Search

Controlling the interfaces and interactions of colloidal nanoparticles (NPs) via tethered molecular moieties is crucial for NP applications in engineered nanomaterials, optics, catalysis, and nanomedicine. Despite a broad range of molecular types explored, there is a need for a flexible approach to rationally vary the chemistry and structure of these interfacial molecules for controlling NP stability in diverse environments, while maintaining a small size of the NP molecular shell. Here, we demonstrate that low-molecular-weight, bifunctional comb-shaped, and sequence-defined peptoids can effectively stabilize gold NPs (AuNPs). The generality of this robust functionalization strategy was also demonstrated by coating of silver, platinum, and iron oxide NPs with designed peptoids. Each peptoid (PE) is designed with varied arrangements of a multivalent AuNP-binding domain and a solvation domain consisting of oligo-ethylene glycol (EG) branches. Among designs, a peptoid (PE5) with a diblock structure is demonstrated to provide a superior nanocolloidal stability in diverse aqueous solutions while forming a compact shell (∼1.5 nm) on the AuNP surface. We demonstrate by experiments and molecular dynamics simulations that PE5-coated AuNPs (PE5/AuNPs) are stable in select organic solvents owing to the strong PE5 (amine)-Au binding and solubility of the oligo-EG motifs. At the vapor-aqueous interface, we show that PE5/AuNPs remain stable and can self-assemble into ordered 2D lattices. The NP films exhibit strong near-field plasmonic coupling when transferred to solid substrates.

Blood-based, or “liquid,” biopsies enable minimally invasive diagnostics but have limits on sensitivity due to scarce cell-free DNA (cfDNA). Improvements to sensitivity have primarily relied on enhancing sequencing technologyex vivo. Here, we sought to augment the level of circulating tumor DNA (ctDNA) detected in a blood draw by attenuating the clearance of cfDNAin vivo. We report a first-in-class intravenous DNA-binding priming agent given 2 hours prior to a blood draw to recover more cfDNA. The DNA-binding antibody minimizes nuclease digestion and organ uptake of cfDNA, decreasing its clearance at 1 hour by over 150-fold. To improve plasma persistence and limit potential immune interactions, we abrogated its Fc-effector function. We found that it protects GC-rich sequences and DNase-hypersensitive sites, which are ordinarily underrepresented in cfDNA. In tumor-bearing mice, priming improved tumor DNA recovery by 19-fold and sensitivity for detecting cancer from 6% to 84%. These results suggest a novel method to enhance the sensitivity of existing DNA-based cancer testing using blood biopsies.

Liquid biopsies are enabling minimally invasive monitoring and molecular profiling of diseases across medicine, but their sensitivity remains limited by the scarcity of cell-free DNA (cfDNA) in blood. Here, we report an intravenous priming agent that is given prior to a blood draw to increase the abundance of cfDNA in circulation. Our priming agent consists of nanoparticles that act on the cells responsible for cfDNA clearance to slow down cfDNA uptake. In tumor-bearing mice, this agent increases the recovery of circulating tumor DNA (ctDNA) by up to 60-fold and improves the sensitivity of a ctDNA diagnostic assay from 0% to 75% at low tumor burden. We envision that this priming approach will significantly improve the performance of liquid biopsies across a wide range of clinical applications in oncology and beyond.

Autoimmune diseases and in particular type 1 diabetes rely heavily on treatments that target the symptoms rather than prevent the underlying disease. One of the barriers to better therapeutic strategies is the inability to detect and efficiently target rare autoreactive T-cell populations that are major drivers of these conditions. Here, we develop a unique artificial antigen-presenting cell (aAPC) system from biocompatible polymer particles that allows specific encapsulation of bioactive ingredients. Using our aAPC, we demonstrate that we are able to detect rare autoreactive CD4 populations in human patients, and using mouse models, we demonstrate that our particles are able to induce desensitization in the autoreactive population. This system provides a promising tool that can be used in the prevention of autoimmunity before disease onset.

