Blood Endotoxin Levels Research Articles

Interaction Between Peroxisome Proliferator-Activated Receptors and Cannabidiol in the Gut of Chickens Applied to Different Challenge Conditions.

Peroxisome proliferator-activated receptors (PPARs) are important targets for cannabidiol (CBD), which mediate many of its biological actions. The hypothesis of the present research assumed that PPARs affect the gut response to different challenge factors in chickens (C. perfringens vs. lipopolysaccharides (LPS) from E. coli), and that CBD can mediate the pathways of this response. The study proved that CBD and the challenge factors significantly affect the expression level of PPARα (p = 0.001) and selected genes determining gut barrier function. A positive correlation was demonstrated between PPARs and genes involved in the formation of tight junctions, immune, and oxidative stress responses in chickens. Dietary supplementation with CBD actively mediated the expression rate of PPARs, but the mechanism of interaction between CBD and PPARs was different depending on the stress factor used. The addition of CBD to the birds' diets did not contribute to reducing intestinal permeability under induced stress conditions nor cause stress, as indicated by the absence of elevated blood cortisol and endotoxin levels. CBD also supported the mechanisms of protecting intestinal cells from the cytotoxic effects in a C. perfringens challenge through the levels of genes involved in oxidative stress. This study indicates the importance of research toward understanding the mechanisms of PPARs as a target for enhancing intestinal barrier function, provides new results on the biological action of CBD in chickens, and shows a constant PPAR association with the jejunum mucosa of birds.

Escin alleviates cerebral ischemia-induced intestinal pyroptosis via the GR-dependent p38 MAPK/NF-κB signaling and NLRP3 inflammasome activation

Cerebral ischemia-induced systemic inflammation and inflammasome-dependent pyroptotic cell death in ileum, causing serious intestinal injury. Glucocorticoid receptor (GR) mediates the effects of glucocorticoids and participates in inflammation. Escin has corticosteroid-like, neuroprotective, and anti-intestinal dysfunction effects. This study aimed to investigate the effect of Escin on the intestinal barrier injury in rats subjected to middle cerebral artery occlusion (MCAO) and on Caco-2 cells exposed to lipopolysaccharides. The MCAO-caused brain injury was evaluated by assessing neurological function, cerebral infarct volume, and plasma corticosterone (Cort) levels. Intestinal injury was evaluated by observing the histopathological changes, assessing the intestinal barrier function, and determining blood FD4, endotoxin and IL-1β levels. The levels of the tight-junction proteins such as claudin-1, occludin, and ZO-1, and proteins involved in the GR/p38 MAPK/NF-κB pathway and NLRP3-inflammasome activation were evaluated using western blotting or immunofluorescence. Administration of Escin suppressed the cerebral ischemia-induced increases in Garcia-test scores and infarct volume, alleviated the injury to the intestinal barrier, and decreased the levels of Cort, endotoxin, and IL-1β. Additionally, Escin upregulated GR and downregulated phospho(p)-p65, p-p38MAPK, NLRP3, GSDMD-N, and cleaved-caspase-1 in the intestine. The effects of Escin could be suppressed by the GR antagonist RU486 or enhanced by the p38 MAPK antagonist SB203580. We revealed details how Escin improves cerebral ischemia-induced intestinal barrier injury by upregulating GR and thereby inhibiting the pyroptosis induced by NF-κB-mediated NLRP3 activation. This study will provide a experimental foundation for the features of glucocorticoid-like activity and the discovery of new clinical application for Escin.

Dental markers of endogenous intoxication in patients with non-alcoholic fatty liver disease

