Protein Trans -Splicing as a Means for Viral Vector-Mediated In Vivo Gene Therapy

Inteins catalyze protein splicing in a fashion similar to how self-splicing introns catalyze RNA splicing. Split-inteins catalyze precise ligation of two separate polypeptides through trans-splicing in a highly specific manner. Here we report a method of using protein trans-splicing to circumvent the packaging size limit of gene therapy vectors. To demonstrate this method, we chose a large dystrophin gene and an adeno-associated viral (AAV) vector, which has a small packaging size. A highly functional 6.3-kb Becker-form dystrophin cDNA was broken into two pieces and modified by adding appropriate split-intein coding sequences, resulting in split-genes sufficiently small for packaging in AAV vectors. The two split-genes, after codelivery into target cells, produced two polypeptides that spontaneously trans-spliced to form the expected Becker-form dystrophin protein in cell culture in vitro. Delivering the split-genes by AAV1 vectors into the muscle of a mouse model of Duchenne muscular dystrophy rendered therapeutic gene expression and benefits

Curcumin-Loaded Mixed Micelles: Preparation, Characterization, and In Vitro

The objective of this study was to prepare curcumin-loaded mixed Soluplus/TPGS micelles (Cur-TPGS-PMs) for oral administration. The Cur-TPGS-PMs showed a mean size of 65.54 ± 2.57 nm, drug encapsulation efficiency over 85%, and drug loading of 8.17%. The Cur-TPGS-PMs were found to be stable in various pH media (pH 1.2 for 2 h, pH 6.8 for 2 h, and pH 7.4 for 6 h). The X-ray diffraction (XRD) patterns illustrated that curcumin was in the amorphous or molecular state within PMs. The In vitro release test indicated that Cur-TPGS-PMs possessed a significant sustained-release property. The cell viability in MCF-7 cells was found to be relatively lower in Cur-TPGS-PM-treated cells as compared to free Cur-treated cells. CLSM imaging revealed that mixed micelles were efficiently absorbed into the cytoplasm region of MCF-7 cells. Therefore, Cur-TPGS-PMs could have the significant value for the chronic breast cancer therapy

Structural Health Monitoring and Fault Diagnosis based on Artificial Immune System

This thesis presents a development of Structural Health Monitoring (SHM) and Fault Diagnosis based on Artificial Immune System (AIS), a biology-inspired method motivated from the Biological Immune System (BIS). Using the antigen to model structural health or damage condition of specific characteristics and the antibody to represent an information system or a database that can identify the specific damage pattern, the AIS can detect structural damage and then take action to ensure the structural integrity. In this study the antibodies for SHM were first trained and then tested. The feature space in training includes the natural frequencies and the modal shapes extracted from the simulated structural response data including both free-vibration and seismic response data. The concepts were illustrated for a 2-DOF linear mass-spring-damper system and promising results were obtained. It has shown that the methodology can be effectively used to detect, locate, and assess damage if it occurred. Consistently good results were obtained for both feature spaces of the natural frequencies and the modal shapes extracted from both response data sets. As the only exception, some significant errors were observed in the result for the seismic response data when the second modal shape was used as the feature space. The study has shown great promises of the methodology for structural health monitoring, especially in the case when the measurement data are not sufficient. The work lays a solid foundation for future investigations on the AIS application for large-scale complex structures

Do tracking by clustering anchors output from region proposal network

Most existing clustering algorithms suffer from the computation of similarity function and the representation of each object. In this paper, we propose a clustering tracker based on region proposal network (RPN-C) to do tracking by clustering anchors output by region proposal network into potential centers. We first cut off the second part of Faster RCNN and then cast clustering algorithms in feature space of anchors, including K-Means, mean shift and density peak clustering strategy in terms of anchors’ centroid and scale information. Without fully connected layers, the RPN-C tracker can lower the computational cost up to 60% and still, it can effectively maintain an accurate prediction for the localization in next frame. To evaluate the robustness of this tracker, we establish a dataset containing over 2000 training images and 7 testing sequences of 8 kinds of fruits. The experimental results on our own datasets demonstrate that the proposed tracker performs excellently both in location of object and the decision of scale and has a strong advantage of stability in the context of occlusion and complicated background

Isolation and Stabilization of a Pheromone in Crystalline Molecular Capsules

The active monomer form of the male-produced pheromone of the Mediterranean fruit fly can be isolated selectively from its equilibrating trimer species by encapsulation within a calixarene pocket built into a hydrogen-bonded framework from guanidinium 4-sulfocalix[4]­arene. Encapsulation of the Δ¹-pyrroline guest significantly perturbs the assembly of the quasihexagonal two-dimensional guanidinium-sulfonate network of the guest-free framework, to the extent that guanidinium ions are excluded from some sites to accommodate the steric requirements of the guest. Nonetheless, single crystal X-ray diffraction reveals the preservation of a layered structrure in which the calixarene capsules stack in an antiparallel configuration. These observations illustrate that the binding of the pheromone monomer by the calixarene is sufficiently strong to overcome the loss of guanidinium-sulfonate hydrogen bonds, which is corroborated by the strong binding constants measured in solution. The solid-state encapsulation stabilizes the otherwise volatile unstable monomer form, suggesting an effective strategy for the storage, application, and controlled release of an important agricultural adjuvant