This thesis investigated the different aspects and mechanisms of leak-before-break (LBB) assessment. The main objective was to improve the understanding of the transition between surface and through wall defects. While existing procedures generally idealise the through-wall crack into a rectangular shape, in reality a crack propagates with a shape depending on the loading. Comparison between the related solutions from established procedures have been undertaken. The apparent variation depending on the solutions used in the assessment has been highlighted. Two different methodologies have been employed to investigate the transition of flaw: (i) non-ideal through-wall and (ii) surface-breaking flaw propagation. The first approach consists of numerical models of non-idealised flaws in order to assess the effect on LBB parameters. For the second approach, experiments have been first carried out to visualise the shape of defect growths. To further study surface-breaking flaws, both experimental and numerical studies were performed. Fatigue tests on deeply notched plates with two crack aspect ratios were carried out. Strain evolutions on the back surface were recorded along the axes parallel and perpendicular to the crack. Numerical models have been prepared to investigate a larger scope. Behaviour of growing surface-breaking defects was examined. Based on the work conducted in this research, the major findings can be summarised as follows: - The existing solutions to carry out a LBB assessment using available procedures were reviewed and discussed. For axial flaws, SIF solutions were found similar and in good agreement with FEA values. Reference stress solutions showed significant difference between BS 7910 and API 579-1/ASME FFS-1. When compared to experimental data, API's solutions were able to distinguish between leak and break cases. - Flaw geometry assumption for through-wall crack yet to become idealised did not always reflect the actual behaviour, especially for COA calculation. In this case, FEA can be used as a good predictive tool for LBB to estimate margins when assessing leak rate. - The experiment using metallic specimens showed that high stress/strain on back surface would provide a good estimate of the crack propagation as it approached break-through. This offers a more accurate monitoring mechanism. Strain-mapping devices such as gauges could be used.

Despite advancements in the analysis of statnamic load testing data, there exists uncertainty with underlying procedural assumptions. Two such assumptions are that the system mass and soil-related damping coefficient remain constant throughout the loading event. These assumptions are the culprit of aberrant predictions of the static capacity at small displacements when the overall displacement is large. An exploration of the assumptions may validate prior existing test results as well as solidify the current analysis process. However, an exploration could also reveal an overestimation or underestimation of portions of the predicted static load responses. The testing program outlined herein consists of a two-phase sequential agenda devoted toward the preparation and familiarization of a new laboratory statnamic device. The first phase involves the development of user guidelines for accurately targeting a desired statnamic test, and the second incorporates the guidelines into a preliminary testing regime specifically targeted at determining a suspected strain-dependant statnamic damping coefficient. The steps taken in this thesis are intended to launch future research endeavors toward obtaining a better understanding of the statnamic damping coefficient.

