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Cultured vascular grafts must be endothelialized to supply a nonthrombogenic lining to the lumen. After seeding of the endothelial cells, our tissue-engineered vessels possess analogs of the medial and endothelial layers present in native artery. Reagents and Materials Bovine aortic ECs, at or below passage 4 to ensure adequate cell proliferation and differentiation 0.25% trypsin-EDTA (See Protocol 12.1) Plastic conical centrifuge tube, 50 ml Endothelial Cell Culture Medium (See Section 2.1) Master ex modular drive pump Enhanced Culture Medium (See Section 2.4) Protocol (See Fig. 12.6b) (a) Trypsinize con uent bovine aortic ECs with 0.25% trypsin-EDTA. (b) Centrifuge cells in a 50-ml conical centrifuge tube for 5 min at 425 g. (c) Resuspend the cell pellet in Endothelial Cell Culture Medium to a density of 3 106 cells/ml. (d) Inject the EC suspension into the vessel lumen through one sidearm of the bioreactor.

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In order to identify enzymes responsible for the postmortem aging of meat, researchers have used the criteria that candidate enzyme systems must: (1) be endogenous to skeletal muscle and have access to substrates, and (2) have the ability to degrade the same proteins that are degraded during the postmortem storage of muscle (Goll et al. 1983, 2003, 2008; Koohmaraie 1996). Of the three major enzyme systems investigated, only calpains meet both criteria. Calpains have access to substrates and have been shown to have limited proteolytic capabilities (Goll et al. 2003), cleaving myo brillar proteins at a speci c number of sites to produce large polypeptide fragments similar to those observed after the postmortem storage of muscle. In contrast, lysosomal cathepsins and proteasomes are capable of exhaustively degrading proteins into small peptides or short amino acid segments but cannot disassemble the myo bril and do not generate the same degradation patterns of myo brillar proteins observed during meat aging. Furthermore, the location of cathepsins in lysosomes is thought to restrict their access to substrates. Thus, the prevailing theory is that the calpain/calpastatin system is the predominant driver of postmortem proteolysis and aging tenderization. While it is widely accepted that proteolysis of key myo brillar proteins by the calpain enzyme system is primarily responsible for increased tenderness during postmortem storage (Koohmaraie et al. 1991; Uytterhaegen et al. 1994; Goll et al. 2003; Koohmaraie and Geesink 2006), it can be argued that calpains alone are not suf cient to fully explain postmortem proteolysis and meat tenderization. Questions regarding the role of calpains in postmortem tenderization initially centered on: (1) the observation that calpain activity

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You create animation in a Macromedia Flash MX document by changing the contents of successive frames. You can make an object move across the Stage, increase or decrease its size, rotate, change color, fade in or out, or change shape. Changes can occur independently of, or in concert with, other changes. For example, you can make an object rotate and fade in as it moves across the Stage. There are two methods for creating an animation sequence in Flash: tweened animation, and frame-by-frame animation. In tweened animation, you create starting and ending frames and let Flash create the frames in between. Flash varies the object s size, rotation, color, or other attributes evenly between the starting and ending frames to create the appearance of movement. See About tweened animation on page 169. In frame-by-frame animation, you create the image in every frame. See About frame-by-frame animation on page 170. To simplify the process of creating tweened animation, you can distribute multiple objects to separate layers. See Distributing objects to layers for tweened animation on page 172. You can use a mask layer to create a hole through which the contents of one or more underlying layers are visible. Using an animated movie clip, you can create a dynamic layer mask. See Using mask layers on page 183. For an interactive introduction to animation, choose Help > Lessons > Creating Tweened Animation.

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are two irreducible and distinct polynomials,

8 Early JM Structure-determined gain-band product of junction triode transistors Proceedings of the IRE, December 1958 p 1924 9 Goldey JM, Ryder RM Are transistors approaching their maximum capabilities Proceedings of the International Solid-State Circuits Conference, February 1963 10 Pritchard RL Frequency response of grounded-base and grounded emitter junction transistors AIEE Winter Meeting, January 1954 11 Webster WM On variation of junction transistor current ampli cation with emitter current Proceeding of the IRE 1954;42:914 20 12 Giacoletto LJ Comparative high frequency operation of junction transistors made of different semiconductor materials RCA Review 1955;16:34 13 Johnson EO, Rose A Simple general analysis of ampli er devices with emitter, control, and collector functions Proceedings of the IRE, March 1959 p 407 14 Johnson EO Whither the tunnel diode AIEE-IRE Electron Device Conference, October 1962 15 Johnson EO Physical limitations on frequency and power parameters of transistors RCA Review 1965;26:163 77 16.

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