The aim of this work was to compare production of endotoxin and to determine susceptibility to antibiotics in two groups of specimens-wild-type strains Ochrobactrum anthropi isolated from the environment and the strains isolated from patients with cystic fibrosis. The determination of the endotoxin produced by the test strains was carried on by using a limulus amebocyte lysate test (LAL test). Determination of ATB sensitivity was accomplished by means of a broth dilution method in a microtiter plate (MIC). No significant difference was found between the group of ochrobacters isolated from the environment and the group of ochrobacters isolated from cystic fibrosis patients. Antibiotic sensitivity testing has indicated that the resistance to tigecycline, trimethoprim/sulfamethoxazole, and gentamicin was slightly higher in strains isolated from cystic fibrosis patients in comparison with strains isolated from the environment. In general, most of the test strains were sensitive to most of the antibiotics tested. Significant resistance has been demonstrated for cefotaxime. Resistance was also found for gentamicin in strains number 4 and 7. The MIC was equal to the breakpoint for this antibiotic (8000 mg/L).
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence MeSH
- cystická fibróza mikrobiologie MeSH
- dítě MeSH
- endotoxiny metabolismus MeSH
- gramnegativní bakteriální infekce mikrobiologie MeSH
- lidé MeSH
- Limulus test MeSH
- mikrobiální testy citlivosti MeSH
- mikrobiologie životního prostředí MeSH
- mladiství MeSH
- Ochrobactrum anthropi účinky léků izolace a purifikace metabolismus MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
- mladiství MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- antibakteriální látky MeSH
- endotoxiny MeSH
Similar to lipopolysaccharide (LPS), a product of Gram-negative bacteria, the signal macromolecules of Gram-positive bacteria lipoteichoic acid (LTA) and peptidoglycan (PGN) possess multiple biological activities. They may be a source of misinterpretation of experimental findings. We have found that not only LPS but also LTA and PGN can be detected by the Limulus amebocyte lysate (LAL) assay. All of them stimulate the high output in vitro nitric oxide (NO) production of in rat peritoneal cells. The onset of the NO enhancement was observed with 25-100pg/ml of LPS and 25-100ng/ml of PGN and LTA. Polymyxin B (PMX), if applied at concentration 10,000-fold higher than that of LPS, can completely inhibit the NO and LAL binding responses of LPS. The NO-stimulatory and LAL-binding properties of LTA and PGN are not eliminated by PMX. Handling of LPS contamination with PMX may be associated with serious problems because it possesses intrinsic biological activity and becomes cytotoxic at concentration >25μg/ml. The present findings suggest a convenient alternative avoiding these issues. As monitored by the NO and LAL assays, even high amounts of LPS as well as PGN and LTA can be removed by molecular mass cutoff microfiltration. All types of the filters (3kDa to 100kDa) are equally effective. It is suggested that the microfiltration procedure may be considered as a preferable, general and easy method of sample decontamination.
- MeSH
- Bacillus subtilis chemie izolace a purifikace MeSH
- buněčné linie MeSH
- centrifugace MeSH
- Escherichia coli chemie izolace a purifikace MeSH
- filtrace * MeSH
- krysa rodu Rattus MeSH
- kyseliny teichoové analýza MeSH
- Limulus test * MeSH
- lipopolysacharidy analýza MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- oxid dusnatý biosyntéza metabolismus MeSH
- peptidoglykan analýza MeSH
- potkani Wistar MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- kyseliny teichoové MeSH
- lipopolysacharidy MeSH
- lipoteichoic acid MeSH Prohlížeč
- oxid dusnatý MeSH
- peptidoglykan MeSH
A chemical analysis was performed of lipid A, isolated by acid hydrolysis of the lipopolysaccharide from the S and R forms of Shigella dysenteriae serovar 1. Differences in the moiety of both lipids and sugars were compared. The lipid portion consisted of a homologous series of fatty acids ranging from C12:0 to C18:0 (predominant homologues, C12:0, C14:0 and C16:0) and the homologous series of 3-hydroxy acids ranging from C12:0 to C16:0 (predominant homologue, 3-OH-C14:0). The major sugar portion consisted of D-glucosamine. The toxicity of lipid A in mice (LD50) ranged between 300-400 micrograms/mouse, and values from the Limulus amebocyte lysate assay were recorded as titres of 10(-5) to 10(-6) mg/ml.
