Poly(beta-hydroxybutyrate-co-beta-hydroxyvalerate) supports in vitro osteogenesis

Studies have demonstrated that polymeric biomaterials have the potential to support osteoblast growth and development for bone tissue repair. Poly( beta- hydroxybutyrate- co- beta- hydroxyvalerate) ( PHBV), a bioabsorbable, biocompatible polyhydroxy acid polymer, is an excellent candidate that, as yet, has not been extensively investigated for this purpose. As such, we examined the attachment characteristics, self- renewal capacity, and osteogenic potential of osteoblast- like cells ( MC3T3- E1 S14) when cultured on PHBV films compared with tissue culture polystyrene ( TCP). Cells were assayed over 2 weeks and examined for changes in morphology, attachment, number and proliferation status, alkaline phosphatase ( ALP) activity, calcium accumulation, nodule formation, and the expression of osteogenic genes. We found that these spindle- shaped MC3T3- E1 S14 cells made cell - cell and cell - substrate contact. Time- dependent cell attachment was shown to be accelerated on PHBV compared with collagen and laminin, but delayed compared with TCP and fibronectin. Cell number and the expression of ALP, osteopontin, and pro- collagen alpha 1( I) mRNA were comparable for cells grown on PHBV and TCP, with all these markers increasing over time. This demonstrates the ability of PHBV to support osteoblast cell function. However, a lag was observed for cells on PHBV in comparison with those on TCP for proliferation, ALP activity, and cbfa- 1 mRNA expression. In addition, we observed a reduction in total calcium accumulation, nodule formation, and osteocalcin mRNA expression. It is possible that this cellular response is a consequence of the contrasting surface properties of PHBV and TCP. The PHBV substrate used was rougher and more hydrophobic than TCP. Although further substrate analysis is required, we conclude that this polymer is a suitable candidate for the continued development as a biomaterial for bone tissue engineering.

Enhancement of drug affinity for cell membranes by conjugation with lipoamino acids II. Experimental and computational evidence using biomembrane models

Lipoamino acids (LAAs) are promoieties able to enhance the amphiphilicity of drugs, facilitating their interaction with cell membranes. Experimental and computational studies were carried out on two series of lipophilic amide conjugates between a model drug (tranylcypromine, TCP) and LAA or alkanoic acids containing a short, medium or long alkyl side chain (C-4 to C-16). The effects of these compounds were evaluated by monolayer surface tension analysis and differential scanning calorimetry using dimyristoylphosphatidylcholine nnonolayers and liposomes as biomembrane models. The experimental results were related to independent calculations to determine partition coefficient and blood-brain partitioning. The comparison of TCP-LAA conjugates with the related series of TCP alkanoyl amides confirmed that the ability to interact with the biomembrane models is not due to the mere increase of lipophilicity, but mainly to the amphipatic nature and the kind of LAA residue. (C) 2005 Elsevier B.V. All rights reserved.

Gravity spun polycaprolactone fibers for applications in vascular tissue engineering:proliferation and function of human vascular endothelial cells

Poly(ε-caprolactone) (PCL) fibers produced by wet spinning from solutions in acetone under low-shear (gravity-flow) conditions resulted in fiber strength of 8 MPa and stiffness of 0.08 Gpa. Cold drawing to an extension of 500% resulted in an increase in fiber strength to 43 MPa and stiffness to 0.3 GPa. The growth rate of human umbilical vein endothelial cells (HUVECs) (seeded at a density of 5 × 104 cells/mL) on as-spun fibers was consistently lower than that measured on tissue culture plastic (TCP) beyond day 2. Cell proliferation was similar on gelatin-coated fibers and TCP over 7 days and higher by a factor of 1.9 on 500% cold-drawn PCL fibers relative to TCP up to 4 days. Cell growth on PCL fibers exceeded that on Dacron monofilament by at least a factor of 3.7 at 9 days. Scanning electron microscopy revealed formation of a cell layer on samples of cold-drawn and gelatin-coated fibers after 24 hours in culture. Similar levels of ICAM-1 expression by HUVECs attached to PCL fibers and TCP were measured using RT-PCR and flow cytometry, indicative of low levels of immune activation. Retention of a specific function of HUVECs attached to PCL fibers was demonstrated by measuring their immune response to lipopolysaccharide. Levels of ICAM-1 expression increased by approximately 11% in cells attached to PCL fibers and TCP. The high fiber compliance, favorable endothelial cell proliferation rates, and retention of an important immune response of attached HUVECS support the use of gravity spun PCL fibers for three-dimensional scaffold production in vascular tissue engineering. © Mary Ann Liebert, Inc.