Objective — to determine some dental markers of endogenous intoxication in the treatment of patients with non‑alcoholic fatty liver disease (NAFLD).
 Materials and methods. Examinations involved 44 patients with verified NAFLD diagnosis (the main group). The control group consisted of 20 somatically healthy subjects, representative in terms of gender and age. Clinical dental examinations were carried out twice, before and after the treatment, with an interval of 10 days. A questionnaire survey was conducted to assess the discipline of oral hygiene care. The level of oral hygiene was studied using the Greene–Vermillion index. The level of endotoxin in venous blood and oral fluid was assessed by immunoenzymatic method. The degree of endogenous intoxication was evaluated based on the structural and optical properties of a drop of oral fluid on a glass slide using an electron microscope. An analysis of correlations between non‑invasive and invasive techniques has been carried out.
 Results. More than half of the patients in the main group (61.3%) cleaned their teeth once a day. The value of the Greene–Vermillion oral hygiene index was equal to 2.3 points, which indicated an unsatisfactory state of hygienic care. In patients of the main group, the level of endotoxemia was equal to 0.9 EU/ml, and the level of LPS in oral fluid was 33.5 U/ml, which exceeded similar parameters in the control group by 2 and 1.6 times, respectively. In the micro preparations of oral fluid facies, 71.0% of patients in the main group had markers of endogenous intoxication (EI) of 3rd grade, 20.4% — EI of 2nd grade, and only 9.0% had EI of 1st grade. After 10 days, according to the re‑questioning data, 72.7% of patients in the main group followed full hygienic care of the oral cavity, and the Greene–Vermillion index decreased to 1.6 points, which was assessed as a satisfactory condition. In the micro preparations of facies of oral fluid, markers of EI of the 2nd grade were observed in 56.8% of the main group and markers of EI of 1st grade in 22.7% of subjects. The number of patients with EI markers of the 3rd grade decreased in 3.5 times compared to the first visit.
 Conclusions. Proper hygienic care of the oral cavity of patients with NAFLD results in the reduction of endogenous intoxication degree.

C-X-C Motif Chemokine Ligand 1 Promotes Colitis by Modulating the Gut Microbiota

Introduction: C-X-C motif chemokine ligand 1 (CXCL1) is a potent neutrophil chemoattractant that plays a pivotal role in recruiting neutrophils during inflammatory conditions. This study explored the role of CXCL1 in modulating the gut microbiota, influencing neutrophil infiltration, and contributing to the development of colitis. Methods: We employed quantitative PCR to assess CXCL1 expression in colon samples. A mouse model of dextran sulfate sodium (DSS)-induced colitis was utilized to explore the progression of colitis in wild-type (WT) and CXCL1-deficient (CXCL1−/−) mice. Results: Colitis attenuation was evident in CXCL1−/− mice. Significant alterations were observed in the gut microbiome, as revealed by 16S rRNA gene sequencing. Furthermore, CXCL1−/− mice exhibited reduced gut permeability and diminished endotoxin levels in peripheral blood following DSS treatment compared to WT mice. In response to DSS treatment, WT mice showed a clear increase in neutrophil infiltration, while CXCL1−/− mice exhibited lower levels of infiltration. Fecal microbiota transplantation (FMT) using stools from CXCL1−/− mice alleviated DSS-induced colitis. Interestingly, FMT from patients with colitis increased CXCL1 and Ly6G expression in the colons of gut-sterilized mice. Clinical data analysis revealed elevated CXCL1 and CD15 expression in patients with colitis, with a positive correlation between the severity of colitis and the expression of CXCL1 and CD15. Conclusion: These findings shed light on the pivotal role of CXCL1 in promoting colitis by modulating the gut microbiota.

Retraction of: Establishing a stable platform for the measurement of blood endotoxin levels in the dialysis population.

Retraction of: Establishing a stable platform for the measurement of blood endotoxin levels in the dialysis population.

Palmitic Acid Modulates Microglial Cell Response to Metabolic Endotoxemia in an In Vitro Study.

Metabolic endotoxemia (ME) is characterized by a 2-3-fold increase in blood endotoxin levels and low-grade systemic inflammation without apparent infection. ME is usually accompanied by metabolic syndrome, characterized by central obesity and hyperlipidemia. According to numerous studies, ME may lead to functional brain disorders, including cognitive decline, depression, and dementia. In the current in vitro study, we aimed to determine the direct and indirect impact of endotoxin (LPS) and palmitic acid (PA), representing saturated fatty acids, on the inflammatory and oxidative stress response in the human microglial HMC3 cells unstimulated and stimulated with IFNγ. The study's results revealed that direct HMC3 cell exposition to endotoxin and PA increased inflammatory response measured as levels of IL-6 and MCP-1 released into the medium and PGE2 levels in cell lysates. Moreover, direct HMC3 cell treatment with PA and LPS induced oxidative stress, i.e., ROS and COX-2 production and lipid peroxidation. On the contrary, an indirect effect of LPS and PA on microglial cells, assessed as the impact of macrophage metabolites, was much lower regarding the inflammatory response, although still associated with oxidative stress. Interestingly, IFNγ had a protective effect on microglial cells, reducing the production of pro-inflammatory mediators and oxidative stress in HMC3 cells treated directly and indirectly with LPS and PA.

