Critical Care Vol 4 Suppl 1 20th International Symposium on Intensive Care and Emergency Medicine
P56 Role of platelet activating factor (PAF) on leukocyte-independent plasma extravasation and mast
cell activation during endotoxemia
A Walther, N Yilmaz, W Schmidt, A Secchi, MM Gebhard*, E Martin and H Schmidt
Department of Anesthesiology, and *Department of Experimental Surgery, Ruprecht-Karls-University, Im Neuenheimer Feld 110,
69120 Heidelberg, Germany
Introduction: Independently from leukocyte adherence,
endothelial factors and mast cell activation promote
microvascular permeability . The platelet activating
factor (PAF) has been shown to play a significant role in
endotoxin-induced leukocyte adherence . The aim of
our study was to investigate if there is also a role of PAF in
mediating leukocyte-independent microvascular perme-
ability changes and activation of mast cells during endo-
toxemia. Therefore, during endotoxemia, microvascular
permeability and mast cell activation were determined
after inhibition of the L-selectin mediated leukocyte-adher-
ence by fucoidin and after inhibition of PAF effects by the
the PAF receptor antagonist BN52021.
Methods: In male Wistar rats, microvascular permeability
(MP), leukocyte adherence (LA) and mast cell activation
(MCA) were determined in mesenteric postcapillary
venules using intravital microscopy at baseline and at 60
and 120 min after the start of a continuous infusion of
endotoxin (ETX; 2 mg/kg/hr, E. coli 026:B6) (group A,
n=10). Leukocyte-endothelial interaction was blocked
using fucoidin (25 mg/kg b.w.). In addition to the proce-
dure in group A, group B (n=10) received BN52021 (5
mg/kg b.w.) after baseline measurements. Group C
(n=10; control group) only received equivalent volumes of
NaCI 0.9%. Statistical analysis was performed using stu-
dent's t-test. A P-value <0.05 was considered significant.
Results: In groups A and B, fucoidin prevented LA, and
so there were no significant differences in LA between
these groups. In group A, MP and MCA significantly
increased starting at 60 min (P<0.05 vs baseline). There
was no significant increase in MP and MCA in groups B
and C. Differences between groups A and B in MP and
MCA were significant at 120 min.
Conclusions: The results of this study demonstrate that
PAF plays a significant role in the initiation of endotoxin-
induced leukocyte-independent plasma extravasation and
mast cell activation.
1. Kubes P: Am J Physiol 1996, 271:H2438.
2. Schmidt H: J Surg Res 1996, 60:29.
P57 TNF-a: a possible mediator of remote tissue injury after viper envenomation
0 Szold, AA Weinbroum, R Ben-Abraham, TE Englender, D Ovadia, S Marmor, M Sorkine, R Flaison and P Sorkine
General Intensive Care and the Departments of Oncology and Pathology, Tel Aviv Sourasky Medical Center and the Sackler
Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel; and Unite des Venins, Institute Pasteur, Paris,Cedex 15, France
Introduction: TNF is a potent inflammatory promoting agent
that can potentiate organ and tissue injury. A possible role
of TNF in causing local tissue damage following snakebite
was recently demonstrated in an intact rat model. We com-
pared the systemic effects of TNF on the hearts and lungs
of rats following an intramuscular injection of a sub-lethal
dose of Vipera asis venom (500 [tg/kg, experimental
groups) to equivalent injections of saline (control group).
Results: Systemic TNF activity, heart rate and blood pres-
sure as well as lung permeability and neutrophil sequestra-
tion were then evaluated in both groups. The venom
caused a significant reduction in heart rate and arterial
blood pressure, and the serum TNF levels peaked after
two hours. These values remained unchanged in the
control group. In contrast, lung microvasculature perme-
ability and neutrophil sequestration were not significantly
different between the experimental and control groups.
Conclusions: This study of the systemic and inflammatory
effects of Vipera aspis venom showed that intramuscular
injection of the poison results in systemic effects that are
possibly mediated in part by TNF. These findings may
have therapeutic implications in the treatment of patients
with severe systemic manifestations after snakebites in
terms of the possible benefits of blocking TNF activity.
P58 Phagocytosis of granulocytes is decreased in most patients with severe sepsis or septic shock
G Fischer, E Barth, R Remmele, LL Moldawer*, EM Schneider, M Georgieff and M Weiss
Department of Anesthesiology, University of Ulm, Germany, and *Department of Surgery, University of Florida, Gainesville, USA
Objectives: To investigate the phagocytotic activity of
granulocytes during septic shock in postoperative/post-
traumatic patients compared to that of healthy volun-
Methods: Over a six month period 19 patients with septic
shock were monitored on a daily basis during their stay in
the intensive care unit. Phagocytotic activity of granulo-
cytes was determined by flow cytometry (PhagotestTM
Orpegen, Heidelberg, Germany). The normal range of
phagocytosis (expressed as mean intensity, Mnl) was
defined by a control group of 11 healthy volunteers.
