Paul B. Spalding

Splanchnic perfusion evaluation during hemorrhage and resuscitation with gastric near-infrared spectroscopy

OBJECTIVE The purpose of this study was to use a prototype side-illuminating near-infrared spectroscopy (NIRS) nasogastric probe to continuously measure changes in gastric tissue oxygen saturation (Sto2) in a pig hemorrhage model. METHODS Swine (n = 12; 6 per group) underwent laparotomy and placement of a gastric NIRS probe, jejunal tonometer, superior mesenteric artery (SMA) flow probe, and a portal vein catheter. Animals underwent hemorrhage (28 mL/kg) t = 0 to 20 minutes (where t = time). Pigs in group I were resuscitated (t = 20-40 minutes) with lactated Ringers solution (84 mL/kg), whereas group II had no resuscitation. RESULTS A significant decrease in mean arterial pressure and SMA flow was observed after hemorrhage. SMA flow significantly correlated in group I with both NIRS Sto2 (r = 0.58, p = 0.0001) and regional CO2 (r = -0.54, p = 0.0001). In group II, superior mesenteric flow correlated with NIRS Sto2 (r = 0.30, p = 0.03), but not regional CO2 (r = -0.23, p = 0.09). CONCLUSION Direct measurement of tissue oxygen saturation with a prototype side-illuminating near-infrared spectroscopy gastric probe appeared to rapidly reflect changes in splanchnic perfusion.

Body surface area prediction in normal, hypermuscular, and obese mice.

BACKGROUND Accurate determination of body surface area (BSA) in experimental animals is essential for modeling effects of burn injury or drug metabolism. Two-dimensional surface area is related to three-dimensional body volume, which in turn can be estimated from body mass. The Meeh equation relates body surface area to the two-thirds power of body mass, through a constant, k, which must be determined empirically by species and size. We found older values of k overestimated BSA in certain mice; thus we determined empirically k for various strains of normal, obese, and hypermuscular mice. MATERIALS AND METHODS BSA was computed from digitally scanned pelts and nonlinear regression analysis was used to determine the best-fit k. RESULTS The empirically determined k for C57BL/6J mice of 9.82 was not significantly different from other inbred and outbred mouse strains of normal body composition. However, mean k of the nearly spheroid, obese lepr(db/db) mice (k = 8.29) was significantly lower than for normals, as were values for dumbbell-shaped, hypermuscular mice with either targeted deletion of the myostatin gene (Mstn) (k = 8.48) or with skeletal muscle specific expression of a dominant negative myostatin receptor (Acvr2b) (k = 8.80). CONCLUSIONS Hypermuscular and obese mice differ substantially from normals in shape and density, resulting in considerably altered k values. This suggests Meeh constants should be determined empirically for animals of altered body composition. Use of these new, improved Meeh constants will allow greater accuracy in experimental models of burn injury and pharmacokinetics.

Involvement of CD44 and the cytoskeletal linker protein ankyrin in human neutrophil bacterial phagocytosis.

The leukocyte CD44 and CD45 cell surface receptors are associated via the linker proteins ankyrin and fodrin with the cytoskeleton, which itself is important in immune cell functions such as adherence, chemotaxis, and phagocytosis. The effects of rat antihuman CD44 and CD45 monoclonal antibodies on phagocytosis of fluoresceinated heat‐killed Staphylococcus aureus 502A by normal human neutrophils (PMNs) during 2 hr incubation in RPMI‐1640 was studied via flow cytometry and confocal microscopy. Flow cytometry was performed using an excitation wavelength of 488 nm, fluorescence being measured at 515–560 nm on 50,000 PMNs per sample. Confocal microscopy was performed on samples after further incubation with rhodamine‐conjugated antiankyrin. Anti‐CD44 resulted in an increase of 27–31% compared to control (P = 0.004) in the proportion of PMNs fluorescing, an increase of 17–24% (P = 0.001) in mean intracellular fluorescence per PMN, and an increase in total PMN fluorescence of 50–58% compared to control (P < 0.001). In contrast, anti‐CD45 had little effect on phagocytosis. Colchicine (a microtubule‐disrupting agent) enhanced, whereas cytochalasin‐D (a microfilament inhibitor) inhibited bacterial phagocytosis; cytochalasin‐D completely abrogated the effect of anti‐CD44 on this PMN function. Hyaluronic acid augmented phagocytosis by an increment similar to that observed with anti‐CD44. Two‐color flow cytometry and confocal microscopy demonstrated that ankyrin always colocalized with ingested fluorescein isothiocyanate (FITC)‐labeled bacteria. These data strongly suggest that CD44 is involved in bacterial phagocytosis, provide further evidence of CD44 receptor linkage to cytoskeletal elements in human leukocytes, and suggest that ankyrin has a significant role in the transport of phagosomes.

The role of Eh, pH and iron in the bactericidal power of human plasma

The bactericidal power of fresh human plasma against Klebsiella pneumoniae and Escherichia coli was extremely sensitive to changes in Eh and pH. At a high Eh (approx. +200 mV) the bacteria were destroyed, but rapid regrowth occurred when the Eh was lowered to approx. -400 mV. Abolition of the bactericidal effect was also produced by adding ferric iron at a high Eh (approx. +200 mV). Lowering the pH to 6.50 reduced or prevented the bactericidal effect. These results are probably related to the availability of iron for bacterial growth, and could be important for understanding the development of infection in injured or diseased tissue.

The bactericidal power of the blood and plasma of patients with burns.

Patients with burns are unusually susceptible to bacterial infections, but so far there is no satisfactory explanation for this lack of resistance. Since resistance to infection involves many different mechanisms, examination of individual components of the immune system may not sufficiently explain the underlying reasons for increased susceptibility. The use of whole blood for antibacterial tests has the advantage that all the immune systems present in that fluid compartment can take part in the bactericidal effect. Tests with Klebsiella pneumoniae and Staphylococcus aureus showed no evidence that the bactericidal power of the blood and plasma of patients with burns was less than that of normal control plasma. This suggests that the solution to the problem of increased susceptibility to infection in patients with burns does not lie with the blood but must be looked for elsewhere.

High-fat diets suppress CD3 and CD25 expression on the surface of murine lymphocytes.

To determine the effects of dietary fats on surface antigen expression, we tested the effects of amount and type of dietary fat on murine lymphocytes. Mice were fed diets with 12 en%, 23 en%, or 47 en% fat containing coconut, olive, safflower, or linseed oil. After 2 wk of ad libitum feeding, the mice were killed and splenic lymphocytes were harvested. Lymphocytes were incubated with fluorescent-tagged monoclonal antibodies and assayed for mean and total surface expression using flow cytometry. Our results show that high-fat (47 en%) diets suppress expression of CD3 and CD25 antigens. We also found that linseed-oil diets suppress expression of CD11a but enhance expression of CD25 antigens. Both CD3 and CD25 are critical for lymphocyte activation, and we conclude that immunosuppression associated with high-fat diets may be associated with suppression of these surface antigens.