➥ T he leading cause of death in intensive care units around the world is the syndrome of exaggerated and prolonged systemic inflam- mation response (SIRS), which if not controlled will lead to an irreversible, often multiple, organ dysfunction and organ failure syndrome (MODS). Host phagocytic cells, predominantly macrophages and neutrophils, and their soluble products, play a central role. Accumulation of neutrophils is often observed, especially in the lungs, 1-3 intestines, 4 liver 5 and kidneys. 6 Neu- trophilic infiltration of distant organs, 7 and espe- cially of the lungs, 1 has been a characteristic finding of patients dying from sepsis. However, the mechanisms responsible for the exuberant inflammation and neutrophil infiltration are not yet fully understood. It has recently been demon- strated that this, at least in part, is due to an inhibition of neutrophil apoptosis, 8 and there is a suggested association with activation of nuclear factor — κB (NF-κB), reduced activity of caspas- es-9 and -3 and maintenance of mitochondrial transmembrane potential. 8 A composition of four lactic acid bacteria (LAB) and four plant fibres (Synbiotic 2000™) has proven extraordinarily effective in reducing inflammation and rate of infection in patients after extensive operations, including transplan- tations, 9 severe acute pancreatitis (Oláh A, per- sonal communication), trauma (Kompan L personal communication) and in critically ill patients (Girling K, personal communication) [Author: are there any published papers to provide evidence for these assertions? Normally we do not reference unpublished material]. The LAB for the composition were identified — and to a large extent selected — for their unique ability to transcribe NF-κB. 10 This study was undertaken in order to investigate if neutrophil infiltration could be inhibited by sup- plementation of these LAB. However, as many critically ill patients cannot eat and it takes sev- eral hours for the LAB to reach the large intes- tine when orally administered, we wanted to explore the efficacy of subcutaneous administra- tion. Subcutanous administration of live LAB was recently shown to attenuate both murine colitis and murine arthritis. 11 M ATERIALS AND M ETHODS The study was conducted at the Experimental Animal Research Laboratory, Faculty of Medicine, Ege University, Ismir, Turkey. Eighty male adult Wistar albino rats weighing 250–300 g were ran- domly selected and divided into eight groups. All the animals had free access to water and a stan- dard rat chew containing 4.5% plant fibres. The lactic acid bacteria for the study are ingredients in a synbiotic composition called Synbiotic 2000 FORTE™ (S2000F), which con- sists of a mixture of four LAB, one from each of the four main genera of lactobacillus; 10 11 of pedi- acoccus pentosaceus 5–33:3, 10 11 of leuconostoc mesenteroides 32–77:1,10 11 of lb paracasei subsp paracasei 19 and 10 11 of lb plantarum 2362, i.e. a total of 40 billion LAB per dose, plus a mixture of four well studied bioactive plant fibres: 2.5 g betaglucan, 2.5 g inulin, 2.5 g pectin and 2.5 g resistant starch, in total 10 g of plant fibres per dose. The composition was determined after extensive studies of >350 human and >180 plant strains by microbiologists from Lund University, Sweden, Åsa Ljungh and Torkel Wadström. 10,12 The LAB to be used in the composition were cho- sen after studies of a series of microbial functions such as ability to produce bioactive proteins, transcribe NF-κB, produce pro- and anti-inflam- matory cytokines, produce antioxidants, and to function and complement each other. The four LAB have partially different functions and demonstrate, in combination, potentiating/syner- gistic effects. S2000F is produced by Medipharm, Kågeröd Sweden and Des Moines, Iowa, USA. Further detaits regarding preparation of the bac- terial solutions are set out below. The animals were randomly divided into eight groups and received treatment as shown in Table 1. Preparation of bacterial solution The preparation procedure was performed at the Bacteriology Laboratory of Celal Bayar Uni- versity, Manisa, Turkey. The four LAB strains were individually prepared. Stock cultures of Lactobacillus casei, Lactobacillus plantarum, Lactobacillus mesenteroides and Pediococcus pen- tosaceus strains were provided by Medipharm, Kågeröd, Sweden. Each bacterial strain was cul- tured by the use of the BACTEC 9120 system (Becton Dickenson, USA) and subcultures made from the BACTEC bottles onto MRS agar (Oxoid, UK). The plates were incubated over- night under micro-aerophilic conditions at 37 0 C. Each bacterial colony was suspended in sterile saline in separate tubes. The bacteria were har- vested after centrifugation, washed three times with saline, and re-suspended into saline with a concentration of 10 11 bacteria per ml. The con- centration of the bacteria was first estimated by spectrophotometry (UV 1601, Shimadzu Corpo- ration, Japan) at 620 nm. The number of viable bacteria was directly determined by enumera- ORIGINAL PAPER SUMMER 2005 20 Subcutaneous administration of live lactobacillus prevents sepsis-induced lung organ failure in rats O Ilkgul* H Aydede* Y Erhan* S Surucuoglu* H GazI* S Vatansever* F Taneli* C Ulman* C Kose* S Bengmark [Author: please supply at least one qualification (e.g. ‘MD’) for each author] *Faculty of Medicine, Celal Bayar University, Manisa, Turkey †Institute of Hepatology, University College London Medical School, London, UK