epiglucan and Shock--Septic

Invasive pulmonary aspergillosis (IPA) is an important cause of morbidity/mortality in critically ill patients with hematological malignancies. New diagnosis strategies include the noninvasive biomarkers 1,3-beta-D-glucan (BDG) and serum galactomannan (GM).. For early detection of IPA, we compared BDG Fungitell assay with GM Platelia assay.. Twenty-two out of 30 patients (74 %) had elevated BDG levels (mean 306 pg/ml) beyond the cutoff of 80 pg/ml. GM levels were elevated in only 3 patients (10 %) over the ODI cutoff of >0.5. Following the BDG/GM and microbiological findings, 10 (34 %) cases were classified as probable IPA and 12 (40 %) as possible IPA. Eight (26 %) were classified as no IPA. An overall sensitivity of 90 % (95 % CI 86-96 %) and specificity of 85 % (95 % CI 79-86 %) was found for the BDG Fungitell assay in IPA. In contrast, an overall sensitivity of 30 % (95 % CI 26-38 %) and specificity of 98 % (95 % CI 94-100 %) was found for the GM Platelia assay. A false-negative rate of 70 % for probable IPA and 85 % for probable/possible IPA was detected for GM. The false-negative rate for BDG was 0 % in cases of probable IPA and 45 % in cases of possible cases.. BDG is a sensitive marker for patients' surveillance at risk of IPA. In patients with hematological malignancies and septic shock, early diagnosis of IPA might be significantly improved by BDG compared to GM, also considering that BDG has the advantage of detecting fungal diseases other than IPA.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; beta-Glucans; Critical Illness; Diagnostic Tests, Routine; Early Diagnosis; Female; Galactose; Hematologic Neoplasms; Humans; Invasive Pulmonary Aspergillosis; Male; Mannans; Middle Aged; Proteoglycans; Sensitivity and Specificity; Serum; Shock, Septic; Young Adult

Gastrointestinal (GI) leakage is believed to exacerbate sepsis and new, validated markers of GI barrier performance might benefit clinical decision-making. Serum (1→3)-β-D-glucan (BG) was evaluated as a potential GI leakage marker. Serum BG was tested in several mouse models of GI leakage, including dextran sulfate solution (DSS) administration, endotoxin (LPS) injection, and cecal ligation and puncture sepsis (CLP). Serum BG titer was also evaluated in patients with sepsis and septic shock, for comparison.With 0.75% DSS administration, BG increased only after oral administration of heat-killed C. albicans, but increased spontaneously with 1.5% DSS. In the LPS and CLP models, BG increased as early as 1 h and at 12 h after LPS administration and surgery, respectively. GI leakage was confirmed by orthogonal validation methods including FITC-dextran oral administration in the DSS, LPS, and CLP models and, in the DSS model, with urine sucralose after oral administration and serum endotoxemia. IL-6 increased in parallel with serum BG. Serum BG or IL-6, at 18 h, anticipated sepsis mortality in the CLP model.Analysis of serum BG from patients with febrile neutropenic sepsis (N = 49) and febrile non-neutropenic sepsis (N = 39) demonstrated BG elevation. Patients with bacterial septic shock had serum BG titers similar to levels observed in invasive fungal disease, regardless of febrile neutropenia. Serum BG was lower in less severe cases of bacterial sepsis. Elevated serum IL-6 was associated with GI leakage and elevated serum BG.Serum BG may have potential as a sepsis/septic shock biomarker and further study in this context is warranted.

Topics: Animals; beta-Glucans; Candida albicans; Disease Models, Animal; Feces; Gastrointestinal Tract; Humans; Male; Mice; Pilot Projects; Proteoglycans; Sepsis; Shock, Septic