Targeted therapy using polymyxin B hemadsorption in patients with sepsis: a post-hoc analysis of the JSEPTIC-DIC study and the EUPHRATES trial

BackgroundPolymyxin B hemadsorption (PMX-HA) reduces blood endotoxin levels, but characteristics of patients with sepsis likely to benefit from PMX-HA are not well known. We sought to identify patient subgroups likely to benefit from PMX-HA.MethodsWe retrospectively identified 1911 patients with sepsis from a retrospective observational study in Japan (the JSEPTIC-DIC study) and 286 patients with endotoxemic septic shock from a randomized controlled trial in North America that restricted patients to those with high endotoxin activity (the EUPHRATES trial). We applied the machine learning-based causal forest model to the JSEPTIC-DIC cohort to investigate heterogeneity in treatment effects of PMX-HA on 28-day survival after adjusting for potential confounders and ascertain the best criteria for PMX-HA use. The derived criteria for targeted therapy by PMX-HA were validated using the EUPHRATES trial cohort.ResultsThe causal forest model revealed heterogeneity in treatment effects of PMX-HA. Since patients having higher treatment effects were more likely to have severe coagulopathy and hyperlactatemia, we identified the potential treatment targets of PMX-HA as patients with PT-INR > 1.4 or lactate > 3 mmol/L. In the EUPHRATES trial cohort, PMX-HA use on the targeted subpopulation (75% of all patients) was significantly associated with higher 28-day survival (PMX-HA vs. control, 68% vs. 52%; treatment effect of PMX-HA, + 16% [95% CI + 2.2% to + 30%], p = 0.02).ConclusionsAbnormal coagulation and hyperlactatemia in septic patients with high endotoxin activity appear to be helpful to identify patients who may benefit most from PMX-HA. Our findings will inform enrollment criteria for future interventional trials targeting patients with coagulopathy and hyperlactatemia.

WITHDRAWN: Evidence Synthesis to Advance Clinical Practice and Scientific Research: A CGH Pillar

WITHDRAWN: Evidence Synthesis to Advance Clinical Practice and Scientific Research: A CGH Pillar

Escin alleviates stress-induced intestinal dysfunction to protect brain injury by regulating the gut-brain axis in ischemic stroke rats

Hyperactivity of HPA axis results in intestinal dysfunction, which may play a role in brain injury caused by ischemic stroke (IS). Escin shows a neuroprotective effect but it may not penetrate blood brain barrier (BBB). Previous work in our laboratory showed that escin ameliorated intestinal injury in animals. The aim of this study is to investigate whether escin attenuates brain injury by improving intestinal dysfunction in middle cerebral artery occlusion (MCAO) rats, to mimic IS. MCAO rats and lipopolysaccharides (LPS)-induced Caco-2 cells were used to evaluate the effects of escin in vivo and in vitro. The results showed that escin could not penetrate BBB but reduced brain infarct volume, improved neurological function, inhibited neuroinflammation, ameliorated intestinal dysfunction and tissue integrity by increasing the expression of the tight junction protein in vivo and in vitro. Escin reduced the increased corticosterone and endotoxin level in blood of MCAO rats, regulated GR/p38 MAPK/NF-κB signaling pathway in ileal tissue and LPS/TLR4/NF-κB signaling pathway in ischemic brain tissue. These findings suggest that escin could attenuate ischemic brain injury by improving intestinal dysfunction, and it may be a promising way to protect brain injury by protecting intestine, instead of targeting the brain directly after IS.

A Human and Animal Based Study Reveals That a Traditionally Fermented Rice Beverage Alters Gut Microbiota and Fecal Metabolites for Better Gut Health

Fermented rice beverages are consumed globally, especially in Southeast Asia. In India, such beverages are consumed by a substantial population of ethnic communities. In this study, the gut bacterial diversity of rice beverage drinkers from Assam, India (n = 27) was compared with that of nondrinkers (n = 21) with the next-generation sequencing (NGS) of fecal metagenomic 16S rDNA, which indicated changes in 20 bacterial genera. Further, mice (n = 6, per treatment group) were gavaged daily for 30 days with different fractions of the beverage, which included rice beverage (RB), soluble (SF), and insoluble fractions (IF) to determine the effects of different components of the beverage. A comparison of gut bacteria at two time points, 0 and 30 days of treatments, suggested changes in 48 bacterial genera across the different treatment groups in mice. Major bacterial changes were suggestive of functional components associated with gut health, as observed in both humans and mice. Next, the Gas Chromatography–Mass Spectrometry (GC–MS) of mice stool after 30 days of treatments showed a total of 68 metabolites, of which hexadecanoic acid, a flavor component of this beverage, was present in the feces of all mouse treatment groups except controls. These metabolites showed treatmentwise clustering in groups in a partial least-squares discriminant analysis (PLS–DA) plot. Blood endotoxin levels were lower in all treatment groups in the mice compared to those of the controls. The findings of the study are suggestive of the gut modulatory effects of the beverage on the basis of the observed features of the bacterial changes.

