A suite of parallel vectors for baculovirus expression

The expression of proteins using recombinant baculoviruses is a mature and widely used technology. However, some aspects of the technology continue to detract from high throughput use and the basis of the final observed expression level is poorly understood. Here, we describe the design and use of a set of vectors developed around a unified cloning strategy that allow parallel expression of target proteins in the baculovirus system as N-terminal or C-terminal fusions. Using several protein kinases as tests we found that amino-terminal fusion to maltose binding protein rescued expression of the poorly expressed human kinase Cot but had only a marginal effect on expression of a well-expressed kinase IKK-2. In addition, MBP fusion proteins were found to be secreted from the expressing cell. Use of a carboxyl-terminal GFP tagging vector showed that fluorescence measurement paralleled expression level and was a convenient readout in the context of insect cell expression, an observation that was further supported with additional non-kinase targets. The expression of the target proteins using the same vectors in vitro showed that differences in expression level were wholly dependent on the environment of the expressing cell and an investigation of the time course of expression showed it could affect substantially the observed expression level for poorly but not well-expressed proteins. Our vector suite approach shows that rapid expression survey can be achieved within the baculovirus system and in addition, goes some way to identifying the underlying basis of the expression level obtained. (c) 2006 Elsevier Inc. All rights reserved.

Basic constraints between two vectors

This chapter deals with the characterization of the basic constraints between two vectors. This issue plays a crucial role in the formulation of constraint equations for mechanical joints. In particular, relations between two parallel and two perpendicular vectors are derived. Moreover, formulation for a vector that connects two generic points is presented. The material described here is developed under the framework of multibody systems formulation for spatial systems.

Alternatives for parallel Krylov subspace basis computation

Numerical methods related to Krylov subspaces are widely used in large sparse numerical linear algebra. Vectors in these subspaces are manipulated via their representation onto orthonormal bases. Nowadays, on serial computers, the method of Arnoldi is considered as a reliable technique for constructing such bases. However, although easily parallelizable, this technique is not as scalable as expected for communications. In this work we examine alternative methods aimed at overcoming this drawback. Since they retrieve upon completion the same information as Arnoldi's algorithm does, they enable us to design a wide family of stable and scalable Krylov approximation methods for various parallel environments. We present timing results obtained from their implementation on two distributed-memory multiprocessor supercomputers: the Intel Paragon and the IBM Scalable POWERparallel SP2. (C) 1997 by John Wiley & Sons, Ltd.

Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.

Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration.

Linear predicted two-pass hexagonal algorithm with parallel implementation for motion estimation

This paper presents a paralleled Two-Pass Hexagonal (TPA) algorithm constituted by Linear Hashtable Motion Estimation Algorithm (LHMEA) and Hexagonal Search (HEXBS) for motion estimation. In the TPA., Motion Vectors (MV) are generated from the first-pass LHMEA and are used as predictors for second-pass HEXBS motion estimation, which only searches a small number of Macroblocks (MBs). We introduced hashtable into video processing and completed parallel implementation. We propose and evaluate parallel implementations of the LHMEA of TPA on clusters of workstations for real time video compression. It discusses how parallel video coding on load balanced multiprocessor systems can help, especially on motion estimation. The effect of load balancing for improved performance is discussed. The performance or the algorithm is evaluated by using standard video sequences and the results are compared to current algorithms.

Two-pass hexagonal algorithm with parallel implementation for video coding

This paper presents a paralleled Two-Pass Hexagonal (TPA) algorithm constituted by Linear Hashtable Motion Estimation Algorithm (LHMEA) and Hexagonal Search (HEXBS) for motion estimation. In the TPA, Motion Vectors (MV) are generated from the first-pass LHMEA and are used as predictors for second-pass HEXBS motion estimation, which only searches a small number of Macroblocks (MBs). We introduced hashtable into video processing and completed parallel implementation. We propose and evaluate parallel implementations of the LHMEA of TPA on clusters of workstations for real time video compression. It discusses how parallel video coding on load balanced multiprocessor systems can help, especially on motion estimation. The effect of load balancing for improved performance is discussed. The performance of the algorithm is evaluated by using standard video sequences and the results are compared to current algorithms.

Improved two-pass hexagonal algorithm with parallel implementation for video coding - art. no. 60630J

This paper presents an improved parallel Two-Pass Hexagonal (TPA) algorithm constituted by Linear Hashtable Motion Estimation Algorithm (LHMEA) and Hexagonal Search (HEXBS) for motion estimation. Motion Vectors (MV) are generated from the first-pass LHMEA and used as predictors for second-pass HEXBS motion estimation, which only searches a small number of Macroblocks (MBs). We used bashtable into video processing and completed parallel implementation. The hashtable structure of LHMEA is improved compared to the original TPA and LHMEA. We propose and evaluate parallel implementations of the LHMEA of TPA on clusters of workstations for real time video compression. The implementation contains spatial and temporal approaches. The performance of the algorithm is evaluated by using standard video sequences and the results are compared to current algorithms.

