Cora K. Ogle

Beneficial effects of aggressive protein feeding in severely burned children.

To determine any potential benefit of feeding increased amounts of protein to hypermetbolic burned patients, 18 children with burns averaging 60% total surface area were randomized into two matched groups and studied serially for at least six weeks: the first group was given a normal diet with a balanced nutritional supplement, and the second group was supplemented with milk whey protein. The normal protein group received 87.1% of their desired caloric intake with 16.5% of calories from protein compared to 77.7% of desired caloric intake with 23.0% of calories from protein for the high protein group. Despite a higher caloric intake, the normal protein group had a worse opsonic index compared to the high protein group (0.42 +/- 0.04 vs. 0.62 +/- 0.05, p < 0.0007), lower levels of C3 (1371 +/- 55 vs. 1585 +/- 64 micrograms/ml, p < 0.01), lower levels of IgG (805 +/- 52 vs. 975 +/- 56 micrograms/ml, p < 0.03), lower levels of transferrin (200 +/- 10 vs. 283 +/- 18 mg/dl, p < 0.0001), lower levels of total serum protein (5.5 +/- 0.1 vs. 6.3 +/- 0.2 g/dl, p < 0.005), more bacteremic days (11% vs. 8%, p < 0.005) and worse survival (5/9--56% vs. 9/9--100%, p < 0.03). Patients receiving the high protein diet had significantly higher plasma levels of valine, lysine, threonine, leucine, aginine, isoleucine, proline, serine, asparagine, tryptophane, and tyrosine. Asparagine levels were significantly (p < 0.01) associated with better neutrophil function and opsonic index. Except for phenylalanine, significant associations were found for serum levels of each of the amino acids with concentrations of one or more serum proteins. These studies provide evidence that many immunologic functions are dependent upon optimal availability of specific amino acids, and that routine diets do not provide sufficient protein to satisfy the needs of seriously burned children.

The importance of lipid type in the diet after burn injury

: The effects of different types of dietary lipids were tested in burned guinea pigs. All diets were identical except for the type of lipid, with total energy intake from lipids equaling 10%. All animals received a 30% total body surface area (TBSA) flame burn and were fed identically by pump-controlled gastrostomy feedings for 14 days. When compared to safflower oil (74% linoleic acid) as well as linoleic acid alone, fish oil (18% eicosapentaenoic acid or EPA) administration resulted in less weight loss, better skeletal muscle mass, lower resting metabolic expenditure, better cell mediated immune responses, better opsonic indices, higher splenic weight, lower adrenal weight, higher serum transferrin, and lower serum C3 levels. With the exception of better cell mediated immune responses in the animals fed linoleic acid, the administration of indomethacin made little difference. These findings can be explained by a reduction in the synthesis of the dienoic prostaglandins that are derived from the omega 6 series of fatty acids, some of which are significantly immunosuppressive. Regulation of dietary lipids may be an important therapeutic advance in nutritional support after burn injury, and controlled trials should be considered.

Effect of Glutamine on Phagocytosis and Bacterial Killing by Normal and Pediatric Burn Patient Neutrophils

Glutamine is essential for the function of lymphocytes and macrophages, where it serves, among other things, as a source of energy. Little information is available concerning the fuel that polymorphonuclear cells use for their metabolic and bactericidal functions. It was the purpose of this study to determine whether glutamine would enhance the in vitro bactericidal function of normal neutrophils and whether the amino acid would restore the observed impaired function in burn patients to or above the normal level. Twelve burn patients with total body surface area burns ranging from 32% to 87% were studied. At various postburn times, neutrophils were isolated and their ability to kill Staphylococcus aureus in the presence and absence of glutamine was determined and compared with that in normal subjects. Glutamine enhanced the bactericidal function of normal neutrophils. In every patient, at all but two postburn times, glutamine caused an improvement in the observed abnormal neutrophil bactericidal function and often restored it to or slightly above the normal level. Glutamine had no effect on the expression of C3b receptors (CR1 or CD35) or on phagocytosis by the cells. This study confirms the beneficial effects of glutamine in at least one arm of the immune system and adds evidence for the possible advantage of including this amino acid in the diets of burn and other trauma patients.

