Natural and synthetic polypeptides that recognize the conserved lipid a binding site of lipopolysaccharides.
Explore the source record for details and available documents.
Acute-Phase Proteins↗
SEARCH · Search PubMed
Results for “Limulus Test”
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
[Infective factors of adult secretory otitis media].
There were histories of the upper expiratory tract infection in 49 out of 86 cases in adult secretory otitis media (SOM). Among them, thirty two cases were administrated antibiotics a week ago when the effusions were collected. The patient's Eustachian tubes in twenty nine cases were clinically ventilated. The endotoxins were positive in 39 out of 86 specimen tested with limulus assay, among these specimen, bacteria were cultured from 11 specimen. The data exibited that the appearance of the SOM is related to the existence of the infective factors in the middle ear cavity. The upper expiratory tract infection before the onset of the middle ear effusions is one of the important causes inducing the disease. The lower rate of bacteria culture than that of endotoxins is related with the administration of antibiotics before the onset of the disease. The administration of antibiotics in the treatment of the middle ear effusions will help to elimilate bacteria in the cavity of the middle ear effusions, improve the ventilating function of the Eustachian tube and make the effusions turn to disappearance.
Adolescent↗
Endotoxin activity measured by limulus assay.
The binding activity of endotoxin (Escherichia coli 0111) to dentin powder pre-treated with 10% sodium hypochlorite and with 3% hydrogen peroxide was investigated. This was carried out for periods of 30, 10 and 1 min. The endotoxin was diluted into 100 ng/ml solutions. Dentin powder suspended in a 34 micrograms/ml protein concentration was used in the study. The limulus amoebocyte lysate test was used to determine the amount of endotoxin binding, which was measured by the microtiter plate reader. The binding activity level of that concentration of endotoxin was significantly lower in dentin powder treated with 10% sodium hypochlorite than in that treated with 3% hydrogen peroxide at 1 and 10 min. Endotoxin binding activity measured the lowest in the 30 min test period.
Dentin↗
Comparison of several control standard endotoxins to the National Reference Standard Endotoxin--an HIMA collaborative study.
A collaborative study, initiated under the auspices of the Health Industry Manufacturers Association (HIMA), was designed to establish the relationship of Escherichia coli O55:B5 endotoxin (the control standard endotoxin of HIMA and the Food and Drug Administration's Office of Medical Devices) to the U.S. National Reference Standard Endotoxin and to two internationally used control standard endotoxins. By using two Limulus amoebocyte lysate test systems, it was established that the E. coli O55:B5 endotoxin lot originally used by HIMA and the Office of Medical Devices to establish Limulus amoebocyte lysate release test criteria for pyrogen testing of medical devices contains approximately 4.5 endotoxin units (EU) per ng. Thus, the 1.0-ng/kg endotoxin dose limit currently established for medical devices is approximately the same as the 5.0-EU/kg endotoxin limit (on an activity basis) established by several other Food and Drug Administration agencies for human and animal parenteral drugs and biological products.
Endotoxins↗
Conventional tube and microplate Limulus amoebocyte lysate procedures for determination of gram-negative bacteria in milk.
A comparison was made of the conventional tube and microplate Limulus amoebocyte lysate assay for detection of gram-negative bacterial lipopolysaccharide in milk. Raw whole milk samples were maintained frozen and portions were examined periodically on alternate days during 13-d storage to evaluate the reproducibility of both Limulus amoebocyte lysate procedures and to determine optimum reaction conditions for the microplate method. One-day-old, raw and locally purchased pasteurized milk samples, held at 7 degrees C, were analyzed during storage to establish the correlation of both procedures with aerobic and modified psychrotrophic plate counts. Vitamin- and mineral-fortified dairy-based products were examined using the microplate Limulus amoebocyte lysate test as a potential indicator of raw material or finished product bacterial quality and possible postprocessing contamination. Statistical analysis of the data collected comparing the conventional tube and the microplate Limulus amoebocyte lysate assay demonstrated no significant difference exists between the methods when either the modified psychrotrophic bacterial count or the aerobic plate count was used to determine gram-negative bacteria in pasteurized or raw milk (P less than .91). The microplate method, which uses half the lysate reagent, was a good indicator of the bacterial quality of milk and fortified dairy products, consistently detecting bacterial levels greater than 10(3) to 10(4)/ml.
Animals↗
Characterization of sodium hypochlorite degradation of beta-glucan in relation to its metabolism in vivo.
