Dual Vector Research Articles

A gene therapy for inherited blindness using dCas9-VPR-mediated transcriptional activation.

Catalytically inactive dCas9 fused to transcriptional activators (dCas9-VPR) enables activation of silent genes. Many disease genes have counterparts, which serve similar functions but are expressed in distinct cell types. One attractive option to compensate for the missing function of a defective gene could be to transcriptionally activate its functionally equivalent counterpart via dCas9-VPR. Key challenges of this approach include the delivery of dCas9-VPR, activation efficiency, long-term expression of the target gene, and adverse effects in vivo. Using dual adeno-associated viral vectors expressing split dCas9-VPR, we show efficient transcriptional activation and long-term expression of cone photoreceptor-specific M-opsin (Opn1mw) in a rhodopsin-deficient mouse model for retinitis pigmentosa. One year after treatment, this approach yields improved retinal function and attenuated retinal degeneration with no apparent adverse effects. Our study demonstrates that dCas9-VPR-mediated transcriptional activation of functionally equivalent genes has great potential for the treatment of genetic disorders.

Abstract CT105: First in human phase I trial of adenoviral vectors expressing Flt3L and HSV1-TK to treat newly diagnosed high-grade glioma by reprogramming the brain immune system

Abstract High grade gliomas (HGG) such as glioblastoma lack effective treatment with poor prognosis of median overall survival (OS) around 14-16 months with standard of care. Initiation of effective immune response against cancer requires functional dendritic cells, which are absent from the central nervous system, resulting in lack of anti-HGG immune responses. An effective anti-glioma immune response can be achieved by combining glioma cytotoxicity with HSV1-TK and valacyclovir, and recruitment of dendritic cells to the brain with Flt3L. This dual approach makes endogenous tumor antigens available to infiltrating dendritic cells in its microenvironment by causing: (i) dendritic cells' infiltration of gliomas, (ii) CD8+, CD4+ T cell immune cytotoxicity and memory, and (iii) the systemic immune system to recognize tumor neoantigens. We report the first in human phase I dose escalation trial of adenoviral vectors expressing HSV1-TK and Flt3L (NCT01811992). Injection of dose escalated HSV1-TK and Flt3L adenovectors (range 1x10^9 vp - 1x10^11 vp) to the tumor bed post-resection of newly diagnosed HGG was followed by two cycles of 14-day course of valacyclovir starting 1-3 days and 10-12 weeks post-op combined with standard of care upfront radiation, concurrent and adjuvant temozolomide. Key inclusion criteria were ages 18-75, KPS ≥70, and suspected newly diagnosed HGG amenable to gross total resection. Enrollment and vector injection occurred after frozen pathology confirmed HGG. Out of 18 patients, six are still alive. The primary endpoint of maximal tolerated dose was not reached and the experimental treatment was well tolerated without dose limiting toxicity. The secondary endpoint of OS is promising with median of 21.9 months (range 5.4-52.7). Five out of six patients (83%) who had re-resection at the time of suspected radiographic progression had treatment effect rather than true progression, and increase in markers for dendritic cells, CD4+ T cells, and macrophages were noted, indicating successful immunity recruitment consistent with pre-clinical findings. Updated survival data, as well as comparison to matched controls, and detailed toxicities will be presented at the time of the meeting. In conclusion, the use of dual adenoviral vectors expressing Flt3L and HSV1-TK is safe and well tolerated in newly diagnosed HGG patients. Our results indicate promising preliminary survival outcome and histological evidence of immune infiltration. Future studies to assess treatment efficacy is warranted. Citation Format: Pedro Lowenstein, Daniel Orringer, Yoshie Umemura, Oren Sagher, Jason Heth, Shawn Hervey-Jumper, Aaron Mammoser, Denise Leung, Ted Lawrence, Mishell Kim, Daniel Wahl, Paul McKeever, Sandra Camelo-Piragua, Andrew Lieberman, Sriram Venneti, Kait Verbal, Karen Sagher, Patrick Dunn, Daniel Zamler, Andrea Comba, David Altshuler, Lili Zhao, Karin Muraszko, Larry Junck, Maria G. Castro. First in human phase I trial of adenoviral vectors expressing Flt3L and HSV1-TK to treat newly diagnosed high-grade glioma by reprogramming the brain immune system [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr CT105.

