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At least 919 records · Page 51Linked to original sources
PubMed1998

Gradual solubilization of Candida cell wall beta-glucan by oxidative degradation in mice.

Candida spp. is a medically important fungi which induces disseminated candidiasis and candidemia in hospitalized immunocompromised patients. The cell wall of Candida is mainly composed of two polysaccharides, mannan and beta-glucan, and at least part of beta-glucan is basically insoluble in H2O or NaOH. We became interested in when and how particulate beta-glucan changes to the soluble form. However, the fate of wall components has not been examined in detail. In this study, modification and solubilization of the cell wall beta-glucan were analyzed in vivo and in vitro. Cells of Candida, intravenously administered to mice (1 mg/mouse), were immediately deposited mainly in liver as determined by 3H-labeled cells. Beta-Glucans were detected in these mice for at least for 6 months by the beta-glucan specific assay. During this period, the insoluble cell wall beta-glucan was gradually solubilized in these organs, probably by oxidative stress of macrophages. Candida cells and particulate beta-glucans were also gradually solubilized in vitro using sodium hypochlorite solution, but part of the cell wall beta-glucan was still insoluble even after treatment with concentrated hypochlorite solution for one day at room temperature. These findings strongly suggested that the fungal cell wall beta-glucans were quite resistant to oxidative metabolism in vivo and in vitro, and thus deposited for quite long period in the host.

Animals↗
PubMed1996

Biological activities of lipopolysaccharide of Burkholderia (Pseudomonas) pseudomallei.

Endotoxic activities of lipopolysaccharide (LPS) isolated from Burkholderia (Pseudomonas) pseudomallei, a causative agent of melioidosis, were investigated. Compared to an enterobacterial LPS (SAE-LPS), B. pseudomallei LPS (BP-LPS) exhibited weaker pyrogenic activity in rabbits, lethal toxicity in galactosamine-sensitized mice and murine macrophage activation, i.e. production of tumor necrosis factor, interleukin-6 and nitric oxide. BP-LPS, on the other hand, exhibited stronger mitogenic activity to murine splenocytes than SAE-LPS; moreover, it stimulated even the splenocytes of LPS-resistant C3H/HeJ mice. Unusual chemical structures in the acid-stable inner core region attached to the lipid A moiety of BP-LPS may be responsible for this strong mitogenic activity.

Animals↗
PubMed1986

Identity of limulus amoebocyte lysate-active root surface materials from periodontally involved teeth.

A new approach has been developed to circumvent the problems of false positive reactions in the Limulus Amoebocyte Lysate (LAL) assay for lipopolysaccharide (LPS) in root surface materials. These LAL-reactive materials include thrombin, thromboplastin, ribonuclease, ribonucleic acid, lipoteichoic acid and peptidoglycan fragments. In the present study, hot phenol/water extraction of these substances followed by ultracentrifugation of the resulting aqueous phases reduced their concentrations to very low levels. Furthermore, the application of Polymyxin B/Sepharose 4B affinity chromatography to these extracts enabled their intrinsic LAL-activity to be determined. Use of these techniques to assay root surface materials has identified LPS as being the major LAL-reactive material present. The mean LPS yield for the periodontally involved teeth was 4.13 micrograms/tooth, representing 2.82 micrograms/root. In contrast, the mean yield of LPS for the periodontally uninvolved teeth was 3.12 ng/tooth.

Adult↗
PubMed1986

The distribution of bacterial lipopolysaccharide (endotoxin) in relation to periodontally involved root surfaces.

The distribution of lipopolysaccharide (LPS) in periodontally involved root surface associated materials has been investigated using improved methods of identification and quantification. 39% of the LPS could be removed by gently washing in water for 1 min and 60% by brushing for 1 min with a slowly rotating bristle brush. The finding that 99% of the LPS can be removed by comparatively gentle procedures suggests that effective root surface debridement may be achieved by methods other than traditional hand instrumentation.

Chromatography, Affinity↗
PubMed1988

Root surface debridement--an in vitro assessment.

