Engineered discoidin domain from factor VIII binds αvβ3 integrin with antibody-like affinity

Alternative scaffolds are non-antibody proteins that can be engineered to bind new targets. They have found useful niches in the therapeutic space due to their smaller size and the ease with which they can be engineered to be bispecific. We sought a new scaffold that could be used for therapeutic ends and chose the C2 discoidin domain of factor VIII, which is well studied and of human origin. Using yeast surface display, we engineered the C2 domain to bind to αvβ3 integrin with a 16 nM affinity while retaining its thermal stability and monomeric nature. We obtained a crystal structure of the engineered domain at 2.1 Å resolution. We have christened this discoidin domain alternative scaffold the “discobody.”

Analysis of the piston ring/liner oil film development during warm-up for an SI-engine

A one-dimensional ring-pack lubrication model developed at MIT is applied to simulate the oil film behavior during the warm-up period of a Kohler spark ignition engine [1]. This is done by making assumptions for the evolution of the oil temperatures during warm-up and that the oil control ring during downstrokes is fully flooded. The ring-pack lubrication model includes features such as three different lubrication regimes, i.e. pure hydrodynamic lubrication, boundary lubrication and pure asperity contact, non-steady wetting of both inlet and outlet of the piston ring, capability to use all ring face profiles that can be approximated by piece-wise polynomials and, finally, the ability to model the rheology of multi-grade oils. Not surprisingly, the simulations show that by far the most important parameter is the temperature dependence of the oil viscosity. This dependence is subsequently examined further by choosing different oils. The baseline oil is SAE 10W30 and results are compared to those using the SAE 30 and the SAE 10W50 oils.

Variability of nanoindentation responses of bone and artificial bone-like composites

Nanoindentation is a popular technique for measuring the intrinsic mechanical response of bone and has been used to measure a single-valued elastic modulus. However, bone is a composite material with 20-80 nm hydroxyapatite plates embedded in a collagen matrix, and modern instrumentation allows for measurements at these small length scales. The present study examines the indentation response of bone and artificial gelatin-apatite nanocomposite materials across three orders of magnitude of lengthscale, from nanometers to micrometers, to isolate the composite phase contributions to the overall response. The load-displacement responses were variable and deviated from the quadratic response of homogeneous materials at small depths. The distribution of apparent elastic modulus values narrowed substantially with increasing indentation load. Indentation of particulate nanocomposites was simulated using finite element analysis. Modeling results replicated the convergence in effective modulus seen in the experiments. It appears that the apatite particles are acting as the continuous ("matrix") phase in bone and nanocomposites. Copyright © 2004 by ASME.

Stejnulxin, a novel snake C-type lectin-like protein from Trimeresurus stejnegeri venom is a potent platelet agonist acting specifically via GPVl

Stejnulxin, a novel snake C-type lectin-like protein with potent platelet activating activity, was purified and characterized from Trimeresurus stejnegeri venom. Under non-reducing conditions, it migrated on a SDS-polyacrylamide gel with an apparent molecular mass of 120 kDa. On reduction, it separated into three polypeptide subunits with apparent molecular masses of 16 kDa (alpha), 20 kDa (beta(1)) and 22 kDa (beta(2)), respectively. The complete amino acid sequences of its subunits were deduced from cloned cDNAs. The N-terminal sequencing and cDNA cloning indicated that beta(1) and beta(2) subunits of stejnulxin have identical amino acid sequences and each contains two N-glycosylation sites. Accordingly, the molecular mass difference between 1 and 2 is caused by glycosylation heterogenity. The subunit amino acid sequences of stejnulxin are similar to those of convulxin, with sequence identities of 52.6% and 66.4% for the U. and beta, respectively. Stejnulxin induced human platelet aggregation in a dose-dependent manner. Antibodies against UNA inhibited the aggregation response to stejnulxin, indicating that activation of alpha(IIb)beta(3) and binding of fibrinogen are involved in stejnulxin-induced platelet aggregation. Antibodies against GPIbalpha or alpha(2)beta(1) as well as echicetin or rhodocetin had no significant effect on stejnulxin-induced platelet aggregation. However, platelet activation induced by stejnulxin was blocked by anti-GPVI antibodies. In addition, stejnulxin induced a tyrosine phosphorylation profile in platelets that resembled that produced by convulxin. Biotinylated stejnulxin bound specifically to platelet membrane GPVI.