Approximate solutions in nonsmooth and nonconvex cone constrained vector optimization

In this paper, we employ some advanced tools of variational analysis to provide new necessary optimality conditions for approximate (weak) Pareto solutions of a nonconvex and nonsmooth cone constrained vector optimization problem. The obtained necessary conditions are exhibited in a fuzzy form and a Fritz-John type. Sufficient optimality conditions for approximate (weak) Pareto solutions of the multiobjective problem are established by using assumptions of (strictly) approximately generalized convexity. Moreover, we address an approximate dual vector problem for the cone constrained vector optimization problem and examine converse and strong dualities for approximate (weak) Pareto solutions.

Baker–Campbell–Hausdorff–Dynkin Formula for the Lie Algebra of Rigid Body Displacements

The paper proposes, for the first time, a closed form of the Baker–Campbell–Hausdorff–Dynkin (BCHD) formula in the particular case of the Lie algebra of rigid body displacements. For this purpose, the structure of the Lie group of the rigid body displacements S E ( 3 ) and the properties of its Lie algebra s e ( 3 ) are used. In addition, a new solution to this problem in dual Lie algebra of dual vectors is delivered using the isomorphism between the Lie group S E ( 3 ) and the Lie group of the orthogonal dual tensors.

“Elder’s Dual Vector Foil” and the “Spectre” of Karl Marx: The Imagination of Social Form in the 21st Century Chinese Science Fictions

“Elder’s Dual Vector Foil” and the “Spectre” of Karl Marx: The Imagination of Social Form in the 21st Century Chinese Science Fictions

Performance Analysis of Dual Space Vector Modulation Technique-Based Quasi Z-Source Direct Matrix Converter

—This article proposes a dual space vector control strategy for quasi Z-source direct matrix converter (QZSDMC) which is made up of quasi Z-source network coupled to a matrix converter (MC). It ensures effective regulation of voltage regulation range and overpowers the voltage transfer ratio restrictions of conventional matrix converter. The QZSDMC can afford buck boost operation by observing the bidirectional operation capability and shoot through duty ratio. Also, it provides continuous input current and eliminates the need for input LC filter. First, the principle of operation of the proposed QZSDMC is presented in detail. Then, a dual space vector pulse width modulation technique for QZSDMC has been analyzed along with the distribution of shoot through time in the zero vector. The proposed topology with the control strategy has been simulated in MATLAB/SIMULINK and the simulation results are verified with the experimental results from the basic prototype machine controlled by FPGA.

CRISPR-Cas9-Mediated In Vivo Gene Integration at the Albumin Locus Recovers Hemostasis in Neonatal and Adult Hemophilia B Mice

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 loaded by vectors could induce high rates of specific site genome editing and correct disease-causing mutations. However, most monogenic genetic diseases such as hemophilia are caused by different mutations dispersed in one gene, instead of an accordant mutation. Vectors developed for correcting specific mutations may not be suited to different mutations at other positions. Site-specific gene addition provides an ideal solution for long-term, stable gene therapy. We have demonstrated SaCas9-mediated homology-directed factor IX (FIX) in situ targeting for sustained treatment of hemophilia B. In this study, we tested a more efficient dual adeno-associated virus (AAV) strategy with lower vector dose for liver-directed genome editing that enables CRISPR-Cas9-mediated site-specific integration of therapeutic transgene within the albumin gene, and we aimed to develop a more universal gene-targeting approach. We successfully achieved coagulation function in newborn and adult hemophilia B mice by a single injection of dual AAV vectors. FIX levels in treated mice persisted even after a two-thirds partial hepatectomy, indicating stable gene integration. Our results suggest that this CRISPR-Cas9-mediated site-specific gene integration in hepatocytes could transform into a common clinical therapeutic method for hemophilia B and other genetic diseases.

3-Hom-Lie Algebras Based on $$\sigma $$-Derivation and Involution

We show that, having a Hom-Lie algebra and an element of its dual vector space that satisfies certain conditions, one can construct a ternary totally skew-symmetric bracket and prove that this ternary bracket satisfies the Hom-Filippov-Jacobi identity, i.e. this ternary bracket determines the structure of 3-Hom-Lie algebra on the vector space of a Hom-Lie algebra. Then we apply this construction to two Hom-Lie algebras constructed on an associative, commutative algebra using $$\sigma $$ -derivation and involution, and we obtain two 3-Hom-Lie algebras.

