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Sawada et al [574 576] characterized the iso-pH 74 MDCK permeabilities of very lipophilic molecules, including chlorpromazine (CPZ) [574] They included 3% wt/vol bovine serum albumin (BSA) on the apical (donor) side, and 01 3% BSA on the basolateral (acceptor) side, and found that plasma protein binding greatly affected the ability of molecules to permeate cellular barriers They observed cell tissue retention of CPZ ranging within 65 85%, depending on the amount of BSA present in the receiving compartment They concluded that the rapid rate of disappearance of lipophilic compounds from the donor compartment was controlled by the unstirred water layer (UWL; see Section 776), a rate that was about the same for most lipophilic compounds; however, the very slow appearance of the compounds in the receiving compartment depended on the rate of desorption from the basolateral side of the membranes, which was strongly in uenced by the presence of serum proteins in the receiving compartment They recommended the use of serum proteins in the receiving compartment, so as to better mimic the in vivo conditions when using cultured cells as in vitro assays Yamashita et al [82] also studied the effect of BSA on transport properties in Caco-2 assays They observed that the permeability of highly lipophilic molecules could be rate limited by the process of desorption off the cell surface into the receiving solution, due to high membrane retention and very low water solubility They recommended using serum proteins in the acceptor compartment when lipophilic molecules are assayed (which is a common circumstance in discovery settings) 744 Effects of Cosolvents, Bile Acids, and Other Surfactants

In deploying your site, you'll rarely (if ever) have any direct control over whether or how clients cache your content You won't know what browser they're using or what kind of intermediate caching systems exist between you and them How, then, should you annotate the content you serve Should you use the Last-Modified or the Expires field Maybe the ETag field instead As usual, it depends on the particulars of your deployment However, I highly suggest that you

Figure 713 shows some of the structures of common bile acids In low ionic strength solutions, sodium taurocholate forms tetrameric aggregates, with critical

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Assume the potential for some caching between you and the client Attempt to support both HTTP 10 and 11 caching models To do that, you should construct your responses so that they contain valid values for the Expires or the Last-Modified field, the ETag field, and the Cache-Control field

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One tricky issue is choosing the Expires or the Last-Modified field It can be argued that Last-Modified is safer than Expires because it limits the server's commitment to the consistency scheme It places the burden on the client to validate the content, and the nature of conditional GET requests is such that the overhead for this validation is insignificant (compared to the content, which may be substantial) True, this approach requires an HTTP request and response and is therefore not as scalability friendly as the Expires approach However, it provides a safe way to encourage caching by both HTTP 10 and 11 clients In contrast, the Expires approach suffers from the genie-out-ofthe-bottle syndrome: Once you release an object with Expires, you have to wait the specified time before assuming that clients will update their copy Such a risk may be too costly depending on the nature of your application Of course, there are clear advantages to Expires First of all, it eliminates the need to make even a conditional request on a resource A smart client or an intermediary can use an Expires tag set to a future date to avoid the need to have the server validate its content

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Expires is really the only option for dynamic content Recall that it's possible for clients and intermediaries to cache GET responses even if they're dynamically generated (from CGI scripts,

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