Invasive fungal disease (IFD) remains a significant cause of morbidity and mortality in critically ill patients.. Examination of 1,3-β-D-glucan (BDG) for IFD and as outcome parameter in immunocompromised critically ill patients with septic shock.. Thirty-two (69 %) out of 46 included patients had BDG beyond the cutoff of >80 pg/ml (mean 320 pg/ml). Twelve (37 %) had findings of Aspergillus spp. in BAL (mean BDG 413 pg/ml). EORTC/MSG guidelines classified these as probable invasive aspergillosis (IA)/IFD. Five (16 %) had candidaemia (mean BDG level 361 pg/ml). Sensitivity of 78 % (95 % CI 58-88 %) and specificity of 68 % (95 % CI 52-77 %) for IFD were found on the BDG Fungitell assay. In detail, a sensitivity of 73 % (95 % 58-84 %) and specificity of 83 % (95 % CI 68-93 %) for IA and a sensitivity of 77 % (CI 95 % 62-87 %) and specificity 53 % (95 % CI 37-73 %) for candidaemia were found. APACHE II, SOFA score and mortality rate were in the elevated BDG group significantly altered (26 vs. 21, p < 0.003; 15 vs. 13, p < 0.006; 72 vs. 50 %, p < 0.004).. 1,3-β-D-glucan assay is helpful for early detection of IFD; moreover, elevated BDG levels can be used as a predictor for outcome in immunocompromised critically ill patients as presented in our study.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; beta-Glucans; Critical Illness; Diagnostic Tests, Routine; Early Diagnosis; Female; Humans; Immunocompromised Host; Invasive Fungal Infections; Male; Middle Aged; Proteoglycans; Sensitivity and Specificity; Shock, Septic; Treatment Outcome; Young Adult

beta-Glucans are glucose polymers with a variety of stimulatory effects on the immune system. The objective of this study was to determine the effect of prophylactic oral administration of soluble Saccharomyces cerevisiae-derived beta-1,3/1,6-glucan (SBG) on the outcome of experimental endotoxaemia and shock-associated organ injury. Male Wistar rats were pretreated with SBG orally (SBGpo, 20 mg/kg/day) for 14 days, subcutaneously (SBGsc, 2 mg/kg/day) for 3 days, or vehicle (placebo). Rats were anaesthetized and subjected to endotoxaemia by intravenous infusion of Escherichia coli lipopolysaccharide (LPS) (6 mg/kg) or saline infusion (sham). We observed significant levels of plasma beta-glucan in the SBGpo group (P<0 x 5), although the SBGsc group had levels approximately 40-fold higher despite a 10-fold lower dose. SBG prophylaxis caused enhanced blood pressure recovery following LPS-induced blood pressure collapse. Oral treatment with SBG attenuated the LPS-induced rise in plasma creatinine levels (P<0 x 05), indicating protection against renal injury. SBG also attenuated the plasma levels of aspartate aminotransferase and alanine aminotransferase (SBGpo, P<0 x 01; SBGsc, P<0 x 01), indicating protection against LPS-induced hepatic injury. A moderate increase in baseline interleukin (IL)-1beta levels was observed in the SBGsc group (P< 0 x 05). In the LPS-challenged rats, plasma levels of proinflammatory cytokines was moderately reduced in both SBG-treated groups compared to placebo. SBG treatment, particularly oral administration, had a striking effect on the haemodynamics of LPS-treated rats, although only a minute fraction of the orally administered beta-glucan translocated to the circulation. Enhanced organ perfusion may thus be responsible for the attenuated levels of indicators of kidney and liver injury seen in SBG-treated rats.

Topics: Administration, Oral; Animals; beta-Glucans; Blood Pressure; Cytokines; Endotoxemia; Injections, Subcutaneous; Lipopolysaccharides; Male; Multiple Organ Failure; Rats; Rats, Wistar; Saccharomyces cerevisiae; Shock, Septic

We have developed an animal model of sepsis in mice by repeatedly administering beta-glucan, a biological response modifier, and indomethacin (IND), a nonsteroidal anti-inflammatory drug. The combination of these drugs induced bacteremia by translocation of the enterobacterial flora, resulting in increasing the number of activated leukocytes, and inducing hyper cytokinemia. In the present study, we examined the effect of antibiotics on beta-glucan and IND-induced septic shock. Treatment with antibiotics inhibited microbial translocation, inhibited contraction of the colon, reduced lipopolysaccharides (LPS)-elicited production of TNF-alpha and IL-6, and finally prolonged survival. However, the efficacy of antibiotics treatment was limited in mice administered IND orally. These findings strongly suggested that the antibiotics controlled the gut-associated action of IND and reduced various symptoms accompanying sepsis.