Particle methods parallel implementations by GP-GPU strategies

This paper outlines the problems found in the parallelization of SPH (Smoothed Particle Hydrodynamics) algorithms using Graphics Processing Units. Different results of some parallel GPU implementations in terms of the speed-up and the scalability compared to the CPU sequential codes are shown. The most problematic stage in the GPU-SPH algorithms is the one responsible for locating neighboring particles and building the vectors where this information is stored, since these specific algorithms raise many dificulties for a data-level parallelization. Because of the fact that the neighbor location using linked lists does not show enough data-level parallelism, two new approaches have been pro- posed to minimize bank conflicts in the writing and subsequent reading of the neighbor lists. The first strategy proposes an efficient coordination between CPU-GPU, using GPU algorithms for those stages that allow a straight forward parallelization, and sequential CPU algorithms for those instructions that involve some kind of vector reduction. This coordination provides a relatively orderly reading of the neighbor lists in the interactions stage, achieving a speed-up factor of x47 in this stage. However, since the construction of the neighbor lists is quite expensive, it is achieved an overall speed-up of x41. The second strategy seeks to maximize the use of the GPU in the neighbor's location process by executing a specific vector sorting algorithm that allows some data-level parallelism. Al- though this strategy has succeeded in improving the speed-up on the stage of neighboring location, the global speed-up on the interactions stage falls, due to inefficient reading of the neighbor vectors. Some changes to these strategies are proposed, aimed at maximizing the computational load of the GPU and using the GPU texture-units, in order to reach the maximum speed-up for such codes. Different practical applications have been added to the mentioned GPU codes. First, the classical dam-break problem is studied. Second, the wave impact of the sloshing fluid contained in LNG vessel tanks is also simulated as a practical example of particle methods

Control of insect vectors and plant viruses in protected crops by novel pyrethroid-treated nets

Long-lasting insecticide-treated nets (LLITNs) constitute a novel alternative that combines physical and chemical tactics to prevent insect access and the spread of insect-transmitted plant viruses in protected enclosures. This approach is based on a slow-release insecticide-treated net with large hole sizes that allow improved ventilation of greenhouses. The efficacy of a wide range of LLITNs was tested under laboratory conditions against Myzus persicae, Aphis gossypii and Bemisia tabaci. Two nets were selected for field tests under a high insect infestation pressure in the presence of plants infected with Cucumber mosaic virus and Cucurbit aphid-borne yellows virus. The efficacy of Aphidius colemani, a parasitoid commonly used for biological control of aphids, was studied in parallel field experiments.

Etiology of cutaneous leishmaniasis and anthropophilic vectors in Juruti, Pará State, Brazil

In a preliminary study in Juruti, a mining municipality in western Pará State, Brazil, 12 out of 21 patients suspected of presenting cutaneous leishmaniasis showed positive PCR (SSUrDNA and G6PD): Leishmania (Viannia) braziliensis (9/12; 75%) and L. (V.) sp. (3/12; 25%). Entomological studies in the same location revealed the presence of 12 different phlebotomine species (n =105). One of the most common species was Lutzomyia (Psychodopygus) complexa (17%) which is both highly anthropophilic and a known vector of L. (V.) braziliensis in other regions of Pará. These preliminary findings should serve to guide future epidemiological surveillance in Juruti.

Epidemiological and ecological aspects related to malaria in the area of influence of the lake at Porto Primavera dam, in western São Paulo State, Brazil

A study was carried out in the area of influence of the Porto Primavera Hydroelectric Power Station, in western São Paulo State, to investigate ecological and epidemiological aspects of malaria in the area and monitor the profile of the anopheline populations following the environmental changes brought about by the construction of the lake. Mosquitoes captured were analyzed by standardized indicator species analysis (ISA) before and during different flooding phases (253 m and 257 m elevations). The local human population was studied by means of parasitological (thin/thick blood smears), molecular (PCR) and serological tests. Serological tests consisted of Enzyme Linked Immunosorbent Assay (ELISA) with synthetic peptides of the circumsporozoite protein (CSP) from classic Plasmodium vivax, P. vivax variants (VK247 and "vivax-like"), P. malariae and P. falciparum and Indirect Immunofluorescence Assay (IFA) with asexual forms of P. vivax, P. malariae and P. falciparum. The results of the entomological survey indicated that, although the Anopheles darlingi population increased after the flooding, the population density remained very low. No malaria, parasite infection or DNA was detected in the inhabitants of the study area. However, there was a low frequency of antibodies against asexual forms and a significant prevalence of antibodies against P. vivax, P. vivax variants, P. falciparum and P. malariae; the presence of these antibodies may result from recent or less recent contact with human or simian Plasmodium (a parallel study in the same area revealed the existence of a sylvatic cycle). Nevertheless, these results suggest that, as in other places where malaria is present and potential vectors circulate, the local epidemiological conditions observed could potentially support the transmission of malaria in Porto Primavera Lake if infected individuals are introduced in sufficient numbers. Further studies are required to elucidate the phenomena described in this paper.

