Microwave-induced release and degradation of airborne endotoxins from Escherichia coli bioaerosol.

Endotoxins are widely distributed toxins in the outer cell-wall membranes of Gram-negative bacteria and other microorganisms. Chronic exposure to endotoxins can induce and exacerbate airway symptoms and diseases. However, the release and degradation of airborne endotoxins from bioaerosol by microwave (MW) irradiation have not yet been reported. This study investigated the distribution and fate of airborne endotoxins during MW irradiation process, as well as the kinetics and thermodynamics of the degradation of airborne endotoxins. Results showed that MW irradiation induced cell lysis, thus considerably increasing the proportion of cells with ruptured membranes. Furthermore, MW irradiation changed the distribution of airborne endotoxins, sharply decreased the concentration of bound endotoxins from 230 EU/m3 to 68 EU/m3, and increased the concentration of free endotoxins from 21 EU/m3 to 122 EU/m3. These results indicated that MW irradiation released endotoxins from cells into the atmosphere. MW irradiation likely degraded endotoxins by exerting thermal effects, which achieved a total endotoxin removal efficiency of as high as 35%. Endotoxin degradation was a first-order reaction and required the activation energy of 26.3 kJ/mol.

Inactivation of airborne bacteria using different UV sources: Performance modeling, energy utilization, and endotoxin degradation.

Airborne bacteria-containing bioaerosols have attracted increased research attention on account of their adverse effects on human health. Ultraviolet germicidal irradiation (UVGI) is an effective method to inactivate airborne microorganisms. The present study models and compares the inactivation performance of three UV sources in the UVGI for aerosolized Escherichia coli. Inactivation efficiency of 0.5, 2.2 and 3.1 logarithmic order was obtained at an exposure UV dose of 370 J/m3 under UVA (365 nm), UVC (254 nm) and UVD (185 nm) sources, respectively. A Beer-Lambert law-based model was developed and validated to compare the inactivation performances of the UV sources, and modeling enabled prediction of inactivation efficiency and analysis of the sensitivity of several parameters. Low influent E. coli concentrations and high UV doses resulted in high energy consumption (EC). The change in airborne endotoxin concentration during UV inactivation was analyzed, and UVC and UVA irradiation showed no marked effect on endotoxin degradation. By contrast, both free and bound endotoxins could be removed by UVD treatment, which is attributed to the ozone generated by the UVD source. The results of this study can provide a better understanding of the air disinfection and airborne endotoxin removal processes.

Oxidative degradation of endotoxin by advanced oxidation process (O3/H2O2 & UV/H2O2).

The presence of endotoxin in water environments may pose a serious public health hazard. We investigated the effectiveness of advanced oxidative processes (AOP: O3/H2O2 and UV/H2O2) in the oxidative degradation of endotoxin. In addition, we measured the release of endotoxin from Escherichia coli following typical disinfection methods, such as chlorine, ozone alone and UV, and compared it with the use of AOPs. Finally, we tested the AOP-treated samples in their ability to induce tumor necrosis factor alpha (TNF-α) in mouse peritoneal macrophages. The production of hydroxyl radical in AOPs showed superior ability to degrade endotoxin in buffered solution, as well as water samples from Korean water treatment facilities, with the ozone/H2O2 being more efficient compared to UV/H2O2. In addition, the AOPs proved effective not only in eliminating E. coli in the samples, but also in endotoxin degradation, while the standard disinfection methods lead to the release of endotoxin following the bacteria destruction. Furthermore, in the experiments with macrophages, the AOPs-deactivated endotoxin lead to the smallest induction of TNF-α, which shows the loss of inflammation activity, compared to ozone treatment alone. In conclusion, these results suggest that AOPs offer an effective and mild method for endotoxin degradation in the water systems.

The effect of gamma sterilization on the insecticidal toxicity of engineered and conventional Bacillus thuringiensis strains.

This study evaluates the effect of gamma radiation on the spore activity, toxicity, and crystal structures of two engineered Bacillus thuringiensis (Bt) strains, TnX and TnY, and the reference Bt strain HD-1. We attempted to identify dosages of cobalt-60 gamma radiation that would inactivate Bt spores but not affect its toxicity. In the radiation dosage range of 10-15 kilogray, no viable spore formation and no significant reduction of the efficiency of Bt against lepidopteran larvae were observed. However, further sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results show that the components of the protoxin are affected by gamma radiation and that some bands are absent after treatment compared with the controls; the change in the protoxin band pattern depends on the type of Bt strain. Furthermore, the spore crystal structure of three Bt strains was studied with scanning electron microscopy and transmission electron microscopy. The results show that there are no changes in the size or shape of the treated Bt spores and crystals compared with the controls. The use of gamma radiation is effective to inactivate the spores of engineered Bt strains while preserving stable Bt toxicity against the target insect larvae.