The ability to coordinate and confine enzymes presents an opportunity to affect their performance and to create chemically active materials. Recent studies show that polymers and biopolymers can be used to scaffold enzymes, and that can lead to the modulated biocatalytic efficiency. Here, we investigated the role of microenvironments on enzyme activity using a well-defined molecular scaffold. An enzyme, glucose oxidase (GOx), was positioned at different locations of a three-dimensional (3D) octahedral DNA scaffold (OS), allowing the enzyme's polyanionic environments to be altered. Using electrical sensing, based on a bipolar junction transistor, we measured directly and in real-time the enzyme's proton generation at these different microenvironments. We found a 200% enhancement of immobilized enzyme over free GOx and about a 30% increase in catalytic rates when the enzyme was moved on the same molecular scaffold to a microenvironment with a higher local concentration of polyanions, which suggests a role of local pH on the enzymatic activity.

Understanding the mechanisms behind crystal nucleation and growth is a fundamental requirement for the design and production of bespoke nanomaterials with controlled sizes and morphologies. Herein, we select gold (Au) nanoparticles as the model system for our study due to their representative applications in biology, electronics and optoelectronics. We investigate the radiation-induced in situ growth of gold (Au) particles using liquid cell transmission electron microscopy (LCTEM) and study the growth kinetics of non-spherical Au structures. Under controlled electron fluence, liquid flow rate and Au3+ ion supply, we show the favoured diffusion-limited growth of multi-twinned nascent Au seed particles into branched structures when using thin liquid cells (100 nm and 250 nm) in LCTEM, whereas faceted structures (e.g., spheres, rods, and prisms) formed when using a 1 µm thick liquid cell. In addition, we observed that anisotropic Au growth could be modulated by Au-binding amyloid fibrils, which we ascribe to their capability of regulating Au3+ ion diffusion and mass transfer in solution. We anticipate that this study will provide new perspectives on the shapecontrolled synthesis of anisotropic metallic nanomaterials using LCTEM.

Versatile methods to organize proteins in space are required to enable complex biomaterials, engineered biomolecular scaffolds, cell-free biology, and hybrid nanoscale systems. Here, we demonstrate how the tailored encapsulation of proteins in DNA-based voxels can be combined with programmable assembly that directs these voxels into biologically functional protein arrays with prescribed and ordered two-dimensional (2D) and three-dimensional (3D) organizations. We apply the presented concept to ferritin, an iron storage protein, and its iron-free analog, apoferritin, in order to form single-layers, double-layers, as well as several types of 3D protein lattices. Our study demonstrates that internal voxel design and inter-voxel encoding can be effectively employed to create protein lattices with designed organization, as confirmed by in situ X-ray scattering and cryo-electron microscopy 3D imaging. The assembled protein arrays maintain structural stability and biological activity in environments relevant for protein functionality. The framework design of the arrays then allows small molecules to access the ferritins and their iron cores and convert them into apoferritin arrays through the release of iron ions. The presented study introduces a platform approach for creating bio-active protein-containing ordered nanomaterials with desired 2D and 3D organizations., Organising proteins in 2D and 3D is needed to develop complex bimolecular materials for a range of applications. Here, the authors report the encapsulation of ferritin and apoferritin in DNA-based voxels with programmed assembly to generate both 2D and 3D protein lattices and demonstrate the retention of protein function.

The evolution of life on earth eventually leads to the emergence of species with increased complexity and diversity. Similarly, evolutionary chemical space exploration in the laboratory is a key step to pursue the structural and functional diversity of supramolecular systems. Here, we present a powerful tool that enables rapid peptide diversification and employ it to expand the chemical space for supramolecular functions. Central to this strategy is the exploitation of palladium-catalyzed Suzuki–Miyaura cross-coupling reactions to direct combinatorial synthesis of peptide arrays in microtiter plates under an open atmosphere. Taking advantage of this in situ library design, our results unambiguously deliver a fertile platform for creating a set of intriguing peptide functions including green fluorescent protein-like peptide emitters with chemically encoded emission colors, hierarchical self-assembly into nano-objects, and macroscopic hydrogels. This work also offers opportunities for quickly surveying the diversified peptide arrays and thereby identifying the structural factors that modulate peptide properties.

