Systemic Interleukin-6 Response Research Articles

Systemic Interleukin-6 Response after Intravesical Instillation of Bacillus Calmette-Guérin and Mitomycin C in Superficial Bladder Cancer.

Interleukin-6 (IL-6) is proposed to play a significant role in pathogenesis of urinary bladder cancer (UBC). This role may be influenced by chemotherapy (mitomycin C; MMC) or immunotherapy (Bacillus Calmette-Guérin; BCG). A case-control study was conducted to determine IL-6 levels in serum of newly diagnosed cases (NDC) of superficial UBC, as well as in patients treated with MMC or BCG intravesical instillation. A total sample of 111 patients (36 NDC, 45 MMC and 30 BCG) was included, as well as a control group of 107 healthy controls (HC). IL-6 was detected by enzyme-linked immunosorbent assay. Results revealed that median levels of IL-6 were significantly elevated in NDC group (15.8 pg/mL; P<0.001) compared to MMC and BCG groups (7.5 and 5.3 pg/mL, respectively) or HC (4.4 pg/mL); while, there were no significant differences between the latter three groups (MMC, BCG and HC). Receiver operating characteristic curve analysis revealed that IL-6 is a very good predictor of UBC in NDC group versus HC (area under the curve=0.885; 95% confidence interval=0.828-0.942; P<0.001; cut-off value=10.5 pg/mL; Youden index=0.62; sensitivity=80.6%; specificity=81.3%). Logistic regression analysis confirmed this significance and IL-6 was associated with a higher risk of UBC (odds ratio=1.18; 95% confidence interval=1.11-1.26; P<0.001). In conclusion, this study indicated that IL-6 level was upregulated in serum of NDC of UBC. Further, IL-6 level was restored to normal levels after intravesical instillation of MMC or BCG.

Biochemical and pharmacological characterization of PhTX-I a new myotoxic phospholipase A2 isolated from Porthidium hyoprora snake venom

This paper reports the biochemical and pharmacological characterization of a new myotoxic PLA2 (EC 3.1.1.4) called PhTX-I, purified from Porthidium hyoprora venom by one step analytical chromatography reverse phase HPLC. The homogeneity of the PhTX-I fraction and its molecular mass were initially evaluated by SDS–PAGE and confirmed by MALDI-TOF spectrometry, indicating a molecular mass of 14.249Da and constituted of a single polipeptidic chain. Amino acid sequence was determined by “de novo sequencing,” in tandem mass spectrometry, belonging to D49-PLA2 enzyme class and exhibiting high identity (44–90%) with other myotoxics PLA2 from snake venoms. The enzymatic investigation showed maximal activity at pH 8 and 35–45°C. This activity was dependent on Ca2+, other cations (Mg2+, Mn2+, Cd2+ and Zn2+) reduced notably the enzymatic activity, suggesting that the arrangement of the catalytic site presents an exclusive structure for Ca2+. Ex vivo, whole venom and PhTX-I PLA2 caused blockade of the neuromuscular transmission in young chick biventer cervicis preparations similar to other isolated snake venom toxins from the Bothrops genus. In vivo, both induced local myotoxicity and systemic interleukin-6 response upon intramuscular injection, additionally, induced moderate footpad edema. In vitro, both induced low cytotoxicity in skeletal muscle myoblasts, however PhTX-I PLA2 was able to lyse myotubes.

Structural and pharmacological characterization of the crotamine isoforms III-4 (MYX4_CROCu) and III-7 (MYX7_CROCu) isolated from the Crotalus durissus cumanensis venom

Two major crotamine isoforms (III-4 and III-7) were obtained combining two chromatographic steps on molecular exclusion chromatography (Sephadex G-75) and ion-exchange column (Protein Pack SP 5PW) of the rattlesnake Crotalus durissus cumanensis venom. The “in vivo” myotoxic effect of the venom, its “in vitro” cytotoxicity in myoblasts and myotubes (C2C12) and the neurotoxic and edema-forming activity were characterized. The molecular masses of the crotamine isoforms were 4907.94 Da (III-4) and 4985.02 Da (III-7) and, as determined by mass spectrometry, both contained six Cys residues. Enzymatic hydrolysis followed by de novo sequencing through tandem mass spectrometry was used to determine the primary structure of both isoforms. III-4 and III-7 isoforms presented a 42-amino acid residues sequence and showed high molecular amino acid sequence identity with other crotamine-like proteins from Crotalus durissus terrificus. In vivo, both crotamine isoforms induced myotoxicty and a systemic interleukin-6 response upon intramuscular injection. These new crotamine isoforms induced low cytotoxicity in skeletal muscle myoblasts and myotubes (C2C12) and both induced a facilitatory effect on neuromuscular transmission in young chick biventer cervicis preparation. Edema-forming activity was also analyzed by injection of the crotamine isoforms into the right paw, since both crotamine isoforms exert a strong pro-inflammatory effect.

