Protein quality of enteral nutrition products is consistent with label claims during shelf life and beyond expiration date.

OBJECTIVE: Eternal formulas were monitored during their shelf life and beyond expiration date to examine protein quality. DESIGN: Protein quality was determined by protein efficiency ratio (PER) bioassays and amino acid analyses. SETTING: A certified laboratory performed the PER tests according to procedures established by the Association of Official Analytical Chemists and recognized by the US Food and Drug Administration (FDA). The amino acid analyses were performed in our laboratory using validated methods. SAMPLES: Commercially available formulas (Ensure, Osmolite HN, TwoCal HN) that contained protein blends of caseinates or caseinates with soybean protein isolate were studied. MAIN OUTCOME MEASURES: Achievement of protein-quality values greater than or equal to 70% of the fresh reference casein value as determined by the PER method would be consistent with adequate protein quality as described by the FDA. Levels of indispensable amino acids that meet or exceed the standards established by the National Research Council of the National Academy of Sciences are considered high-quality proteins. Levels of amino acids throughout shelf life were compared with published label claims. RESULTS: Amino acid analyses, which included measurement of tryptophan and total sulfur amino acids, revealed that both fresh and outdated products met or exceeded standards for proteins of high biologic value and were consistent with label claims. The PER values ranged from 90% to 96% of the control diet for fresh product and 82% to 87% for products evaluated after expiration. CONCLUSION: The enteral products studied provide high-quality protein throughout the shelf life of the product.

Enteral formula composition does not affect response to lethal infectious challenge in mice.

The effects of enteral formulations on the response of mice to infectious challenge with Listeria monocytogenes, influenza A or Candida albicans were studied to test the efficacy of specialized ingredients. CF-1 outbred female mice (12-15 g) were fed nonpurified diet (Purina No. 5002) or commercially available liquid formulas: Osmolite HN, Perative or Impact. There were no differences between the groups fed the liquid formulas with regards to mean survival time or percentage of survivors in any of these models of infection. Examination of spleens from the groups challenged with L. monocytogenes, lungs from mice infected with Influenza A and kidneys from the groups challenged with C. albicans revealed no differences in cure rate of survivors. Pre-feeding periods of up to 8 d before infection produced similar results for mice fed enteral formulations compared to nonpurified diet. Contrary to previous reports, the use of Impact did not improve resistance to disease in mice challenged with lethal doses of L. monocytogenes, as compared with mice fed Osmolite HN. Additionally, mice fed Impact, Perative, or nonpurified diet responded similarly to challenge with L. monocytogenes, C. albicans or influenza A. The results indicate that these acute lethal animal models of infectious challenge may be of limited use to distinguish effects of modified nutrient composition of enteral formulas.

Comparison of specialized and standard enteral formulas in trauma patients.

STUDY OBJECTIVE: To compare selected nutrition and immunologic markers and infection in trauma patients receiving a specialized enteral formula with those receiving standard enteral therapy. DESIGN: Prospective, randomized clinical trial. SETTING: Level 1 trauma center at a county government hospital. PATIENTS: Forty-one consecutive patients with major trauma who required enteral nutrition support. Thirty-seven patients completed the study. Four patients (two in each group) were excluded, as additional operative procedures prevented initiation of enteral feedings within 7 days of injury. INTERVENTIONS: Nineteen patients fed the specialized enteral formula received supplemental arginine, linolenic acid, beta-carotene, and hydrolyzed protein for up to 10 days. Eighteen control patients received standard enteral nutrition. MEASUREMENTS AND MAIN RESULTS: After study entry, patients who received the specialized enteral formula had fewer infections than those receiving standard enteral nutrition (3/19 vs 10/18; p < 0.05). The change in nitrogen balance was significantly better (p < 0.05) from day 1 (-11.8 +/- 1.8 g/day) to day 5 (-5.9 +/- 2.0 g/day) for the group who received the specialized formula compared with the group who received standard enteral nutrition (-7.3 +/- 1.7 g/day to -7.4 +/- 2.8 g/day). Similarly, the change in C-reactive protein serum concentration was significantly better (p < 0.05) from day 1 (18.0 +/- 2.1 mg/dl) to day 5 (11.8 +/- 1.5 mg/dl) in the group who received the specialized formula compared with the group who received standard enteral nutrition (17.6 +/- 1.2 mg/dl to 14.4 +/- 1.7 mg/dl). The CD4:CD8 ratio increased more in the group who received the specialized formula, although this difference did not reach statistical significance. CONCLUSION: Trauma patients who received the specialized enteral formula demonstrated a decreased incidence of infection and increased improvements in nitrogen balance and other indexes of stress. Additional clinical trials demonstrating positive patient outcomes are necessary before these specialized enteral formulas are used as the standard of practice in critically ill patients.

