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Allow tissue fragments to settle Discard the turbid supernate Add 9 ml DNase I, 28 g/ml, to digest traces of DNA that impair sedimentation of tissue fragments and separation from isolated cells DNase digestion is facilitated by triturating 25 times with a wide-bore pipette Allow the tissue fragments to settle, and discard the supernate Add 10 ml trypsin for the next cycle of digestion, this time for about 5 min Collect the supernates of this trypsin digestion and of the following trituration with DNase I in 50-ml tubes containing 2 ml fetal calf serum (FCS) to inactivate the trypsin, and keep on ice Repeat this digestion cycle until nearly complete digestion of the heart tissue is obtained Over time, gradually reduce the volumes of trypsin and DNase I to 75 ml trypsin and 65 ml DNase I After most tissue fragments are disaggregated, the remainder is discarded.

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P(Y = y) =

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Centrifuge the 50-ml collection tubes (generally 4 6 tubes) at 50 g for 15 min at 4 C, and discard the supernate Resuspend the cell pellet in 2 ml complete medium and pool all the pellets in one of the tubes Rinse the other tubes with 2 ml medium each and pool Add 250 l DNase I stock solution per 30-ml cell suspension, triturate the suspension 25 , and centrifuge as above Discard the supernate and resuspend the cell pellet in 32 ml medium Filter the cell suspension through a 100- m mesh lter into four 15-cm culture dishes Incubate the dishes for 1 h in the CO2 incubator During this time, broblasts and other noncardiac myocytes adhere, whereas cardiac myocytes remain oating Transfer nonadherent cells to a 50-ml tube, sediment by centrifugation, resuspend the pellet in fresh complete medium, and count cells Adjust the cell concentration to 6.

1.20%

4 106 cells/ml Cell yield per animal is generally 2 3 106 viable cells/heart as judged by trypan blue exclusion..

95% PI (42.318, 67.574)

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The signi cance of the entropy rate of a stochastic process arises from the AEP for a stationary ergodic process. We prove the general AEP in Section 16.8, where we show that for any stationary ergodic process, 1 log p(X1 , X2 , . . . , Xn ) H (X) n (4.24)

(12.7) (12.8)

where lmax is the maximum codeword length and a(m) is the number of source sequences x k mapping into codewords of length m. But the code is uniquely decodable, so there is at most one sequence mapping into each code m-sequence and there are at most D m code m-sequences. Thus, a(m) D m , and we have

+ I)!

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4. Decide on your color palette, and stick with it. If your colors are navy, burgundy, and cream, don t make your links green. 5. Anti-alias your display fonts. The mark of a professional site can sometimes be as simple as the look of the fonts. 6. Be target-aware. Many sites make the mistake of providing hundreds of links to other sites. It s hard enough to get people to come to your site why send them away If you link to other sites, use the target =_blank attribute-value pair to open the link in a new window. This way, visitors can return to your site easily. Opening links in a separate window also solves anomalies that occur when your site has frames and you link to a site that also has frames. 7. People read from left to right and from top to bottom. This seems obvious, but inexperienced designers seem to have trouble remembering this. Put your most important elements at the top left of the page. If you don t grab the reader s attention there, you re never going to get them to read more. 8. Think carefully about bells and whistles. Animations, marquees, DHTML, and so forth are all great, when used in moderation. Don t pull all your tricks out of the bag and plop them all on one page. If your content isn t good enough to stand alone, rethink it.

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