Rice endosperm is cost-effective for the production of recombinant griffithsin with potent activity against HIV

Protein microbicides containing neutralizing antibodies and antiviral lectins may help to reduce the rate of infection with human immunodeficiency virus (HIV) if it is possible to manufacture the components in large quantities at a cost affordable in HIV‐endemic regions such as sub‐Saharan Africa. We expressed the antiviral lectin griffithsin (GRFT), which shows potent neutralizing activity against HIV, in the endosperm of transgenic rice plants (Oryza sativa), to determine whether rice can be used to produce inexpensive GRFT as a microbicide ingredient. The yield of (OS)GRFT in the best‐performing plants was 223 μg/g dry seed weight. We also established a one‐step purification protocol, achieving a recovery of 74% and a purity of 80%, which potentially could be developed into a larger‐scale process to facilitate inexpensive downstream processing. (OS)GRFT bound to HIV glycans with similar efficiency to GRFT produced in Escherichia coli. Whole‐cell assays using purified (OS)GRFT and infectivity assays using crude extracts of transgenic rice endosperm confirmed that both crude and pure (OS)GRFT showed potent activity against HIV and the crude extracts were not toxic towards human cell lines, suggesting they could be administered as a microbicide with only minimal processing. A freedom‐to‐operate analysis confirmed that GRFT produced in rice is suitable for commercial development, and an economic evaluation suggested that 1.8 kg/ha of pure GRFT could be produced from rice seeds. Our data therefore indicate that rice could be developed as an inexpensive production platform for GRFT as a microbicide component

Optimal Population Codes for Space: Grid Cells Outperform Place Cells

Rodents use two distinct neuronal coordinate systems to estimate their position: place fields in the hippocampus and grid fields in the entorhinal cortex. Whereas place cells spike at only one particular spatial location, grid cells fire at multiple sites that correspond to the points of an imaginary hexagonal lattice. We study how to best construct place and grid codes, taking the probabilistic nature of neural spiking into account. Which spatial encoding properties of individual neurons confer the highest resolution when decoding the animal’s position from the neuronal population response? A priori, estimating a spatial position from a grid code could be ambiguous, as regular periodic lattices possess translational symmetry. The solution to this problem requires lattices for grid cells with different spacings; the spatial resolution crucially depends on choosing the right ratios of these spacings across the population. We compute the expected error in estimating the position in both the asymptotic limit, using Fisher information, and for low spike counts, using maximum likelihood estimation. Achieving high spatial resolution and covering a large range of space in a grid code leads to a trade-off: the best grid code for spatial resolution is built of nested modules with different spatial periods, one inside the other, whereas maximizing the spatial range requires distinct spatial periods that are pairwisely incommensurate. Optimizing the spatial resolution predicts two grid cell properties that have been experimentally observed. First, short lattice spacings should outnumber long lattice spacings. Second, the grid code should be self-similar across different lattice spacings, so that the grid field always covers a fixed fraction of the lattice period. If these conditions are satisfied and the spatial “tuning curves” for each neuron span the same range of firing rates, then the resolution of the grid code easily exceeds that of the best possible place code with the same number of neurons

Biochemical Properties of a Decoy Oligodeoxynucleotide Inhibitor of STAT3 Transcription Factor.

Cyclic STAT3 decoy (CS3D) is a second-generation, double-stranded oligodeoxynucleotide (ODN) that mimics a genomic response element for signal transducer and activator of transcription 3 (STAT3), an oncogenic transcription factor. CS3D competitively inhibits STAT3 binding to target gene promoters, resulting in decreased expression of proteins that promote cellular proliferation and survival. Previous studies have demonstrated antitumor activity of CS3D in preclinical models of solid tumors. However, prior to entering human clinical trials, the efficiency of generating the CS3D molecule and its stability in biological fluids should be determined. CS3D is synthesized as a single-stranded ODN and must have its free ends ligated to generate the final cyclic form. In this study, we report a ligation efficiency of nearly 95 percent. The ligated CS3D demonstrated a half-life of 7.9 h in human serum, indicating adequate stability for intravenous delivery. These results provide requisite biochemical characterization of CS3D that will inform upcoming clinical trials