Topics: Animals; Anti-Bacterial Agents; Bacterial Translocation; beta-Glucans; Body Weight; Colon; Cytokines; Disease Models, Animal; Drug Therapy, Combination; Indomethacin; Liver; Male; Mice; Shock, Septic

We have developed a septic shock model in mice by sequential administration of beta-glucan, a biological response modifier, and indomethacin (IND), a nonsteroidal anti-inflammatory drug. Lethality was significantly related to the translocation of gut flora to various organs and mal-adjustment of the cytokine network. In the present study, we have examined the effect of antibiotics on this model to further clarify meanings of microbial flora. Schizophyllan (SPG), antitumor beta-glucan for clinical use, obtained from the culture filtrate of Schizophyllum commune, was used to induce sepsis. Lincomycin (LCM), imipenem (IPM), cilastatine (CS), and ampicillin (ABPC) were used for antibiotics treatment. The survival rate of SPG/IND-treated mice was significantly increased by administering LCM or ABPC/IPM/CS, and the effect was more significant by LCM. In in vitro spleen cell culture, LCM decreased proinflammatory cytokine production. Moreover, prednisolone, immune suppresser treatment improved survival of SPG/IND-treated mice. These findings suggest that LCM is an effective antibiotic in this endogenous septic model by modulating gut microbial flora and, at least a part, by regulating cytokine production of leukocytes.

Topics: Animals; Anti-Bacterial Agents; Anti-Inflammatory Agents, Non-Steroidal; beta-Glucans; Blood Glucose; Cells, Cultured; Colon; Colony Count, Microbial; Cytokines; Feces; Gastrointestinal Tract; Indomethacin; Lincomycin; Male; Mice; Mice, Inbred ICR; Prednisolone; Schizophyllum; Shock, Septic; Spleen

Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 --> 3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p < 0.05) induced cytokine production IL-6 > TNF alpha > IL-1 beta > GM-CSF > IL-10 > IFN gamma. The induction of these cytokines was significantly (p < 0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P < 0.05) LPS and SA induced cytokines. PTx further augmented (p > 0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti-inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent.

Topics: beta-Glucans; Cell Culture Techniques; Cytokines; Gram-Negative Bacterial Infections; Gram-Positive Bacterial Infections; GTP-Binding Proteins; Humans; Inflammation; Lipopolysaccharides; Monocytes; Proteoglycans; Shock, Septic; Signal Transduction; Staphylococcal Infections; Staphylococcus aureus

We have previously shown that repeated administration of nonsteroidal anti-inflammatory drugs (NSAIDs) to mice treated with beta-glucan, a biological response modifier, induced severe lethality. The lethality would be strongly related to the translocation of enterobacterial flora to the peritoneal cavity and disruption of the cytokine network. Reports suggest that nitric oxide (NO) can have an effective or detrimental role in septic shock. In the present study, we examined the effect of NO, an inflammatory mediator, on beta-glucan/indomethacin (IND)- induced septic shock by inhibiting its synthesis with N(G)-nitro-L-arginine methyl ester (L-NAME), a nonselective NO synthase (NOS) inhibitor. Nitrite concentration was used as an indicator of NO generation. Mortality in beta-glucan/IND-treated mice was increased by administering L-NAME. Numbers of bacteria in various organs of mice treated with beta-glucan/IND rose significantly within a couple of days of the administration of L-NAME. Additionally, TNF-alpha, IL-1beta, and IL-6 concentrations were enhanced in peritoneal exuded cells in culture. These results suggest a significant loss of the bactericidal activity of macrophages on the administration of a NOS inhibitor which enhanced the rate of enterobacterial invasion to the peritoneal cavity, resulting in systemic inflammatory response syndrome. The production of NO, therefore, provides a protective effect in beta-glucan/IND-induced sepsis.