Root planing has been advocated to render periodontally involved root surfaces 'biologically compatible' with the surrounding soft tissues and thus promote healing. However, recent work has shown that only small amounts of cytotoxic material are likely to be incorporated within the root surfaces, thereby questioning the rationale for the traditional emphasis that is placed upon the removal of 'diseased cementum'. This investigation assessed by limulus amoebocyte lysate (LAL) assay and Polymyxin B affinity chromatography the extent of residual lipopolysaccharide (LPS) following root surface instrumentation in vitro. A conservative regime was carried out, consisting of 15 instrument strokes per surface designed to ensure complete overlapping of the strokes. This harvested varying amounts of LPS from 18 single-rooted teeth while leaving behind less than 0.24 ng of LPS per tooth in the majority (72%) of cases. This finding endorses the growing belief that extensive root planing may not be warranted.

Chromatography, Affinity↗
PubMed1990

The assessment of ultrasonic root surface debridement by determination of residual endotoxin levels.

Recent demonstrations of the superficial nature and ease of removal of root-surface-associated cytotoxic materials suggest that extensive root-surface instrumentation is not warranted clinically. This in vitro investigation determined the detoxifying effects of a conservative regime of ultrasonic root debridement, using the Limulus amoebocyte lysate assay for lipopolysaccharide (LPS) as an indication of the presence or absence of cytotoxic materials. 20 extracted periodontally-involved single-rooted teeth, with no clinically detectable calculus deposits, were debrided with a Cavitron TF-10 tip. Light pressure, resulting from a force application of approximately 50 g, lasting for 0.8 s/mm2 of root surface was used and complete overlapping instrumentation ensured. The finding of LPS levels of less than 2.5 ng per root in 19 out of 20 teeth after such debridement was comparable to LPS levels found on healthy, uninvolved control teeth, and thus endorses the growing belief that root surface cleanliness can be readily achieved.

Dental Prophylaxis↗
PubMed1991

Assessment of ultrasonic debridement of calculus-associated periodontally-involved root surfaces by the limulus amoebocyte lysate assay. An in vitro study.

This investigation assessed the effectiveness of an in vitro ultrasonic debridement regime on periodontally involved teeth with visually-detectable calculus deposits, using the LAL assay for lipopolysaccharides (LPS) as a marker of bacterial products. 34 single rooted teeth with extensive amounts of calcified accretions were selected, of which 10 served as uninstrumented controls for the estimation of existing LPS levels. The periodontally involved roots of the 24 experimental teeth were debrided with a Cavitron TF-10 tip for a limited time using light pressure and overlapping strokes which ensured complete surface instrumentation without reference to the presence of calculus. Marked reductions in root surface area coverage by calculus were nevertheless achieved as assessed planimetrically from photographic records of the teeth before and after debridement. The root surfaces of each tooth were then stripped away and the harvested material subjected to 'Westphal' extraction and LAL assay for LPS. The resulting LPS yields from the experimental teeth ranged from less than 0.08 to 22.387 ng compared to 1,900 to 29,200 ng from the uninstrumented controls. The extremely small residual LPS yields from the experimental teeth not only confirm the efficacy of ultrasonic debridement but, when coupled with the lack of a relationship with the widely varying amounts of the remaining calculus, support the contention that bacterial plaque rather than calculus has the greater pathogenic potential. These results reinforce the growing impression of the superficial location of bacterial toxic products associated with periodontally involved root surfaces.

Adolescent↗
PubMed1995

Lipopolysaccharides from periodontal pathogens prime neutrophils for enhanced respiratory burst: differential effect of a synthetic lipid a precursor IVA (LA-14-PP).

When neutrophils are incubated with bacterial lipopolysaccharide (LPS), they become primed for enhanced release of superoxide anion (O2-) in response to stimulation by FMLP. We investigated the human neutrophil-priming activity of LPS from the periodontal pathogens, Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi) and Actinobacillus actinomycetemcomitans (Aa) in comparison with that of LPS from Escherichia coli (E. coli). The optimum conditions for LPS to prime neutrophils were assessed for every LPS and found to be as follows: Neutrophils were incubated with LPS in the presence of 10% heat-inactivated plasma and 1 mM EDTA at 37 degrees C for 30 min and then stimulated with 1 microM FMLP at 37 degrees C for 7 min. Under these conditions, half-maximum priming was observed at 6.2 ng/ml Pg-LPS, 45 ng/ml Pi-LPS, 1.5 ng/ml Aa-LPS and 1.5 ng/ml E. coli-LPS. The priming activity of each LPS was neutralized by polymyxin B. Anti-CD14 monoclonal antibody inhibited priming by all LPS. The priming by Aa-LPS and E. coli-LPS was inhibited by LA-14-PP, a synthetic lipid A precursor IVA, but that by Pg-LPS and Pi-LPS was not. Priming by tumor necrosis factor alpha was not affected by polymyxin B, anti-CD14 antibody or LA-14-PP. Gelation of Limulus amebocyte lysate occurred at 10 pg/ml Pg-LPS, 30 pg/ml Pi-LPS, 3 pg/ml Aa-LPS and 3 pg/ml E. coli-LPS. Thus LPS from different periodontal pathogens primed neutrophils with different efficacy.(ABSTRACT TRUNCATED AT 250 WORDS)