- MeSH
- glukosamin analýza MeSH
- LD50 MeSH
- Limulus test MeSH
- lipid A chemie toxicita MeSH
- mastné kyseliny analýza MeSH
- myši inbrední ICR MeSH
- myši MeSH
- sacharidy analýza MeSH
- sérotypizace MeSH
- Shigella dysenteriae chemie klasifikace MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- glukosamin MeSH
- lipid A MeSH
- mastné kyseliny MeSH
- sacharidy MeSH
The lipopolysaccharides, lipopolysaccharide-protein complexes and their lipids A, isolated from Shigella dysenteriae 1, exhibited lethal toxicity (LD50 300-400 micrograms per mouse), pyrogenicity (0.01-1.0 microgram), activity in the Limulus test (10(-3)-10(-12) mg ml-1) and produced a positive local Shwartzman reaction. It was found that these substances bound to Al(OH)3 were capable of successfully protecting mice from challenge with both homologous and heterologous infections (up to 90% of mice survived a challenge with LD100), whereas immunization with substances with no biological carrier exhibited very low or zero protective effects. Furthermore, the positive cross-reactivity of lipids A in the agar immunodiffusion test confirms the wide applicability for vaccination purposes.
- MeSH
- bacilární dyzentérie prevence a kontrola MeSH
- bakteriální vakcíny izolace a purifikace toxicita MeSH
- endotoxiny izolace a purifikace toxicita MeSH
- hydroxid hlinitý MeSH
- imunodifuze MeSH
- králíci MeSH
- LD50 MeSH
- Limulus test MeSH
- myši inbrední ICR MeSH
- myši MeSH
- pyrogeny MeSH
- Shigella dysenteriae klasifikace imunologie MeSH
- Shwartzmanův fenomén MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- Názvy látek
- bakteriální vakcíny MeSH
- endotoxiny MeSH
- hydroxid hlinitý MeSH
- pyrogeny MeSH
- MeSH
- bakteriální infekce vrozené diagnóza MeSH
- chybná diagnóza MeSH
- lidé MeSH
- Limulus test * MeSH
- novorozenec MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
The interferon (IFN)-inducing activity of detoxified lipopolysaccharide (LPS) was tested in rabbits treated with LPS preparation derived from Escherichia coli, Salmonella typhi, Salmonella enteritidis and Shigella dysenteriae serovar 1. Of the detoxification procedures used, alkaline hydrolysis, hydroxylaminolysis, formalization, treatment with sodium deoxycholate and the radiodetoxification (fast or slow) methods had no appreciable effects on the IFN-inducing potential of LPS. In contrast, acetylation or prolonged alkaline hydrolysis of LPS resulted in up to a 9-fold reduction of IFN-induction capacity and effects of Cu++ or Fe++ cations bound to LPS were clearly inhibitory (Fe more than Cu).
- MeSH
- acetylace MeSH
- endotoxiny izolace a purifikace farmakologie MeSH
- Escherichia coli metabolismus MeSH
- hydrolýza MeSH
- interferony biosyntéza krev MeSH
- králíci MeSH
- Limulus test MeSH
- lipopolysacharidy izolace a purifikace fyziologie MeSH
- metabolická inaktivace * MeSH
- Salmonella metabolismus MeSH
- Shigella metabolismus MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- srovnávací studie MeSH
- Názvy látek
- endotoxiny MeSH
- interferony MeSH
- lipopolysacharidy MeSH
- MeSH
- Bacteroides fragilis MeSH
- endotoxiny analýza MeSH
- Escherichia coli MeSH
- Limulus test * MeSH
- lipopolysacharidy analýza MeSH
- Listeria monocytogenes MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- endotoxiny MeSH
- lipopolysacharidy MeSH