Initial Estimates On Shipping’s Cost Impacts and Emissions for a Range of Policy Options - A Prototype Model

Shipping was estimated, in 2007, to be responsible for 3.3% of anthropogenic CO2 emissions. Scenarios for future growth in transport demand suggest that this share could substantially increase in the next 40 years, and without regulation the growth in emissions associated with that demand growth would be uncontrolled. Modelling can be used to understand the potential trajectories of emissions from the shipping industry and its potential development and impacts under foreseeable economic scenarios. Modelling can also be used to estimate the response (in terms of changes to those emissions trajectories and impacts on the industry) due to hypothetical regulation and policies. This paper proposes methods for conceptualizing the different components of the shipping industry for these purposes

Aristonectes quiriquinensis, sp. nov., a new highly derived elasmosaurid from the upper Maastrichtian of central Chile

This paper describes a new species of elasmosaurid plesiosaur, Aristonectes quiriquinensis, sp. nov., based on a partial skeleton recovered from upper Maastrichtian beds of the Quiriquina Formation of central Chile. The material described here consists of two skeletons, one collected near the village of Cocholgue, and a second juvenile specimen from Quiriquina Island. Prior to these finds, Aristonectes was viewed as a monospecific genus, including only the enigmatic Aristonectes parvidens, the holotype of which consists of an incomplete skull and incomplete postcranium. Other material referred to the genus includes an incomplete juvenile skull and other postcranial material from the upper Maastrichtian of Antarctica, as well as a partial skull from the Quiriquina Formation of central Chile. The relationships of Aristonectes have been controversial, with competing theories assigning the genus to Cryptoclididae, Elasmosauridae, and Aristonectidae; however, there is a developing consensus that Aristonectes is a derived elasmosaurid, and this paper gives strong evidence for this view. Comparison of the specimen here studied with the holotype of A. parvidens demonstrates that A. quiriquinensis is a distinct species. The completeness of the adult skeleton allows the first confident size estimates for adult Aristonectes. It is a large plesiosaurian with a relatively large skull with numerous homodont teeth, a moderately long and laterally compressed neck, and relatively narrow trunk, with slender and elongate forelimbs. The two specimens are restricted to the upper Maastrichtian of central Chile, posing questions concerning the austral circumpolar distribution of different elasmosaurids towards the end of the Cretaceous.Fil: Otero, Rodrigo A.. Universidad de Chile; ChileFil: Soto Acuña, Sergio. Museo Nacional de Historia Natural de Santiago; Chile. Universidad de Chile; ChileFil: O'Keefe, Frank Robin. Marshall University; Estados UnidosFil: O'gorman, Jose Patricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. División Paleontología Vertebrados; ArgentinaFil: Stinnesbeck, Wolfgang. Heidelberg University; AlemaniaFil: Suárez, Mario E.. Museo Nacional de Historia Natural de Santiago; Chile. Universidad de Chile; ChileFil: Rubilar-Rogers, David. Museo Nacional de Historia Natural de Santiago; Chile. Universidad de Chile; ChileFil: Salazar, Christian. Museo Nacional de Historia Natural de Santiago; Chile. Universidad de Chile; ChileFil: Quinzio Sinn, Luis Arturo. Universidad de Concepción; Chil

Tracheostomy timing and the duration of weaning in patients with acute respiratory failure