Construction of an expression vector mediated by the dual promoter for prokaryotic and mammalian cell expression system.

The aim of this study was to construct an expression vector mediated by the dual promoter that can simultaneously drive the recombinant protein production in eukaryotic and prokaryotic cells. The prokaryotic T7 promoter and ribosome binding site (RBS) was cloned downstream of CMV promoter in the eukaryotic expression vector pIRES-neo, and T7 termination sequence was inserted upstream of neomycin phosphotransferase gene to generate the dual promoter vector. The enhanced green fluorescent protein (eGFP) gene was used as reporter gene. Then, the resultant vector was transfected into Chinese hamster ovary (CHO) cells and transformed into Escherichia coli (E. coli) BL21, and the eGFP expression levels were analyzed by fluorescence microscopy, flow cytometry and Western blot, respectively. Fluorescence microscopy revealed that the eGFP was expressed in both CHO cells and E. coli BL21. Flow cytometry showed that the eGFP expression level had no significant difference between the dual promoter vector and control vector in transfected CHO cells. Western blot analysis indicated the eGFP expressed in transformed E. coli. In conclusion, a prokaryotic-eukaryotic double expression vector was successfully constructed, which has potential applications in rapid cloning and expression of recombinant proteins in both prokaryotic and eukaryotic expression systems.

Assessment of AAV Dual Vector Safety in theAbca4-/- Mouse Model of Stargardt Disease.

PurposeAdeno-associated viral (AAV) gene therapy treatment for Stargardt disease currently requires a dual vector approach owing to the size of the ATP-binding cassette transporter family member gene (ABCA4). The nature of the dual vector system creates the potential for adverse events. Here we have investigated an overlapping adeno-associated viral ABCA4 dual vector system for signs of toxicity in Abca4−/− mice as a prelude to dual vector first in human clinical trials.Methods Abca4−/− mice received a subretinal injection of a 1:1 5′:3′ dual vector mix; 5′ vector only; 3′ vector only; a GFP reporter vector; or diluent only (sham). All vectors were adeno-associated virus-8 Y733F. Mice were subsequently assessed for signs of toxicity as measured by loss in retinal structure by optical coherence tomography and retinal function by electroretinography up to 6 months after injection.ResultsSubretinal delivery of the dual vector system and its comprising parts induced no structural or functional changes relative to paired uninjected eyes beyond those observed in the sham control cohort. Histologic changes were limited to the superior retina where the injection was performed. Electroretinography analysis confirmed the dual vector system inferred no functional changes beyond those observed in the sham control cohort.ConclusionsAn optimized overlapping dual vector system for the treatment of Stargardt disease shows no additional signs of toxicity beyond those observed from a sham injection.Translational RelevanceThis presentation of safety of a dual vector system for the treatment of Stargardt disease encourages its future use in clinical trial.

UDP-glucose pyrophosphorylase gene affects mycelia growth and polysaccharide synthesis of Grifola frondosa

To elucidate potential roles of UDP-glucose pyrophosphorylase (UGP) in mycelial growth and polysaccharide synthesis of Grifola frondosa, a putative 2036-bp UDP-glucose pyrophosphorylase gene gfugp encoding a 53.17-kDa protein was cloned and re-annotated. Two dual promoter RNA silencing vectors of pAN7-iUGP-P-dual and pAN7-iUGP-C-dual were constructed to down-regulate gfugp expression by targeting its promoter or conserved functional sequences, respectively. Results showed that silence of gfugp promoter sequence had a higher down-regulating efficiency with slower mycelial growth and polysaccharide production than those of conserved sequence. The monosaccharide compositions/percentages of mycelial and exo-polysaccharides significantly changed with the increase of galactose and arabinose contents possibly due to block of UDP-glucose supply by gfugp silence and alteration of sugar metabolism via up-regulation of UDP-glucose-4-epimerase (gfuge) and UDP-xylose-4-epimerase (gfuxe) transcription. Our findings would provide a reference to know the biosynthesis pathway of mushroom polysaccharides and improve their production by metabolic regulation.