Autoimmune diseases and in particular type 1 diabetes rely heavily on treatments that target the symptoms rather than prevent the underlying disease. One of the barriers to better therapeutic strategies is the inability to detect and efficiently target rare autoreactive T-cell populations that are major drivers of these conditions. Here, we develop a unique artificial antigen presenting cell (aAPC) system from biocompatible polymer particles that allows specific encapsulation of bioactive ingredients. Using our aAPC we demonstrate that we are able to detect rare autoreactive CD4 populations in human patients and using mouse models we demonstrate that our particles are able to induce desensitization in the autoreactive population. This system provides a promising tool that can be used in the prevention of autoimmunity before disease onset.

Due to the central role of DNA, its interactions with inorganic salts and small organic molecules are important. For example, such interactions play important roles in various fundamental cellular processes in living systems and are involved in many DNA-damage related diseases. Strategies to improve the sensitivity of existing techniques for studying DNA interactions with other molecules would be appreciated in situations where the interactions are too weak. Here we report our development and demonstration of bent DNA bows for amplifying, sensing, and detecting the interactions of 14 inorganic salts and small organic molecules with DNA. With the bent DNA bows, these interactions were easily visualized and quantified in gel electrophoresis, which were difficult to measure without bending. In addition, the strength of the interactions of DNA with the various salts/molecules were quantified using the modified Hill equation. This work highlights the amplification effects of the bending elastic energy stored in the DNA bows and the potential use of the DNA bows for quantitatively measuring DNA interactions with small molecules as simple economic methods, it may also pave the way for exploiting the bent DNA bows for other applications such as screening DNA-interacting molecules and drugs.

Significance DNA nanotechnology provides a structural toolkit for the fabrication of programmable DNA nano-constructs; however, their use in biomedical applications is challenging due the limited structural integrity in complex biological fluids. Here, we report a class of tailorable molecular coatings, peptoids, which can efficiently stabilize three-dimensional wireframed DNA constructs under a variety of biomedically relevant conditions, including magnesium-ion depletion and presence of degrading nuclease. Furthermore, we show that peptoid-coated DNA constructs offer a controllable anticancer drug release and an ability to display functional biomolecules on the DNA surfaces. Our study demonstrates an approach for building multifunctional and environmentally robust DNA-based molecular structures for nanomedicine and biosensing., DNA nanotechnology has established approaches for designing programmable and precisely controlled nanoscale architectures through specific Watson−Crick base-pairing, molecular plasticity, and intermolecular connectivity. In particular, superior control over DNA origami structures could be beneficial for biomedical applications, including biosensing, in vivo imaging, and drug and gene delivery. However, protecting DNA origami structures in complex biological fluids while preserving their structural characteristics remains a major challenge for enabling these applications. Here, we developed a class of structurally well-defined peptoids to protect DNA origamis in ionic and bioactive conditions and systematically explored the effects of peptoid architecture and sequence dependency on DNA origami stability. The applicability of this approach for drug delivery, bioimaging, and cell targeting was also demonstrated. A series of peptoids (PE1–9) with two types of architectures, termed as “brush” and “block,” were built from positively charged monomers and neutral oligo-ethyleneoxy monomers, where certain designs were found to greatly enhance the stability of DNA origami. Through experimental and molecular dynamics studies, we demonstrated the role of sequence-dependent electrostatic interactions of peptoids with the DNA backbone. We showed that octahedral DNA origamis coated with peptoid (PE2) can be used as carriers for anticancer drug and protein, where the peptoid modulated the rate of drug release and prolonged protein stability against proteolytic hydrolysis. Finally, we synthesized two alkyne-modified peptoids (PE8 and PE9), conjugated with fluorophore and antibody, to make stable DNA origamis with imaging and cell-targeting capabilities. Our results demonstrate an approach toward functional and physiologically stable DNA origami for biomedical applications.