Results: Phagocytosis of healthy volunteers was in the
range of 34 MnI and 149 Mnl. All survivors and 7 of 10
nonsurvivors (i.e. 84% of all patients) had a lower phago-
cytotic activity on more than 50% of days in septic shock
compared to that of healthy volunteers. No patient had a
phagocytosis greater than 149 Mnl, the upper value of the
Conclusions: Phagocytosis is decreased in most patients
during septic shock compared to healthy volunteers. Thus
diminished phagocytotic activity may contribute to an
impaired mechanism of bacterial elimination or a reduced
Number of patients with more than 50% of days during septic
shock within a distinct range of phagocytosis during septic
shock compared to the normal range.
compared to normal Below Within Above
range (<34) (34-149) (>149) Total
Survivors 9 0 0 9
Nonsurvivors 7 3 0 10
Total 16 3 0 19
resolution of infection in these patients. Moreover, phago-
cytotic activity of granulocytes does not discriminate sur-
vivors and nonsurvivors of septic shock.
P59 Triiodothyronine (T3) ameliorates the cytokine storm in rats with sepsis
T Yokoe*, M Negishi*, Y lino* and Y Morishitat
*Department of Emergency and Critical Care Medicine, tSecond Department of Surgery, Gunma University School of Medicine,
3-39-15 Showa-machi Maebashi Maebashi, Gunma 371-8511, Japan
We reported that T3 eliminated acidosis and improved sur-
vival rate in a rat model of sepsis. The aim of this study is
to clarify the effect of T3 on cytokine levels.
Male Sprague-Dawley rats, weighing 350-420 g, were
ligated in the cecum with puncture (CLP method) under
anesthesia with pentobarbital (40 mg/kg). After CLP, 50
ml/kg of saline was injected subcutaneously for fluid
resuscitation. Rats were assigned two groups; no treat-
ment group (control group, n=15) and a T3 treated group
(n=10). The T3 treated group was given 3 ng/hr of T3
using a osmotic pump embedded subcutaneously. Sur-
vival rate, levels of IL-1 P, TNF-Q, IL-6, IL-8, and IL-10 were
studied 24 h after surgery.
Three animals died in the control group, whereas no
animals died in T3 treated group. There was no significant
difference on mean value of ILI-P and TNF-( between two
groups. Mean levels of IL-6 and IL-8 in the control group
were 237.4 pg/ml and 5342.7 pg/ml, respectively,
however, those in the T3 group were suppressed to the
level of 183.5 pg/ml and 55.2 pg/ml, respectively
(P<0.05). Mean IL-10 level (6.0 pg/ml) in the T3 treated
group was lower than that (72.4 pg/ml) in the control
It is concluded that T3 prevents the cytokine storm and
improves the survival rate in this rat model of sepsis.
P60 Interleukin-17 stimulates intraperitoneal neutrophil infiltration through the release of the
chemokine GROa from peritoneal mesothelial cells
J Witowski*t, K Pawlaczykt, A Breborowiczt, A Scheuren*, M Kuzlan-Pawlaczykt, J Wisniewskat, A Polubinskat, H Friess*,
GM Gahl*, U Frei* and A Jorres*
*Nephrology and Medical Intensive Care, UK Charit6, Campus Virchow-Klinikum, Augustenburger Platz 1, D-13353 Berlin,
Germany; tPathophysiology, University Medical School, Poznan, Poland; and *Visceral and Transplant Surgery, University of
Bern, Inselspital, Bern, Switzerland
Introduction. IL-17 is a newly discovered cytokine impli-
cated in the regulation of hematopoiesis and inflammation.
Since IL-1 7 production is restricted to activated T lympho-
cytes the effects exerted by IL-1 7 may help to understand
the contribution of T cells to the inflammatory response.
We investigated the role of IL-17 in leukocyte recruitment
into the peritoneal cavity.
Methods: Leukocyte infiltration in vivo was assessed in BalB/
CJ mice. Effects of IL-17 on chemokine generation in vitro
were examined in human peritoneal mesothelial cells (HPMC).
Results: Intraperitoneal administration of IL-1 7 resulted in
a selective recruitment of neutrophils into the peritoneum
and increased levels of KC chemokine (murine homologue
of human GROQ). Pre-treatment with anti-KC antibody
significantly reduced the IL-1 7-driven neutrophil accumula-
tion. Primary cultures of HPMC expressed IL-17 receptor
mRNA. Exposure of HPMC to IL-17 led to a dose- and
time-dependent induction of GROa mRNA and protein.
Combination of IL-17 together with TNFQ resulted in an
increased stability of GROa mRNA and synergistic