The Gulf of Mexico commercial reef fish fishery has experienced significant management changes and disturbance in recent years, including transitioning two major fisheries from a traditional open access system into a limited entry individual fishing quota (IFQ) system in 2007 and 2010. Also in 2010, the Deepwater Horizon oil spill (DWH) released an estimated 4.9 million barrels of oil into the Gulf (~206 million U.S. gallons), and is still the largest U.S. environmental disaster to date. Emergency fishing closures initiated shortly after the oil spill began were successful in keeping tainted seafood from reaching markets. However, effects of DWH closures on fisher decision making, fishery productivity, and distribution of fishing effort all remain poorly understood. Understanding the range and magnitude of fishers’ responses to perturbations — including regulatory change and human-induced environmental disasters — is critical for designing effective management and disaster response policies that can meet biological, ecological, economic, social, and sustainability objectives. This work characterized the spatial and temporal patterns of productivity and fishing effort for the Gulf of Mexico (GoM) commercial reef fish fishery. Patterns of productivity and effort distribution were used to examine the response of fishers to management change and large-scale disturbance, namely the DWH fishing closures. Fisheries-dependent logbook trip reports were used to quantify revenue and catch-per-unit-effort (CPUE) patterns from 2000-2014. Novel to fisheries work in the GoM, complementary vessel monitoring systems (VMS) satellite tracking data were used to quantify high-resolution spatial distribution patterns over time, relative to the DWH fishing closures. A general linear modeling (GLM) approach was also used to examine which variables may have contributed to resilience of fishers after DWH closures. Results suggested that this fishery was largely resilient to the DWH fishing closures in 2010, although exact outcomes varied by region. Overall fleet-level productivity steadily increased over time, but regional patterns were based on major species in catch. Productivity in the western GoM was consistently highest over time, and trips in the west and central GoM were dominated by Red snapper (Lutjanus campechanus) and Vermilion snapper (Rhomboplites aurorubens). Trips in the east were dominated by Red grouper (Epinephelus morio) and Gag grouper (Mycteroperca microlepis). Shifts in spatial distribution to new productive fishing grounds or reduced competition via fewer vessels or trips may explain the increases in productivity observed over the study period. Consolidation in the fleet was apparent, with fewer individual vessels and fewer total trips over time. However, the rate of vessel drop out after DWH (5%) was far below the annual background attrition rate of ~14-20%. Relative productivity patterns inside vs. outside the boundaries of fishing closures did not change over time, and there were even some increases in productivity observed during and after DWH in the eastern GoM. Yet, vessels that dropped out after DWH were concentrated in the north-central and eastern GoM. Distribution of fishing grounds before and after DWH were highly similar, and there were increases in effort along the outer West Florida Shelf. Variability in revenue and CPUE, CPUE magnitude, and magnitude of grouper landings were significant predictors of dropping out of the fishery in the GLMs. Synergies with the Red snapper or Grouper-Tilefish IFQs may have “primed” the fishery for resilience by eliminating inconsistent or marginal fishers before the oil spill, and may further explain some of the spatially varying patterns of productivity and attrition after 2010. Resilience was likely also enhanced by the more than $2 billion in emergency compensation payments made to captains, crew, and vessel owners for lost fishing income and assistance with oil remediation efforts. This work stands to make a significant contribution to our understanding of how the DWH oil spill impacted fisheries and communities in the GoM. The results add to a growing body of literature suggesting that the acute population- and ecosystem-level impacts of the DWH oil spill were not as strong or severe as initially anticipated. This work also stands to make contributions to the broader understanding of how this fishery has performed in the wake of recent management change and major environmental disturbance.