Single session of Nd:YAG laser intracanal irradiation neutralizes endotoxin in dental root dentin.

Endotoxins released in the dental root by Gram-negative microorganisms can be neutralized by calcium hydroxide, when this medication is applied inside the root canal for at least seven days. However, several clinical situations demand faster root canal decontamination. Thus, for faster endotoxin neutralization, endodontists are seeking additional treatments. The in vitro study tested whether or not intracanal Nd:YAG laser irradiation would be able to neutralize endotoxin within the human dental root canal in a single session. Twenty-four human teeth with one root were mounted between two chambers. After conventional endodontic treatment, root canals were contaminated with Escherichia coli endotoxin. Then they were irradiated or not (controls) in contact mode with an Nd:YAG laser (1.5 W, 15 Hz, 100 mJ and pulse fluency of 124 J/cm2). The endotoxin activity was measured using the limulus lysate technique and data were statistically compared (p≤0.05). The concentration of active endotoxin measured in the negative control group was significantly lower than that of the positive control group (p=0.04). The concentrations of endotoxin in both irradiated groups were significantly lower than that of the positive control group (p=0.027) and similar to that of negative control group (p=0.20). A single session of intracanal Nd:YAG laser irradiation is able to neutralize endotoxin in the dental root tissues.

Melanin pigment formation and increased UV resistance in Bacillus thuringiensis following high temperature induction.

The pigment melanin is well known to protect against the damaging effects of UV radiation. In this study, we show that thirty-five of thirty-seven tested Bacillus thuringiensis strains possess the potential to produce melanin in the presence of L-tyrosin at elevated temperature (42 degrees C). These findings offer a method of protecting insecticidal toxins produced by B. thuringiensis from UV degredation and may therefore have important applications in the field of crop protection. Toxicity assays on Heliothis armigera suggested that the insecticidal activity of B. thuringiensis that produced melanin was significantly higher after UV irradiation than when melanin was not produced.

Removal of bacterial endotoxin from root surface with Er:YAG laser.

PURPOSE: To determine the potential of 2.94 microm Er:YAG laser radiation to remove bacterial endotoxin from root surfaces. MATERIALS AND METHODS: 40 extracted teeth were divided into four groups of 10 samples each. A 16 mm2 area of the root surface on each sample was inoculated with an aliquot of 7 microl of a lipopolysaccharide suspension at a concentration of 50 IU/ml. LPS was derived from a non-oral Escherichia coli strain (E. coli 055:B5). Source of laser radiation was an Er:YAG laser emitting pulsed infrared radiation at a wavelength of 2.94 microm, with a pulse duration of 250 micros, and a pulse repetition rate of 15 pps. Three specimen groups were irradiated with 105 laser pulses at a radiation energy of 60 mJ, 100 mJ and 140 mJ. One specimen group was untreated (control). The LPS concentration with each sample was determined using a chromogenic, quantitative Limulus-amoebocyte-lysate assay. Statistical analysis was ANOVA and Scheffé-test. RESULTS: Mean LPS yield from the untreated control samples was 50.1 (+/- 35.9) IU/ml. Following laser irradiation the average LPS on the root surfaces was 19.86 (+/- 14.4) IU/ml at 60 mJ, 12.86 (+/- 8.1) IU/ml at 100 mJ and 8.58 (+/- 4.9) IU/ml at 140 mJ.

Endotoxin inactivation in water by using medium-pressure UV lamps.

Deionized water was spiked with various concentrations of endotoxin and exposed to UV irradiation from medium-pressure UV lamps to assess endotoxin inactivation. It was found that endotoxin inactivation was proportional to the UV dose under the conditions examined. The inactivation rate was determined to be approximately 0.55 endotoxin unit/ml per mJ/cm(2) of irradiation delivered.

In vitro effect of laser irradiation on cementum-bound endotoxin isolated from periodontally diseased roots.