A semi-analytical approach for the flexural analysis of in-plane functionally graded magneto-electro-elastic plates

This paper carries out the semi-analytical flexural analysis of functionally graded magneto-electro-elastic plates with material coefficients mathematically expressed as arbitrary functions in terms of the in-plane coordinates utilizing the scaled boundary finite element method (SBFEM) in association with the precise integration method (PIM). The introduced technique can be exploited to explore the deformable behaviors of magneto-electro-elastic plates with various geometrical shapes, thickness-to-span ratios and boundary constraints. Only five variables including elastic displacement components through the x ,y and z directions, electric potential and magnetic potential are served as the primary unknowns without importing a priori assumptions on variations of the 3D multi-physical quantities. Furthermore, the mechanical, electric, and magnetic fields along the transverse direction of the plate can be analytically formulated. Moreover, the developed methodology only demands to discretize a surface of plate, which is profitable to simplify the 3D plate structure into a 2D problem and lower the calculation effort. Aided by the transformation between the scaled boundary and Cartesian coordinate system, the principle of virtue work and the dual vector technique, the SBFEM governing matrix equation for the in-plane functionally graded magneto-electro-elastic plate is depicted as a first order ordinary differential one and its corresponding general solution is analytically modelled as an exponential matrix according to the z-coordinate. As a convenient and highly accurate approach by dint of the 2N algorithm, the PIM is employed to construct the stiffness equation from the exponential matrix and compute the mechanical, electric and magnetic components. Finally, several numerical examples are provided to verify the high accuracy, applicability and effectiveness of the proposed method and manifest the effects of boundary conditions, thickness-to-span ratios and external loading forms on distributions of the multi-physical coupling quantities in functionally graded magneto-electro-elastic plates with in-plane material stiffness.

Fast prediction mode selection and CU partition for HEVC intra coding

Compared with the predecessor H.264/advanced video coding, high-efficiency video coding (HEVC) is a new video coding standard with nearly double coding efficiency under the same coding quality. However, the computing complexity of HEVC increases sharply. To solve this problem, a fast algorithm for intra prediction mode selection based on mode grouping is proposed by reducing the number of modes entering rough mode decision. Moreover, the dual support vector machine is proposed to efficiently select the coding unit (CU) size, which is based on texture features of CU and sub-CUs including content complexity and direction complexity. By using the new CU size selection algorithm, the encoder can confirm the split for complex CU and terminate the split for simple CU in advance, so as to reduce the computing complexity of CU size selection. The experimental results show that by employing the two fast algorithms in intra coding, it can save 42.80% encoding time, with 0.98% increment in bit rate and 0.018 dB loss of peak-signal-to-noise ratio of luminance, compared with the reference software x265-1.7.

Modified Alphavirus-Vesiculovirus Hybrid Vaccine Vectors for Homologous Prime-Boost Immunotherapy of Chronic Hepatitis B.

Virus-like vesicles (VLV) are hybrid vectors based on an evolved Semliki Forest virus (SFV) RNA replicon and the envelope glycoprotein (G) from vesicular stomatitis virus (VSV). Previously, we showed that VLV can be used to express protein antigens and generate protective antigen-specific CD8+ T cells. This report describes VLV vectors designed for enhanced protein expression and immunogenicity. Expressing hepatitis B virus (HBV) middle S antigen (MHBs) from VLV using a dual subgenomic promoter significantly increased MHBs-specific CD8+ T cell and antibody production in mice. Furthermore, envelope glycoprotein switch from VSV Indiana to the glycoprotein of Chandipura virus enabled prime-boost immunization and further increased responses to MHBs. Therapeutic efficacy was evaluated in a mouse model of chronic HBV infection initiated by HBV delivery with adeno-associated virus. Mice with lower or intermediate HBV antigen levels demonstrated a significant and sustained reduction of HBV replication following VLV prime-boost immunization. However, mice with higher HBV antigen levels showed no changes in HBV replication, emphasizing the importance of HBV antigenemia for implementing immunotherapies. This report highlights the potential of VLV dual promoter vectors to induce effective antigen-specific immune responses and informs the further development and evaluation of hybrid viral vaccine platforms for preventative and therapeutic purposes.