Purification and cloning of a novel C-type lectin-like protein with platelet aggregation activity from Trimeresurus mucrosquamatus venom

TMVA, a novel C-type lectin-like protein that induces platelet aggregation in a dose-dependent manner, was purified from the venom of Trimeresurus mucrosquamatus. It consists of two subunits, alpha (15,536 Da) and beta (14,873 Da). The mature amino acid sequences of the a (135 amino acids) and beta subunits (123 amino acids) were deduced from cloned cDNAs. Both of the sequences show great similarity to C-type lectin-like venom proteins, including a carbohydrate recognition domain. The cysteine residues of TMVA are conserved at positions corresponding to those of flavocetin-A and convulxin, including the additional Cys135 in the alpha subunit and Cys3 in the beta subunit. SDS-PAGE, mass spectrometry analysis and amino acid sequence showed that native TMVA exists as two convertible multimers Of (alphabeta)(2) and (alphabeta)(4) with molecular weights of 63,680 and 128,518 Da, respectively. The (alphabeta)(2) complex is stabilized by an interchain disulfide bridge between the two alphabeta-heterodimers, whereas the stabilization of the (alphabeta)(4) complex seems to involve non-covalent interactions between the (alphabeta)(2) complexes. (C) 2002 Elsevier Science Ltd. All rights reserved.

Non-reflective boundary conditions for non-uniform mean velocity inlet and outlet flows

The numerical solution of problems in unbounded physical space requires a truncation of the computational domain to a reasonable size. As a result, the conditions on the artificial boundaries are generally unknown. Assumptions like constant pressure or velocities are only valid in the far field and lead to spurious reflections if applied on the boundaries of the truncated domain. A number of attempts have been made over the past decades to design conditions that prevent such reflections. One approach is based on characteristics. The standard analysis assumes a spatially uniform mean flow field but this is often impractical. In the present paper we show how to extend the formulation to the more general case of a non-uniform mean velocity field. A number of test cases are provided and our results compare favourably with other boundary conditions. In principle the present approach can be extended to include non-uniformities in all variables.

Linear quadratic game and non-cooperative predictive methods for potential application to modelling driver-AFS interactive steering control

This paper is concerned with the modelling of strategic interactions between the human driver and the vehicle active front steering (AFS) controller in a path-following task where the two controllers hold different target paths. The work is aimed at extending the use of mathematical models in representing driver steering behaviour in complicated driving situations. Two game theoretic approaches, namely linear quadratic game and non-cooperative model predictive control (non-cooperative MPC), are used for developing the driver-AFS interactive steering control model. For each approach, the open-loop Nash steering control solution is derived; the influences of the path-following weights, preview and control horizons, driver time delay and arm neuromuscular system (NMS) dynamics are investigated, and the CPU time consumed is recorded. It is found that the two approaches give identical time histories as well as control gains, while the non-cooperative MPC method uses much less CPU time. Specifically, it is observed that the introduction of weight on the integral of vehicle lateral displacement error helps to eliminate the steady-state path-following error; the increase in preview horizon and NMS natural frequency and the decline in time delay and NMS damping ratio improve the path-following accuracy. © 2013 Copyright Taylor and Francis Group, LLC.

Structural analysis of plate-type fuel assemblies and development of a non-destructive method to assess their integrity

This work is concerned with the structural behaviour and the integrity of parallel plate-type nuclear fuel assemblies. A plate-type assembly consists of several thin plates mounted in a box-like structure and is subjected to a coolant flow that can result in a considerable drag force. A finite element model of an assembly is presented to study the sensitivity of the natural frequencies to the stiffness of the plates' junctions. It is shown that the shift in the natural frequencies of the torsional modes can be used to check the global integrity of the fuel assembly while the local natural frequencies of the inner plates can be used to estimate the maximum drag force they can resist. Finally a non-destructive method is developed to assess the resistance of the inner plates to bear an applied load. Extensive computational and experimental results are presented to prove the applicability of the method presented. © 2013 Elsevier B.V. All rights reserved.

