There are many classics fairy tales, the Grimm's Fairy Tales, Hans Christian Andersen fairy tale, Arabian Nights, etc. which are well known by every child across the world. These stories not only are interesting, but also have educational value. The rapid development of mobile devices and the increase in their use by preschool children, revealing how technology plays an important role in this generation. Technology makes content more engaging through interaction, particularly e-book. Instead of being bounded by limitations of traditional paper books, multimedia applications open up a new world in reading. In view of this, the main purpose of this research is to develop a set of interactive learning applications, based on the fairy tales as learning background for preschool children. To achieve this purpose, the methodology adopted multiplatform game development in C# with Unity, in Windows 10. The Android 5.0.2 version of the smart phone system was used as the implementation platform. This Interactive Application includes the interactive learning prospect that allows the incorporation of animation, moving pictures and sounds which extend the ability to present materials that encourage children’s interaction with the subject matter. In addition, this system adds interaction features with readers and discussion questions, including puzzles, multiple-choice, true-false questions, sums, etc. This application provides a thinking platform for the children. It helps in the child to develop the reasoning, logic, imaginative skills and capacity for independent thinking.

PURPOSE:Our aim was to study the effect of three-dimensional (3D) environment and overexpression of human fibroblast growth factor 2 (FGF-2) on meniscal fibrochondrocytes in vitro.METHODS:Human meniscal fibrochondrocytes were transfected with expression plasmid vectors carrying the Photinus pyralis luciferase gene, the Escherichia coli β-galactosidase gene or a human FGF-2 cDNA. Modified fibrochondrocytes were cultivated in 3D alginate hydrogel or cell pellets or in 2D monolayer culture.RESULTS:The levels of luciferase activity showed a peak at day two and returned to baseline levels by day 11, regardless of the type of cultivation. Both 3D environments supported the secretion of human FGF-2 protein upon FGF-2 transfection. Overexpression of human FGF-2 by genetically modified human meniscal fibrochondrocytes stimulated proliferation but not glycosaminoglycan synthesis only in 3D culture. Culture in alginate spheres resulted in a larger difference in cell numbers compared with pellet cultures.CONCLUSIONS:Three-dimensional alginate spheres are well suited for the culture of genetically modified human meniscal fibrochondrocytes. These data are of value for cell-based approaches to meniscal repair using genetically modified human meniscal fibrochondrocytes overexpressing human FGF-2.

Purpose Our aim was to evaluate whether nonviral vectors can genetically modify primary human juvenile and adult meniscal fibrochondrocytes at low toxicity in vitro and to test the hypothesis that transfected human meniscal fibrochondrocytes transplanted into longitudinal defects and onto human medial meniscus explant cultures are capable of expressing transgene products in vitro. Methods Eighteen nonviral gene transfer systems were examined to identify the best suited method for an efficient transfection of primary cultures of juvenile and adult human meniscal fibrochondrocytes using luciferase and lacZ reporter gene constructs and then transplanted to meniscus explant cultures. Results Gene transfer systems FuGENE 6, GeneJammer, TurboFectin 8, calcium phosphate co-precipitates and GeneJuice led to minimal toxicity in both cell types. Nanofectin 2 and JetPEI resulted in maximal luciferase activity in both cell types. Maximal transfection efficiency based on X-gal staining following lacZ gene transfer was achieved using Lipofectamine 2000, revealing a mean transfection efficiency of 8.6 % in human juvenile and of 8.4 % in adult meniscal fibrochondrocytes. Transfected, transplanted meniscal fibrochondrocytes adhered to the meniscal tissue and continued to express the transgene for at least five days following transfection. Conclusions Nonviral gene transfer systems are safe and capable of transfecting both juvenile and adult human meniscal fibrochondrocytes, which, when transplanted to meniscal tissue in vitro, permit the expression of selected transgenes to be maintained. These results are of value for combining gene therapy and cell transplantation approaches as a means to enhance meniscal repair.

A new speech enhancement architecture using convolutive blind signal separation (CBSS) and subspace-based speech enhancement is presented. The spatial and spectral information are integrated to enhance the target speech signal and suppress both interference noise and background noise. Real-world experiments were carried out in a noisy office room. Experimental results demonstrate the superiority of the proposed architecture.

Multimodal human-robot interaction integrates various physical communication channels for face-to-face interaction. However, face-to-face interaction is not the only communication method. The proposed framework achieves more flexible communications such as communication to others at a distance. Intimate and loose interactions are categorized as ubiquitous multimodal human-robot interaction. Therefore, this work presents a framework design for human-robot interaction using ubiquitous voice control and face localization and authentication implemented for intimate and loose interactions. The simulation results demonstrate the practicality of the proposed framework.