Unmasking of GDP binding sites on hamster brown adipose tissue mitochondria and uncoupling protein.

1. A rapid unmasking of GDP binding sites on brown adipose tissue (BAT) mitochondria was observed when hamsters acclimatized to 28 degrees C were exposed to a temperature of 4 degrees C for 2 hr. 2. No rapid unmasking of GDP binding sites was observed when hamsters housed at 22 degrees C were briefly exposed to 4 degrees C. 3. The amount of GDP bound to BAT mitochondria from hamsters increased during 2 weeks of exposure to 4 degrees C, but did not change between 2 weeks and 30 days of cold exposure. 4. Incubation of mitochondria with 10 mM Mg2+ prior to the GDP binding assay increased the subsequent GDP binding to BAT mitochondria from hamsters housed at 28, 22 or 4 degrees C, albeit to different degrees. 5. The amount of GDP bound to uncoupling proteins isolated from untreated and Mg(2+)-treated mitochondria of hamsters and rats was measured. Scatchard analyses of the binding of GDP to purified uncoupling protein indicate that increases in the number of binding sites due to Mg2+ treatment of mitochondria do not change the affinity of the protein for GDP.

The concentration of uncoupling protein correlates with Mg2+ activated mitochondrial binding of GDP.

Rats were housed at 4 degrees C for periods of up to 26 days. As little as 2 h of cold exposure caused an increase in the binding of [3H]GDP to mitochondria from brown adipose tissue. Incubation of mitochondria in vitro with 10 mM Mg2+ caused a marked increase in the subsequent binding of GDP to mitochondria from rats housed at 28 degrees C and a smaller increase in that from rats exposed to 4 degrees C for 2 h. Chronic exposure to cold led to an even greater increase in the amount of GDP bound to mitochondria incubated with Mg2+. The time course for the increase in the concentration of uncoupling protein was compared with that for GDP binding to mitochondria with and without Mg2+ treatment. The concentration of uncoupling protein appears to be correlated with the GDP-binding values for mitochondria treated with Mg2+ (r = 0.70) but not with the GDP binding to untreated mitochondria (r = 0.36). Therefore, the binding of GDP to untreated mitochondria may represent thermogenic activity at the time of death, whereas that after treatment with Mg2+ may more closely reflect total thermogenic capacity of the mitochondrion.

Immunochemical detection and quantitation of brown adipose tissue uncoupling protein.

A polyclonal antisera against rat brown adipose tissue mitochondrial uncoupling protein was used to examine mitochondrial samples from liver and white and brown adipose tissue from several mammalian species. A sodium dodecyl sulfate--polyacrylamide gel electrophoretic separation of proteins combined with an immunochemical method allowed for visualization of antigen--antibody complexes on nitrocellulose blots. Hamster, cavy, monkey, and mouse brown adipose tissue mitochondrial samples cross-reacted with the antisera. Mitochondria prepared from white fat obtained from young swine and sheep contained two closely migrating, antigenically active proteins. Hepatic mitochondria samples did not contain antigenically active protein. Reflectance densitometry was used for quantitation of the uncoupling protein in various mitochondrial samples. In rats fed diets low in protein, there appears to be a dissociation between the concentration of uncoupling protein and the number of nucleotide binding sites as given by the [3H]GDP binding assay. These results are indicative of a physiological activation of the uncoupling protein.

Evidence for unmasking of rat brown-adipose-tissue mitochondrial GDP-binding sites in response to acute cold exposure. Effects of washing with albumin on GDP binding.

Rats, previously acclimated to 29 degrees C, were moved into the cold (4 degrees C) for 2 h. Scatchard analysis of GDP binding to the brown-adipose-tissue mitochondria of these animals showed a 2.3-fold increase in the number of high-affinity sites and a 1.5-fold increase in the number of low-affinity sites compared with binding in animals maintained at 29 degrees C. Immunochemical determination showed no increase in the amount of mitochondrial uncoupling protein during this period. This strongly suggests an unmasking of existing GDP-binding sites before a detectable increase in synthesis of uncoupling protein can occur. Washing with albumin increased the number of GDP-binding sites of brown-adipose-tissue mitochondria from both warm-housed and cold-exposed animals to the same extent. This indicates that the effects of washing with albumin and cold exposure are independent and additive.