Aggregatibacter actinomycetemcomitans↗
PubMed1979

Electron microscopical study of Neisseria meningitidis releasing various amounts of free endotoxin.

A study has been made of the ultrastructure of four strains of Neisseria meningitidis which liberate varying amounts of free endotoxin in a chemically-defined, protein-free medium. The two strains which did not release detectable or only sparse amounts of free, filtrable endotoxin were rather uniform in cell size. Their cells appeared to be intact and showed a low tendency to aggregate, In addition cells of these strains showed only sporadic loose, trilaminar membranes and blebs, and free membranous structures were sparse in the medium. The endotoxin releasing strains liberated a high yield of free structures from the outer cell wall into the medium. These structures may represent the lipopolysaccharide (LPS).

Bacteriological Techniques↗
PubMed1989

Endotoxin liberation studied by biological and chemical methods. Chemical characterization of six meningococcal lipopolysaccharides.

Release of endotoxin (or lipopolysaccharides, LPS) from four meningococcal strains was studied with a chemical and a biological technique. Two strains were endotoxin-liberating (E+; 270E+ and 840E+) and two had no or low endotoxin release E-; 270E- and 840E-). LPS was quantitated by gas chromatography (GC) of LPS-specific hydroxy fatty acid, in parallel with assay of endotoxin by Limulus Amebocyte Lysate (LAL), in cell suspensions of equal O.D. and in filtered samples. The GC and LAL methods showed a reasonably good agreement in the determination of LPS in filtrates, which had distinctly higher levels (approx. 10-100 times) for the E+ strains than the E- strains, in accordance with earlier LAL studies. This difference was not due to overproduction of LPS in the E+ strains, since all four strains had the same level of LPS (by GC) in cell suspensions of equal O.D. Here the agreement between the GC and LAL methods was substantially less, with lower values by LAL for the two E-strains. The chemical composition of purified LPS was determined by methanolysis and GC for the four strains and for two additional strains 247 and 714 with a high degree of genetic similarity with strains 270E- and 840E-, respectively. Amounts of unphosphorylated L-glycero-D-mannoheptose and 2-keto-3-deoxyoctonic acid were the same in all 6 LPS. Otherwise distinct differences were found between LPS of the 6 strains. LPS of the two E+ strains formed one group with about 2.4 mol of galactose (gal), 1.4 mol of glucose (glc) and 2.8 mol of glucosamine (glcN) in the carbohydrate chain. Another group, LPS of all the E- strains except 270E-, had 1.1 mol of gal, 2.8 mol of glc and 1.3 mol of glcN in the LPS chain. LPS 270E- also had 1.3 mol of glcN but deviated strongly form all other LPS by a complete lack of gal and glc. On the basis of genetic evidence strain 270E- is regarded as a "rough" LPS mutant of strain 247. The atypical chemistry of LPS 270E- may explain an observed hydrophobicity of this LPS, and it may be related to the previously described sulfonamide sensitivity. Whether the chemical difference observed for LPS of the E+ and E- strains is a mere coincidence remains to be elucidated by detailed studies of more strains of known tendency of endotoxin liberation.

Chemical Phenomena↗
PubMed2006

Endotoxin adsorption therapy for septic shock using polymyxin B-immobilized fibers (PMX): evaluation by high-sensitivity endotoxin assay and measurement of the cytokine production capacity.