Dual Extreme Learning Machines-Based Spatiotemporal Modeling for Nonlinear Distributed Thermal Processes

Many industrial thermal processes belong to distributed parameter systems (DPSs), which have two coupled nonlinear blocks. Dual least square support vector machines (LS-SVM) has been proposed to model such systems. However, due to the use of two LS-SVM, this method often leads to heavy computation and long learning time, which does not suit for online application. In this study, a dual extreme learning machine (ELM)-based spatiotemporal modeling method is proposed for such two nonlinearities embedded DPSs. Firstly, the KL method is applied to reduce the dimension of the original system and obtain the spatial basis functions (BFs). Then, dual ELM is designed to match the two nonlinear structures. Finally, through the reconstruction of space–time synthesis, the approximate spatiotemporal distribution model of the original system is obtained. In addition, simulations on a curing process is studied to confirm the effectiveness of the proposed method.

Fourier transform, quantum mechanics and quantum field theory on the manifold of general relativity

A proof is given for the Fourier transform for functions in a quantum mechanical Hilbert space on a non-compact manifold in general relativity. In the (configuration space) Newton–Wigner representation, we discuss the spectral decomposition of the canonical operators and give a proof of the Parseval–Plancherel relation and the Born rule for linear superposition. We then discuss the representations of pure quantum states and their dual vectors and construct the Fock space and the associated quantum field theory for Bose–Einstein and Fermi–Dirac statistics.

Particle motion measurements near a rocky shore off Cabo Frio Island.

This paper describes the lessons learned from the experiment BIOCOM'19 carried out in January 2019, in a shallow water bay off the island of Cabo Frio (RJ, Brazil). A dual accelerometer vector sensor hydrophone was deployed for two days, near a rocky shore covered with a significant benthic fauna. The results show that the frequency band above approximately 1.5 kHz is mostly associated with invertebrate biological noise and that the acoustic and the particle motion fields have a similar behavior, following the usual dawn-dusk activity pattern, and a coherent directivity content. At low frequencies, below ∼300 Hz, the acoustic pressure and the particle acceleration fields have significantly different spectral content along time. Many of these differences are due to anthropogenic noise sources related with nearby boating activity, while during quiet periods, they may be attributed to the biological activity from the rocky shore.

NEW APPROACHES ON DUAL SPACE

In this paper, we give how to define the basic concepts of differential geometry on Dual space. For this, dual tangent vectors that have p as dual point of application are defined. Then, the dual analytic functions defined by Dimentberg are examined in detail, and by using the derivative of the these functions, dual directional derivatives and dual tangent maps are introduced.

What Is So Special about Quantum Clicks?

This is an elaboration of the “extra” advantage of the performance of quantized physical systems over classical ones, both in terms of single outcomes as well as probabilistic predictions. From a formal point of view, it is based on entities related to (dual) vectors in (dual) Hilbert spaces, as compared to the Boolean algebra of subsets of a set and the additive measures they support.

Chloroplast Dual Divergent Promoter Plasmid for Heterologous Protein Expression in Tetraselmis suecica (Chlorophyceae, Chlorodendrales).

The eukaryotic green microalga Tetraselmis suecica is commonly used for aquaculture purposes because of its high stress tolerance and ease of culture in a wide spectrum of environments; they are therefore suitable candidates for biotechnology applications. To date, no data are available regarding chloroplast transformation vectors based on specific endogenous promoters and homologous targeting regions. We report on the identification of Tetraselmis suecica genes encoding the ribulose bisphosphate carboxylase/oxygenase large subunit protein, the photosystem II D1 protein and the ATP synthase CF1-beta subunit protein together with their untranslated regions (5'UTR, 3'UTR). The full-length ORFs of the putative genes with their regulatory sequences were obtained. We were also able to identify the downstream 3' end of the large subunit ribosomal RNA gene (23S) along with the 5S RNA end-to-end with the psbA gene on the complementary strand. The intergenic region between these genes appears to be a good target site for the integration of target proteins. Moreover, we identified a back-to-back promoter region among the rbcL and atpB genes. To assess the bidirectionality activities of both promoters, a dual reporter vector was constructed for Tetraselmis suecica transformation containing the cat and TurboGFP genes driven by the 5'rbcL/5'atpB divergent promoter. The vector included the 23S-5S and psbA nucleotide sequences as flanking regions. These flanking regions provided suitable insertionsites within the chloroplast genome forcassette integration via homologous recombination.Simultaneous expression of the chloramphenicol-resistant conferring gene and the gene coding for TurboGFP driven by 5'rbcL/5'atpB showed a potent natural bidirectional promoter as a reliable genetic tool.