Haemostasis is a complex physiological process involving cellular and plasma protein components that interact to keep the blood fluid under normal conditions and prevent blood loss after vessel injury by promoting clot formation. Primary haemostasis encompasses the activation and aggregation of platelets and is supported by secondary haemostasis, in which the coagulation factors of the plasma interact in a complex series of reactions. Secondary haemostasis is initiated by the exposure of tissue factor (TF) to the blood after vessel injury. TF forms a complex with activated factor VII (FVIIa), which in turn activates factor X (FXa) and ultimately results in fibrin formation. The TF-FVIIa complex and FXa are tightly regulated by tissue factor pathway inhibitor (TFPI), a trivalent Kunitz-type protease inhibitor. The endothelium, consisting of endothelial cells (ECs), constitutes the inner lining of all blood vessels. As such, it is in constant contact with the blood and plays a major role in haemostasis by synthesising and storing both pro- and anti- coagulant substances, including TF and TFPI. Release of TFPI from ECs is increased after exposure to both unfractionated and low molecular weight heparins, though the mechanisms are not clearly defined. TFPI circulates in plasma, predominantly bound to lipoproteins, though the effect of the three major lipoproteins [low density (LDL), very low density (VLDL) and high density (HDL)] on the release of TFPI from ECs is not well established. Furthermore, previous studies have not systematically investigated the effect of these lipoproteins on both TF and TFPI. The initial aim of this project was to establish assays for the measurement of TF activity and TFPI antigen to supplement the TFPI activity assay that is well established in our laboratory. / These assays were then used to determine the effects of heparin and the major lipoproteins on the expression of TF and the release of TFPI on/from ECs. Human umbilical vein endothelial cells (HUVECs) were used as the EC model because their collection and isolation is well established and they have biochemical and physiological properties representative of in vivo conditions. A TF activity assay, based on a previously published method, was successfully modified and validated for the measurement of cell surface TF (standard curve R2 = 0.997). Despite exhaustive attempts, adaptation of this assay for plasma TF was unsuccessful, raising doubts regarding the plasma fractionation procedure of the originally published assay [Fukuda, C., Iijima, K. and Nakamura, K. (1989). "Measuring tissue factor (factor III) activity in plasma." Clinical Chemistry 35(9): 1897‐1900]. A novel insect cell expression system was used to produce well defined recombinant TFPI standards for use in TFPI activity and antigen assays. For the first time, truncated TFPI variants, containing the first Kunitz domain only, the first and second Kunitz domains only, and the first through third Kunitz domains minus the carboxyl terminus, were successfully produced in insect cells, though the full length molecule was not. Possible reasons for this include codon bias, protein instability and/or the signal peptide used. An ELISA to measure TFPI antigen was designed using a monoclonal anti‐TFPI antibody directed against the N‐terminus for protein capture and a polyclonal anti‐ TFPI antibody for detection. The assay was successfully optimised (standard curve R2 = 0.978, intra‐assay CV = 4.8%), however it produced inaccurate results (normal range = 498.7 ± 156.3 ng/mL), probably due to the antibody combination used. / TF and TFPI activity assays were used to determine the effect of both unfractionated and low molecular weight heparins (UFH and LMWH, respectively) on the release of TFPI and the expression of TF from/on ECs. A significant increase in the secretion of functional TFPI from ECs due to heparin (0 U/ml vs 1 and 10 U/mL) was demonstrated only in the presence of serum (UFH: 9.0 mU/mL vs 18.3 and 18.4 mU/mL, p < 0.0001; LMWH: 8.8 mU/mL vs 13.3 and 21.4 mU/mL, p < 0.05), suggesting, for the first time, that a component of serum is required for the heparin‐dependent release of TFPI. The effect of LDL, VLDL and HDL on the release of TFPI and the expression of TF from/on ECs was also investigated. All three lipoprotein fractions increased the secretion of functional TFPI after one hour incubation (LDL: 12.5 μg/mL, p < 0.01; 25 μg/mL, p < 0.05; VLDL: 50 μg/mL, p < 0.01; HDL: 50 μg/mL, p < 0.05). This is the first data to demonstrate a HDL‐dependent increase in released TFPI. After 24 hours, both LDL and VLDL decreased levels of secreted functional TFPI (LDL: 25 μg/mL, p < 0.01; 50 μg/mL, p < 0.01; VLDL: 12.5 μg/mL, p < 0.01), probably due to the oxidation and subsequent association of both lipoprotein species with TFPI. Surprisingly, both LDL and VLDL decreased cell surface TF, though this effect was not dose dependent. These results suggest that the major lipoproteins have a short term anticoagulant effect which is reversed in the longer term due to lipid oxidation. In summary, this thesis describes the successful adaptation of a chromogenic assay for the measurement of cell surface TF activity and the production of truncated TFPI variants. / Both will be used for the measurement of TF and TFPI, their association with thrombus formation and propagation, and investigations into potential therapeutic applications of TFPI. The results presented in this thesis extend the current knowledge on the expression and release of TF and TFPI on/from ECs by heparin, highlighting the importance of serum in the heparin dependent release of TFPI in vitro. Furthermore, it describes for the first time the effects of the major lipoprotein fractions on TFPI release and TF expression. The data support novel mechanisms by which LDL and VLDL are procoagulant, and HDL anticoagulant. This study provides a foundation for future research of the TF pathway in cellular models, which is critical in increasing the understanding of the pathogenesis and treatment of thrombotic disease. vitro. Furthermore, it describes for the first time the effects of the major lipoprotein fractions on TFPI release and TF expression. The data support novel mechanisms by which LDL and VLDL are procoagulant, and HDL anticoagulant. This study provides a foundation for future research of the TF pathway in cellular models, which is critical in increasing the understanding of the pathogenesis and treatment of thrombotic disease.