ABSTRACT Heliothis zea nudivirus 1 (HzNV-1 or Hz-1 virus), previously regarded as a nonoccluded baculovirus, recently has been placed in the Nudivirus genus. This virus generates HzNV-1 HindIII-I 1 (hhi1) and many other transcripts during productive viral infection; during latent viral infection, however, persistency-associated gene 1 (pag1) is the only gene expressed. In this report, we used transient expression assays to show that hhi1 can trigger strong apoptosis in transfected cells, which can be blocked, at least partially, by the inhibitor of apoptosis genes Autographa californica iap2 (Ac-iap2) and H. zea iap2 (Hz-iap2). In addition to these two genes, unexpectedly, pag1, which encodes a noncoding RNA with no detectable protein product, was found to efficiently suppress hhi1-induced apoptosis. The assay of pro-Sf-caspase-1 processing by hhi1 transfection did not detect the small P12 subunit at any of the time intervals tested, suggesting that hhi1 of HzNV-1 induces apoptosis through alternative caspase pathways.

Detection engines capable of inspecting packet payloads for application-layer network information are urgently required. The most important technology for fast payload inspection is an efficient multipattern matching algorithm, which performs exact string matching between packets and a large set of predefined patterns. This paper proposes a novel Enhanced Hierarchical Multipattern Matching Algorithm (EHMA) for packet inspection. Based on the occurrence frequency of grams, a small set of the most frequent grams is discovered and used in the EHMA. EHMA is a two-tier and cluster-wise matching algorithm, which significantly reduces the amount of external memory accesses and the capacity of memory. Using a skippable scan strategy, EHMA speeds up the scanning process. Furthermore, independent of parallel and special functions, EHMA is very simple and therefore practical for both software and hardware implementations. Simulation results reveal that EHMA significantly improves the matching performance. The speed of EHMA is about 0.89-1,161 times faster than that of current matching algorithms. Even under real-life intense attack, EHMA still performs well.

Background Baculovirus is an effective vector for gene delivery into primary chondrocytes and repeated baculovirus transduction (i.e. supertransduction) appears to be promising for prolonging transgene expression, but how supertransduction may influence baculovirus-mediated gene delivery is unknown.Methods We first investigated whether prior baculovirus transduction suppressed the ensuing transgene expression mediated by the supertransduced baculovirus, and then examined whether baculovirus triggered the expression of various cytokines. Whether interferon-alpha and -beta (IFN-alpha/beta) suppressed the transgene expression as well as the pivotal step responsible for the attenuated transgene expression were examined.Results Baculovirus transduction of chondrocytes elicited an immediate yet transient expression of IFN-alpha/beta, which repressed the transgene expression in a dose-dependent manner. The attenuation was observed for transgene expression driven by different promoters and resulted neither from internalization or nuclear import of baculovirus. Moreover, the attenuation was alleviated if supertransduction was performed when IFN-alpha/beta responses diminished. Baculovirus transduction also triggered the expression of tumor necrosis factor-alpha and interleukin (IL)-6, but not IL-1 beta. Despite the induction of these responses, supertransduction of chondrocytes with a baculovirus expressing bone morphogenetic protein-2 successfully enhanced the chondrogenic differentiation and matrix synthesis.Conclusions Baculovirus transduction of primary chondrocytes elicits antiviral effects that suppress transgene expression. Nonetheless, baculovirus supertransduction comprises a feasible approach to extend transgene expression for cartilage engineering. Copyright (C) 2009 John Wiley & Sons, Ltd.

This work presents a novel approach to acoustic and phoneme modeling in order to recognize ubiquitous mixed-language speech. The conventional approaches to perform multilingual speech recognition are the usage of a multilingual phone set. A confusion matrix combining acoustic between every two phonetic is built for phonetic unit clustering. In this work, we are interested in speaker independent voice command recognition. The IPA representation is adapted for phonetic unit modeling. The EAT is applied to construct speaker independent acoustic models. The experimental results show that the proposed method can perform 70~80% lexicon recognition accuracy.

This work presents a robust environmental sound recognition system for home automation. Specific home automation services can be activated based on identified sound classes. Additionally, when the sound category is human speech, such speech can be recognized for detecting human intentions as in conventional research on home automation. To attain this ambitious goal, this study uses two key techniques: signal-to-noise ratio-aware subspace-based signal enhancement and sound recognition with independent component analysis mel-frequency cepstral coefficients and a frame-based multiclass support vector machines, respectively. Simulations and an experiment in a real-world environment are given to illustrate the performance of the proposed robust sound recognition system.

