Bacterial endotoxins (lipopolysaccharides (LPSs)) are important mediators of inflammatory processes induced by Gram-negative microorganisms. LPSs are the key inducers of septic shock due to a Gram-negative bacterial infection; thus, the structure and functions of LPSs are of specific interest. Often, highly purified bacterial endotoxins must be isolated from small amounts of biological material. Each of the currently available methods for LPS extraction has certain limitations. Herein, we describe a rapid and simple microscale method for extracting LPSs. The method consists of the following steps: ultrasonic destruction of the bacterial material, LPS extraction via heating, LPS purification with organic solvents, and treatment with proteinase K. LPSs that were extracted by using this method contained less than 2-3% protein and 1% total nucleic acid. We also demonstrated the structural integrity of the O-antigen and lipid A via the sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) methods, respectively. We demonstrated the ability of the extracted LPSs to induce typical secretion of cytokines and chemokines by primary macrophages. Overall, this method may be used to isolate purified LPSs with preserved structures of both the O-antigen and lipid A and unchanged functional activity from small amounts of bacterial biomass.

BACKGROUND: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a leading cause of end-stage liver disease associated with increased mortality and cardiovascular disease. Obesity and diabetes are the most important risk factors of MASLD. It is well-established that obesity-associated insulin resistance leads to a situation of tissue lipotoxicity characterized by an accumulation of excess fat in non-fat tissues such as the liver, promoting the development of MASLD, and its progression into metabolic dysfunction-associated steatohepatitis.
METHODS: Here, we aimed to review the impact of disrupted intestinal permeability, antimicrobial proteins and bacterial endotoxin in the development and progression of MASLD.
CONCLUSIONS: Recent studies demonstrated that obesity- and obesogenic diets-associated alterations of intestinal microbiota along with the disruption of intestinal barrier integrity, the alteration in antimicrobial proteins and, in consequence, an enhanced translocation of bacterial endotoxin into bloodstream might contribute to this pathological process through to impacting liver metabolism and inflammation.

Ultrasonic-assisted activated carbon separation (UACS) was first employed to improve product quality by regulating adsorption rate and removing bacterial endotoxin from salvia miltiorrhizae injection. The adsorption rate was related to three variables: activated carbon dosage, ultrasonic power, and pH. With the increase of activated carbon dosage from 0.05 % to 1.0 %, the adsorption rates of salvianolic acids and bacterial endotoxin increased simultaneously. The adsorption rates at which bacteria endotoxins increased from 52.52 % to 97.16 % were much higher than salvianolic acids. As the ultrasonic power increased from 0 to 700 W, the adsorption rates of salvianolic acids on activated carbon declined to less than 10 %, but bacterial endotoxin increased to more than 87 %. As the pH increased from 2.00 to 8.00, the adsorption rate of salvianolic acid dropped whereas bacterial endotoxin remained relatively stable. On the basis of response surface methodology (RSM), the optimal separation conditions were established to be activated carbon dose of 0.70 %, ultrasonic power of 600 W, and pH of 7.90. The experimental adsorption rates of bacterial endotoxin were 94.15 %, which satisfied the salvia miltiorrhizae injection quality criterion. Meanwhile, salvianolic acids\' adsorption rates were 1.92 % for tanshinol, 4.05 % for protocatechualdehyde, 2.21 % for rosmarinic acid, and 3.77 % for salvianolic acid B, all of which were much lower than conventional activated carbon adsorption (CACA). Salvianolic acids\' adsorption mechanism on activated carbon is dependent on the component\'s molecular state. Under ideal separation conditions, the molecular states of the four salvianolic acids fall between 1.13 % and 6.60 %. The quality of salvia miltiorrhizae injection can be improved while maintaining injection safety by reducing the adsorption rates of salvianolic acids to less than 5 % by the use of ultrasound to accelerate the desorption mass transfer rate on the activated carbon surface. When activated carbon adsorption was used in the process of producing salvia miltiorrhizae injection, the pH of the solution was around 5.00, and the proportion of each component\'s molecular state was tanshinol 7.05 %, protocatechualdehyde 48.93 %, rosmarinic acid 13.79 %, and salvianolic acid B 10.28 %, respectively. The loss of useful components was evident, and the corresponding activated carbon adsorption rate ranged from 20.74 % to 41.05 %. The average variation rate in plasma His and IgE was significant (P < 0.05) following injection of 0.01 % activated carbon, however the average variation rate of salvia miltiorrhizae injection was dramatically decreased with the use of UACS and CACA (P > 0.05). The ultrasonic at a power intensity of 60 W/L and the power density of 1.20 W/cm2 may resolve the separation contradiction between salvianolic acids and bacterial endotoxin, according to experiments conducted with UACS at different power intensities. According to this study, UACS has a lot of potential applications in the pharmaceutical manufacturing industry and may represent a breakthrough in the field of ultrasonic separation.

