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PubMedDate not supplied

Comparison of the binding of gram-negative bacterial endotoxin by polymyxin B sulphate, colistin sulphate and colistin sulphomethate sodium.

Polymyxins are cyclic polypeptide antibiotics. In addition to their bactericidal activity they bind lipid A and neutralize the biological effects of bacterial endotoxin. We have studied the three available polymyxin preparations: polymyxin B sulphate (PB), colistin sulphate (CS) and colistin sulphomethate sodium (CMS), and compared their endotoxin binding capacity at equivalent therapeutic dosage. Each polymyxin was bound to a column of Sepharose 4B and challenged with 5 micrograms of endotoxin from Escherichia coli O127:B8. Recovery of endotoxin in the eluate was measured by a quantitative Limulus lysate microassay. PB and CS bound 94% of the challenge dose, CMS 89% and the control column (Sepharose alone) 24%. These results suggest that parenteral CMS (the least toxic polymyxin) retains useful anti-endotoxin capacity, and that in neutropenic patients, oral polymyxin may exert both anti-endotoxin and antimicrobial effects.

Colistin↗
PubMed1983

Endotoxaemia in septic shock: clinical and post mortem correlations.

The Limulus amoebocyte lysate assay was used as one of a series of laboratory and clinical investigations on a group of 31 patients suffering from septic shock in order to assess the clinical significance of this assay for the detection of circulating endotoxin in clinical gram-negative sepsis. Four patients with cardiogenic shock served as controls. Endotoxin was detected in the bloodstream of all patients with septic shock during the 24 h following referral and was not detected in the control patients. Eventual clinical recovery was associated with the disappearance of endotoxin from the peripheral blood. Blood cultures were unhelpful as a prognostic indicator in these critically ill patients. A quantitative assay of endotoxin in blood may allow a more precise relationship with the clinical manifestations of major sepsis.

Adolescent↗
PubMed1993

Anti-lipopolysaccharide activity of histatins, peptides from human saliva.

Histatins are histidine-rich polypeptides secreted in human saliva. They were found to inhibit lipopolysaccharide (LPS)-mediated gelation of Limulus amoebocyte lysate, and to reverse the anti-complement action of LPS or lipid A. Histatins also gave precipitate bands in agarose gels with various LPS. The results indicate that histatins neutralized the activity of LPS by binding to the lipid A moiety of LPS.

Escherichia coli↗
PubMed1988

Detection of gram-negative bacteria in urine by the chromogenic limulus assay.

The value of the chromogenic limulus assay for detection of gram-negative bacteria in urine was determined. The assay was performed in microtiter plates at room temperature. In 311 consecutively collected urine samples from patients with suspected urinary tract infection, the assay was positive in all 35 samples containing greater than or equal to 10(5) bacteria/ml. No false positive or false negative results were obtained. Four of six urine specimens from patients with gonococcal infection were positive in the assay, whereas samples from patients with chlamydial infection did not yield a positive result. The assay is a rapid and reliable method for detection of urinary tract infection caused by gram-negative bacteria when greater than or equal to 10(5) bacteria/ml are present.

Bacteriuria↗
PubMed1986

Release of endotoxin from bacteria exposed to ciprofloxacin and its prevention with polymyxin B.

An in vitro system for studying the phenomenon of antibiotic-induced release of endotoxin from gram-negative bacteria is described. Endotoxin was measured by a quantitative limulus lysate microassay. Using a strain of Escherichia coli, ciprofloxacin was shown to cause an immediate and sustained increase in endotoxin release compared to control cultures whereas gentamicin did not, despite an equally rapid bactericidal effect. The potential value of the anti-endotoxin effect of polymyxin B was examined using a polymyxin B resistant strain of Klebsiella aerogenes. Ciprofloxacin alone caused endotoxin release, but when given with polymyxin B significantly less endotoxin was detected. These results indicate that in antibiotic-induced endotoxin release, the rate of cell death is less important than the site of antibiotic effect, and suggest that ciprofloxacin may act directly on the cell wall as well as on DNA gyrase. It is possible that polymyxin B could be used to reduce the extent of endotoxin release, and that this would be of clinical benefit.

Cell Wall↗
PubMed1980

Comparative study of biological activities and mitogenic effect of extracts from Haemophilus influenzae type a.

Two preparations designated as PWE and NCE were obtained from H, influenzae type a by treatment with either phenol water or hypertonic NaCl citrate respectively. The protein content of NCE was three fold higher than for PWE. Electroimmunodiffusion against H. influenzae antiserum disclosed an immunological relationship between NCE and PWE. The toxicity for mice was lower for both antigenic extracts (LD50 PWE: 47 +/- 7 mg/kg; LD50 NCE: 943 +/- 5 mg/kg) than for E. coli LPS. Limulus assay showed that both H. influenzae extracts had a weaker potency (100 to 1000 times less) than E. coli LPS. Sanarelli and Shwartzman reactions failed for PWE and NCE. The blast transformation of DBA/2 and nude mouse spleen cells were strikingly different for the three preparations: PWE, NCE, and E. coli LPS. Neither extract from H. influenzae cells showed the biological properties of a classical endotoxin. We suggest that they are endotoxin-like substances.

Animals↗
PubMed1992

Endotoxin tolerance and polymyxin B modify liver damage and cholestasis induced by a single dose of alpha-naphthylisothiocyanate in the rat.