Because of its low sensitivity, the conventional measurement method for endotoxin (ET) is not the most appropriate for monitoring the effect of ET adsorption therapy. Thus, the efficacy of ET adsorption therapy was investigated using a newly developed high-sensitivity ET assay method. The changes in the cytokine production capacity of whole blood were also examined. We treated 24 peritonitis patients who had developed postoperative septic shock with ET adsorption therapy using a column of polymyxin B-immobilized fibers (PMX) and their serum ET levels were measured using the high-sensitivity ET assay based on the kinetic turbidimetric Limulus assay. In addition, the changes in the tumor necrosis factor-(TNF-alpha) production capacity of whole blood following lipopolysaccharide (LPS) stimulation and clinical outcome in the study patients were also examined. The 28-day mortality rate was 12%. PMX-direct hemoperfusion (PMX-DHP) was associated with elevation of the mean arterial pressure and urine output, reduction in the mean dose requirement of vasopressor agents, and recovery from the shock state in all the patients. The PaO2/FIO2 ratio also showed significant improvement. Using the high-sensitivity ET assay, ET was detected in the blood of 20 out of the 24 patients (80%) before the PMX-DHP, and a significant reduction in the ET level was noted after the PMX-DHP. The TNF-alpha production capacity of whole blood, which was found to be lower in the septic shock patients than in healthy subjects, was significantly increased after PMX-DHP. Elimination of ET by PMX-DHP in septic shock patients was confirmed by the high-sensitivity ET assay. PMX-DHP is thus considered to be a useful adjuvant therapeutic technique in the treatment of septic shock. Also, PMX-DHP might alleviate the immunosuppression associated with severe sepsis.

Adsorption↗
PubMedDate not supplied

Endotoxin concentrations measured by a chromogenic assay in portal and peripheral venous blood in ten dogs with portosystemic shunts.

A chromogenic Limulus amebocyte lysate assay was used to measure portal and peripheral venous endotoxin concentrations in ten medically managed dogs undergoing surgery for correction of a single extrahepatic portosystemic shunt. In all dogs, both peripheral and portal venous blood samples were obtained at the time of surgical manipulation of the anomalous vessel. In six dogs, peripheral venous samples were obtained an average of 8.0 months after surgery. Five physically normal dogs without biochemical or histologic evidence of liver disease served as controls. Data analysis failed to demonstrate significant differences in peripheral and portal venous endotoxin concentrations between the control and study groups. Postoperatively five of six dogs showed a measurable reduction in peripheral venous endotoxin concentration over intraoperatively obtained values, but the differences were not statistically significant (P = 0.06). Based on results of this study it was concluded that systemic endotoxemia was not present in dogs with a single extrahepatic portosystemic shunt that were medically stable prior to surgery.

Animals↗
PubMed1989

Plasma endotoxin concentrations in experimental and clinical equine subjects.

Endotoxin (LPS) was quantitated in experimental subjects and in horses with naturally occurring gastrointestinal strangulation obstruction and/or septicaemic diseases to establish the fate of LPS and the clinical usefulness of the Limulus amoebocyte lysate (LAL) assay. The assay was validated for sensitivity (10 pg/ml), recovery (90 to 106 per cent), intra-assay precision (CV = 5.5 per cent) inter-assay precision (CV = 11 per cent), and stability of diluted, heat treated, frozen samples (at least 90 days). Plasma concentrations of LPS after sublethal (3 micrograms/kg) jugular or portal vein bolus injections of LPS rose to 4000 pg/ml and 1500 pg/ml respectively followed by a rapid phase of clearance. Peak plasma concentrations of LPS, associated with slow portal infusion, were lower than peak values associated with bolus injections, remained elevated during the infusion (2 h), but rapidly decreased after infusion was stopped. Thirty seven horses with 38 episodes of naturally occurring gastrointestinal or septicaemic disease were assayed for LPS. Eight episodes involving gastrointestinal disease and eight involving septicaemic disease were positive for LPS. It is concluded that the LAL assay is sensitive and reliable for detecting LPS in equine plasma and it may have clinical value for establishing the severity of endotoxaemia or for distinguishing between septic and non-septic conditions. Problems of rapid clearance of LPS from plasma, low concentrations, the possibility of sample contamination, and the time and method of sample procurement remain to be addressed.

Animals↗