Endurance exercise can disturb intestinal epithelial integrity, leading to increased systemic indicators of cell injury, hyperpermeability, and pathogenic translocation. However, the interaction between exercise, diet, and gastrointestinal disturbance still warrants exploration. This study examined whether a 6-day dietary intervention influenced perturbations to intestinal epithelial disruption in response to a 25-km race walk. Twenty-eight male race walkers adhered to a high carbohydrate (CHO)/energy diet (65% CHO, energy availability = 40 kcal·kg FFM-1·day-1) for 6 days prior to a Baseline 25-km race walk. Athletes were then split into three subgroups: high CHO/energy diet (n = 10); low-CHO, high-fat diet (LCHF: n = 8; <50 g/day CHO, energy availability = 40 kcal·kg FFM-1·day-1); and low energy availability (n = 10; 65% CHO, energy availability = 15 kcal·kg FFM-1·day-1) for a further 6-day dietary intervention period prior to a second 25-km race walk (Adaptation). During both trials, venous blood was collected pre-, post-, and 1 hr postexercise and analyzed for markers of intestinal epithelial disruption. Intestinal fatty acid-binding protein concentration was significantly higher (twofold increase) in response to exercise during Adaptation compared to Baseline in the LCHF group (p = .001). Similar findings were observed for soluble CD14 (p < .001) and lipopolysaccharide-binding protein (p = .003), where postexercise concentrations were higher (53% and 36%, respectively) during Adaptation than Baseline in LCHF. No differences in high CHO/energy diet or low energy availability were apparent for any blood markers assessed (p > .05). A short-term LCHF diet increased intestinal epithelial cell injury in response to a 25-km race walk. No effect of low energy availability on gastrointestinal injury or symptoms was observed.

Surrogate viruses theoretically provide an opportunity to study the viral spread in an indoor environment, a highly needed understanding during the pandemic, in a safe manner to humans and the environment. However, the safety of surrogate viruses for humans as an aerosol at high concentrations has not been established. In this study, Phi6 surrogate was aerosolized at high concentration (Particulate matter2.5: ∼1018 μg m-3) in the studied indoor space. Participants were closely followed for any symptoms. We measured the bacterial endotoxin concentration of the virus solution used for aerosolization as well as the concentration in the room air containing the aerosolized viruses. In addition, we measured how the bacterial endotoxin concentration of the sample was affected by different traditional virus purification procedures. Despite the purification, bacterial endotoxin concentration of the Phi6 was high (350 EU/ml in solution used for aerosols) with both (two) purification protocols. Bacterial endotoxins were also detected in aerosolized form, but below the occupational exposure limit of 90 EU/m3. Despite these concerns, no symptoms were observed in exposed humans when they were using personal protective equipment. In the future, purification protocols should be developed to reduce associated bacterial endotoxin levels in enveloped bacterial virus specimens to ensure even safer research use of surrogate viruses.

The liver is exposed to gut-derived bacterial endotoxin via portal circulation, and recognizes it through toll-like receptor 4 (TLR4). Endotoxin lipopolysaccharide (LPS) stimulates the self-ubiquitination of ubiquitin ligase TRAF6, which is linked to scaffold with protein kinase TAK1 for auto-phosphorylation and subsequent activation. TAK1 activity is a signal transducer in the activating pathways of transcription factors NF-κB and AP-1 for production of various cytokines. Here, we hypothesized that TRAF6-TAK1 axis would be implicated in endotoxin-induced liver disease. Following exposure to endotoxin LPS, TLR4-mediated phosphorylation of TAK1 and transcription of cell-death cytokine TNF-α were triggered in Kupffer cells but not in hepatocytes as well as TNF receptor-mediated and caspase-3-executed apoptosis was occurred in D-galactosamine (GalN)-sensitized hepatocytes under co-culture with Kupffer cells. Treatment with pyridinylmethylene benzothiophene (PMBT) improved endotoxin LPS-induced hepatocyte apoptosis in GalN-sensitized C57BL/6 mice via suppressing NF-κB- and AP-1-regulated expression of TNF-α in Kupffer cells, and rescued the mice from hepatic damage-associated bleeding and death. As a mechanism, PMBT directly inhibited Lys 63-linked ubiquitination of TRAF6, and mitigated scaffold assembly between TRAF6 and the TAK1-activator adaptors TAB1 and TAB2 complex in Kupffer cells. Thereby, PMBT interrupted TRAF6 ubiquitination-induced activation of TAK1 activity in the TLR4-mediated signal cascade leading to TNF-α production. However, PMBT did not directly affect the apoptotic activity of TNF-α on GalN-sensitized hepatocytes. Finally, we propose chemical inhibition of TRAF6-TAK1 axis in Kupffer cells as a strategy for treating liver disease due to gut-derived endotoxin or Gram-negative bacterial infection.