Sepsis and endotoxemia stimulate intestinal interleukin-6 production

BACKGROUND Endotoxemia stimulates tumor necrosis factor (TNF) and interleukin-1 (IL-1) production in mucosa of the small intestine, but the effect on IL-6 production is not known. Intestinal IL-6 may be especially important, considering its role in the acute phase response. We tested the influence of endotoxemia and sepsis in mice on intestinal IL-6 and IL-6 messenger RNA (mRNA) levels. METHODS Mice were injected with lipopolysaccharide (LPS 10 mg/kg) or saline solution. In some experiments animals were pretreated with indomethacin (5 mg/kg) or N-nitro-L-arginine (NNA, 100 mg/kg) before LPS injection. In other experiments, sepsis was induced by cecal ligation and puncture (CLP); controls were sham operated. Serum and jejunal mucosa were harvested at intervals during 16 hours, and IL-6 levels were determined by enzyme-linked immunosorbent assay. IL-6 mRNA was detected by polymerase chain reaction. RESULTS Endotoxemia and sepsis increased serum and mucosal IL-6 and IL-6 mRNA, with maximum levels noted at 1 and 4 hours after LPS and at 8 hours after CLP. Pretreatment of endotoxemic mice with indomethacin or NNA blunted the increase in mucosal IL-6. CONCLUSIONS Results suggest that sepsis and endotoxemia stimulate IL-6 production in small intestinal mucosa and that this response may be transcriptionally regulated. The effect of endotoxemia may be partly mediated by prostaglandins and nitric oxide. The results also suggest that the intestinal mucosa may be a participant in the cytokine response, rather than just a passive bystander.

THE 1994 LINDBERG AWARD: The Production of Tumor Necrosis Factor, Interleukin-1, Interleukin-6, and Prostaglandin E2 by Isolated Enterocytes and Gut Macrophages: Effect of Lipopolysaccharide and Thermal Injury

Increasing evidence shows that cells other than immune cells have the potential for producing cytokines and arachidonate metabolites. It was the purpose of this study to determine whether isolated enterocytes could produce tumor necrosis factor, interleukin-1, interleukin-6, and prostaglandin E2, to compare the production with that of isolated gut macrophages, and to determine whether a difference existed in the production of these mediators after thermal injury. Guinea pigs received a 30% total body surface area burn and were killed 24 hours after injury. Isolated enterocytes and related intestinal macrophages (5 x 10(5) cells/ml) were cultured for 24 hours in the presence and absence of endotoxin, and the supernatants were assayed for the mediators. An increase was seen in production of interleukin-6 by enterocytes and by macrophages after thermal injury. In general enterocytes and gut macrophages produced about the same amounts of the different mediators. In contrast to macrophages from other tissues, enterocytes did not produce more prostaglandin E2 after stimulation with lipopolysaccharide, and with one exception gut macrophages did not produce larger amounts of mediators after stimulation with lipopolysaccharide. Enterocytes may be a significant source of immunomediator production and could contribute to the inflammatory response.

Heterogeneity of kupffer cells and splenic, alveolar, and peritoneal macrophages for the production of TNF, IL-1, AND IL-6

Kupffer cells and alveolar, splenic, and peritoneal macrophages from normal rats were incubated for various periods of time in the presence of LPS, and the culture supernatants were analyzed for IL-6, IL-1, and TNF. There was very little difference in the amounts of the cytokines produced by the macrophages when stimulated with 0.01–10μ/ml of LPS. The shapes of the time course curves for the production of the cytokines by the different types of macrophages were generally similar, although only Kupffer cells continued to produce IL-6 throughout the entire incubation period and splenic macrophages showed a lag period in the production of IL-1. Kupffer cells produced more IL-6 than that produced by the other populations of macrophages, and alveolar macrophages produced more IL-1 compared to that produced by splenic cells. Kupffer cells and peritoneal macrophages produced more IL-6 in 24 h than in 6 h of culture, and splenic macrophages produced more IL-1 in 24 compared to 6 h of culture. Alveolar macrophages produced more TNF than that produced by the other populations of cells but only when integrated over the entire incubation period. These results confirm and extend the observed functional heterogeneity of macrophages obtained from different tissues of the same animal. This study and future studies will lead to a better understanding of the role of cytokines in the inflammatory response.

Consumptive Opsoninopathy: Possible Pathogenesis in Lethal and Opportunistic Infections

Serum levels of properdin, Factor B and C3 and the ability of these sera to opsonize E. coli 075 were measured in 17 patients with surgical infections ranging in severity from mild to fatal. There was good direct correlation between severity of infection, serum levels of properdin and C3, and the ability of the serum to support opsonization. The levels of Factor B were not significantly reduced when measured by radial immunodiffusion, but Immunoelectrophoresis showed conversion. Restoration of full opsonic activity was accomplished only by the addition of a combination of C3, Factor B, and properdin in excess. The findings provide evidence that severe bacterial infection causes a consumption of opsonic proteins which may result in a reduced ability of the patients serum to opsonize bacteria and thereby further increase susceptibility to infection.

