahymena thermophila. The recombinant antigen was secreted into the culture medium and purified by monoclonal antibody (mAb) affinity chromatography. The vaccine was immunogenic in MF1 mice, eliciting high antibody titers against both N- and C-terminal components. Sera from immunized animals reacted strongly with P. falciparum parasites from three antigenically different strains by immunofluorescence assays, confirming that the antibodies produced are able to recognize parasite antigens in their native form. Epitope mapping of serum reactivity with a peptide library derived from all three MSP-1 Block 2 serotypes confirmed that the MSP-1 Block 2 hybrid component of the vaccine had effectively targeted all three serotypes of this polymorphic region of MSP-1. This study has successfully demonstrated the use of Tetrahymena thermophila as a recombinant protein expression platform for the production of malaria vaccine antigens

Unconfined Aquifer Flow Theory - from Dupuit to present

Analytic and semi-analytic solution are often used by researchers and practicioners to estimate aquifer parameters from unconfined aquifer pumping tests. The non-linearities associated with unconfined (i.e., water table) aquifer tests makes their analysis more complex than confined tests. Although analytical solutions for unconfined flow began in the mid-1800s with Dupuit, Thiem was possibly the first to use them to estimate aquifer parameters from pumping tests in the early 1900s. In the 1950s, Boulton developed the first transient well test solution specialized to unconfined flow. By the 1970s Neuman had developed solutions considering both primary transient storage mechanisms (confined storage and delayed yield) without non-physical fitting parameters. In the last decade, research into developing unconfined aquifer test solutions has mostly focused on explicitly coupling the aquifer with the linearized vadose zone. Despite the many advanced solution methods available, there still exists a need for realism to accurately simulate real-world aquifer tests

Centromere Plasmid: A New Genetic Tool for the Study of Plasmodium falciparum

The introduction of transgenes into Plasmodium falciparum, a highly virulent human malaria parasite, has been conducted either by single crossover recombination or by using episomal plasmids. However, these techniques remain insufficient because of the low transfection efficiency and the low frequency of recombination. To improve the genetic manipulation of P. falciparum, we developed the centromere plasmid as a new genetic tool. First, we attempted to clone all of the predicted centromeres from P. falciparum into E. coli cells but failed because of the high A/T contents of these sequences. To overcome this difficulty, we identified the common sequence features of the centromere of Plasmodium spp. and designed a small centromere that retained those features. The centromere plasmid constructed with the small centromere sequence, pFCEN, segregated into daughter parasites with approximately 99% efficiency, resulting in the stable maintenance of this plasmid in P. falciparum even in the absence of drug selection. This result demonstrated that the small centromere sequence harboured in pFCEN could function as an actual centromere in P. falciparum. In addition, transgenic parasites were more rapidly generated when using pFCEN than when using the control plasmid, which did not contain the centromere sequence. Furthermore, in contrast to the control plasmid, pFCEN did not form concatemers and, thus, was maintained as a single copy over multiple cell divisions. These unique properties of the pFCEN plasmid will solve the current technical limitations of the genetic manipulation of P. falciparum, and thus, this plasmid will become a standard genetic tool for the study of this parasite

In-Line estimation of the standard colour index of citrus fruits using a computer vision system developed for a mobile platform