This study aimed to evaluate the genomic features of novel Kenyan virulent phage isolates infecting carbapenemase-producing Klebsiella pneumoniae and to determine the safety of their lysates using mice model in a preclinical study. The genomics showed that the Klebsiella phages vB_KpM_CPRSA and vB_KpM_CPRSB belonged to the genus Slopekvirus with a similarity index of less than 92% compared to the most closest relative species. Their genomes did not contain antimicrobial resistance and toxin genes. Then endotoxin levels in the Klebsiella phage lysates were statistically significant (p value ˃ 0.05). The serum activities of aspartate aminotransferase, alanine aminotransferase and urea in the group of balb/c mice injected with bacteriophage lysates through the intravenous route were higher compared to that of the intranasal route. Unexpectedly, there was mild congestion of the central veins of kidneys and liver without damage to renal tubules and hepatocytes and a lack of physical discomfort and pain in the mice. Our study isolated and characterised Klebsiella phages against carbapenem-resistant K. pneumoniae, which are promising therapeutic agents for the treatment of respiratory tract infections using the topical mode of administration as the preferred route of bacteriophage delivery.

Endotoxins as the outer membrane of most Gram-Negative Bacteria (GNB) and typical toxic biochemical produced by microorganisms are identified as one of the emerging pollutants. These microbial by-products are harmful compounds that can be present in various environments including air, water, soil, and other ecosystems which were discussed in detail in this review. Environmental and occupational exposure caused by endotoxin occurs in water and wastewater treatment plants, industrial plants, farming, waste recovery, and composting facilities. Even though the health risk related to endotoxin injection in intravenous and dialysis are well identified, the harmful effects of ingestion, inhalation, and other way of exposure are not well quantified and there is insufficient information on the potential health risks of endotoxins exposure in water environments, and another exposure. Because of limited studies, the outbreaks of diseases related to endotoxins in the various source of exposure not been well documented. Endotoxin removal from different environments are investigated in this review. The results of various studies have shown that conventional treatment methods have been unable to remove endotoxins from water and wastewater, therefore, monitoring the effectiveness of these processes in controlling this contaminant and also using the appropriate removal method is essential. However, management of water and wastewater treatment processes and the use of advanced processes such as Advanced Oxidation Processes (AOPs) can be effective in monitoring and reducing endotoxin levels during water and wastewater treatment. One of the limitations of endotoxin monitoring is the lack of sufficient information to develop monitoring levels. In addition, the lack of guidelines and methods of controlling them at high levels may cause irreparable disaster.

