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increased resolution is necessary to observe isotopic peaks when the charge state of the molecular ion is increased; the resolution provided by most instrumentation allows the observation of isotopic peaks only from singly to triply charged molecular ions Isotope peaks observed for proteins are mainly produced by the 12C and 13C mass difference of $1 mass unit Once one molecular ion charge state is determined, the other charge states are readily assigned by spectral observation ESI, MALDI, and FAB are usually used in the positive-ion mode, especially for protein MW determination; therefore the proton mass must be used to calculate the ion mass, as shown in Table 157 155.

Round-robin DNS is useful in rack-and-stack clusters, or shared-nothing clusters where you achieve redundancy by building out multiple instances of web servers (we cover rack-and-stack, or horizontal, clustering in 14). Each web server has its own filesystem, its own IP address and hostname, and its own interface to the world. DNS does the heavy lifting by associating multiple IP addresses with the virtual hostname; for example, you may assign four IP addresses to www.blah.com so that incoming traffic will go (fairly evenly) to all four addresses over time. Some implementations of DNS do this automatically, circulating through all available IP addresses for a given hostname. In other configurations, you re going to need to get a copy of the publicly available load-balancing BIND daemon (lbbindd). When a client gets an IP address via round-robin DNS, it will use it until the DNS lifetime has expired. Sites using round-robin DNS typically shrink the DNS lifetime to several minutes, forcing clients to rebind the hostname to an address more regularly and reducing the average window of an outage. Done well, round-robin DNS is a low-impact and low-budget solution with its own good and bad points, which are shown in Table 9.1

Figure 4.16. Example dashboard tracking compliance with Sarbanes-Oxley ( Axway, 2006)

We discuss the cost of downtime in other places around the book, but no discussion of the business impact of downtime and availability is complete

PROTEIN IDENTIFICATION The protein molecular mass is insuf cient information for identi cation, but it is adequate to con rm identity; therefore, MS is one of the preferred techniques for characterization and quality control of recombinant proteins and other biomolecules In the same way, it has been used to study posttranslational modi cations (like glycosylation and disul de bonding pattern), and other processes that can modify protein mass11 For identi cation purposes, two basic approaches have been used In the rst, a puri ed protein is enzymatically digested, the product peptide mixture is analyzed and a mass spectral pattern is obtained This pattern, called the MS ngerprint , is used to search in internet-available protein or DNA databases17 Information about protein origin and an estimate of its MW are required in order to improve the chances of a correct match.

without mentioning it once again. The Meta Group, an industry consulting firm, estimates the cost to a business of total system downtime at just over $1 million an hour. A University of Minnesota study says that a 48-hour outage would put 20 percent of the Fortune 500 out of business. And a study from Contingency Planning Research Inc. says that the average time to recover a business s systems after a disaster is four days. The University of Minnesota study also said that of companies that suffer a major outage from a disaster and do not have a working recovery plan, just 43 percent of them will ever resume operations. Of that 43 percent, just 29 percent of those will still be in business two years later. Relax, we ve done the math for you: 87 percent of companies that have a disaster and do not have an adequate plan will be out of business within 2 years. So, how long will it take your enterprise to recover Do you have a plan

Once you have learned what you need to know, the next step is to begin to put together a calm and rational case that explains in nontechnical terms what the vulnerabilities, risks, and costs are. The case must include a discussion of the risks of inaction.

The search algorithm then theoretically digests all appropriate proteins in the database with the speci ed enzyme, and matches the theoretical and experimental masses The best matches are ranked, and a con dence parameter is usually calculated18 The second approach uses peptide sequence tags to identify the peptide, and therefore the protein This method is based on the assumption that in a MS/MS peptide spectrum some consecutive amino acid residues can usually be clearly identi ed In a typical protocol, the peptides obtained from a protein tryptic digest are.