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The amount of HDV RNA in the culture medium of cells treated with the indicated amount of FTI-277 was quantitated using a phosphorimager and plotted as percentage of the untreated control (0 mM) (black bars right panel) To assess for nonspeci c effects of the inhibitor, prior to total RNA extraction, XTT assays were performed to monitor cell metabolism (grey bars right panel) and supernatant HBV surface antigen (HBsAg) levels were determined to monitor for general protein expression and secretion (empty bars right panel) [Part (a) reproduced with permission from the Journal Virology25] (b) In vivo treatment of HDV viremia with the prenylation inhibitor FTI-2153 HBV transgenic mice were hydrodynamically transfected with an HDV genome-encoding construct to establish HDV viremia29 Following transfection, the mice were treated with carrier alone (solid circles) or carrier plus FTI-2153 (open circles) for the indicated number of days prior to sacri ce.

It is important that compounds that cause primary skin irritation be tested for skin sensitization at concentrations low enough that the two effects are not confused Phototoxicity tests are designed to evaluate the combined dermal effects of light (primarily ultraviolet [UV] light) and the chemical in question Tests have been developed for both phototoxicity and photoallergy In both cases the light energy is believed to cause a transient excitation of the toxicant molecule, which, on returning to the lower energy state, generates a reactive, free-radical intermediate In phototoxicity these organic radicals act directly on the cells to cause lesions, whereas in photoallergy they bind to body proteins These modi ed proteins then stimulate the immune system to produce antibodies, because the modi cations cause them to be recognized as foreign or nonself proteins.

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We refer to such a charging scheme as transaction pricing Transaction pricing is widely used in the telephone network (with the notable exception of local networks in North America), where switches and links are sized to ensure that congestion occurs only exceptionally The price must be set at a value allowing the network operator to recover the cost of investment Differential pricing according to the time of day is used to smooth out the demand pro le to some extent but this is not generally viewed as a congestion control mechanism Choice between at rate pricing, congestion pricing, and transaction pricing depends among other things on their ability to assure the economic viability of the network provider Congestion pricing is intended to optimize the use of a network, not to recover the cost of installed infrastructure, which is regarded as a ``sunk cost'' in the economic optimization.

These tests are basically modi cations of the tests for primary irritation and sensitization except that, following application of the test chemical, the treated area is irradiated with UV light The differences between the animals treated and irradiated and those treated and not irradiated is a measure of the phototoxic effect..

Serum HDV RNA was quantitated by RT PCR and normalized for transfection ef ciency by quantitation of total HDV liver RNA See text for details [Part (b) reproduced with permission from Journal of Clinical Investigation29].

If the network is well provisioned and always offers good quality of service, for example, costs must be entirely recovered by at rate access charges Transaction pricing has proved successful for telephone network operators, but then so has at rate pricing in the case of North American local networks Transaction pricing has the advantage of distributing the cost of shared network resources in relation to usage In addition to being appealing from a fairness point of view, this is in line with the trend in telecommunications for ``unbundling'' and cost related pricing A second major issue is the complexity of implementing the different schemes Any move from at rate pricing appears as a major change for the Internet, requiring accounting and billing systems at least as complex as those of the telephone network The cost of such systems must be weighed against any expected improvements in ef ciency.

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