Baculovirus has emerged as a new gene delivery vector thanks to a number of advantages. This study demonstrated that baculovirus conferred efficient gene delivery and mediated expression of growth factors (TGF-β1, IGF-1 and BMP-2) to therapeutic levels in rabbit chondrocytes. Interestingly, the cellular response to growth factor stimulation was dependent on the cell passage. The highly de-differentiated passage 5 (P5) chondrocytes failed to respond to the stimulation by either growth factor. The de-differentiated P3 cells also failed to maintain the chondrocyte phenotype, but baculovirus-mediated BMP-2 expression remarkably reversed the de-differentiation and enhanced the aggrecan and collagen II production in 2D and 3D cultures, as evidenced by cell morphology, histological staining and gene expression analyses. Baculovirus-mediated TGF-β1 expression modestly enhanced the cartilage-specific matrix production, although to a lesser extent. Intriguingly, IGF-1, a well-known chondroinductive protein, failed to stimulate the P3 cells likely due to the loss of IGF-1 receptor expression. In summary, this study proved for the first time the potentials of baculovirus in modulating the differentiation status of chondrocytes in the context of cartilage tissue engineering, but also highlighted the importance of selecting appropriate cell passage and growth factor for genetic manipulation.

Conjugating a single glycosaminoglycan (GAG) species such as chondroitin-6-sulfate (CSC) to chitosan is beneficial to chondrocyte culture and extracellular matrix (ECM) production, but whether fabrication of 3D chitosan scaffolds with additional minor GAG species such as dermatan sulfate (DS) further improves the ECM production is unknown. In this study, Response Surface Methodology (RSM) was employed to design CSC/DS/chitosan scaffolds of various formulations for cartilage engineering and to investigate the roles of individual GAG species in cartilage formation. The CSC/DS formulation affected neither the physical properties of scaffolds nor cell adhesion, but influenced cell morphology, GAGs and collagen production and chondrocytic gene expression. The linear effects elucidated by RSM analysis suggested that within the level range higher CSC levels favored GAGs and collagen production, whereas lower DS levels were desired for these responses. Nonetheless, the quadratic effects of DS and two-way interactions between CSC and DS also contributed to the GAGs and collagen production. Accordingly, the optimal formulation, as predicted by RSM and validated by experiments, comprised 2.8 mg CSC and 0.01 mg DS per scaffold. This study confirmed the importance of DS in cartilage tissue engineering and implicated the feasibility of rational CSC/DS/chitosan scaffold design with the aid of RSM.

Background Baculovirus transduction of cultured mammalian cells is typically performed by incubating the cells with virus using culture medium (e.g. Dulbecco's modified Eagle's medium (DMEM)) as the surrounding solution. However, we previously uncovered that DMEM hinders the baculovirus-mediated gene transfer.Methods In this study, we systematically explored the influences of promoter and medium constituents on the transduction efficiency by using different recombinant viruses and surrounding solutions for transduction, followed by flow cytometric analyses. Whether the key medium component impeded baculovirus binding to the cells and subsequent virus entry was investigated by immunofluorescence/confocal microscopy and quantitative real-time polymerase chain reaction (Q-PCR).Results We demonstrated that the poorer transduction by using DMEM as the surrounding solution is independent of the promoter. Examination of the medium constituents group by group revealed that the balanced salt solution suppresses the baculovirus transduction. By omitting individual salt species in the balanced salt solution, we surprisingly uncovered that NaHCO3, a common buffering agent, exerts the inhibitory effects in a concentration-dependent manner. Intriguingly, NaHCO3 did not debilitate the baculovirus, nor did it inhibit virus binding to the cells. Instead, NaHCO3 inhibited baculovirus transduction by reducing the intracellular virus number.Conclusions To our best knowledge, this is the first report unraveling the significance of NaHCO3 in gene transfer. Our finding suggests that baculovirus-mediated gene transfer can be readily enhanced by omitting NaHCO3 from the medium during the transduction period. Copyright (C) 2007 John Wiley & Sons, Ltd.

We have previously demonstrated that baculovirus can efficiently transduce human mesenchymal stem cells (MSCs) and MSCs-derived adipogenic, chondrogenic, and osteogenic progenitors without compromising the differentiation capacity. Remarkably, the transgene expression level and duration varied widely with the differentiation states at which the progenitors were transduced. However, whether the variation was a general phenomenon and what caused the variation were unclear. Here we demonstrated that transduction of the MSCs and MSC-derived progenitors using baculoviruses carrying egfp driven by CMV, EF-1alpha or CAG promoter resulted in a general trend of varied expression, that is, the chondrogenic progenitors allowed for the poorest expression while the adipogenic progenitors conferred the best expression. Quantification of the nuclear and cytoplasmic egfp gene copy numbers by quantitative real-time PCR revealed that the varied expression did not arise from the discrepancies in gene delivery efficiency nor was it due to the disparities in nuclear transport efficiency. In contrast, the transcription levels paralleled the overall expression levels. These data suggested that although the egfp genes could be efficiently delivered into the nuclei of chondrogenic progenitors, they were not transcribed as well as they were in the adipogenic progenitors. In conclusion, the rapidly altering cellular transcription machinery in the course of differentiation progression predominantly led to the varied expression levels.