Soluble (SSG, beta-1,3-D-glucan obtained from the culture filtrates of a fungus, Sclerotinia sclerotiorum IFO 9395) and particulate (zymosan) beta-glucans were oxidized by sodium hypochlorite (NaClO), and the oxidized products were analyzed by gel filtration and ion-exchange chromatographies and by limulus G-test to study the metabolism of beta-glucans in vivo. By oxidative degradation, SSG was gradually oxidized to anionic polymers, which decreased the molecular weight and reduced the content of the sidechain at the same time. Zymosan, a particle from fungi cell wall, was easily solubilized to a high molecular weight polysaccharide by oxidative degradation. The elution profiles on ion exchange columns and the limulus G-test reactivity of the products were similar to those of polysaccharides obtained by in vivo experiments using SSG and zymosan. These results suggest that oxidative degradation is the main metabolic pathway of beta-glucans in vivo, and that the sidechains would be oxidized faster than the main chain.
Ascomycota↗
Determination of endotoxins on hypodermic needles by means of a chromogenic Limulus amoebocyte lysate assay. Development of a test model.
A test model was developed in order to describe the determination of endotoxins on hypodermic needles in a reliable and reproducible manner. As in all in-vitro experiments one has to be very careful about extrapolating data to in-vivo situations. However by choosing a hydrophilic (Salmonella typhimurium ATCC 14028) and a hydrophobic bacterium (Acinetobacter calcoaceticus var. anitratus RR 8212/113), one could hope to obtain a quite representative idea about the extraction of contaminating gram-negative micro-organisms. Using the proposed extraction procedure and a chromogenic LAL assay as detection system, it was possible to achieve a quantitative idea about the extraction of endotoxins on hypodermic needles. Extracting needles at 19 degrees C during 83 to 98 min produced the highest yield as to the detection of endotoxins from hydrophilic strains. For the detection of endotoxins from hydrophobic strains, the highest yield values were obtained when the extraction was performed at temperatures between 64 and 80 degrees C during 96 to 120 min. However it seems necessary to perform the extractions at least three times in order to obtain reproducible results. In conclusion, using the extraction procedure as described, it is possible to measure endotoxin-like activity, on the inner and outer surface of hypodermic needles in a simple but accurate way, using water as extraction fluid and a chromogenic Limulus Amoebocyte Lysate assay as detection system.
Acinetobacter↗
Augmentation of endotoxin fever by recombinant human beta interferon in rabbits.
Nonpyrogenic amounts of endotoxin (0.1 to 1 ng/kg), hardly detectable by conventional Limulus amoebocyte lysate tests, could produce a fever of around 1 degree C when injected with a nonpyrogenic dose (6 X 10(5) U/kg) of recombinant human beta interferon (IFN-beta) in rabbits. Release of endogenous IFN and tumor necrosis factor by endotoxin was also dramatically increased by recombinant human IFN-beta, and their levels in the blood were closely correlated with the increase of body temperature. These data suggest, if the synergism between IFN and endotoxin also operates in the homologous system (human IFN-human cells), that contaminating endotoxin in IFNs, even if not detectable by Limulus amoebocyte lysate test, can contribute to IFN fever to a considerable extent in humans.
Animals↗
Teicoplanin reduces in-vitro reactivity and murine lethality of Salmonella minnesota R595 lipopolysaccharide.
Three different tests were performed to investigate the effect of teicoplanin on lipopolysaccharide (LPS). After incubation for 3 h with teicoplanin, LPS from Salmonella minnesota R595 showed reduced reactivity in the metachromatic dimethyl-methylene blue assay and the limulus amoebocyte lysate test. In addition, galactosamine-sensitized mice had an increased survival rate, from 29% to 72%, when teicoplanin was pre-incubated for 3 h with the LPS to be injected intraperitoneally. The results suggest that teicoplanin may have a neutralizing effect on LPS.
Animals↗
Detection of endotoxin in media from sterile corneal organ cultures.
Lipopolysaccharides (LPS) are elements of the cell wall of gram-negative bacteria. They are also called endotoxins and are known to stimulate various inflammatory reactions by interaction with cytokines and macrophages. Additionally, they may have a direct effect on other cells. As contamination of sterile organ-culture media with bacterial substances may influence the donor-tissue prognosis, we investigated a series of culture media drawn from organ culture for the presence of endotoxin. A total of 341 samples of sterile organ-culture media from 3 different cornea banks using either organ culture or the refrigerated-storage technique were tested for endotoxin. The assay was performed using the Limulus amebocyte-lysate test. A level of endotoxin above the background threshold was found in 99/341 (29%) culture media. The incidence of endotoxin ranged from 14% up to 50%, depending on the cornea bank and culture system used. Endotoxin detected in sterile corneal organ cultures probably derives from nonreplicating bacterial postmortem donor-tissue contamination. The presence of endotoxin-positive cultures varied between the eye banks but was not related to a given storage method. As endotoxin may directly influence graft viability or trigger inflammatory host responses, these findings may have significance for the clinical results of corneal grafting.