Ocular fundus image can provide information on pathological changes caused by local ocular diseases and early signs of certain systemic diseases. Automated analysis and interpretation of fundus images has become a necessary and important diagnostic procedure in ophthalmology. Among the features in ocular fundus image are the optic disc, fovea (central vision area), lesions, and retinal vessels. These features are useful in revealing the states of diseases in the form of measurable abnormalities such as length of diameter, change in color, and degree of tortuosity in the vessels. In addition, retinal vessels can also serve as landmarks for image-guided laser treatment of choroidal neovascularization. Thus, reliable methods for blood vessel detection that preserve various vessel measurements are needed. In this paper, we will examine the pathological issues in the analysis of retinal vessels in digital fundus images and give a survey of current image processing methods for extracting vessels in retinal images with a view to categorize them and highlight their differences and similarities. We have also implemented two major approaches using matched filter and mathematical morphology respectively and compared their performances. Some prospective research directions are identified. / Singapore-MIT Alliance (SMA)

No description available.

Tissue engineering is a rapidly growing field which seeks to provide alternatives to organ transplantation in order to address the increasing need for transplantable tissues. One huge hurdle in this effort is the provision of thick tissues; this hurdle exists because currently there is no way to provide prevascularized or rapidly vascularizable scaffolds. To design thick, vascularized tissues, scaffolds are needed that can induce vessels which are similar to the microvasculature found in normal tissues. Angiogenic biomaterials are being developed to provide useful scaffolds to address this problem. In this thesis angiogenic and cell signaling and adhesion factors were incorporated into a biomimetic poly(ethylene glycol) (PEG) hydrogel system. The composition of these hydrogels was precisely tuned to induce the formation of differing vessel morphology. To sensitively measure induced microvascular morphology and to compare it to native microvessels in several tissues, this thesis developed an image-based tool for quantification of scale invariant and classical measures of vessel morphology. The tool displayed great utility in the comparison of native vessels and remodeling vessels in normal tissues. To utilize this tool to tune the vessel response in vivo , Flk1::myr-mCherry fluorescently labeled mice were implanted with Platelet Derived Growth Factor-BB (PDGF-BB) and basic Fibroblast Growth Factor (FGF-2) containing PEG-based hydrogels in a modified mouse corneal angiogenesis assay. Resulting vessels were imaged with confocal microscopy, analyzed with the image based tool created in this thesis to compare morphological differences between treatment groups, and used to create a linear relationship between space filling parameters and dose of growth factor release. Morphological parameters of native mouse tissue vessels were then compared to the linear fit to calculate the dose of growth factors needed to induce vessels similar in morphology to native vessels. Resulting induced vessels did match in morphology to the target vessels. Several other covalently bound signals were then analyzed in the assay and resulting morphology of vessels was compared in several studies which further highlighted the utility of the micropocket assay in conjunction with the image based tool for vessel morphological quantification. Finally, an alternative method to provide rapid vasculature to the constructs, which relied on pre-seeded hydrogels encapsulated endothelial cells was also developed and shown to allow anastamosis between induced host vessels and the implanted construct within 48 hours. These results indicate great promise in the rational design of synthetic, bioactive hydrogels, which can be used as a platform to study microvascular induction for regenerative medicine and angiogenesis research. Future applications of this research may help to develop therapeutic strategies to ameliorate human disease by replacing organs or correcting vessel morphology in the case of ischemic diseases and cancer.