Enteral Feeding in Burn Hypermetabolism: Nutritional and Metabolic Effects of Different Levels of Calorie and Protein Intake

Enteral nutrition was provided by continuous pump-controlled gastrostomy tube feeding for 14 days in 97 guinea pigs bearing a 30% full thickness burn. Seven defined combinations of caloric and protein intake were studied. With a caloric intake of 175 kcal/kg/day, equaling the measured energy expenditure, the animals receiving 10% of calories as protein had a significantly greater postburn weight loss (p less than 0.05) and muscle mass depletion (p less than 0.05), and a significantly lower muscle nitrogen concentration (p less than 0.05), serum albumin level (p less than 0.01) and liver nitrogen content (p less than 0.01). With the same caloric intake but with more than 20% of calories as protein, the weight loss and the muscle wasting were reduced, but not abolished, and the serum albumin level and liver nitrogen content were normalized. Also with the diets containing 200 kcal/kg/day the muscle tissue depletion could not be abolished. However, with this caloric intake, the animals given 20% of calories as protein had a lower weight loss and a higher serum albumin level (p less than 0.01), but also a greater fat infiltration of the liver (p less than 0.01). At both levels of caloric intake, the nitrogen balance correlated significantly with the level of nitrogen intake but did not correlate with the changes of body weight. The incidence of diarrhea was lowest in animals fed 20% protein calories at a caloric intake of 175 kcal/kg/day. All things considered, the best metabolic and nutritional results were obtained with diets containing 20 to 30% of calories as protein and providing a caloric intake that paralleled the measured energy expenditure.

Increased synthesis of secreted hepatic proteins during abdominal sepsis.

To study the effect of intraabdominal sepsis on hepatic protein synthesis, male Sprague-Dawley rats underwent celiotomy with either cecal ligation and puncture (CLP) or sham operation. Eight and sixteen hours later total hepatic protein synthesis was measured by flooding dose technique. Specific synthetic rates of structural or secreted hepatic proteins were further studied 16 hr after CLP in an isolated perfused liver model. Total hepatic protein synthesis was significantly elevated at 16 hr (59 +/- 6%/day vs 37 +/- 6%/day, P less than 0.05), but not 8 hr post-CLP. Structural hepatic protein synthesis was unchanged after CLP; however, the synthetic rates of the acute-phase secretory proteins alpha 1-acid glycoprotein, transferrin and complement component C3 were significantly increased 16 hr after CLP. However, the albumin synthetic rate was not increased during sepsis. We conclude that sepsis causes augmentation of hepatic protein synthesis primarily to increase acute-phase proteins for host defense.

γδ T cells mitigate the organ injury and mortality of sepsis

Sepsis is a difficult condition to treat and is associated with a high mortality rate. Sepsis is known to cause a marked depletion of lymphocytes, although the function of different lymphocyte subsets in the response to sepsis is unclear. γδ T cells are found largely in epithelial‐rich tissues, and previous studies of γδ T cells in models of sepsis have yielded divergent results. In the present study, we examined the function of γδ T cells during sepsis in mice using cecal ligation and puncture (CLP). Mice deficient in γδ T cells had decreased survival times and increased tissue damage after CLP compared with wild‐type mice. Furthermore, bacterial load was increased in γδ T cell‐deficient mice, yet antibiotic treatment did not change mortality. Additionally, we found that recruitment of neutrophils and myeloid suppressor cells to the site of infection was diminished in γδ T cell‐deficient mice. Finally, we found that circulating levels of IFN‐γ were increased, and systemic levels of IL‐10 were decreased in γδ T cell‐deficient mice after CLP compared with wild‐type mice. γδ T cell‐deficient mice also had increased intestinal permeability after CLP compared with wild‐type mice. Neutralization of IFN‐γ abrogated the increase in intestinal permeability in γδ T cell‐deficient mice. The intestines taken from γδ T cell‐deficient mice had decreased myeloperoxidase yet had increased tissue damage as compared with wild‐type mice. Collectively, our data suggest that γδ T cells modulate the response to sepsis and may be a potential therapeutic target.