A single oral dose of alpha-naphthylisothiocyanate (ANIT) induces intrahepatic cholestasis and endotoxemia in the rat. To assess if a pathogenic relationship between endotoxin and ANIT-induced liver injury could be postulated, rats were pretreated by either induction of endotoxin tolerance, or with the anti-endotoxin agent polymyxin B. A single oral dose (10 or 20 mg/100 g body wt) of ANIT was then given to ascertain whether these methods of modifying endotoxicity would protect the animals against ANIT damage. Both pretreatments significantly reduced the incidence of endotoxemia after ANIT administration, as detected by either lead acetate enhancement method or the Limulus gelation test (LGT). The lethality of a single 20 mg/100 g body wt dose of ANIT was reduced from 55% to 15% by polymyxin B administration, and to 10% by an endotoxin-tolerant state. Moreover, when 10 mg/100 g body wt ANIT was given none of the animals died in 10 days, and the serum levels of bilirubin, alkaline phosphatase (AlPh), gamma-glutamyl transferase (gamma-GT), and transaminases (evaluated 1, 2, and 5 days after treatments) were significantly lower in the endotoxin-tolerant or polymyxin B administered rats; this biochemical protection was mirrored in the lack of histological alteration. The results demonstrate that the modification of endotoxicity offers significant protection against acute liver damage induced by ANIT. Thus the development of endotoxemia may play a pathogenic role in ANIT-induced liver injury. This conclusion is supportive of the hypothesis that endotoxins are necessary for the hepatotoxic agent to exert its full effects.

1-Naphthylisothiocyanate↗
PubMed1988

Endogenous endotoxemia in patients with liver cirrhosis--a quantitative analysis of endotoxin in portal and peripheral blood.

In order to investigate the mechanism of endogenous endotoxemia (that is, endotoxemia observed in the absence of infection) in patients with liver cirrhosis, the concentration of endotoxin in the portal (PO-Et) and peripheral blood (PE-Et) from fifty three patients undergoing abdominal surgery was simultaneously measured by a quantitative endotoxin assay. The PE-Et of the patients with liver cirrhosis (19.8 +/- 20.2 pg/ml, n = 23) was significantly elevated, when compared with that of the patients without liver cirrhosis (9.2 +/- 5.1 pg/ml, n = 30), and was close to the normal range of PE-Et obtained from thirty healthy volunteers (7.2 +/- 4.1 pg/ml, n = 30). The PO-Et was also higher in the patients with liver cirrhosis than in the patients without liver cirrhosis. Moreover, PO-Et was significantly higher than PE-Et in all the patients (p less than 0.05). The per cent difference in the endotoxin concentration between the portal and peripheral blood (percentage of delta Et) was significantly decreased in the cirrhotic patients, especially in those with esophageal varices, which was well correlated with the phagocytic activity of the reticuloendothelial system (RES) determined by the clearance of iron colloid. The endogenous endotoxemia is thus likely to be due to the impaired clearance of endogenous endotoxin in portal blood, resulting from both the decreased phagocytic activity of RES in the liver and the coexisting porta-systemic bypass.

Endotoxins↗
PubMed1989

A study on limulus amebocyte lysate (LAL) reactive material derived from dialyzers.

The amebocytes of horseshoe crab (Limulus) hemolymph contain a coagulation system highly sensitive to bacterial endotoxins. Limulus amebocyte lysate (LAL) reactive material derived from cuproammonium rayon membranes, however, is not an endotoxin and acts upon the alternative pathway in the coagulation cascade found in Limulus amebocyte lysate. This study confirmed these facts by using the coagulation system of Limulus without factor G, which is a substrate of the alternative pathway. LAL reactive material lingered in the circulation for a relatively long time. In acute hemodialysis, its plasma concentration increased by an average of 100 pg/ml with each dialysis and eventually reached a plateau of approximately 300 pg/ml. In patients with chronic renal failure under regular hemodialysis, the mean level of LAL reactive material was 330.0 +/- 8.0 pg/ml before hemodialysis which increased by 70.6 +/- 20.7 pg/ml after four hours of hemodialysis.

Acute Kidney Injury↗
PubMed1986

Superoxide anion generating capacity of polymorphonuclear cells in patients with liver cirrhosis.

The superoxide anion generating capacity of polymorphonuclear cells (PMNs) in patients with liver cirrhosis and the effect of lipopolysaccharide on rat PMNs were examined. Superoxide anion generating capacity of PMNs was measured as luminol-dependent photon emission (chemiluminescence) during phagocytosis of peptide in vitro. Chemiluminescence of PMNs from patients with liver cirrhosis was significantly enhanced compared with normal healthy volunteers, and endotoxemia was detected in 3 out of 20 cases of liver cirrhosis by the limulus gelatin test. Serial studies revealed that chemiluminescence of PMNs and endotoxin in plasma decreased after administration of polymyxin B (3 X 10(6) u/day). Chemiluminescence of rat PMNs was also markedly enhanced after the injection of lipopolysaccharide, and persisted for more than 8 days even though endotoxemia was not detected. These results indicate that the enhancement of chemiluminescence by PMNs is related to endotoxins spilling over from the liver in patients with liver cirrhosis.

Animals↗