Neurotoxic, Myotoxic and Cytolytic Activities of the New Basic PLA2 Isoforms BmjeTX-I and BmjeTX-II Isolated from the Bothrops marajoensis (Marajó Lancehead) Snake Venom

The BmjeTX-I and BmjeTX-II isoforms of PLA(2) were purified from Bothrops marajoensis venom by ion-exchange chromatography and reverse phase HPLC. Both isoforms showed a molecular mass of 13808.89 Da (BmjeTX-I) and 13863.97 Da (BmjeTX-II) determined by based on the determined primary structures and SDS-PAGE and confirmed experimentally by MALDI-TOF mass spectrometry. Multiple alignment of BmjeTX-I and BmjeTX-II isoforms of PLA(2) show high degree of homology with basic PLA(2) myotoxins from other Bothrops venoms. Ex vivo, both isoforms caused a blockade of the neuromuscular transmission in young chick biventer cervicis preparations in a similar way to other Bothrops species. In chick preparations, contractures to exogenous acetylcholine (55 and 110 microM) or KCl (13.4 mM) were unaltered after complete blockade for the both isoforms BmjeTX-I and BmjeTX-II of PLA(2). These results, which strongly suggested a presynaptic mechanism of action for these toxins. In mice, both isoforms induced myonecrosis and a systemic interleukin-6 response upon intramuscular injection. Both isoforms BmjeTX-I and BmjeTX-II of PLA(2) also induced moderate marked paw edema, evidencing the local increase in vascular permeability. Since both isoforms of PLA(2) exert a strong proinflammatory effect, the enzymatic hydrolysis of phospholipids might be relevant for this phenomenon and produced cytotoxicity in murine skeletal muscle C2C12 myoblasts and myotubes.

Concentrated ambient fine particles and not ozone induce a systemic interleukin-6 response in humans

Epidemiological studies have established significant associations between ambient pollutants, including fine particulate matter (PM2.5) and ozone (O3), and cardiopulmonary morbidity and mortality. One mechanism that has been proposed is a pulmonary/systemic inflammatory response. Although controlled human exposure studies have examined the independent inflammatory responses of PM2.5 and O3, no studies have previously examined their joint effects. The study objective was to examine the independent and combined associations between ambient PM2.5 and O3 and acute respiratory/inflammatory responses. Using their concentrated ambient particle (CAP) facility for PM2.5, the authors studied 10 mild asthmatic and 13 nonasthmatic individuals. The 2-h exposures included CAP (range 48–199 μg/m3) and filtered air (FA), with/without O3 (120 ppb), in a randomized block design. Response measures included pulmonary function and inflammatory indices in induced sputum (interleukin [IL]-6, cytology) and blood (IL-6, tumor necrosis factor [TNF]-α) measured before and after exposures. Three hours post exposure, there was an increase in blood levels of IL-6, but only after CAP alone exposures; the IL-6 increase was associated with increasing PM2.5 mass concentration (p = .005). Some individuals switched to shallow breathing during CAP+O3, possibly accounting for an attenuation of the resultant blood IL-6 response. Asthmatic and nonasthmatic responses were similar. There were no adverse changes in pulmonary function or other inflammatory measures. The study demonstrated an acute IL-6 response to PM2.5, providing evidence to support the epidemiological findings of associations between ambient levels of particles and cardiopulmonary morbidity and mortality.