INTRODUCTION: The effect of various airway management strategies, such as the timing of tracheostomy, on liberation from mechanical ventilation (MV) is uncertain. We tested the hypothesis that tracheostomy, when performed prior to active weaning, does not influence the duration of weaning or of MV in comparison with a more selective use of tracheostomy. PATIENTS AND METHODS: In this observational prospective cohort study, surgical patients requiring ≥ 72 hours of MV were followed prospectively. Patients undergoing tracheostomy prior to any active weaning attempts (early tracheostomy [ET]) were compared with patients in whom initial weaning attempts were made with the endotracheal tube in place (selective tracheostomy [ST]). RESULTS: We compared the duration of weaning, the total duration of MV and the frequency of fatigue and pneumonia. Seventy-four patients met inclusion criteria. Twenty-one patients in the ET group were compared with 53 patients in the ST group (47% of whom ultimately underwent tracheostomy). The median duration of weaning was shorter (3 days versus 6 days, P = 0.05) in patients in the ET group than in the ST group, but the duration of MV was not (median [interquartile range], 11 days [9–26 days] in the ET group versus 13 days [8–21 days] in the ST group). The frequencies of fatigue and pneumonia were lower in the ET group patients. DISCUSSION: Determining the ideal timing of tracheostomy in critically ill patients has been difficult and often subjective. To standardize this process, it is important to identify objective criteria to identify patients most likely to benefit from the procedure. Our data suggest that in surgical patients with resolving respiratory failure, a patient who meets typical criteria for a trial of spontaneous breathing but is not successfully extubated within 24 hours may benefit from a tracheostomy. Our data provide a framework for the conduct of a clinical trial in which tracheostomy timing can be assessed for its impact on the duration of weaning. CONCLUSION: Tracheostomy prior to active weaning may hasten liberation from ventilation and reduce complications. However, this does not reduce the overall duration of MV

Spitz and wingless, emanating from distinct borders, cooperate to establish cell fate across the engrailed domain in the drosophila epidermis

A key step in development is the establishment of cell type diversity across a cellular field. Segmental patterning within the Drosophila embryonic epidermis is one paradigm for this process. At each parasegment boundary, cells expressing the Wnt family member Wingless confront cells expressing the homeoprotein Engrailed. The Engrailed-expressing cells normally differentiate as one of two alternative cell types. In investigating the generation of this cell type diversity among the 2-cell-wide Engrailed stripe, we previously showed that Wingless, expressed just anterior to the Engrailed cells, is essential for the specification of anterior Engrailed cell fate. In a screen for additional mutations affecting Engrailed cell fate, we identified anterior open/yan, a gene encoding an inhibitory ETS-domain transcription factor that is negatively regulated by the Rasl-MAP kinase signaling cascade. We find that Anterior Open must be inactivated for posterior Engrailed cells to adopt their correct fate. This is achieved by the EGF receptor (DER), which is required autonomously in the Engrailed cells to trigger the Ras1-MAP kinase pathway. Localized activation of DER is accomplished by restricted processing of the activating ligand, Spitz. Processing is confined to the cell row posterior to the Engrailed domain by the restricted expression of Rhomboid. These cells also express the inhibitory ligand Argos, which attenuates the activation of DER in cell rows more distant from the ligand source. Thus, distinct signals flank each border of the Engrailed domain, as Wingless is produced anteriorly and Spitz posteriorly. Since we also show that En cells have the capacity to respond to either Wingless or Spitz, these cells must choose their fate depending on the relative level of activation of the two pathways.Louise O’Keefe, Scott T. Dougan, Limor Gabay, Erez Raz, Ben-Zion Shilo and Stephen DiNard

Use of immunoglobulin-loaded protein A-bearing staphylococci as a primary solid phase immunoadsorbent in radioimmunoassay.

Protein A-bearing formalinized heat-inactivated Staphylococcus aureus bind rabbit 125I-IgG with high capacity of IgG for antigen. The affinity of immobilized IgG for antigen is equal to the affinity of soluble antibody, and the capacity for antigen approximates the capacity of soluble antibody for antigen. 125I-IgG bound to high affinity sites on bacteria is not substantially displaced in the presence of human serum after 4 h at 4 degrees C, but rabbit immunoglobulin can displace bound rabbit IgG. Protein A-bearing bacteria which have adsorbed IgG free from protease activity in antiserum provide a stable, sensitive, primary solid phase adsorbent with unique features for radioimmunoassay. Sedimentation characteristics of bacteria permit facile separation of bound from free ligand by centrifugation of the primary antibody. Immunoglobulin-loaded staphylococci can adsorb ligand from serum, simultaneously purifying and concentrating ligands for measurement in a single subsequent step. Immunoglobulin-bearing bacteria can be used directly in biological fluids containing immunoglobulin, such as serum and culture media. Improved economy of staphylococci, low nonspecific binding, high functional capacity, stability, and unique characteristics of radioimmunoassay using immunoglobulin-loaded staphylococci compare favorably with conventional solid phase adsorbents