Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations

Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B₄ (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.     

Phenolic Phytoalexins in Rice: Biological Functions and Biosynthesis

Phytoalexins are inducible secondary metabolites possessing antimicrobial activity against phytopathogens. Rice produces a wide array of phytoalexins in response to pathogen attacks and environmental stresses. With few exceptions, most phytoalexins identified in rice are diterpenoid compounds. Until very recently, flavonoid sakuranetin was the only known phenolic phytoalexin in rice. However, recent studies have shown that phenylamides are involved in defense against pathogen attacks in rice. Phenylamides are amine-conjugated phenolic acids that are induced by pathogen infections and abiotic stresses including ultra violet (UV) radiation in rice. Stress-induced phenylamides, such as N-trans-cinnamoyltryptamine, N-p-coumaroylserotonin and N-cinnamoyltyramine, have been reported to possess antimicrobial activities against rice bacterial and fungal pathogens, an indication of their direct inhibitory roles against invading pathogens. This finding suggests that phenylamides act as phytoalexins in rice and belong to phenolic phytoalexins along with sakuranetin. Phenylamides also have been implicated in cell wall reinforcement for disease resistance and allelopathy of rice. Synthesis of phenolic phytoalexins is stimulated by phytopathogen attacks and abiotic challenges including UV radiation. Accumulating evidence has demonstrated that biosynthetic pathways including the shikimate, phenylpropanoid and arylmonoamine pathways are coordinately activated for phenolic phytoalexin synthesis, and related genes are induced by biotic and abiotic stresses in rice.     

An adaptive image steganographic scheme based on Noise Visibility Function and an optimal chaotic based encryption method

Steganography is the science of hiding secret message in an appropriate digital multimedia in such a way that the existence of the embedded message should be invisible to anyone apart from the sender or the intended recipient. This paper presents an irreversible scheme for hiding a secret image in the cover image that is able to improve both the visual quality and the security of the stego-image while still providing a large embedding capacity. This is achieved by a hybrid steganography scheme incorporates Noise Visibility Function (NVF) and an optimal chaotic based encryption scheme. In the embedding process, first to reduce the image distortion and to increase the embedding capacity, the payload of each region of the cover image is determined dynamically according to NVF. NVF analyzes the local image properties to identify the complex areas where more secret bits should be embedded. This ensures to maintain a high visual quality of the stego-image as well as a large embedding capacity. Second, the security of the secret image is brought about by an optimal chaotic based encryption scheme to transform the secret image into an encrypted image. Third, the optimal chaotic based encryption scheme is achieved by using a hybrid optimization of Particle Swarm Optimization (PSO) and Genetic Algorithm (GA) which is allowing us to find an optimal secret key. The optimal secret key is able to encrypt the secret image so as the rate of changes after embedding process be decreased which results in increasing the quality of the stego-image. In the extracting process, the secret image can be extracted from the stego-image losslessly without referring to the original cover image. The experimental results confirm that the proposed scheme not only has the ability to achieve a good trade-off between the payload and the stego-image quality, but also can resist against the statistics and image processing attacks.     

Model checking temporal knowledge and commitments in multi-agent systems using reduction

Though modeling and verifying Multi-Agent Systems (MASs) have long been under study, there are still challenges when many different aspects need to be considered simultaneously. In fact, various frameworks have been carried out for modeling and verifying MASs with respect to knowledge and social commitments independently. However, considering them under the same framework still needs further investigation, particularly from the verification perspective. In this article, we present a new technique for model checking the logic of knowledge and commitments (CTLKC⁺). The proposed technique is fully-automatic and reduction-based in which we transform the problem of model checking CTLKC⁺ into the problem of model checking an existing logic of action called ARCTL. Concretely, we construct a set of transformation rules to formally reduce the CTLKC⁺ model into an ARCTL model and CTLKC⁺ formulae into ARCTL formulae to get benefit from the extended version of NuSMV symbolic model checker of ARCTL. Compared to a recent approach that reduces the problem of model checking CTLKC⁺ to another logic of action called GCTL¹, our technique has better scalability and efficiency. We also analyze the complexity of the proposed model checking technique. The results of this analysis reveal that the complexity of our reduction-based procedure is PSPACE-complete for local concurrent programs with respect to the size of these programs and the length of the formula being checked. From the time perspective, we prove that the complexity of the proposed approach is P-complete with regard to the size of the model and length of the formula, which makes it efficient. Finally, we implement our model checking approach on top of extended NuSMV and report verification results for the verification of the NetBill protocol, taken from business domain, against some desirable properties. The obtained results show the effectiveness of our model checking approach when the system scales up.     