Due to the central role of DNA, its interactions with inorganic salts and small organic molecules are important for understanding various fundamental cellular processes in living systems, deciphering the mechanism of many diseases related to DNA damages, and discovering or designing inhibitors and drugs targeting DNA. However, there is still a need for improved sensitivity to detect these interactions, especially in situations where expensive sophisticated equipment is not available. Here we report our development and demonstration of bent DNA bows for amplifying, sensing, and detecting the interactions of 14 inorganic salts and small organic molecules with DNA. With the bent DNA bows, these interactions were easily visualized and quantified in gel electrophoresis, which were difficult to measure without bending. In addition, the strength of the interactions of DNA with the various salts/molecules were quantified using the modified Hill equation. This work highlights the amplification effects of the bending elastic energy stored in the DNA bows and the potential use of the DNA bows for quantitatively measuring DNA interactions with small molecules as simple economic methods; it may also pave the way for exploiting the bent DNA bows for other applications such as monitoring water quality and screening DNA-targeting molecules and drugs.

Supramolecular self-assembly is an important process that enables the conception of complex structures mimicking biological motifs. Herein, we constructed helical fibrils through chiral self-assembly of nucleobase--peptide conjugates (NPCs), where achiral nucleobases are helically displayed on the surface of fibrils, comparable to polymerized nucleic acids. Selective binding between DNA and the NPC fibrils was observed with fluorescence polarization. Taking advantage of metal--nucleobase recognition, we highlight the possibility of deposition/assembly of plasmonic nanoparticles onto the fibrillar constructs. In this approach, the supramolecular chirality of NPCs can be adaptively imparted to metallic nanoparticles, covering them to generate structures with plasmonic chirality that exhibit significantly improved colloidal stability. The self-assembly of rationally designed NPCs into nanohelices is a promising way to engineer complex, optically diverse nucleobase-derived nanomaterials.

A method is developed for growing three-dimensional hierarchic structures of porous graphenic carbon film/hollow carbon nanorods where porous graphenic carbon film is first synthesized followed by, growth of carbon nanorods. By annealing an amorphous carbon layer deposited underneath a nickel thin film at elevated temperatures, the porous graphenic carbon film forms on top via carbon diffusion and precipitation from the grain boundaries of the nickel film. The porosity of the graphenic carbon film is determined by the surface voids of the nickel film resulting from grain coalescence during annealing. Hollow carbon nanorods can then be grown on the pore edges of the porous graphenic carbon film by chemical vapor deposition without catalysts. It is speculated that the dangling bonds of the carbon atoms on the pore edges of the graphene layers might be responsible for the nucleation of the hollow carbon nanorods. The microstructures and growth mechanisms of both porous graphenic carbon film and hollow carbon nanorods are characterized and discussed in detail.

Recent development in fastener industry has heightened the need for steel wire coil. The wire usually has to be annealed to improve its cold formability. The quality of spheroidizing annealed wire affects the forming quality of screws. Various parameters affect the quality of spheroidized annealing such as spheroidized annealing temperature, prolonged heating time, furnace cooling temperature. The effects of spheroidized annealing parameters affect the quality characteristics, such as tensile strength, hardness and ductility. A series of experimental tests on AISI 10B21 steel wire is carried out in a vacuum drying oven and Taguchi method is used to obtain optimum spheroidized annealing conditions to improve the mechanical properties of steel wires for cold forming. The results show experimentally that spheroidized annealing temperature is the main factor to influence the mechanical properties of AISI 10B21 steel wires.

Amyloid fibrillation is a nucleation-dependent process known be involved in the development of more than 20 progressive and chronic diseases. The detection of amyloid formation at the nucleation stage can greatly advance early diagnoses and treatment of diseases. In this work, we developed a new assay for the early detection of amylin fibrillation using the biarsenical dye, 4,5-bis(1,3,2-dithiarsolan-2-yl)fluorescein (FlAsH), which could recognise the tetracysteine motifs and transform from non-fluorescent form into strongly fluorescent complexes. Due to the close proximity of two cysteine residues within the hydrophilic domain of amylin, a non-contiguous tetracysteine motif can form upon amylin dimerisation or oligomerisation, which can be recognised by FlAsH and emit strong fluorescence. This enables us to report the nucleation-growth process of amylin without modification of the protein sequence. We showed that the use of this assay not only allowed the tracking of initial nucleation events, but also enabled imaging of amyloid fibrils and investigating the effects of amyloid inhibitor/modulator toward amylin fibrillation.

