No abstract

1. Erythrocytes are known to haemolyse in vitro at 48-50 degrees C. We hypothesized that erythrocytes might be damaged at much lower temperatures if they are incubated for prolonged periods. Erythrocytes from healthy human donors (n = 7) were incubated at 37, 40, 42, 44, 46 or 48 degrees C for 4-48 h. The haemolytic percentage and osmotic fragility were then measured by a modification of the method of Parpart et al. 2. Significant haemolysis and increased fragility were not observed at any temperatures after incubation for 4 h. However, the haemolytic percentage increased after incubation for 24 h at 44 degrees C (9.1 +/- 4.9%, P less than 0.01), 46 degrees C (52.4 +/- 14.1%, P less than 0.01) and 48 degrees C (98.0 +/- 2.6%, P less than 0.01) and after incubation for 48 h at 42 degrees C (9.8 +/- 4.5%, P less than 0.01) when compared with the values before heating (1.1 +/- 0.9%). The osmotic fragility also increased after incubation for 24 h at and above 42 degrees C. 3. Although heat-induced haemolysis and an increase in fragility have not been known to occur below 48 degrees C, these were quite apparent after incubation for 24-48 h at only 42 degrees C. This suggests that with regard to thermal effects, it is important to consider not only the temperature but also the duration of heating.

Perioperative changes of neutrophil function were investigated in 28 patients who underwent major surgery, particularly focusing on the potential capacity for superoxide and leukotriene production, which seem to be important in host defence. The superoxide-producing capacity of neutrophils, which was examined using N-formyl-peptide or phorbol myristate acetate as an attractant, significantly decreased postoperatively to 55 and 69 per cent of the pre-operative values, respectively. The leukotriene-producing capacity of neutrophils, which was stimulated with calcium ionophore A23187 in the presence of arachidonic acid, significantly changed postoperatively. The leukotriene B4 (LTB4) production increased together with an increment of production of 6-trans 6-trans LTB4, however, increased postoperatively to 1.2 times the (LTC4) production decreased postoperatively. The total production of leukotriene A4 (LTA4) metabolites consisting of LTB4, LTC4, and 6-trans LTB4, however, increased postoperatively to 1.2 times the pre-operative values. This indicates that neutrophils in the postoperative period have a higher capacity for LTA4 production but a lower capacity for superoxide production than those in the pre-operative period.

Antithrombin (AT) reveals its antiinflammatory activity by promoting endothelial release of prostacyclin (PGI(2)) in vivo. Since neuroinflammation is critically involved in the development of ischemia/reperfusion (I/R)-induced spinal cord injury (SCI), it is possible that AT reduces the I/R-induced SCI by attenuating the inflammatory responses. We examined this possibility using rat model of I/R-induced SCI in the present study. AT significantly reduced the mortality and motor disturbances by inhibiting reduction of the number of motor neurons in animals subjected to SCI. Microinfarctions of the spinal cord seen after reperfusion were markedly reduced by AT. AT significantly enhanced the I/R-induced increases in spinal cord tissue levels of 6-keto-PGFIalpha, a stable metabolite of PGI2. AT significantly inhibited the I/R-induced increases in spinal cord tissue levels of TNF-alpha, rat interleukin-8 and myeloperoxidase. In contrast,Trp(49) -modified AT did not show any protective effects. Pretreatment with indomethacin significantly reversed the protective effects of AT. An inactive derivative of factor Xa, which selectively inhibits thrombin generation, has been shown to fail to reduce SCI. Taken together, these observations strongly suggested that AT might reduce I/R-induced SCI mainly by the antiinflammatory effect through promotion of endothelial production of PGI(2). These findings also suggested that AT might be a potential neuroprotective agent.

The use of cryopreserved aortic allografts in cardiovascular surgery is widespread and has resulted in excellent outcomes. However, it is controversial whether cryopreservation suppresses the antigenicity of tissue. We designed experimental models to study whether the cryopreservation process alters antigenicity in comparison with that found in fresh and glutaraldehyde treated tissues. Fresh, cryopreserved, and glutaraldehyde treated thoracic aorta from Brown Norway rats were subcutaneously implanted into Lewis rats. Inflammatory cells infiltrating around the grafts were measured on days 7, 14, 28, and 56 after implantation. The glutaraldehyde treated grafts showed significantly less infiltration than the fresh or cryopreserved grafts (p < 0.005). No significant difference was detected between the fresh and cryopreserved grafts. Another study examined the effect of modifications of the aortic allograft on subsequent allogeneic skin graft antigenicity. Subcutaneous implantation of fresh, cryopreserved, and glutaraldehyde treated aortic grafts from Brown Norway into Lewis rats resulted in subsequent skin graft rejection at 4.4+/-0.7, 5.1+/-0.8, and 6.6+/-2.1 days, respectively. There was no significant difference between the fresh and cryopreserved groups; whereas skin grafts in the glutaraldehyde group survived longer than those in the cryopreserved group. These results indicate that cryopreservation had no significant influence on antigenic suppression of arterial allografts.