The final publication is available at Springer via http://dx.doi.org/10.1007/s11947-012-1015-2A key aspect for the consumer when it comes to deciding on a particular product is the colour. In order to make fruit available to consumers as early as possible, the collection of oranges and mandarins begins before they ripen fully and reach their typical orange colour. As a result, they are therefore subjected to certain degreening treatments, depending on their standard colour citrus index at harvest. Recently, a mobile platform that incorporates a computer vision system capable of pre-sorting the fruit while it is being harvested has been developed as an aid in the harvesting task. However, due to the restrictions of working in the field, the computer vision system developed for this machine is limited in its technology and processing capacity compared to conventional systems. This work shows the optimised algorithms for estimating the colour of citrus in-line that were developed for this mobile platform and its performance is evaluated against that of a spectrophotometer used as a reference in the analysis of colour in food. The results obtained prove that our analysis system predicts the colour index of citrus with a good reliability (R2 = 0.925) working in real time. Findings also show that it is effective for classifying harvested fruits in the field according to their colour. © 2012 Springer Science+Business Media New York.This work was partially funded by the INIA through research project RTA2009-00118-C02-01 with the support of European FEDER funds, and by the project PAID-05-11-2745, Vicerectorat d'Investigacio, Universitat Politecnica de Valencia.Vidal, A.; Talens Oliag, P.; Prats-Montalbán, JM.; Cubero García, S.; Albert Gil, FE.; Blasco Ivars, J. (2013). In-Line estimation of the standard colour index of citrus fruits using a computer vision system developed for a mobile platform. Food and Bioprocess Technology. 6(12):3412-3419. https://doi.org/10.1007/s11947-012-1015-2S34123419612Arzate-Vázquez, I., Chanona-Pérez, J. J., Perea-Flores, M. J., Calderón-Domínguez, G., Moreno-Armendáriz, M. A., Calvo, H., Godoy-Calderón, S., Quevedo, R., & Gutiérrez-López, G. (2011). Image processing applied to classification of avocado variety Hass (Persea americana Mill.) during the ripening process. Food and Bioprocess Technology, 4(7), 1307–1313.Blasco, J., Aleixos, N., Cubero, S., Gómez-Sanchis, J., & Moltó, E. (2009). Automatic sorting of satsuma (Citrus unshiu) segments using computer vision and morphological features. Computers and Electronics in Agriculture, 66, 1–8.Campbell, B. L., Nelson, R. G., Ebel, C. E., Dozier, W. A., Adrian, J. L., & Hockema, B. R. (2004). 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Cambridge: Woodhead Publishing.Jiménez-Cuesta MJ, Cuquerella J & Martínez-Jávega JM (1981) Determination of a color index for citrus fruit degreening. In Proc. of the International Society of Citriculture, Vol. 2, 750-753Kang, S. P., East, A. R., & Trujillo, F. J. (2008). Colour vision system evaluation of bicolour fruit: A case study with ‘B74’ mango. Postharvest Biology and Technology, 49, 77–85.Lang, C., & Hübert, T. (2011). A colour ripeness indicator for apples. Food and Bioprocess Technology, 5(8), 3244–3249.López-Camelo, A. F., & Gómez, P. A. (2004). Comparison of color indexes for tomato ripening. Horticultura Brasileira, 22(3), 534–537. http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-05362004000300006 .López-García, F., Andreu-García, A., Blasco, J., Aleixos, N., & Valiente, J. M. (2010). Automatic detection of skin defects in citrus fruits using a multivariate image analysis approach. 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The conservation value of human-modified landscapes for the world's primates

Land-use change pushes biodiversity into human-modified landscapes, where native ecosystems are surrounded by anthropic land covers (ALCs). Yet, the ability of species to use these emerging covers remains poorly understood. We quantified the use of ALCs by primates worldwide, and analyzed species' attributes that predict such use. Most species use secondary forests and tree plantations, while only few use human settlements. ALCs are used for foraging by at least 86 species with an important conservation outcome: those that tolerate heavily modified ALCs are 26% more likely to have stable or increasing populations than the global average for all primates. There is no phylogenetic signal in ALCs use. Compared to all primates on Earth, species using ALCs are less often threatened with extinction, but more often diurnal, medium or large-bodied, not strictly arboreal, and habitat generalists. These findings provide valuable quantitative information for improving management practices for primate conservation worldwide

Effective-Range Expansion of the Neutron-Deuteron Scattering Studied by a Quark-Model Nonlocal Gaussian Potential

The S-wave effective range parameters of the neutron-deuteron (nd) scattering are derived in the Faddeev formalism, using a nonlocal Gaussian potential based on the quark-model baryon-baryon interaction fss2. The spin-doublet low-energy eigenphase shift is sufficiently attractive to reproduce predictions by the AV18 plus Urbana three-nucleon force, yielding the observed value of the doublet scattering length and the correct differential cross sections below the deuteron breakup threshold. This conclusion is consistent with the previous result for the triton binding energy, which is nearly reproduced by fss2 without reinforcing it with the three-nucleon force.Comment: 21 pages, 6 figures and 6 tables, submitted to Prog. Theor. Phy