Novel biodegradable fibres for applications in tissue engineering and drug delivery

The preparation and characterisation of novel biodegradable polymer fibres for application in tissue engineering and drug delivery are reported. Poly(e-caprolactone) (PCL) fibres were produced by wet spinning from solutions in acetone under low shear (gravity flow) conditions. The tensile strength and stiffness of as-spun fibres were highly dependent on the concentration of the spinning solution. Use of a 6% w/v solution resulted in fibres having strength and stiffness of 1.8 MPa and 0.01 GPa respectively, whereas these values increased to 9.9 MPa and 0.1 GPa when fibres were produced from 20% w/v solutions. Cold drawing to an extension of 500% resulted in further increases in fibre strength (up to 50 MPa) and stiffness (0.3 GPa). Hot drawing to 500% further increased the fibre strength (up to 81 MPa) and stiffness (0.5 GPa). The surface morphology of as-spun fibres was modified, to yield a directional grooved pattern by drying in contact with a mandrel having a machined topography characterised by a peak-peak separation of 91 mm and a peak height of 30 mm. Differential scanning calorimetery (DSC) analysis of as-spun fibres revealed the characteristic melting point of PCL at around 58°C and a % crystallinity of approximately 60%. The biocompatibility of as-spun fibres was assessed using cell culture. The number of attached 3T3 Swiss mouse fibroblasts, C2C12 mouse myoblasts and human umbilical vein endothelial cells (HUVECs) on as-spun, 500% cold drawn, and gelatin coated PCL fibres were observed. The results showed that the fibres promoted cell proliferation for 9 days in cell culture and was slightly lower than on tissue culture plastic. The morphology of all cell lines was assessed on the various PCL fibres using scanning electron microscopy. The cell function of HUVECs growing on the as-spun PCL fibres was evaluated. The ability HUVECs to induce an immune response when stimulated with lipopolysaccaride (LPS) and thereby to increase the amount of cell surface receptors was assessed by flow cytometry and reverse transcription-polymerase chain reaction (RT-PCR). The results showed that PCL fibres did not inhibit this function compared to TCP. As-spun PCL fibres were loaded with 1 % ovine albumin (OVA) powder, 1% OVA nanoparticles and 5% OVA nanoparticles by weight and the protein release was assessed in vitro. PCL fibres loaded with 1 % OVA powder released 70%, 1% OVA nanoparticle released 60% and the 5% OVA nanoparticle released 25% of their protein content over 28 days. These release figures did not alter when the fibres were subjected to lipase enzymatic degradation. The OVA released was examined for structural integrity by SDS-PAGE. This showed that the protein molecular weight was not altered after incorporation into the fibres. The bioactivity of progesterone was assessed following incorporation into PCL fibres. Results showed that the progesterone released had a pronounced effect on MCF-7 breast epithelial cells, inhibiting their proliferation. The PCL fibres display high fibre compliance, a potential for controlling the fibre surface architecture to promote contact guidance effects, favorable proliferation rate of fibroblasts, myoblasts and HUVECs and the ability to release pharmaceuticals. These properties recommended their use for 3-D scaffold production in soft tissue engineering and the fibres could also be exploited for controlled presentation and release of biopharmaceuticals such as growth factors.

Modelling the gastric epithelium for testing of new chemical entities

A cell culture model of the gastric epithelial cell surface would prove useful for biopharmaceutical screening of new chemical entities and dosage forms. A successful model should exhibit tight junction formation, maintenance of differentiation and polarity. Conditions for primary culture of guinea-pig gastric mucous epithelial cell monolayers on Tissue Culture Plastic (TCP) and membrane insects (Transwells) were established. Tight junction formation for cells grown on Transwells for three days was assessed by measurement of transepithelial resistance (TEER) and permeability of mannitol and fluorescein. Coating the polycarbonate filter with collagen IV, rather with collagen I, enhanced tight junction formation. TEER for cells grown on Transwells coated with collagen IV was close to that obtained with intact guinea-pig gastric epithelium in vitro. Differentiation was assessed by incorporation of [3H] glucosamine into glycoprotein and by activity of NADPH oxidase, which produces superoxide. Both of these measures were greater for cells grown on filters coated with collagen I than for cells grown on TCP, but no major difference was found between cells grown on collagens I and IV. However, monolayers grown on membranes coated with collagen IV exhibited apically polarized secretion of mucin and superoxide. The proportion of cells, which stained positively for mucin with periodic Schiff reagent, was greater than 95% for all culture conditions. Gastric epithelial monolayers grown on Transwells coated with collagen IV were able to withstand transient (30 min) apical acidification to pH 3, which was associated with a decrease in [3H] mannitol flux and an increase in TEER relative to pH 7.4. The model was used to provide the first direct demonstration that an NSAID (indomethacin) accumulated in gastric epithelial cells exposed to low apical pH. In conclusion, guinea-pig epithelial cells cultured on collagen IV represent a promising model of the gastric surface epithelium suitable for screening procedures.