BACKGROUND: In a previous study, we evaluated the in vivo effects of an Nd:YAG laser on periodontal disease by measuring crevicular interleukin (IL)-1beta levels before and after laser application. It was found that laser therapy was less effective than traditional scaling and root planing. These results might be due to incomplete removal of microbial residues and cementum-bound endotoxin on root surfaces by the laser. In this study, we explored the in vitro effectiveness of an Nd:YAG laser for the elimination of cementum-bound endotoxin by measuring IL-1beta changes in stimulated monocytes. METHODS: Fresh human monocytes were harvested from adults without periodontitis and grown in RPMI 1640 medium. Diseased cementum particles were collected and prepared from teeth with untreated periodontitis and were irradiated with 5 levels of laser energy. Cementum particles were subjected to endotoxin testing by a limulus amebocyte lysate (LAL) assay and then were incubated with cultured monocytes. Production of IL-1beta in stimulated monocytes was measured by enzyme-linked immunosorbent assay and quantified by spectrophotometry. RESULTS: The endotoxin unit (EU) of diseased cementum was 18.4 EU/mg, which seemed to be remarkably lower than that of common periodontal pathogens including Porphyromonas gingivalis (381) at 15,300 EU/mg/ml, Prevotella intermedia (ATCC 25611) at 227 EU/mg/ml, and Fusobacterium nucleatum (ATCC 25586) at 1,987 EU/mg/ml. Monocytes subjected to stimulation by diseased cementum particles without laser irradiation produced 124 to 145 pg/ml IL-1beta, 9- to 18-fold higher than that of unstimulated monocytes (7.07 to 15.95 pg/ml). Diseased cementum particles after irradiation with various energy levels of the Nd:YAG laser could still stimulate monocytes to secrete 89 to 129 pg/ml IL-1beta. No statistically significant difference was found in the production of IL-1beta induced by diseased-bound cementum with or without laser irradiation. CONCLUSIONS: The Nd:YAG laser varying from 50 mJ, 10 pps to 150 mJ, 20 pps, for 2 minutes, did not seem to be effective in destroying diseased cementum endotoxin.

Protection from UV-B damage of mosquito larvicidal toxins from Bacillus thuringiensis subsp. israelensis expressed in Anabaena PCC 7120.

A transgenic strain of the nitrogen-fixing filamentous cyanobacterium Anabaena PCC 7120 protected expressed delta-endotoxin proteins of Bacillus thuringiensis subsp. israelensis from damage inflicted by UV-B, a sunlight component that penetrates Earth's ozone layer. This organism, which serves as a food source to mosquito larvae and could multiply in their breeding sites, may solve the environment-imposed limitations of B. thuringiensis subsp. israelensis as a mosquito biological control agent.

[A new possibility for enhancing natural immunity: a radiation-detoxified endotoxin]. / A természetes ellenállóképesség fokozásának új lehetósége: a sugárzással méregtelenített endotoxin.

Using ionizing radiation the author and co-workers produced a detoxified endotoxin preparation (Tolerin) which seems to be a suitable product for the increase of natural immunity (nonspecific resistance)-including activation of bone marrow in immunosuppressions, immunodeficiencies-protection against various types of shocks-radiation injury, septic/endotoxic shock, etc.- and increase of immunogen effect of antigens (e.g. inactivated virus vaccines) as an immunoadjuvant in human beings and experimental animals.

[In vitro synergic lymphocyte stimulation in HIV-1 infected patients using inosine derivatives and radio-detoxified endotoxin]. / HIV-1 fertözöttek lymphocytáinak in vitro együttes serkenthetösége inozin származékokkal és sugárdetoxikált endotoxinnal.