Potential vectors and hosts of rickettsia spp: epidemiological studies in the Vale do Paraíba, state of Rio de Janeiro/Brazil

FAPESP, CNPq, CAPES

Status of Early 19th-Century Names Authored in Parallel by Wied and Schinz for South American Reptiles and Amphibians, with Designations of Three Nomina Protecta

Prince Maximilian zu Wied's great exploration of coastal Brazil in 1815-1817 resulted in important collections of reptiles, amphibians, birds, and mammals, many of which were new species later described by Wied himself The bulk of his collection was purchased for the American Museum of Natural History in 1869, although many ""type specimens"" had disappeared earlier. Wied carefully identified his localities but did not designate type specimens or type localities, which are taxonomic concepts that were not yet established. Information and manuscript names on a fraction (17 species) of his Brazilian reptiles and amphibians were transmitted by Wied to Prof. Heinrich Rudolf Schinz at the University of Zurich. Schinz included these species (credited to their discoverer ""Princ. Max."") in the second volume of Das Thierreich ... (1822). Most are junior objective synonyms of names published by Wied. However, six of the 17 names used by Schinz predate Wied's own publications. Three were manuscript names never published by Wied because he determined the species to be previously known. (1) Lacerta vittata Schinz, 1822 (a nomen oblitum) = Lacerta striata sensu Wied (a misidentification, non Linnaeus nec sensu Merrem) = Kentropyx calcarata Spix, 1825, herein qualified as a nomen protectum. (2) Polychrus virescens Schinz, 1822 = Lacerta marmorata Linnaeus, 1758 (now Polychrus marmoratus). (3) Scincus cyanurus Schinz, 1822 (a nomen oblitum) = Gymnophthalmus quadrilineatus sensu Wied (a misidentification, non Linnaeus nec sensu Merrem) = Micrablepharus maximiliani (Reinhardt and Lutken, ""1861"" [1862]), herein qualified as a nomen protectum. Qualifying Scincus cyanurus Schinz, 1822, as a nomen oblitum also removes the problem of homonymy with the later-named Pacific skink Scincus cyanurus Lesson (= Emoia cyanura). The remaining three names used by Schinz are senior objective synonyms that take priority over Wied's names. (4) Bufo cinctus Schinz, 1822, is senior to Bufo cinctus Wied, 1823; both, however, are junior synonyms of Bufo crucifer Wied, 1821 = Chaunus crucifer (Wied). (5) Agama picta Schinz, 1822, is senior to Agama picta Wied, 1823, requiring a change of authorship for this poorly known species, to be known as Enyalius pictus (Schinz). (6) Lacerta cyanomelas Schinz, 1822, predates Teius cyanomelas Wied, 1824 (1822-1831) both nomina oblita. Wied's illustration and description shows cyanomelas as apparently conspecific with the recently described but already well-known Cnemidophorus nativo Rocha et al., 1997, which is the valid name because of its qualification herein as a nomen protectum. The preceding specific name cyanomelas (as corrected in an errata section) is misspelled several ways in different copies of Schinz's original description (""cyanom las,"" ""cyanomlas,"" and cyanom""). Loosening, separation, and final loss of the last three letters of movable type in the printing chase probably accounts for the variant misspellings.

Harmonic identification using parallel neural networks in single-phase systems

In this paper, artificial neural networks are employed in a novel approach to identify harmonic components of single-phase nonlinear load currents, whose amplitude and phase angle are subject to unpredictable changes, even in steady-state. The first six harmonic current components are identified through the variation analysis of waveform characteristics. The effectiveness of this method is tested by applying it to the model of a single-phase active power filter, dedicated to the selective compensation of harmonic current drained by an AC controller. Simulation and experimental results are presented to validate the proposed approach. (C) 2010 Elsevier B. V. All rights reserved.

Integrating structural and input design of a 2-DOF high-speed parallel manipulator: A flexible model-based approach

This paper discusses the integrated design of parallel manipulators, which exhibit varying dynamics. This characteristic affects the machine stability and performance. The design methodology consists of four main steps: (i) the system modeling using flexible multibody technique, (ii) the synthesis of reduced-order models suitable for control design, (iii) the systematic flexible model-based input signal design, and (iv) the evaluation of some possible machine designs. The novelty in this methodology is to take structural flexibilities into consideration during the input signal design; therefore, enhancing the standard design process which mainly considers rigid bodies dynamics. The potential of the proposed strategy is exploited for the design evaluation of a two degree-of-freedom high-speed parallel manipulator. The results are experimentally validated. (C) 2010 Elsevier Ltd. All rights reserved.