Evaluating and improving the performance of video content distribution in lossy networks

The contributions in this research are split in to three distinct, but related, areas. The focus of the work is based on improving the efficiency of video content distribution in the networks that are liable to packet loss, such as the Internet. Initially, the benefits and limitations of content distribution using Forward Error Correction (FEC) in conjunction with the Transmission Control Protocol (TCP) is presented. Since added FEC can be used to reduce the number of retransmissions, the requirement for TCP to deal with any losses is greatly reduced. When real-time applications are needed, delay must be kept to a minimum, and retransmissions not desirable. A balance, therefore, between additional bandwidth and delays due to retransmissions must be struck. This is followed by the proposal of a hybrid transport, specifically for H.264 encoded video, as a compromise between the delay-prone TCP and the loss-prone UDP. It is argued that the playback quality at the receiver often need not be 100% perfect, providing a certain level is assured. Reliable TCP is used to transmit and guarantee delivery of the most important packets. The delay associated with the proposal is measured, and the potential for use as an alternative to the conventional methods of transporting video by either TCP or UDP alone is demonstrated. Finally, a new objective measurement is investigated for assessing the playback quality of video transported using TCP. A new metric is defined to characterise the quality of playback in terms of its continuity. Using packet traces generated from real TCP connections in a lossy environment, simulating the playback of a video is possible, whilst monitoring buffer behaviour to calculate pause intensity values. Subjective tests are conducted to verify the effectiveness of the metric introduced and show that the results of objective and subjective scores made are closely correlated.

Understanding enterprise behaviour:a feasibility study

This paper describes work conducted as a joint collaboration between the Virtual Design Team (VDT) research group at Stanford University (USA) , the Systems Engineering Group (SEG) at De Montfort University (UK) and Elipsis Ltd . We describe a new docking methodology in which we combine the use of two radically different types of organizational simulation tool. The VDT simulation tool operates on a standalone computer, and employs computational agents during simulated execution of a pre-defined process model (Kunz, 1998). The other software tool, DREAMS , operates over a standard TCP/IP network, and employs human agents (real people) during a simulated execution of a pre-defined process model (Clegg, 2000).

Connection of Network Sensors to Distributed Information Measurement and Control System for Education and Research

The development of the distributed information measurement and control system for optical spectral research of particle beam and plasma objects and the execution of laboratory works on Physics and Engineering Department of Petrozavodsk State University are described. At the hardware level the system is represented by a complex of the automated workplaces joined into computer network. The key element of the system is the communication server, which supports the multi-user mode and distributes resources among clients, monitors the system and provides secure access. Other system components are formed by equipment servers (CАМАC and GPIB servers, a server for the access to microcontrollers MCS-196 and others) and the client programs that carry out data acquisition, accumulation and processing and management of the course of the experiment as well. In this work the designed by the authors network interface is discussed. The interface provides the connection of measuring and executive devices to the distributed information measurement and control system via Ethernet. This interface allows controlling of experimental parameters by use of digital devices, monitoring of experiment parameters by polling of analog and digital sensors. The device firmware is written in assembler language and includes libraries for Ethernet-, IP-, TCP- и UDP-packets forming.

Model and performance of a no-reference quality assessment metric for video streaming

Video streaming via Transmission Control Protocol (TCP) networks has become a popular and highly demanded service, but its quality assessment in both objective and subjective terms has not been properly addressed. In this paper, based on statistical analysis a full analytic model of a no-reference objective metric, namely pause intensity (PI), for video quality assessment is presented. The model characterizes the video playout buffer behavior in connection with the network performance (throughput) and the video playout rate. This allows for instant quality measurement and control without requiring a reference video. PI specifically addresses the need for assessing the quality issue in terms of the continuity in the playout of TCP streaming videos, which cannot be properly measured by other objective metrics such as peak signal-to-noise-ratio, structural similarity, and buffer underrun or pause frequency. The performance of the analytical model is rigidly verified by simulation results and subjective tests using a range of video clips. It is demonstrated that PI is closely correlated with viewers' opinion scores regardless of the vastly different composition of individual elements, such as pause duration and pause frequency which jointly constitute this new quality metric. It is also shown that the correlation performance of PI is consistent and content independent. © 2013 IEEE.