Restoration of immune functions through promoting cell cycle might delay acquired immunodeficiency syndrome development. Therefore, stimulation of peripheral lymphocytes of human immunodeficiency virus-1 infected patients in successive clinical stages was studied by phytohaemagglutinin and other stimulants. In vitro blastogenesis was quantitated by 3H-thymidine uptake. Stimulation by phytohaemagglutinin decreased in patients with AIDS related complex to 63.1%, with AIDS to 13.6% of control values. Small amount of recombinant interleukin-2 or indomethacin solely not promoting lymphocytes, increased response to phytohaemagglutinin minimally. Alone ineffective methyl-ester and methyl-phosphonate inosine derivatives augmented phytohaemagglutinin-response of controls and patients with AIDS related complex by approx. 1.5-fold, but the effect in the case of AIDS patients was minimal. Radio-detoxified endotoxin alone or in combination with phytohaemagglutinin stimulated lymphocytes of both controls and patients with AIDS related complex slightly. Lymphocyte stimulation of patients with AIDS related complex was augmented in concentration-dependent manner, and by synergic effect it approached phytohaemagglutinin-stimulated blastogenesis of controls. Anergy due to human immunodeficiency virus-1 infection damages synchronisation of secondary messenger systems induced on cell surface receptors, therefore their selective influence by recombinant interleukin-2 or indomethacin is less efficient. Inosine derivatives promote cell cycle by inhibiting cyclic adenosine 3',5'-monophosphate production. In the early stage of virus infection, radio-detoxified endotoxin might bind to receptors of immature T cells and facilitate cell cycle through cyclic guanosine 3',5'-monophosphate stimulation. The clinical trials of radio-detoxified endotoxin (Tolerin) have already been launched.

Radiodetoxified endotoxin-induced tolerance alters monocyte but not neutrophil CD11b and CD18 expression in response to lipopolysaccharide.

OBJECTIVES: To test the hypothesis that pretreatment with radiodetoxified endotoxin (RDE) may mitigate the deleterious effects of subsequent infection, in part by modifying leukocyte adhesion receptor expression, and to investigate the cellular mechanisms of endotoxin tolerance induced by RDE. DESIGN: To assess the effect of RDE pretreatment on mortality from bacterial peritonitis, rats were implanted with an intraperitoneal, barium-fecal inoculum at intervals of 0, 1, 3, and 5 days after RDE injection. Experiments were then conducted to test the effect on leukocyte adhesion receptor expression. Two groups of mice received saline solution, and one group, RDE. After 72 hours, one group received saline solution (saline/saline group), the others, lipopolysaccharide (LPS) (saline/LPS and RDE/LPS groups). Peripheral leukocytes were obtained 1 hour after injection and were analyzed for CD11b and CD18 expression by flow cytometry. SETTING: Laboratory animal study. RESULTS: Survival rates were not improved in rats that were pretreated with RDE 0 and 24 hours before inoculum (0% and 7%, respectively). In rats that were pretreated 72 hours and 120 hours before inoculum, 47% (P < .01) and 60% (P < .01) survived, respectively. CD18 expression on polymorphonuclear leukocytes increased twofold in the RDE/LPS (mean +/- SEM, 300.3 +/- 32.9) and the saline/LPS (mean +/- SEM, 360.4 +/- 59.9) groups compared with controls (mean +/- SEM, 176.4 +/- 18.9) (P < .05). CD11b expression on polymorphonuclear leukocytes increased threefold in the RDE/LPS (mean +/- SEM, 91.3 +/- 8.1) and the saline/LPS (mean +/- SEM, 89.8 +/- 11.4) groups compared with controls (mean +/- SEM, 32.1 +/- 1.8) (P < .05). CD18 expression on monocytes decreased in the saline/LPS group (mean +/- SEM, 134.2 +/- 14.2) and was unchanged in the RDE/LPS group (mean +/- SEM, 200.2 +/- 17.2) compared with controls (mean +/- SEM, 217.6 +/- 16.5) (P < .05). CD11b expression on monocytes decreased in the saline/LPS group (mean +/- SEM, 25.8 +/- 2.2) and was unchanged in the RDE/LPS group (mean +/- SEM, 36.4 +/- 0.9) compared with controls (mean +/- SEM, 39.7 +/- 3.9) (P < .05). CONCLUSIONS: Radiodetoxified endotoxin reduces mortality rates from bacterial peritonitis when given at least 72 hours prior to a bacterial inoculum. Tolerance to subsequent LPS challenge is associated with an abrogation of the reduced peripheral monocyte CD11b and CD18 expression observed in native LPS-stimulated mice but is not associated with changes in polymorphonuclear leukocyte CD11b and CD18 expression. The mechanism of the observed RDE-induced monocyte hyporesponsiveness to LPS and its possible protective effect is uncertain and requires further investigation.

The rate of endotoxin destruction during water treatment using a combination of ozone and ultraviolet radiation.

The rate of destruction of endotoxin in water samples by a combination of UV and ozone has been investigated. After an initial non-linear phase the rate of destruction followed 1st order kinetics with a calculated D value of between six and ten min on endotoxin levels as high as 100 EU/mL. Measuring endotoxin destruction rates has proved an effective way of validating the UV-ozone water treatment process.