Spray coating process variable and property analysis of UV-curable polyurethane acrylate coating on polycarbonate substrate

A series of spray coating experiments were conducted on an UV-curable, polyurethane-modified, acrylate-based coating formulation with the aim to control defects, coating thickness, and thickness variation. Statistical approaches including design of experiment, residual examination, analysis of variance, and t-test were used in designing the experiments and analyzing data. Viscosity of formulation, atomizing pressure, liquid feeding pressure, distance between nozzle and substrate, and travel speed of substrate were the process variables studied. The ranges of process variables that gave defect-free coating were identified and used in the subsequent experiments to determine process variables and interactions that had significant contribution to the changes in coating thickness and thickness variation. All process variables studied were found to have contribution to the change in coating thickness, but they showed no significant contribution to the variation of coating thickness. No interaction displayed significant contribution. Confirmation tests performed on extra samples prepared with varying coating thicknesses indicated a good agreement with the experimental results. Additional samples were tested for total transmittance, transmission haze, adhesion, surface roughness, hardness, scratch hardness, abrasion resistance, and durability to attack of car wash chemicals. Spray coated samples showed slight improvement in the total transmittance over the uncoated samples, while maintaining the transmission haze and exhibiting rougher surfaces. Only samples with thin coatings were found to possess sufficient adhesion to the substrate. These thin coatings gave improved hardness, scratch hardness, and durability to car wash attack to the level comparable to commercial coated polycarbonate headlamp lenses, whilst giving better abrasion resistance.     

Complexities of digital technology use and the teaching and learning of function

Many argue that digital technologies have the potential to enhance the teaching and learning of mathematics. However, the availability of technology is not sufficient to realise this potential. The study reported takes a detailed approach to investigate the utility of the particular offerings of the available technologies in the teaching and learning of a specific area of mathematics, functions. Sixteen affordances identified in the data are described. The complexity of the process involved in resolving a situation where particular affordances would be useful so as they are perceived and enacted is detailed. Finally, a grounded theory framework arising from the data analysis from this study that can be used to explain, predict and guide action in other digital environments is presented.     

An investigation into the cause of loss of containment from the supply of mini-bulk lubricants

An Intermediate Bulk Container (IBC) was punctured during its handling, releasing oil onto soil at an environmentally-sensitive region of Australia. The telehandler did not pierce the plastic of the IBC directly (as was expected) but rather one of the tynes had caught on the underside of the metal base plate, despite numerous controls being in place at time of spill, revealing a previously unreported mechanism for a fluid spill from handling of petroleum hydrocarbons. The diverse investigation team used a root cause analysis (RCA) technique to identify the underlying cause: the inspection process was inadequate with contributing factors of not using a spotter and design of IBC did not anticipate conditions. Engineering controls were put in place as part of the change management process to help prevent spills from occurring from piercing from telehandler tynes on the current project site.     

Automating data exchange in process choreographies

Communication between organizations is formalized as process choreographies in daily business. While the correct ordering of exchanged messages can be modeled and enacted with current choreography techniques, no approach exists to describe and automate the exchange of data between processes in a choreography using messages. This paper describes an entirely model-driven approach for BPMN introducing a few concepts that suffice to model data retrieval, data transformation, message exchange, and correlation – four aspects of data exchange. For automation, this work utilizes a recent concept to enact data dependencies in internal processes. We present a modeling guideline to derive local process models from a given choreography; their operational semantics allows to correctly enact the entire choreography from the derived models only including the exchange of data. Targeting on successful interactions, we discuss means to ensure correct process choreography modeling. Finally, we implemented our approach by extending the camunda BPM platform with our approach and show its feasibility by realizing all service interaction patterns using only model-based concepts.     

Hybrid make-to-stock and make-to-order systems: a taxonomic review

Production planning and control (PPC) systems that employ aspects from both make-to-order (MTO) and make-to-stock (MTS) production control are known as hybrid MTS/MTO systems. While both MTO and MTS separately have been studied extensively, their combined use has received less attention. However, the literature on this topic is growing and this paper shows that the review performed in this paper is an important addition to the field. We categorise relevant literature according to a novel taxonomy and show that hybrid MTS/MTO production control can be used in different contexts. In addition, an overview of the modelling techniques and methods used in these papers is provided. Based on the reviewed literature, relevant research questions and directions for future research are identified. Finally, it is shown that hybrid MTS/MTO production control is prevalent in practice by discussing research with industrial applications. The paper contains an overview of research on hybrid MTS/MTO production control to be used as reference for researchers active in the field, and provides managerial insights and directions for future research on this topic.     

The Role of Cell Cycle Regulators in Cell Survival—Dual Functions of Cyclin-Dependent Kinase 20 and p21Cip1/Waf1

The mammalian cell cycle is important in controlling normal cell proliferation and the development of various diseases. Cell cycle checkpoints are well regulated by both activators and inhibitors to avoid cell growth disorder and cancerogenesis. Cyclin dependent kinase 20 (CDK20) and p21^Cip1/Waf1 are widely recognized as key regulators of cell cycle checkpoints controlling cell proliferation/growth and involving in developing multiple cancers. Emerging evidence demonstrates that these two cell cycle regulators also play an essential role in promoting cell survival independent of the cell cycle, particularly in those cells with a limited capability of proliferation, such as cardiomyocytes. These findings bring new insights into understanding cytoprotection in these tissues. Here, we summarize the new progress of the studies on these two molecules in regulating cell cycle/growth, and their new roles in cell survival by inhibiting various cell death mechanisms. We also outline their potential implications in cancerogenesis and protection in heart diseases. This information renews the knowledge in molecular natures and cellular functions of these regulators, leading to a better understanding of the pathogenesis of the associated diseases and the discovery of new therapeutic strategies.