Dynamics of filopodium-like protrusion and endothelial cellular motility on onedimensional extracellular matrix fibrils

Endothelial filopodia play key roles in guiding the tubular sprouting during angiogenesis. However, their dynamic morphological characteristics, with the associated implications in cell motility, have been subjected to limited investigations. In this work, the interaction between endothelial cells and extracellular matrix fibrils was recapitulated in vitro, where a specific focus was paid to derive the key morphological parameters to define the dynamics of filopodium-like protrusion during cell motility. Based on one-dimensional gelatin fibrils patterned by near-field electrospinning (NFES), we study the response of endothelial cells (EA.hy926) under normal culture or ROCK inhibition. It is shown that the behaviour of temporal protrusion length versus cell motility can be divided into distinct modes. Persistent migration was found to be one of the modes which permitted cell displacement for over 300 μm at a speed of approximately 1 μm min-1. ROCK inhibition resulted in abnormally long protrusions and diminished the persistent migration, but dramatically increased the speeds of protrusion extension and retraction. Finally, we also report the breakage of protrusion during cell motility, and examine its phenotypic behaviours. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

The first non-mammalian CXCR3 in a teleost fish: Gene and expression in blood cells and central nervous system in the grass carp (Ctenopharyngodon idella)

A novel fish chemokine receptor gene, chemokine (C-X-C motif) receptor 3 (CXCR3)-like was isolated from the grass carp Ctenopharyngodon idella , with its full-length genomic sequence. The cDNA of grass carp CXCR3-like (gcCXCR3-like) consists of 1261 bp with a 49bp 5'-UTR and a 189 bp 3'-UTR. An open reading frame of 1023 bp encodes a 341-amino acid peptide, with seven transmembrane helices. The deduced amino acid sequence showed the same sequence identities (37.8%) with its counterparts in goat and human. The gcCXCR3-like gene consists of two exons, with one intervening intron, spaced over approximately 2 kb of genomic sequence. Phylogenetic analyses clearly demonstrated that the gcCXCR3-like resembles the CXCR3s of other vertebrates. Real-time PCR analysis showed that gcCXCR3-like was expressed in all tested organs except heart and the expression level of gcCXCR3-like was highest in brain. Flow cytometric analyses showed the positive rate of labelled leukocytes from the healthy grass carp was 17.3%, and the labelled leukocytes were divided into three types by cell sorting. Immunohistochemical localization revealed that gcCXCR3-like expressed in whole brain regions including cerebel, diencephalon, medulla oblongata, optic lobe, and rhinencephalon, and that the labelled leukocytes are actually populations of monocyte and/or phagocyte, lymphocyte and the granulocyte. It is considered that fish CXCR expression and their function may need to be investigated in both nervous and immune systems. (c) 2006 Elsevier Ltd. All rights reserved.