Quality assessment of video transmission in heterogeneous networks as applied to real-time telecare

This work looks into video quality assessment applied to the field of telecare and proposes an alternative metric to the more traditionally used PSNR based on the requirements of such an application. We show that the Pause Intensity metric introduced in [1] is also relevant and applicable to heterogeneous networks with a wireless last hop connected to a wired TCP backbone. We demonstrate through our emulation testbed that the impairments experienced in such a network architecture are dominated by continuity based impairments rather than artifacts, such as motion drift or blockiness. We also look into the implication of using Pause Intensity as a metric in terms of the overall video latency, which is potentially problematic should the video be sent and acted upon in real-time. We conclude that Pause Intensity may be used alongside the video characteristics which have been suggested as a measure of the overall video quality. © 2012 IEEE.

Exploring the effect of buffer behaviour on perceived video quality

In this work we deal with video streams over TCP networks and propose an alternative measurement to the widely used and accepted peak signal to noise ratio (PSNR) due to the limitations of this metric in the presence of temporal errors. A test-bed was created to simulate buffer under-run in scalable video streams and the pauses produced as a result of the buffer under-run were inserted into the video before being employed as the subject of subjective testing. The pause intensity metric proposed in [1] was compared with the subjective results and it was shown that in spite of reductions in frame rate and resolution, a correlation with pause intensity still exists. Due to these conclusions, the metric may be employed in layer selection in scalable video streams. © 2011 IEEE.

Regional growth, structural change, and natural disaster

Hurricanes, earthquakes, floods, and other serious natural hazards have been attributed with causing changes in regional economic growth, income, employment, and wealth. Natural disasters are said to cause; (1) an acceleration of existing economic trends; (2) an expansion of employment and income, due to recovery operations (the so-called silver lining); and (3) an alteration in the structure of regional economic activity due to changes in "intra" and "inter" regional trading patterns, and technological change.^ Theoretical and stylized disaster simulations (Cochrane 1975; Haas, Cochrane, and Kates 1977; Petak et al. 1982; Ellson et al. 1983, 1984; Boisvert 1992; Brookshire and McKee 1992) point towards a wide scope of possible negative and long lasting impacts upon economic activity and structure. This work examines the consequences of Hurricane Andrew on Dade County's economy. Following the work of Ellson et al. (1984), Guimaraes et al. (1993), and West and Lenze (1993; 1994), a regional econometric forecasting model (DCEFM) using a framework of "with" and "without" the hurricane is constructed and utilized to assess Hurricane Andrew's impact on the structure and level of economic activity in Dade County, Florida.^ The results of the simulation exercises show that the direct economic impact associated with Hurricane Andrew on Dade County is of short duration, and of isolated sectoral impact, with impact generally limited to construction, TCP (transportation, communications, and public utilities), and agricultural sectors. Regional growth, and changes in income and employment reacted directly to, and within the range and direction set by national economic activity. The simulations also lead to the conclusion that areal extent, infrastructure, and sector specific damages or impacts, as opposed to monetary losses, are the primary determinants of a disaster's effects upon employment, income, growth, and economic structure. ^

Adding concurrent data transfer to transport layer

In recent years, the internet has grown exponentially, and become more complex. This increased complexity potentially introduces more network-level instability. But for any end-to-end internet connection, maintaining the connection's throughput and reliability at a certain level is very important. This is because it can directly affect the connection's normal operation. Therefore, a challenging research task is to improve a network's connection performance by optimizing its throughput and reliability. This dissertation proposed an efficient and reliable transport layer protocol (called concurrent TCP (cTCP)), an extension of the current TCP protocol, to optimize end-to-end connection throughput and enhance end-to-end connection fault tolerance. The proposed cTCP protocol could aggregate multiple paths' bandwidth by supporting concurrent data transfer (CDT) on a single connection. Here concurrent data transfer was defined as the concurrent transfer of data from local hosts to foreign hosts via two or more end-to-end paths. An RTT-Based CDT mechanism, which was based on a path's RTT (Round Trip Time) to optimize CDT performance, was developed for the proposed cTCP protocol. This mechanism primarily included an RTT-Based load distribution and path management scheme, which was used to optimize connections' throughput and reliability. A congestion control and retransmission policy based on RTT was also provided. According to experiment results, under different network conditions, our RTT-Based CDT mechanism could acquire good CDT performance. Finally a CWND-Based CDT mechanism, which was based on a path's CWND (Congestion Window), to optimize CDT performance was introduced. This mechanism primarily included: a CWND-Based load allocation scheme, which assigned corresponding data to paths based on their CWND to achieve aggregate bandwidth; a CWND-Based path management, which was used to optimize connections' fault tolerance; and a congestion control and retransmission management policy, which was similar to regular TCP in its separate path handling. According to corresponding experiment results, this mechanism could acquire near-optimal CDT performance under different network conditions.