High crystallinity mesoporous TiO 2 hollow spheres (MHS-TiO 2 ) were prepared using the mesoporous carbon hollow sphere template. The MHS-TiO 2 contains mainly nanostructured anatase. The mesopore of the MHS-TiO 2 has a pore opening in the range of 400–600 nm. The refined extended X-ray absorption fine structure spectra indicate that the MHS-TiO 2 possesses less the 1st-shell Ti–O coordination numbers than the nano-TiO 2 . More surface active species (A 2 ((Ti=O)O 4 )) on the MHS-TiO 2 are also observed by the component fitted X-ray absorption near edge structure spectroscopy. The MHS-TiO 2 photoanode has a better DSSC conversion efficiency than the nano-TiO 2 one by at least 40%. Note that the N3 dye molecules are accessible to the mesopores of the MHS-TiO 2 , and the loading time for N3 can be reduced by at least 70% if compared with those of the nano-TiO 2 .

A comprehensive understanding of the mechanisms of interaction between proteins or peptides and nanomaterials is crucial for the development of nanomaterial-based diagnostics and therapeutics. In this work, we systematically explored the interactions between citrate-capped gold nanoparticles (AuNPs) and islet amyloid polypeptide (IAPP), a 37-amino acid peptide hormone co-secreted with insulin from the pancreatic islet. We utilized diffusion-ordered spectroscopy, isothermal titration calorimetry, localized surface plasmon resonance spectroscopy, gel electrophoresis, atomic force microscopy, transmission electron microscopy (TEM), and molecular dynamics (MD) simulations to systematically elucidate the underlying mechanism of the IAPP–AuNP interactions. Because of the presence of a metal-binding sequence motif in the hydrophilic peptide domain, IAPP strongly interacts with the Au surface in both the monomeric and fibrillar states. Circular dichroism showed that AuNPs triggered the IAPP conformational transition from random coil to ordered structures (α-helix and β-sheet), and TEM imaging suggested the acceleration of IAPP fibrillation in the presence of AuNPs. MD simulations revealed that the IAPP–AuNP interactions were initiated by the N-terminal domain (IAPP residues 1–19), which subsequently induced a facet-dependent conformational change in IAPP. On a Au(111) surface, IAPP was unfolded and adsorbed directly onto the Au surface, while for the Au(100) surface, it interacted predominantly with the citrate adlayer and retained some helical conformation. The observed affinity of AuNPs for IAPP was further applied to reduce the level of peptide-induced lipid membrane disruption.

In viral proteins, labile Zn-sites, where Zn(2+) is crucial for maintaining the native protein structure but the Zn-bound cysteines are reactive, are promising drug targets. Here, we aim to (i) identify labile Zn-sites in viral proteins using guidelines established from our previous work and (ii) assess if clinically safe Zn-ejecting agents could eject Zn(2+) from the predicted target site and thus inhibit viral replication. As proof-of-concept, we identified a labile Zn-site in the hepatitis C virus (HCV) NS5A protein and showed that the antialcoholism drug, disulfiram, could inhibit HCV replication to a similar extent as the clinically used antiviral agent, ribavirin. The discovery of a novel viral target and a new role for disulfiram in inhibiting HCV replication will enhance the therapeutic armamentarium against HCV. The strategy presented can also be applied to identify labile sites in other bacterial or viral proteins that can be targeted by disulfiram or other clinically safe Zn-ejectors.

The evidence of improved survival in patients of colorectal cancer (CRC) receiving multidisciplinary team (MDT) care remains inconclusive.All patients with incident CRC but no prior cancer history in 2005-2008 were included and followed till 2010. A logistic regression model was used to predict the associated factors to participate in the MDT care model. The propensity score method was included under Cox proportional hazards model to reduce potential bias and to conduct survival analyses.In total, 25,766 patients were included; the mean follow-up period was 35.1 months. The factors associated with participating in MDT included receiving treatments at regional hospitals, at private hospitals, and stage III cancer (all p values0.001). The favorable survival factors included participating in MDT (HR=0.91, p=0.001), age of 45-75, top-ranked income group, receiving treatments at district hospitals, or at hospitals or with doctors that had higher service volumes (all p values0.05). Regarding individual stages, the risk of mortality was significantly lower at stage IV (HR=0.88, p=0.002).Colorectal cancer patients with participation in MDT have a lower mortality risk; the improvements of survival exist in all colorectal cancer patients, especially in those with stage IV disease.