Modelling of settlement induced building damage

This thesis focuses on the modelling of settlement induced damage to masonry buildings. In densely populated areas, the need for new space is nowadays producing a rapid increment of underground excavations. Due to the construction of new metro lines, tunnelling activity in urban areas is growing. One of the consequences is a greater attention to the risk of damage on existing structures. Thus, the assessment of potential damage of surface buildings has become an essential stage in the excavation projects in urban areas (Chapter 1). The current damage risk assessment procedure is based on strong simplifications, which not always lead to conservative results. Object of this thesis is the development of an improved damage classification system, which takes into account the parameters influencing the structural response to settlement, like the non-linear behaviour of masonry and the soil-structure interaction. The methodology used in this research is based on experimental and numerical modelling. The design and execution of an experimental benchmark test representative of the problem allows to identify the principal factors and mechanisms involved. The numerical simulations enable to generalize the results to a broader range of physical scenarios. The methodological choice is based on a critical review of the currently available procedures for the assessment of settlement-induced building damage (Chapter 2). A new experimental test on a 1/10th masonry façade with a rubber base interface is specifically designed to investigate the effect of soil-structure interaction on the tunnelling-induced damage (Chapter 3). The experimental results are used to validate a 2D semi-coupled finite element model for the simulation of the structural response (Chapter 4). The numerical approach, which includes a continuum cracking model for the masonry and a non-linear interface to simulate the soil-structure interaction, is then used to perform a sensitivity study on the effect of openings, material properties, initial damage, initial conditions, normal and shear behaviour of the base interface and applied settlement profile (Chapter 5). The results assess quantitatively the major role played by the normal stiffness of the soil-structure interaction and by the material parameters defining the quasi-brittle masonry behaviour. The limitation of the 2D modelling approach in simulating the progressive 3D displacement field induced by the excavation and the consequent torsional response of the building are overcome by the development of a 3D coupled model of building, foundation, soil and tunnel (Chapter 6). Following the same method applied to the 2D semi-coupled approach, the 3D model is validated through comparison with the monitoring data of a literature case study. The model is then used to carry out a series of parametric analyses on geometrical factors: the aspect ratio of horizontal building dimensions with respect to the tunnel axis direction, the presence of adjacent structures and the position and alignment of the building with respect to the excavation (Chapter 7). The results show the governing effect of the 3D building response, proving the relevance of 3D modelling. Finally, the results from the 2D and 3D parametric analyses are used to set the framework of an overall damage model which correlates the analysed structural features with the risk for the building of being damaged by a certain settlement (Chapter 8). This research therefore provides an increased experimental and numerical understanding of the building response to excavation-induced settlements, and sets the basis for an operational tool for the risk assessment of structural damage (Chapter 9).

Interference Effects in Hyperfine Induced 2s2p P-3(0), P-3(2) -> 2s(2) S-1(0) Transitions of Be-like Ions

In this paper, we explored the characteristics of the interference effects between perturbative states in hyperfine induced 2s2p P-3(0), P-3(2) -> 2s(2) S-1(0) transitions of Be-like ions. It was found that the interference effects non-monotonically change with increasing atomic number Z in these two transitions. The strongest interference effect is near Z = 9 for 2s2p P-3(0), -> 2s(2) (1)S(0)transition and near Z = 7 for the other.