The baculovirus expression vector system has been commonly employed as a rapid and inexpensive approach for recombinant protein production in insect cells (1, 2). Despite the wide applications, baculovirus infection results in insect cell death and lysis in a few days post-infection, which can lead to suboptimal protein processing late in the infection phase. In addition, some proteins that are synthesized initially as large inactive precursor proteins, such as peptide hormones, are not efficiently processed in insect cells (3). Furthermore, glycosylation in insect cells differs in many aspects from that in mammalian cells. In particular, insect cells generally fail to sialylate recombinant N-glycoproteins due to the absence of sialyltransferase activities and CMP-sialic acids (4, 5). The resultant glycoproteins, lacking sialic acids, may thus have altered immunogenicity and extremely short half-lives in vivo. In the mid-1990s, Hofmann et al. (6) and Boyce & Bucher (7) first reported that recombinant baculoviruses harboring a mammalian expression cassette could efficiently transduce mammalian cells. Their data indicated a strong preference of baculovirus to enter hepatocytes of different origins. Later, Shoji et al. (8) showed that cells that are not transduced by a baculovirus expressing β-galactosidase under the control of a cytomegalovirus (CMV) promoter can be transduced efficiently by a baculovirus expressing the same reporter protein under the control of the stronger CAG promoter, comprising the β-actin promoter and CMV immediate-early enhancer. These pioneering studies lead to the discovery of a growing list of permissive cells originating from human, rodent, porcine, bovine, fish (for reviews, see 3 and 9) and even chicken (10). Due to the high efficiency of baculovirus-mediated gene delivery to various nondividing and primary cells, increasing efforts have been directed towards employing baculoviruses for in vitro and in vivo gene delivery. In recent years, baculoviruses have emerged as a new expression vector in mammalian cells, know as the BacMam system, thanks to the high gene delivery efficiency, its nonreplicative nature and low cytotoxicity inside mammalian cells, its large genome (~130 kb) capable of harboring multiple genes or large inserts,

Acoustic characteristics have played an essential role in biometrics. In this article, we introduce a robust, text-independent speaker identification/verification system. This system is mainly based on a subspace-based enhancement technique and probabilistic support vector machines (SVMs). First, a perceptual filterbank is created from a psycho-acoustic model into which the subspace-based enhancement technique is incorporated. We use the prior SNR of each subband within the perceptual filterbank to decide the estimator's gain to effectively suppress environmental background noises. Then, probabilistic SVMs identify or verify the speaker from the enhanced speech. The superiority of the proposed system has been demonstrated by twenty speaker data taken from AURORA-2 database with added background noises

In this paper, a new subspace-based speech enhancement algorithm is presented. First, we construct a perceptual filterbank from psycho-acoustic model and incorporate it in the subspace-based enhancement approach. This filterbank is created through a five-level wavelet packet decomposition. The masking properties of the human auditory system are then derived based on the perceptual filterbank. Finally, the prior SNR and the masking threshold of each critical band are taken to decide the attenuation factor of the optimal linear estimator. Five different types of in-car noises in TAICAR database were used in our evaluation. The experimental results demonstrated that our approach outperformed conventional subspace and spectral subtraction methods.

In this paper, a new subspace-based speech enhancement algorithm for in-car human computer speech interaction is presented. We first incorporate a perceptual filterbank which is derived from psycho-acoustic model with signal subspace approach to effectively suppress in-car noises of engine. Second, for real-time applications, a new subspace tracking algorithm is derived by modifying PASTd algorithm to solve the data dependent hazard of tacking algorithm. Six different types of in-car noises in TAICAR database are used in our evaluation. The experimental results demonstrate that our approach is superior to conventional subspace and spectral subtraction methods.

In this paper, we present a robust text-independent speaker recognition system. The proposed system mainly includes an SNR-aware subspace-based enhancement technique and probabilistic support vector machines (SVMs). First, we construct a perceptual filterbank from psycho-acoustic model and incorporate it with the subspace-based enhancement approach. The prior SNR of each subband within the perceptual filterbank is taken to decide the estimator's gain to effectively suppress environmental background noises. Next, this study uses probabilistic SVMs to identify the speaker from the enhanced speech. The superiority of the proposed system has been demonstrated by twenty speaker recognition from AURORA-2 database with in-car noises