Systemic Interleukin-6 Response to Respiratory Syncytial Virus Infection

Systemic Interleukin-6 Response to Respiratory Syncytial Virus Infection

Biological and biochemical characterization of new basic phospholipase A 2 BmTX-I isolated from Bothrops moojeni snake venom

BmTX-I, an Asp49 phospholipase A 2, was purified from Bothrops moojeni venom after only one chromatographic step using reverse-phase HPLC on μ-Bondapak C-18 column. A molecular mass of 14238.71 Da was determined by MALDI-TOF mass spectrometry. Amino acid analysis showed a high content of hydrophobic and basic amino acids as well as 14 half-cysteine residues. The BmTX-I PLA 2 had a sequence of 121 residues of amino acids: DLWQFNKMIK KEVGKLPFPF YGAYGCYCGW GGRGEKPKDG TDRCCFVHDC CYKKLTGCPK WDDRYSYSWK DITIVCGEDL PCEEICECDR AAAVCFYENL GTYNKKYMKH LKPCKKADYP C and p I value 7.84, and showed a high degree of homology with basic Asp49 PLA 2 myotoxins from other Bothrops venoms. BmTX-I presented PLA 2 activity in the presence of a synthetic substrate and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0 and 35–45 °C. Maximum PLA 2 activity required Ca 2+ and in the presence of Mg 2+, Cd 2+ and Mn 2+ it was reduced in presence or absence of Ca 2+. Crotapotin from Crotalus durissus colillineatus rattlesnake venom has significantly inhibited ( P<0.05) the enzymatic activity of BmTX-I. In vitro, the whole venom and BmTX-I caused a blockade of the neuromuscular transmission in young chick biventer cervicis preparations in a similar way to other bothrops species. In mice, BmTX-I and the whole venom-induced myonecrosis and a systemic interleukin-6 response upon intramuscular injection. Edema-forming activity was also analyzed through injection of the venom and the purified BmTX-I into the subplantar region of the right footpad. Since BmTX-I exert a strong proinflammatory effect; the enzymatic phospholipids hydrolysis might be relevant for these phenomena.

Urinary Albumin:Creatinine Ratio (ACR) and the Prediction of Postoperative Complications After Abdominal Aortic Aneurysm Repair

IntroductionOpen repair of abdominal aortic aneurysm (AAA) requires aortic clamping. This results in an ischaemia-reperfusion injury (IRI) which can lead to the development of the systemic inflammatory response syndrome (SIRS) and multiple organ failure (MOF). We investigated the use of urinary albumin:creatinine ratio (ACR) as a simple predictor of the development of complications (SIRS) postoperatively. MethodsForty-four patients undergoing elective infrarenal AAA repair and 10 control patients undergoing major abdominal surgery had fresh urine samples taken before, immediately after and 24h after the procedure. Urinary ACR was calculated on all samples, and daily SIRS scores were calculated for all patients postoperatively. Systemic interleukin-6 (IL-6) levels were measured intraoperatively to measure the cytokine response to surgery. ResultsAAA patients demonstrated a characteristic pattern of ACR levels during the three time points, with a significant increase in the ACR immediately postoperatively and with normalisation by 24h (P<0.001 Wilcoxon signed ranks test). In comparison, control patients did not demonstrate any changes in their ACR (P=0.45 Wilcoxon signed ranks test) suggesting the increased ACR in AAA patients to occur as a result of IRI. ACR did not correlate with the development of SIRS postoperatively or with the systemic IL-6 response. ConclusionsInfrarenal AAA repair is associated with a temporary and reversible renal injury. ACR could not, however, be used as a predictor of complications postoperatively.

Structural and functional characterization of myotoxin I, a Lys49 phospholipase A 2 homologue from the venom of the snake Bothrops atrox

A new myotoxin was isolated from the venom of Bothrops atrox from Colombia. B. atrox myotoxin I is a homodimer, with a subunit molecular mass of 13,826, and a p I of 8.9. Its complete nucleotide sequence was obtained by cDNA cloning, indicating a mature product of 122 residues that belongs to the family of Lys49 phospholipase A 2 (PLA 2) homologues, a subgroup of catalytically inactive proteins within the group IIA. Accordingly, the toxin was devoid of phospholipase and anticoagulant activities, in vitro. In mice, it induced conspicuous local myonecrosis, edema, and a systemic interleukin-6 response. In vitro, it was cytolytic upon myoblasts, and weakly bactericidal. The toxin showed highest homology with other Lys49 PLA 2s, both in its primary and three-dimensional modeled structure, although with an evident difference in the C-terminal region. Unlike Lys49 proteins of American crotalids having 121 residues, this toxin presents an insertion (Asn) between positions 118 and 119. Despite several substitutions within the C-terminal region 115–129 between B. atrox myotoxin I and B. asper myotoxin II, antibodies against synthetic peptide 115–129 of the latter were strongly cross-reactive to the former, indicating the antigenic conservation of this site, known to be critical for the membrane-damaging activities of Lys49 myotoxins.