东营凹陷辛镇复杂断块油藏形成机制和剩余油分布规律研究

In this paper, the complex faulted-block oil reservoir of Xinzhen area in Dongying depression is systematically studied from basic conditions forming faulted-block oil and gas reservoir integrating geology, seismic, logging and reservoir engineering information and computer; guided by petroleum geology, geomechanics, structural geology and geophysics and other theories. Based on analysis of background condition such as regional strata, structure and petroleum geology, structural research on geometry, kinemaitcs and dynamics, oil-controlling fault research on the seal features, sealing mechanism and sealing pattern, and research on enrichment rules and controlling factors of complex faulted-block oil reservoir are carried out to give out the formation mechanics of oil reservoir of Xinzhen complex faulted-block oil reservoir. As a result, the reservoir formation pattern is established. At the same time, through dissecting the characteristics and hydrocarbon enrichment law of complex faulted-block oil reservoir, and studying its distribution law of remaining oil after entering extra high water-cut period, a set of technologies are formed to predict complex faulted-block oil reservoir and its remaining oil distribution and to enhance oil recovery (EOR). Based on the time relationship between migration of hydrocarbon and trap formation, accumulating period of Xinzhen oil reservoir is determined. The formation of Xinzhen anticlinal trap was prior to the primary migration. This is favorable to formation of Xinzhen anticlinal hydrocarbon reservoir. Meanwhile, because anticline top caving isn't at the sane time as that of moving or faulted-trap forming inner anticline, oil and gas migrated many times and Xinzhen complex faulted-block oil reservoir formed from ES_3~(upper) to EG. Accumulating law and controlling factors of complex faulted-block reservoir are analyzed from many aspects such as regional structure background controlling hydrocarbon accumulating, plastic arch-open structure controlling oil-bearing series and reservoir types, sealing-opening of fault controlling hydrocarbon distribution and structure pattern controlling enriched trap types. Also, we established the structure pattern in Xinzhen a'ea: the arch-open of underlying strata cause expanding fracture. The main block groups developed here are shovel-like normal fault block group in the north area of Xinzhen and its associated graben block group. Block groups dominate the formation and distribution of reservoirs. We studied qualitatively and quantitatively the sealing characteristics, sealing history and sealing mechanism of faults, too. And, the sealing characteristics are evaluated and the distribution pattern of hydrocarbon controlled by faults is researched. Due to movement intensity of big faults, deep falling of downthrown block, high degree of repture and development of fracture, shallow layers close to the downthrown block of secondary faults are unfavorable to hydrocarbon accumulation. This is confirmed by the exploration practice in Xinzhen anticline. In terms of the downthrown blocks of sencondary contemporaneous faults lied in the south and north area of Xinzhen, hydrocarbon is poor close to fracture belt, while it is relatively abundant in tertiary companion faults. Because of long-term movement of faults that control hydrocarbon, fi'om ES3 to EG, six set of oil-bearing series formed. And their opening causes the inhomogeneity in hydrocarbon abundance among each block--in two flanks of anticline reservoirs are abundant while in the axial area, oil and gas are sporadic. There the sealing characteristics control oil-bearing area of oil/gas accumulation and the height of oil reservoir. Longitudinally, oil and gas are enriched in dip-flat areas in mid-plane of faults. It is established that there are four types of accumulating patterns in complex faulted-block oil reservoirs in Xinzhen. The first is accumulating pattern of lithologic oil reservoirs in E~S_3~(mid-lowwer), that is, self-generating-self-reserving-self-covering lithologic trap pattern. The second is drag-anticline accumulating pattern in Xinzhen. The structure traps are drag anticlines formed by the contemporaneous faults of the second basement in the north of Xinzhen, and the multiple source rocks involve Ek_2, Es_4, Es_3 and Es_1 members. The reservoirs are fluvial-delta sandstones of the upper member of Shahejie formation and Guantao formation, covered by regional thick mudstone of the upper member of Guantao formation and MingHuazhen formation. The third is the accumulating pattern of reverse listric fault, the third-degree fault of Xinzhen anticline limb and the reservoirs form reservoir screened by reverse listric faults. The forth is accumulating pattern of crossing faults which form closing or semi-closing faulted-blocks that accumulate hydrocarbon. The technologies of predicting remaining oil in complex faulted-block reservoir during the mid and late development stage is formed. Remaining oil in simple large faulted-blocks enriches in structural high, structural middle, structural low of thick bottom water reservoirs, points near bent edge-fault oftertiary faults and part the fourth ones with big falling displacement, microstructure high place of oil-sandbodies and areas where local well pattern isn't perfect. While that in small complex faulted-blocks enriches near small nose, small high point, angle of small faults, small oil-bearing faulted-blocks without well and areas with non-perfect well pattern. The technologies of enhancing recovery factor in complex faulted-block reservoir during the mid and late development stage is formed as follows: fine reservoir description, drilling adjust wells, designing directional wells, sub-dividing layer series of development, improving flooding pattern, changing water-injection direction and enhancing swept volume, cyclic waterflooding and gas-injection, etc. Here, directional wells include directional deflecting wells, lateral-drilling wells, lateral-drilling horizontal wells and horizontal wells. The results of this paper have been used in exploration and development of Shengli oilfield, and have achieved great social and economic profit, especially in predicting distribution of complex faulted-block reservoir, remaining oil distribution during middle and late stage of development, and in EOR. Applying the achievement of fault-closure research, new hydrocarbon-bearing blocks are discovered in flanks of Dongying central uplift and in complex blocks with proved reserves 15 million tons. With the study of remaining oil distribution law in complex faulted-block reservoirs, recovery factors are increased greatly in Dongxin, Xianhe and Linpan complex faulted-block reservoirs and accumulated oil production increment is 3 million tons.

A role for LH in the regulation of expression of mRNAs encoding components of the insulin-like growth factor (IGF) system in the ovine corpus luteum

P.M. Hastie and W. Haresign (2006). A role for LH in the regulation of expression of mRNAs encoding components of the insulin-like growth factor (IGF) system in the ovine corpus luteum. Animal Reproduction Science, 96(1-2), 196-209. Sponsorship: DEFRA RAE2008