A simple and selective assay for detecting methylglyoxal (MGO), a metabolite associated with diabetes, was developed by combining a bio-inspired chemical reaction with the anti-aggregation of gold nanoparticles. This assay could detect MGO at as low as 1 μM by the naked eye and 0.05 μM by UV-vis spectrometry, within the clinical range marking oxidative stress in diabetes, and demonstrated high selectivity over other physiologically relevant ketones and aldehydes.

Objectives We report two siblings, a boy and a girl, with Cornelia de Lange syndrome (CdLS), born to unaffected parents, and attempt to delineate the underlying molecular mechanism leading to familial recurrence. Methods Nipped-B-like (NIPBL) gene mutations were screened using in denaturing high-performance liquid chromatography and sequencing in peripheral blood samples, from one of the affected siblings and her parents, as well as from a sperm sample from the father. Results A heterozygous missense NIPBL mutation, D2433G, was identified in the peripheral blood sample of the affected girl, but not in the peripheral blood samples of her parents. The D2433G mutation was also found in the sperm sample of the father. Conclusion Gonadal mosaicism represents an underappreciated feature of the inheritance pattern of CdLS. To our knowledge, the girl represents the first CdLS patient whose father was documented to have a population of mutant sperm. When a sperm analysis indicates the presence of a mutant allele, it may be reasonable to offer prenatal genetic testing to the family in subsequent pregnancies, given that the sensitivity of fetal ultrasonography is relatively low. Copyright © 2006 John Wiley & Sons, Ltd.

Summary Objectives The aim of this study was to investigate the association between multidisciplinary team (MDT) management and survival of oral cavity cancer patients using a nationwide database in Taiwan. Materials and methods A nationwide cohort study was conducted between 2005 and 2008. The follow-up end point was 2010. Claims data of oral cavity cancer patients were retrieved from the Taiwan Cancer Registry Database. Secondary data were obtained from the Taiwan’s National Health Insurance Research Database. Among 19,766 newly diagnosed oral cavity cancer patients, we identified 16,991 patients who underwent treatment between 2004 and 2008 for further analyses. Results Overall survival was compared between patients who received MDT management ( n = 3324) and those who did not ( n = 13,367). Hazard ratios (HR) of death in patients with MDT management were also analyzed. Patients with MDT management had a lower risk of death when compared with that of patients without MDT management (HR: 0.94, 95% confidence intervals (CI): 0.89–1.00; P = 0.032). The effect of MDT management on survival was stronger for male patients than for female patients (HR: 0.94, 95% CI: 0.89–1.00; P = 0.040 versus HR: 0.98, 95% CI: 0.75–1.27; P = 0.866). In addition, the effect of MDT management was strong among patients with a Charlson Comorbidity Index between 4 and 6, in those without coexisting catastrophic illness/injury, and in patients with stage IV diseases. Conclusion Survival rates in oral cavity cancer patients with MDT management appeared to be marginally better than those of patients without MDT management.

Plastic caps are the most commonly seen bottle caps used in beverage and food containers. They are widely used to seal freshness of beverage or liquids in bottles. Threads are usually grooved inside the caps for easy twist-off caps and sealing rings prevent the liquids from bacterial infection. Companies print logos or pictures on the top surface of plastic cap, such that the quality of printing also indirectly affects the customers purchase. Inspection of plastic caps, including the surface printing, thread, and sealing ring, is a great issue during the caps production currently. The objective of this study is to use machine vision to inspect the defect of the sealing area and the printing surface of a plastic cap. An automated inspection system, which includes two CCD camera, lighting source, sensors, and a cap transporter, is constructed, and a digital image processing software is designed to learn good caps and screen out the defective ones. The experimental results show that the proposed inspection system can self-learn the features of a good surface printing, and effectively detect the defective caps under very few parameters setting, while the major defects in the sealing ring and thread area such as malformation, contamination, overfill, incomplete, scratches, can be successfully identified under the rate of 1,200 piece per minute.