Cornea↗
Designs for the evaluation of the endotoxin assay as a diagnostic test.
The comprehensive evaluation of the usefulness of a new diagnostic test should include a hierarchy of assessments ranging from reliability and validity to impact on clinical decision making and outcome. This paper reviews some of the design methodology appropriate for evaluation of an endotoxin assay such as the Limulus Amebocyte Lysate test. Primary focus is on the design and analysis of studies aimed at establishing diagnostic validity. However, suggestions are made for the design of clinical studies of management change and to determine the impact on patient outcome. Results from such studies must be combined into a cost-utility calculation eventually to determine societal benefit.
Bacterial Infections↗
Non-cultural detection of Neisseria gonorrhoeae in cervical and vaginal washings.
Genetic transformation, an indirect sandwich enzyme-linked immunosorbent assay (ELISA) and the Limulus amoebocyte assay were used to indicate the presence of products of Neisseria gonorrhoeae in vaginal and uterine cervical aspirates from 37 women attending a Department of Genito-Urinary Medicine. In parallel with these tests, qualitative and quantitative assessments of the microbial content of aspirates were made. There was wide variation in the numbers of gonococci cultured. The mean viable count for cervical aspirates was 1 x 10(6) cfu/ml and the range was (5 x 10(3))-(8 x 10(6)) cfu/ml; the mean count for vaginal aspirates was 8.4 x 10(4) cfu/ml and the range (1 x 10(2))-(1 x 10(6)) cfu/ml. Viable counts of organisms other than gonococci in vaginal aspirates were two to tenfold greater than the corresponding counts for cervical aspirates. Of 20 patients with gonorrhoea confirmed by conventional diagnostic cultures, aspirates from 15 (75%) gave a positive transformation result, and 12 (60%) a positive ELISA result; 16 (84.2%) out of 19 of these aspirates tested by the Limulus lysate assay were positive at a dilution of 1 in 100.
Cervix Uteri↗
Endotoxin neutralization with rabbit antisera to Escherichia coli J5 and other gram-negative bacteria.
To study the mechanisms of protection against endotoxin challenge offered by antisera to smooth and rough gram-negative organisms, we have developed an assay to quantitate endotoxin neutralization based on inhibition of the Limulus amoebocyte lysate test. Dilutions of different bacterial lipopolysaccharides (LPSs) were incubated with hyperimmune rabbit sera against Escherichia coli O113, E. coli O18, and rough mutants E. coli J5 and Salmonella minnesota Re595 and were then combined with limulus lysate. The gelation reaction induced by LPS in the lysate was monitored spectrophotometrically, and the concentration of LPS resulting in a 50% lysate response was determined and correlated with antibody titers measured by enzyme-linked immunosorbent assay. Antisera to smooth organisms neutralized homologous LPS markedly and heterologous LPSs only minimally relative to neutralization by preimmune serum. Neutralization of homologous LPS occurred immediately without preincubation of serum and LPS. Antisera to rough mutants neutralized more heterologous LPS than did antisera to smooth organisms. However, this heterologous neutralization required preincubation of serum and LPS and did not appear to be correlated with antibody concentrations. We conclude that antisera to LPS rapidly neutralize the biological activity of the homologous LPS, as detected by limulus lysate, and that neutralization is at least in part antibody mediated. Antisera to rough-mutant organisms slowly neutralized the activity of heterologous LPSs, but this effect appeared not to be correlated with concentrations of antibody to the LPS of the rough mutant, as measured by enzyme-linked immunosorbent assay.
Animals↗
Leakage evaluation of root end filling materials using endotoxin.
Mineral trioxide aggregate (MTA) has been shown to possess excellent sealing ability when tested with dye, bacteria, and a fluid filtration technique. Endotoxin, a component of the cell wall of Gram-negative bacteria, has been proposed to play a role in the pathogenesis of periradicular lesions. This study used a modified Limulus Amebocyte Lysate test for the presence of endotoxin as a tracer and compared the sealing ability of Super-EBA, IRM, amalgam, and MTA. The results showed that MTA permitted less endotoxin leakage than IRM and amalgam at 1, 2, 6, and 12 wk (p < 0.05), and leaked less than Super-EBA at 2 and 12 wk (p < 0.05).
Aluminum Compounds↗
The pyrogenicity of the synthetic adjuvant muramyl dipeptide and two structural analogues.