Topics: Animals; beta-Glucans; Enzyme-Linked Immunosorbent Assay; Indomethacin; Interleukin-6; Liver; Male; Mice; Mice, Inbred ICR; NG-Nitroarginine Methyl Ester; Nitric Oxide; Shock, Septic; Sizofiran; Tumor Necrosis Factor-alpha

Growing evidence supports the role of transcription factor activation in the pathophysiology of inflammatory disorders, sepsis, ARDS, SIRS, and shock. Kinase mediated phosphorylation of IkappaBalpha is a crucial step in the NFkappaB activation pathway. We investigated IKBalpha phosphorylation in murine liver and lung extracts after cecal ligation and puncture (CLP) in the presence and absence of a glucan ligand. ICR mice were subjected to CLP. Unoperated and sham-operated mice served as the controls. Glucan phosphate (50 mg/kg) was administered 1 h before or 15 min after CLP. CLP increased hepatic and pulmonary levels of phospho-IkappaBalpha by 48-192%. Pre- or post-treatment with glucan phosphate decreased (P < 0.05) tissue phospho-IkappaBalpha levels in CLP mice. Phospho-IkappaBalpha in the glucan-CLP group were not significantly different from the unoperated controls. To investigate mechanisms we examined IKKbeta kinase activity, IkappaBalpha phosphorylation and degradation, and NFkappaB activity in a murine macrophage cell line, J774a.1, treated with LPS (1 microg/mL) and/or glucan phosphate (1 microg/mL) for up to 120 min. The glucan ligand blunted LPS-induced IKKbeta kinase activity, phosphorylation and degradation of IkappaBalpha, and NFkappaB nuclear binding activity. The data indicate that one mechanism by which (1-->3)-beta-D-glucan may alter the response to endotoxin or polymicrobial sepsis involves modulation of IKK3 kinase activity with subsequent decreases in IkappaBalpha phosphorylation and NFkappaB activation.

Topics: Animals; beta-Glucans; Cell Line; Cell Nucleus; Cells, Cultured; Cytosol; DNA-Binding Proteins; Gene Expression Regulation; Glucans; I-kappa B Kinase; I-kappa B Proteins; Intestinal Perforation; Ligands; Lipopolysaccharides; Liver; Lung; Macrophages; Male; Mice; Mice, Inbred ICR; NF-kappa B; NF-KappaB Inhibitor alpha; Phosphorylation; Protein Processing, Post-Translational; Protein Serine-Threonine Kinases; Sepsis; Shock, Septic

Soluble carboxymethyl-b-1,3-glucan (CMG), a possible ligand for scavenger receptors, has macrophage-activating action but lacks the granulomatose inflammatory side effect: it is a promising immunomodulator that may mitigate the severity of sepsis. This motivated us to study in rats the effect of CMG (25 mg/kg), injected into the tail vein at 48 and 24 h prior to the administration of 5 mg/kg Escherichia coli 0127.B8 endotoxin on survival, hemodynamic condition, and, in vitro, on the chemiluminescence of PMNs and macrophages, and on macrophagal tumor necrosis factor (TNF) production. Acetylated low density lipoprotein (AcLDL) clearance in vivo and in vitro binding to macrophages was used to study scavenger receptor function. In the nonpretreated group 9 of 10 rats died during the first 24 h after endotoxin, but all CMG-pretreated rats survived. CMG-pretreatment prevented severe decreases in cardiac output and blood pressure after endotoxin. Chemiluminescence of macrophages and PMNs from CMG-pretreated rats was about two times less (p < .05) than that from nonpretreated ones; the endotoxin induced TNF production by macrophages also decreased. Pretreatment with CMG increased, but coinjection of CMG and AcLDL decreased the AcLDL clearance, while coinjection of endotoxin and AcLDL decreased the survival rate. In vitro AcLDL uptake by macrophages decreased after coinjection with CMG. Our results thus showed that CMG was protective in rat endotoxin shock, which seemed partly connected with enhancement of endotoxin clearance through scavenger receptors and to decreased TNF production.

Topics: Animals; beta-Glucans; Glucans; Hemodynamics; Iodine Radioisotopes; Kidney; Leukocytes; Lipoproteins, LDL; Liver; Luminescent Measurements; Macrophages; Male; Membrane Proteins; Rats; Rats, Wistar; Receptors, Immunologic; Receptors, Lipoprotein; Receptors, Scavenger; Scavenger Receptors, Class B; Shock, Septic; Survival Rate; Tumor Necrosis Factor-alpha