The aim of this study is to investigate the relationship between online gaming behavior and special underlying social group structures through the use of player information of a certain MMORPG. Data were collected from the logs of an MMORPG in 2011, recording five types of gaming behaviors, player gender, avatar gender and avatar occupation types to investigate whether different avatars affect gaming behavior. This study includes 1,727 male players and 656 female players. The results indicate that the six-cluster model has the goodness of model fit. Different clusters differ in the five different types of gaming behaviors and the choice of avatar gender and occupations. This result shows that different avatar roles will influence playersi¦ gaming behaviors.

Exergames enable players to interact with games through body movement. With various positive effects of exergaming such as physical, psychosocial and cognitive benefits, exergames are regarded as a desirable tool for promoting physical education. However, there is little research about exergaming experience with flow state, enjoyment, and physical fitness. In this study, The Exergames Flow State Questionnaire (EFSQ) and the Physical Fitness Self-assessment Questionnaire (PFSAQ) were designed and developed with good reliability (£\=0.97; £\=0.92). The results of correlation analysis show that more frequent play of exergames and longer use of the exergame consoles may increase the incidence of flow state and enjoyment. Additionally, the frequency of playing exergames is significantly correlated to PFSAQ, indicating that more often the users play the exergames, the more sense of health physical fitness they will have. The findings of this investigation could be used in physical education planning. Meanwhile, EFSQ and PFSAQ can be used as alternative tools to measure flow state and physical fitness.

There are more than 47 million players playing the MMORPGs (Massively Multi-player Online Role-Playing Games) in the world. One of the characteristics of MMORPGs is that players interact with others through games and virtual communities. Do players of different genders and ages spend the same time while they are playing an MMORPG? This study collected 3, 241 playersi¦ gaming logs from an MMORPG game for 78 days and analyzed playersi¦ playing time by their genders and ages. The findings show that there is a significant difference in game playing time between male and female players at the age from 23 to 40. There is also an interaction effect between genders and age groups in the game playing time.

The MMORPG is an interactive multiplayer online role-playing game. Players interact with each other in the virtual community of MMORPG having different behaviors due to the level of involvement and interaction. Players and players have different interaction in the communities through the game. The level of participation and interaction of players cause different behaviors in the game. Research data comes from those players who created their roles from 18pm to 24pm at the first day the game world started and their behaviors were continuity recording for 7 days. The results of the study found that players who attended to the virtual community were willing to spend more time and pay more money on the online game. Moreover, positive correlation was found between time and money players spent in the MMORPG. Furthermore, there is no difference between players' behavior in the game in different roles of genders.

This study uses the mathematical model of game options to analyze when the distributors of the two countries choose to invest in the domestic or international marketing channel, they will consider competitors' behavior and invest in the marketing channels of the two countries based on the optimal time point. The results will provide distributors with the references of evaluating the influence of the potential channel value on the firm value and deciding whether to enter the international marketing market when they want to invest in the multi-national marketing channel in the future.

We report two siblings, a boy and a girl, with Cornelia de Lange syndrome (CdLS), born to unaffected parents, and attempt to delineate the underlying molecular mechanism leading to familial recurrence.Nipped-B-like (NIPBL) gene mutations were screened using in denaturing high-performance liquid chromatography and sequencing in peripheral blood samples, from one of the affected siblings and her parents, as well as from a sperm sample from the father.A heterozygous missense NIPBL mutation, D2433G, was identified in the peripheral blood sample of the affected girl, but not in the peripheral blood samples of her parents. The D2433G mutation was also found in the sperm sample of the father.Gonadal mosaicism represents an underappreciated feature of the inheritance pattern of CdLS. To our knowledge, the girl represents the first CdLS patient whose father was documented to have a population of mutant sperm. When a sperm analysis indicates the presence of a mutant allele, it may be reasonable to offer prenatal genetic testing to the family in subsequent pregnancies, given that the sensitivity of fetal ultrasonography is relatively low.