The formation of nano–sized precipitates is considered to be the origin of hardening and embrittlement of ferritic steel used as structural material for pressure vessels of nuclear reactors, since these nanoclusters hinder the motion of dislocations within the grains of the polycrystalline bcc–Fe matrix. Previous investigations showed that these small precipitates are coherent and may consist of Cu, Ni, other foreign atoms, and vacancies. In this work a combination of on–lattice simulated annealing based on Metropolis Monte Carlo simulations and off–lattice relaxation by Molecular Dynamics is applied in order to determine the structure, energetics and thermodynamics of coherent clusters in bcc–Fe. The most recent interatomic potentials for Fe–Cu–Ni alloys are used. The atomic structure and the formation energy of the most stable configurations as well as their total and monomer binding energy are calculated. Atomistic simulation results show that pure (vacancy and copper) as well as mixed (vacancy-copper, copper-nickel and vacancy-copper-nickel) clusters show facets which correspond to the main crystallographic planes. Besides facets, mixed clusters exhibit a core-shell structure. In the case of v_lCu_m, a core of vacancy cluster coated with copper atoms is found. In binary Cum_Ni_n, Ni atoms cover the outer surface of copper cluster. Ternary v_lCu_mNi_n clusters show a core–shell structure with vacancies in the core coated by a shell of Cu atoms, followed by a shell of Ni atoms. It has been shown qualitatively that these core–shell structures are formed in order to minimize the interface energy between the cluster and the bcc-Fe matrix. Pure nickel consist of an agglomeration of Ni atoms at second nearest neighbor distance, whereas vacancy-nickel are formed by a vacancy cluster surrounded by a nickel agglomeration. Both types of clusters are called quasi-cluster because of their non-compact structure. The atomic configurations of quasiclusters can be understood by the peculiarities of the binding between Ni atoms and vacancies. In all clusters investigated Ni atoms may be nearest neighbors of Cu atoms but never nearest neighbors of vacancies or other Ni atoms. The structure of the clusters found in the present work is consistent with experimental observations and with results of pairwise calculations. In agreement with experimental observations and with recent results of atomic kinetic Monte Carlo simulation it is shown that the presence of Ni atoms promotes the nucleation of clusters containing vacancies and Cu. For pure vacancy and pure copper clusters an atomistic nucleation model is established, and for typical irradiation conditions the nucleation free energy and the critical size for cluster formation have been estimated. For further application in rate theory and object kinetic Monte Carlo simulations compact and physically–based fit formulae are derived from the atomistic data for the total and the monomer binding energy. The fit is based on the structure of the clusters (core-shell and quasi-cluster) and on the classical capillary model.

It has been postulated that the control of thrombus formation on molecularly engineered surfaces is an important step in developing clinically durable small-diameter vascular prostheses. This has led to designing a membrane-mimetic assembly that contains physiological regulators of blood coagulation, thrombomodulin (TM) and heparin, to provide strategies for generating actively antithrombogenic surfaces. The membrane-mimetic construct contains polymeric phospholipid monolayer on an alkylated polyelectrolyte multilayer supported by planar substrate such as glass or silicone. When incorporated with TM, the model platform exhibited the biological function by catalyzing activation of protein C. Surface TM activity was extensively investigated at physiologic shear rates (50 sec-1 and 500 sec-1). Significantly, reaction rates become saturated at TM surface densities greater than or equal to ~ 800 fmole/cm2 due to due to a transport limitation. Based on the similar membrane-mimetic construct, a functional heparinized surface was designed as an alternative anticoagulant system. Immobilization of heparin onto membrane-mimetic surfaces was achieved through biotin-streptavidin binding specificity. Activity of surface heparin to facilitate thrombin inactivation was investigated at shear rates of 50 and 500 sec-1. Significantly, rate of thrombin decay becomes saturated when the surface coverage of heparin is higher than 4.4 pmole of heparin per cm2. We further investigated the effects of surface bound TM and heparin on tissue factor (TF) -induced thrombin generation in a flow model. Specifically, TF positioned over a 2 x 6 mm2 upstream region as a trigger for thrombin generation and TM and/or heparin positioned over the remaining downstream (34 x 6 mm2) portion of the test film. Compared to TF alone surface, thrombin generation was profoundly reduced in the presence of surface bound TM and/or heparin. Significantly, thrombin production was maximally inhibited more than 85% in the presence of TM and heparin, possibly due to anticoagulant synergism of both anticoagulants. We believe that current membrane-mimetic systems can potentially create actively antithrombogenic surfaces.

According to the author¡¦s professional observations of the 3PL industrial situations and problems, the main issue of this paper is to present real cases providing solutions on 3PL industrial pitfalls. In this paper, the pitfalls of ¡§Third party logistics¡¨ industry actually focus on the ¡§NVOCC¡¨ services, which indicate the vicious competitions of price for these decades. Due to the low entry barrier and low cost requirements; 3PL industry originally is under a keen competitive environment. Plus, because of the different price structure of underlying carrier during the recent decades, for 3PLs, the vicious cycle of NVOCC becomes more and more serious. Therefore, in this paper, author analyzes two practical cases based on existing literature review. The main purpose here is finding out the efficient suggestions of solutions on breaking this vicious cycle of competition by improving the core competitiveness from both internal services offering and external logistic network.