Systemic Interleukin-6 Response to Colorectal Surgery Originates from the Bowel

Background: Surgical trauma evokes a systemic cytokine response which is enhanced in patients with colorectal cancer. The aim of this study was to locate the origin of the systemic cytokine response to colorectal surgery. Methods: The concentrations of interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α) were analysed in systemic and mesenteric venous blood in 12 patients operated on with colorectal resections due to cancer or benign lesions. Immunohistochemical staining and analysis of tissue concentrations of IL-6 and TNF-α in homogenates from tumours and benign specimens were performed. Results: Mesenteric venous blood contained higher concentrations of IL-6 compared to systemic venous blood after resection, but not before. Tissue concentration of IL-6 was higher in the tumours compared to the benign specimens and immunohistochemistry revealed an abundance of IL-6 and TNF-α in malignant epithelium compared to benign mucosa. Conclusion: The higher concentration of IL-6 in venous blood from the mesenterium of the resected colonic segment compared to systemic levels, indicates that the bowel is the source of the IL-6 response to surgical trauma in colorectal surgery.

Central beta-amyloid peptide-induced peripheral interleukin-6 responses in mice.

beta-Amyloid peptides (Abetas) share with lipopolysaccharide, a potent pro-inflammatory agent, the property of stimulating glial cells or macrophages to induce various inflammatory mediators. We recently reported that central administration of lipopolysaccharide induces peripheral interleukin-6 responses via both the central and peripheral norepinephrine system. In this study, the effect of intracerebroventricular injection of various synthetic Abetas on plasma interleukin-6 levels was examined in mice. Abeta(1-42) dose-dependently increased plasma interleukin-6 levels: 'aged' Abeta(1-42) was more effective than fresh, whereas Abeta(42-1) had no effect. 'Aged' Abeta(1-42) (205 pmol/mouse i.c.v.)-induced plasma interleukin-6 peaked at 2 h post injection, which is earlier than the peak time of the Abeta(1-42)-induced brain interleukin-6, tumor necrosis factor-alpha and interleukin-1beta levels, which was 4, 4 and 24 h, respectively. Among various peripheral organs, Abeta(1-42) (205 pmol/mouse i.c.v.) significantly increased interleukin-6 mRNA expression in lymph nodes and liver. Abeta(1-42) (205 pmol/mouse i.c.v.) significantly increased norepinephrine turnover in both hypothalamus and spleen. Either central or peripheral norepinephrine depletion effectively inhibited the Abeta(1-42)-induced peripheral interleukin-6 response. Pretreatment with prazosin (alpha(1)-adrenergic antagonist), yohimbine (alpha(2)-adrenergic antagonist), and ICI-118,551 (beta(2)-adrenergic antagonist), but not with betaxolol (beta(1)-adrenergic antagonist), inhibited Abeta(1-42)-induced plasma interleukin-6 levels. These results demonstrate that centrally administered Abeta(1-42) effectively induces the systemic interleukin-6 response which is mediated, in part, by central Abeta(1-42)-induced activation of the central and the peripheral norepinephrine systems.

Portal endotoxin and cytokine responses during abdominal aortic surgery

Interleukin-6 (IL-6) is a major marker of tissue injury. Surgery induces a systemic IL-6 response related to the magnitude of the operation. An exaggerated IL-6 response is associated with the development of major complications after aortic aneurysm repair. Endotoxemia is a potent stimulus for cytokine release and may occur during aortic surgery. This study attempted to examine whether the observed IL-6 response during abdominal aortic surgery is related to the development of portal endotoxemia. Serial samples of inferior mesenteric and systemic venous blood were obtained before, during, and after aortic cross-clamping in eight patients undergoing elective aortic aneurysm surgery. Portal endotoxemia was detected in only five of the eight patients during the surgery, and there was no correlation with the IL-6 response. Both portal and systemic IL-6 levels rose during the course of surgery, but, by the time of abdominal wound closure, the mean portal IL-6 levels were eight times higher than the systemic IL-6 levels. These results suggest that the portal circulation draining the colon is a major source of the previously observed systemic IL-6 response to aortic surgery. The IL-6 response is not clearly related to portal endotoxemia but may reflect occult cellular injury in the colon occurring during surgery.