OBJECTIVE: We evaluated bacterial endotoxin adhesion, superficial micromorphology and mechanical properties of latex and non-latex intermaxillary orthodontic elastics.
METHODS: To quantify the adhered bacterial endotoxin, elastics were divided into 5 groups: experimental (n = 12) latex and non-latex elastics, previously contaminated by an endotoxin solution, negative control (n = 6) latex and non-latex elastics without contamination, and positive control (n = 6) stainless steel specimens (metallic replicas), contaminated by an endotoxin solution. In parallel, the structural micromorphology (n = 6) and surface roughness of latex and non-latex intermaxillary orthodontic elastics were assessed using confocal laser microscopy. Force degradation (g) and deformation of the internal diameter change (mm) were also evaluated. Structural micromorphology, surface roughness (µm), force degradation (g) and internal diameter (mm) change were evaluated at time 0 and after 24 and 72 h in a deformation test. Data were analyzed by the Shapiro-Wilk, Kruskal-Wallis, Dunn, ANOVA and Bonferroni tests (α = 5%).
RESULTS: Endotoxin adhered similarly to both types of elastics with scores of 3 (> 1.0 EU/mL). The surface microstructure of both types of elastics showed irregularities and porosities at all times. Initially, the latex elastics had a higher surface roughness (p < 0.001) than the non-latex ones. After 24 h loading, surface roughness of the latex elastics was significantly reduced (p < 0.001), while after 72 h, the values were similar for both types (p > 0.05). The non-latex elastics had significantly higher force generation values (p < 0.05) at 0, 24 and 72 h compared with the latex elastics, although there was a significant reduction (p < 0.001) in force over time for both elastics. Despite similar initial values, non-latex elastics had a significantly larger internal diameter (p < 0.001) after the loading periods of 24 and 72 h compared with the latex elastics.
CONCLUSIONS: Both elastics showed high affinity with endotoxin and microstructural irregularities of their surface. The non-latex elastics generated higher force values but demonstrated greater deformation of the internal diameter after loading.
UNASSIGNED: ZIELSETZUNG: Wir untersuchten die bakterielle Endotoxinadhäsion, die oberflächliche Mikromorphologie und die mechanischen Eigenschaften von latexhaltigen und nicht-latexhaltigen intermaxillären kieferorthopädischen Elastics.
METHODS: Zur Quantifizierung des anhaftenden bakteriellen Endotoxins wurden die Elastics in 5 Gruppen eingeteilt: experimentelle (n = 12) Latex- und Nicht-Latex-Elastics, die zuvor mit einer Endotoxinlösung kontaminiert worden waren, Negativkontrolle (n = 6) Latex- und Nicht-Latex-Elastics ohne Kontamination und Positivkontrolle (n = 6) Edelstahlproben (metallische Nachbildungen), die mit einer Endotoxinlösung kontaminiert waren. Parallel dazu wurden die strukturelle Mikromorphologie (n =6 ) und die Oberflächenrauheit von intermaxillären Elastics aus Latex und aus Nicht-Latex mittels konfokaler Lasermikroskopie untersucht. Ebenfalls bewertet wurden der Kraftabbau (g) und die Verformung des Innendurchmessers (mm). Die strukturelle Mikromorphologie, die Oberflächenrauheit (µm), der Kraftabbau (g) und die Änderung des Innendurchmessers (mm) wurden zum Zeitpunkt 0 sowie nach 24 und 72 h in einem Verformungstest bewertet. Die Daten wurden mit den Tests Shapiro-Wilk, Kruskal-Wallis, Dunn, ANOVA und Bonferroni (α = 5%) analysiert.
UNASSIGNED: Das Endotoxin haftete an beiden Arten von Elastics in ähnlicher Weise mit Scores von 3 (> 1,0 EU/ml). Die Oberflächenmikrostruktur beider Arten von Elastics zeigte zu jedem Zeitpunkt Unregelmäßigkeiten und Porositäten. Zu Beginn wiesen die Latex-Elastics eine höhere Oberflächenrauheit auf (p < 0,001) als die Nicht-Latex-Elastics. Nach 24 h Belastung war die Oberflächenrauheit der Latex-Elastics deutlich geringer (p < 0,001), während nach 72 h die Werte für beide Typen ähnlich waren (p < 0,05). Die Nicht-Latex-Elastics wiesen nach 0, 24 und 72 h signifikant höhere Werte für die Krafterzeugung auf (p < 0,05) als die Latex-Elastics, auch wenn die Kraft bei beiden Elastics im Laufe der Zeit signifikant abnahm (p < 0,001). Trotz ähnlicher Ausgangswerte wiesen die Nicht-Latex-Elastics nach den Belastungszeiten von 24 und 72 h einen signifikant größeren Innendurchmesser (p < 0,001) auf als die Latex-Elastics.
UNASSIGNED: Beide Elastics zeigten eine hohe Affinität zu Endotoxin und mikrostrukturelle Unregelmäßigkeiten auf ihrer Oberfläche. Die latexfreien Elastics erzeugten höhere Kraftwerte, wiesen aber nach der Belastung eine größere Verformung des Innendurchmessers auf.

Background and aims: Periodontitis is an inflammatory-infectious disease. Identifying markers of systemic exposure of periodontitis might be of interest to study its interaction with other conditions. Soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) is upregulated during bacterial infections. Our aim was therefore to investigate whether periodontitis and its treatment are associated with bacterial endotoxin and sTREM-1. Methods: Fifty patients with severe periodontitis and 50 age-matched controls were included in a case-control study (all never smokers). A secondary analysis of a previously published intervention study was performed, in which included 69 patients with severe periodontitis were randomized to receive either intensive (IPT) or control periodontal therapy (CPT) and monitored over 6 months. Serum levels of bacterial endotoxin and sTREM-1 were determined at one time point (case-control study) and at baseline, 1 day, 1 and 6 months after periodontal treatment (intervention study). Results: Severe periodontitis was associated with elevated circulating endotoxin levels when cases (22.9 ± 2.2 EU/ml) were compared to controls (3.6 ± 0.5 EU/ml, p < 0.001) and with sTREM-1 levels (1302.6 ± 47.8 vs. 870.6 ± 62.0 pg/ml, p < 0.001). A positive correlation was observed between sTREM-1 and endotoxin levels (r = 0.4, p < 0.001). At 6 months after treatment, IPT significantly decreased serum levels of sTREM-1 compared to CPT (adjusted mean difference of 500.2 pg/ml, 95% CI: 18.9-981.4; p = 0.042). No substantial differences were noted in endotoxin levels at any time point after treatment between groups. Conclusions: Severe periodontitis is linked to increased circulating endotoxin and sTREM-1 levels and following IPT a reduction in sTREM-1 levels is observed.