An approach for design and analysis of User-centric Communication Middleware

Today, the development of domain-specific communication applications is both time-consuming and error-prone because the low-level communication services provided by the existing systems and networks are primitive and often heterogeneous. Multimedia communication applications are typically built on top of low-level network abstractions such as TCP/UDP socket, SIP (Session Initiation Protocol) and RTP (Real-time Transport Protocol) APIs. The User-centric Communication Middleware (UCM) is proposed to encapsulate the networking complexity and heterogeneity of basic multimedia and multi-party communication for upper-layer communication applications. And UCM provides a unified user-centric communication service to diverse communication applications ranging from a simple phone call and video conferencing to specialized communication applications like disaster management and telemedicine. It makes it easier to the development of domain-specific communication applications. The UCM abstraction and API is proposed to achieve these goals. The dissertation also tries to integrate the formal method into UCM development process. The formal model is created for UCM using SAM methodology. Some design errors are found during model creation because the formal method forces to give the precise description of UCM. By using the SAM tool, formal UCM model is translated to Promela formula model. In the dissertation, some system properties are defined as temporal logic formulas. These temporal logic formulas are manually translated to promela formulas which are individually integrated with promela formula model of UCM and verified using SPIN tool. Formal analysis used here helps verify the system properties (for example multiparty multimedia protocol) and dig out the bugs of systems.

Leveraging symbiotic relationships for emulation of computer networks

The lack of analytical models that can accurately describe large-scale networked systems makes empirical experimentation indispensable for understanding complex behaviors. Research on network testbeds for testing network protocols and distributed services, including physical, emulated, and federated testbeds, has made steady progress. Although the success of these testbeds is undeniable, they fail to provide: 1) scalability, for handling large-scale networks with hundreds or thousands of hosts and routers organized in different scenarios, 2) flexibility, for testing new protocols or applications in diverse settings, and 3) inter-operability, for combining simulated and real network entities in experiments. This dissertation tackles these issues in three different dimensions. First, we present SVEET, a system that enables inter-operability between real and simulated hosts. In order to increase the scalability of networks under study, SVEET enables time-dilated synchronization between real hosts and the discrete-event simulator. Realistic TCP congestion control algorithms are implemented in the simulator to allow seamless interactions between real and simulated hosts. SVEET is validated via extensive experiments and its capabilities are assessed through case studies involving real applications. Second, we present PrimoGENI, a system that allows a distributed discrete-event simulator, running in real-time, to interact with real network entities in a federated environment. PrimoGENI greatly enhances the flexibility of network experiments, through which a great variety of network conditions can be reproduced to examine what-if questions. Furthermore, PrimoGENI performs resource management functions, on behalf of the user, for instantiating network experiments on shared infrastructures. Finally, to further increase the scalability of network testbeds to handle large-scale high-capacity networks, we present a novel symbiotic simulation approach. We present SymbioSim, a testbed for large-scale network experimentation where a high-performance simulation system closely cooperates with an emulation system in a mutually beneficial way. On the one hand, the simulation system benefits from incorporating the traffic metadata from real applications in the emulation system to reproduce the realistic traffic conditions. On the other hand, the emulation system benefits from receiving the continuous updates from the simulation system to calibrate the traffic between real applications. Specific techniques that support the symbiotic approach include: 1) a model downscaling scheme that can significantly reduce the complexity of the large-scale simulation model, resulting in an efficient emulation system for modulating the high-capacity network traffic between real applications; 2) a queuing network model for the downscaled emulation system to accurately represent the network effects of the simulated traffic; and 3) techniques for reducing the synchronization overhead between the simulation and emulation systems.