The pyrogenic efect of the synthetic adjuvant N-acetylmuramyl-L-alanine-D-isoglutamine, also known as muramyl dipeptide (MDP), was studied in rabbits. MDP induced biphasic fevers in rabbits, but two structural analogues, N-acetylmuramyl-L-alanine-D-glutamic acid (MDPA) and the dimethylester of MDPA, were 10 times less pyrogenic. This finding was supported by studies in which MDP and its analogues released leukocytic pyrogen (LP) from rabbit phagocytic cells in vitro. In addition, MDP released LP from human phagocytes. Human phagocytes, however, required a 10-fold greater concentration of MDP than did rabbit cells. The structural analogues were similarly less effective than the parent molecule in releasing LP from human cells. All preparations of MDP were negative in the limulus amebocyte lysate test and failed to show pyrogenic cross-tolerance with bacterial endotoxin. Thus MDP, which is a pyrogenic molecule, is also able to release LP from rabbit phagocytes and to a lesser degree from human phagocytes, but does not cause gelation of limulus amebocyte lysate.
Acetylmuramyl-Alanyl-Isoglutamine↗
Limulus lysate assay in detection of gonorrhea in women from a low-incidence population.
An evaluation of the limulus amebocyte lysate assay for detection of gonococcal endotoxin in cervical secretions was undertaken in 48 women from an ambulatory population with a low incidence of gonococcal infection. When cervical secretions were diluted 1 : 2,000, positive limulus amebocyte lysate assay results were obtained in four women (100%) with culture-proved gonococcal infection, and negative results were obtained in 25 of 44 women (57%) with culture-negative specimens. The limulus amebocyte lysate assay was not sufficiently specific in a low-incidence, largely asymptomatic population to be recommended as a rapid diagnostic procedure (60 minutes) in such a group. Standardization of sampling technique, size, and reagent preparation and elimination of vaginal contamination of cervical secretions may produce a limulus amebocyte lysate test with more consistent performance characteristics in varied populations. Since the predictive value of a test varies with the prevalence of the disease in the populations tested, a test proposed for use in a population with a low likelihood of disease must be tested in such a population.
Adolescent↗
[Detection of the endotoxin of Gram-negative bacteria in human blood].
Recent new data on the important role played by lipopolysaccharides (endotoxin) of Gram negative bacteria in physiology and pathogenesis of the most important human infectious and noninfectious diseases testify to the necessity of wide clinical trials of different methods for LPS detection in blood and other physiological fluids. Among presently available diagnostic methods for endotoxinemia detection, the highly sensitive LAL (Limulus Amebocyte Lysate) test in various modifications is most widely used. The LAL test is known to be non-specific, however many drawbacks of this test have been successfully overcome. The results of clinical studies on the determination of the LPS activity in the systemic blood stream and antibody titers to its most common determinants, as well as the reserves of endotoxin binding with granulocytes give grounds for optimistic evaluation of the future studies on the role of LPS in human physiology and pathology. In clinical practice both positive sides and drawbacks of the presently known methods for LPS detection, including the LAL test, must be borne in mind for the complex evaluation of endotoxinemia levels.
Blood↗
Direct cytotoxic action of Shiga toxin on human vascular endothelial cells.
To help explain a role of the Shiga toxin family in hemorrhagic colitis and hemolytic-uremic syndrome in humans, it has been hypothesized that these toxins cause direct damage to the vascular endothelium. We now report that Shiga toxin purified from Shigella dysenteriae 1 does indeed have a direct cytotoxic effect on vascular endothelial cells in cultures. Human umbilical vein endothelial cells (HUVEC) in confluent monolayers were reduced 50% by 10(-8) M Shiga toxin after a lag period of 48 to 96 h. In comparison, nonconfluent HUVEC were reduced 50% by 10(-10) M Shiga toxin within a 24-h period. These data suggest that dividing endothelial cells are more sensitive to Shiga toxin than are quiescent cells in confluent monolayers. Both confluent and nonconfluent HUVEC specifically bound 125I-Shiga toxin. However, in response to the toxin, rates of incorporation of [3H]leucine into protein were more severely reduced in nonconfluent cells than in confluent cells. Toxin inhibition of protein synthesis preceded detachment of cells from the substratum. The specific binding of 125I-Shiga toxin to human endothelial cells and the cytotoxic response were both toxin dose dependent and neutralized by anti-Shiga toxin antibody. Heat-denatured Shiga toxin was without the cytotoxic effect. In addition, the complete culture system contained less than 0.1 ng of bacterial endotoxin per ml, as measured by the Limulus amoebocyte lysate test.
Bacterial Toxins↗