The mechanism of sunlight-mediated inactivation of Bacillus thuringiensis crystals.

Detailed photostability studies were carried out using purified delta-endotoxin crystals from Bacillus thuringiensis subspecies HD-1 and HD-73. The mechanism and time course of sunlight inactivation was investigated by: (a) monitoring the tryptophan damage in the intact crystals by Raman spectroscopy, (b) amino acid analysis and (c) biological assays using insects. The results demonstrate that, for purified HD-1 or HD-73 crystals, the 300-380 nm range of the solar spectrum is largely responsible for bringing about crystal damage and consequent loss of toxicity. Purified Bacillus thuringiensis crystals that were exposed to fermentation liquor after cell lysis were more quickly degraded by sunlight than were crystals from cells that were lysed in water. This effect is attributed to adsorption of chromophores by crystals exposed to the fermenter liquor and the subsequent ability of these chromophores to act as photosensitizers. The importance of a photosensitization mechanism in crystal degradation was further emphasized by irradiating Bacillus thuringiensis crystals in vacuo. The latter crystals were found to be less damaged (20% tryptophan loss after 24 h irradiation by the solar spectrum) compared with crystals from the same sample irradiated in air (60% (60% tryptophan loss). Other methods of decreasing exposure of the crystals to oxygen, e.g. by using glycerol as a humectant, were also found to be successful in controlling photodamage. The results concerning photodegradation support a photosensitization mechanism involving the presence of exogenous (and possibly endogenous) chromophores which create singlet oxygen species upon irradiation by light.

[Changes in immunological indicators in patients with surgical endotoxemia after UV-irradiation of blood]. / Dinamika immunologicheskikh pokazatelei u bol'nykh khirurgicheskim éndotoksikozom na fone UF-oblucheniia krovi.

Changes in the values of the immune status during UV autohemotherapy were studied in 22 patients with surgical endotoxicosis of various genesis. Maximum tension of immunogenesis was found in all patients. The time course of changes in the general pool of rosette-forming and separate populations of neutrophils, as well as in the index of immunocompetent cell ratios are evidence of the high informativeness of these tests in determining the severity of the immune system dysfunctions.

Effect of bacterial endotoxin on placentation of rats.

The effect of bacterial endotoxin on placentation in rats was studied on 160 CFY pregnant rats. Based on this experiment, it was concluded that (i) the endotoxin (1 mg/animal i.p.) inhibited placentation (in 90% of animal). (ii) The endotoxin-induced fetopathy almost exclusively resulted in abortion. (iii) The fetuses reacted to endotoxin with relatively the same degrees of susceptibility. (iv) The growth of surviving fetuses seemed to be undisturbed. (v) Endotoxin-induced damages in mothers first of all depend on the individual susceptibility of these pregnant animals and (vi) the endotoxin tolerance induced by radio-detoxified endotoxin (TOLERIN) significantly protects both the mothers and the fetuses against endotoxin challenge.

Response of humans to gamma-irradiated reference Escherichia coli endotoxin.

This study assesses the effect of gamma-irradiation of endotoxin given intravenously to healthy human volunteers. The national reference standard endotoxin derived from E. coli was placed in aqueous medium in sterile-sealed ampoules and divided into four groups. One group received endotoxin with no radiation while the other three received endotoxin with gamma-irradiation at doses of 0.18, 0.36, or 1.08 Mrad. These doses of radiation cause characteristic alterations to the endotoxin molecule, primarily to the O-polysaccharide moiety. Each of the four different preparations of endotoxin was given intravenously to four volunteers at a concentration of 4 ng/kg. The responses for clinical symptoms, cortisol, and growth hormone were significantly and progressively reduced by increasing the irradiation to the endotoxin. Most strikingly, no clinical symptoms were noted with the endotoxin exposed to the highest dose of radiation (1.08 Mrad). Fever, vital signs, white blood cell count, and differential exhibited no statistically significant differences among the groups, but the kinetics of change were altered by increasing doses of gamma-irradiation. Irradiated endotoxin was significantly more effective in decreasing the platelet count than untreated endotoxin. The fever index correlated significantly with maximum temperature, change in temperature, white blood cell count index, mature neutrophil count index, and the cortisol index. Thus, there is dissociation of biological activities for endotoxin in humans due to molecular changes primarily in the O-polysaccharide moiety from exposure to gamma-irradiation.