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1.2. Advantages and Shortcomings Lipid-based transfection offers many advantages over other transfection systems to introduce genes of interest into primary articular chondrocytes. The high transfection ef ciency avoids the need for cell selection that might result in undesired phenotypic drift, as observed after prolonged monolayer culture of articular chondrocytes [Benya et al., 1978]. The very low toxicity avoids the need to change the cell culture medium after transfection. In addition, lipid-based transfections can be carried out in the presence of serum. The main shortcoming of lipid-based transfection is its relatively high cost, especially when large-scale transfections are required. 1.3. Critical Factors for Ef cient Transfection Several factors greatly in uence the success of lipid-based transfection with FuGENE 6. The starting point is to identify the optimal quantity of DNA per cell. For chondrocytes cultured in a 24-well plate, 1 g of DNA per well is commonly

Anonymous 2008 Fresh vs Frozen Steaks, USDA Prime Beef and Black Angus, Online Buy steak for mail order delivery MyButchercom Accessed 14 October 2008 http://wwwmybutchercom/stores/xq/ xfm/Store_id634/page_id61/qx/storehtm ASHRAE 2006 ASHRAE Handbook Refrigeration Atlanta, Ga: American Society of Heating, Refrigerating and Air-Conditioning Engineers Bech-Jacobsen, K, and L B gh-S rensen 1984 TTTStudies of different retail packed pork products In Thermal Processing and Quality of Foods, edited by P Zeuthen, J C Cheftel, C Eriksson, M Jul, H Leniger, P Linko, G Varela, and G Vos London: Elsevier Applied Science Bendall, J R 1972 The in uence of rate of chilling on the development of rigor and cold shortening In Meat Chilling Why and How Meat Research Institute Symposium No 2, C L Cutting, 31 36 Langford, UK: Meat Research Institute Bendall, J R 1974.

nuclear reactor meltdown at Three Mile Island (TMI) in 1979, psychological assessments revealed that local TMI residents reported more symptoms of distress and intrusive thoughts and continued to have higher blood pressure, heart rate, norepinephrine, and cortisol levels than control subjects who lived 80 miles away, up to ve years after the accident (Davidson & Baum, 1986) TMI residents also had fewer B lymphocytes, T-suppressor/cytotoxic lymphocytes and NK cells, as well as evidence for reactivation of latent HSV (McKinnon, Weisse, Reynolds, Bowles, & Baum, 1989) In the aftermath of the Northridge earthquake, local residents similarly showed a decrease in T cell numbers, blastogenic responses, and NKCC (Solomon, Segerstrom, Grohr, Kemeny, & Fahey, 1997) Chronic stress can have signi cant clinical consequences As previously mentioned, caregivers and TMI residents showed evidence for reactivation of latent herpes viruses (Glaser & Kiecolt-Glaser, 1997; McKinnon et al, 1989).

Following in uenza vaccination, caregivers were less likely to achieve a four-fold increase in antibody titers than controls (Kiecolt-Glaser, Glaser, et al, 1996; Vedhara et al, 1999), which suggests greater susceptibility or more serious illness in the event of exposure to in uenza virus Caregivers also took 24% longer to heal a standardized punch biopsy wound (Kiecolt-Glaser, Marucha, Malarkey, Mercado, & Glaser, 1995) and reported a greater number and duration of illness episodes, with more physician visits than control subjects (Kiecolt-Glaser & Glaser, 1991) The immune dysregulation associated with caregiving may be especially relevant for older adults, as cellular immunity declines with age (Bender, Nagel, Adler, & Andres, 1986; Murasko, Weiner, & Kaye, 1987), and is associated with greater morbidity and mortality, especially due to infectious diseases (Murasko, Gold, Hessen, & Kaye, 1990; Wayne, Rhyne, Garry, & Goodwin, 1990).

However, even in younger populations, longer term stress (greater than one month) has been associated with immune dysregulation and increased susceptibility to infection by a common cold virus (Cohen et al, 1998) The studies mentioned support the argument that immunological dysregulation associated with chronic stress does not necessarily undergo habituation over time Rather, these effects appear to be present for the duration of the stressor, and in some cases, persist even after the stressor is no longer present (Esterling, Kiecolt-Glaser, Bodnar, & Glaser, 1994)..

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The snags and snares of freezing rapidly after slaughter In Meat Freezing: Why and How Meat Research Institute Symposium No 3, edited by C L Cutting Langford, UK: Meat Research Institute Bernholdt, H F 1974 Merchandising frozen meats and consumer attitudes in purchasing and preparation In Meat Freezing Why and How Meat Research Institute Symposium No 3, edited by C L Cutting Langford, UK: Meat Research Institute Buts, B, M Casteels, E Claeys, and D Demeyer 1986 Effects of electrical stimulation, followed by moderate cooling, on meat quality characteristics of veal Longissimus dorsi Meat Science 18:271 279 Cornforth, D 1994 Color Its basis and importance In Advances in Meat Research Series, Volume 9, Quality Attributes and Their Measurement in Meat, Poultry and Meat Products, A M Pearson, and T R Dutson London: Blackie Academic and Professional Critchell, J T, and J Raymond 1912.