Bacterial endotoxins: biological properties and mechanisms of action

Endotoxins (lipopolysaccharides, LPS) are agents of pathogenicity of Gram-negative bacteria, implicated in the development of Gram-negative shock. Endotoxin reacts with lipopolysaccharide-sensitive cells producing endogenous mediators such as tumour necrosis factor alpha (TNFalpha). Macrophages are cells mediating the toxic activities of LPS and TNFalpha is the primary mediator of the lethal action of endotoxin. This review article discusses the various mechanisms by which endotoxin hypersensitivity in bacteria-sensitized animals develops. The paper concludes with a discussion on the possible protective effect of carnitine congeners against the lethal action of LPS.     

Simple method for analysis of diquat in biological fluids and tissues by high-performance liquid chromatography

A simple HPLC method has been described to quantify diquat in biological fluids and tissues. This method permits separation and quantification of diquat from blood, bile, urine, liver and kidney. It does not require special pretreatment of the samples prior to analysis, nor a specially prepared analytical column. Various concentrations of diquat were added (10–300 nmol/ml or g) to fluids or tissues. Analysis of blank samples revealed no substrates that interfere with diquat elution. Excellent recovery (95–105%) was obtained. Diquat (120 μmol/kg, i.v.) was injected to rats and quantified in bile, blood and liver. Concentration of diquat was higher in blood and bile than liver. Therefore, this method is applicable for quantification of diquat in toxicological samples, and may be used to determine structurally similar compounds such as paraquat.     

Simple and reliable method of doxycycline determination in human plasma and biological tissues

Over recent years there has been a resurgence in the use of doxycycline in clinical practice, which does not depend on its antibacterial properties. This paper describes a method of determination of doxycycline in human plasma and atheromatous tissue using high-performance liquid chromatography (HPLC), and a cheap commercially available extraction system. Doxycycline is extracted in the mobile phase and injected directly into the HPLC system, avoiding time consuming drying up steps. A limit of detection of 0.125 μg/ml of plasma, and a relative standard deviation of 3% was achieved, making the method very reliable and useful for assays within the usual therapeutic range. The method has also been applied to the extraction of a mixture of tetracyclines from plasma and atherma with equal efficacy, making it useful for assays of this class of drugs in veterinary practice and assays of food contaminants.     

Simple method for the collection of pancreatic islets by the use of Ficoll-Conray gradient.

Simple method for the collection of large numbers of viable pancreatic islets from normal rats is described. This method involves collagenase digestion of the pancreas, followed by the use of Ficoll-Conray discontinuous gradient for the separation of isolated islets from unwanted acinal debris. Ficoll-Conray A solution was prepared by mixing undialyzed 12.5% Ficoll and 33.4% Conray in the ratio 2 : 1, to make a specific gravity of 1.095 and osmolarity of 396 mOsm/l. The solution B, C, and D, with specific gravities 1.084, 1.072, and 1.048, respectively were obtained by diluting A solution with distilled water. Using Ficoll-Conray gradient, about 200 viable islets which maintained excellently their morphology and function, were collected consistently from a young adult rat pancreas.     

The use of monolaurates for removing endotoxins from preparations of cell-free pertussis vaccine]

The possibility of using monolaurates for removing endotoxin from acellular pertussis vaccines developed at the Laboratory of Immunomodulators, Mechnikov Research Institute for Vaccines and Sera (Moscow), has been studied. Monolaurates PEG-400 and PEG-600 obtained, respectively, from Fluka Chemie AG and Ferak GmbH (Germany) have been used. The use of monolaurate PEG-600 ensures the decrease of the toxicity of acellular pertussis vaccines by 2.34-6.3 times.     

Hygienic evaluation of repurification schemes for waste waters containing complexes for organic substances

Sanitary-chemical and sanitary-toxicological methods were used to study two repurification schemes for biologically purified waste waters from a petrochemical industrial complex. These repurification schemes were, (1) filtration through quartz sand, adsorption to activated charcoal, chlorination; (2) coagulation, filtration through quartz sand, adsorption to activated charcoal, chlorination. Both repurification schemes considerably improved the composition and properties of the waste waters in terms of organoleptic and sanitary-chemical indices. Scheme 1 also considerably lowered the toxic properties of the waste waters and Scheme 2 abolished them completely. Provided that the corresponding sanitary norms are observed, the use of repurification Scheme 1 would be economically reasonable where repurified waste water is recirculated in the industrial plant. Repurification Scheme 2 is recommended where purified waste water is disposed into low-capacity reservoirs.     

Rapid biological and chemical analyses of bacterial endotoxins separated by preparative thin-layer chromatography

Methods have been elaborated for rapid location of biologically active bacterial endotoxin components on preparative thin-layer chromatography (tlc) by Limulus lysate clotting assay. The separation of the components from silicic acid is not necessary. Further assays were established for the semiquantitative estimation of endotoxic activites (Shwartzman reactivity, chick embryo lethality, and non-specific tumor resistance enhancement) without complete removal of silicic acid.Quantitative chemical analytical procedures were also elaborated to determine the molar ratios of the biologically active components separated and detected in the above tlc system. These included phosphorus, hexosamine, heptose, 2-keto-3-deoxyoctonate, and long-chain carboxylic acid content measurements. Here again, the chemical determinations were carried out in the presence of silicic acid.