Time-Related Evidence of Intestinal Oxidative Stress in Obstructive Jaundice-Induced Rats

Introduction: Obstructive jaundice is known to affect intestinal permeability and facilitate bacterial translocation through related mechanisms. This study was conducted to evaluate the alterations concerning blood biochemistry and levels of several markers of oxidative stress (OS) in blood and intestinal mucosa caused by obstructive jaundice and how these fluctuate over time, in order to further explore the possibility of intervening in the OS path in future experiments. Methods: A total of 54 albino Wistar rats were randomly divided into three groups (control, sham operated, and bile duct ligation) and sacrificed at specific time intervals (12 h and 2, 7, and 14 days). The intestinal barrier function was evaluated by measuring endotoxin levels in portal, aortic, and peripheral blood. Also, basic biochemical parameters were simultaneously measured in peripheral blood. Tissue samples collected from the terminal ileum were homogenized for determining the OS markers, lipid peroxidation, and protein-free radical-induced oxidation. Results: We designed this experiment to examine the alterations in enteric mucosa primarily in relation to OS in a period of 14 days. During this time period, we investigated in specific time intervals not only OS fluctuations but also other liver function parameters, as well as CRP and endotoxin levels. The alterations were monitored in relation to time after bile duct ligation. Conclusion: Bile duct ligation in rats causes OS versus post-ligation time progression of the common bile duct. OS was increased by ∼50% compared to control/sham and peaked at 7 days and at least up to 14 days post-ligation. This phenomenon was accompanied with a deranging of liver function after ligation, as anticipated, but not in all measured parameters; biochemical and endotoxin levels followed the same pattern.

Calycosin Improves Intestinal Mucosal Barrier Function after Gastrectomy in Rats through Alleviating Bacterial Translocation, Inflammation, and Oxidative Stress.

Objective Calycosin is the main bioactive extract of Astragali Radix with anti-inflammation, antioxidant, and anticancer properties. Here, our study evaluated the protective effects and mechanisms of calycosin on intestinal mucosal barrier under gastrectomy. Methods After receiving gastrectomy, the rats were administrated with 20 mg/kg, 40 mg/kg, or 80 mg/kg calycosin. Endotoxin, bacterial translocation, and intestinal bacterial flora were assayed. Intestinal injury was detected via hematoxylin and eosin staining. Tight junction indicators (occludin, claudin, and ZO-1) and apoptotic proteins (Bax, Bcl-2, and cleaved caspase 3) were examined in intestinal tissues. Inflammatory indicators (IL-1β, IL-6, and TNF-α) were examined in serum or intestinal specimens via ELISA. Apoptosis was assessed via TUNEL staining. IgA + B cells in intestinal tissues and sIgA in intestinal lumen were examined through immunohistochemistry and ELISA, respectively. Oxidative stress indicators (TSH, SOD, CAT, GSH-Px, and MDA) were also detected via ELISA. Results Our results showed that calycosin administration decreased endotoxin levels in peripheral blood, intestine, and portal vein blood; lowered the bacterial translocation ratio; and regained the balance among intestinal bacterial flora (comprising bifidobacterium, lactic acid bacillus, enterobacter, enterococcus, aerobic bacteria, and anaerobic bacteria) in the rats with gastrectomy. After calycosin treatment, intestinal mucosal damage of the rats with gastrectomy was ameliorated, with the increase in expression of tight junction proteins. Additionally, calycosin reduced intestinal inflammation, apoptosis, secretion of sIgA, and oxidative stress in the rats with gastrectomy. Conclusion Altogether, our findings demonstrate that calycosin may improve intestinal mucosal barrier function under gastrectomy via reducing bacterial translocation, inflammation, and oxidative stress.

Патогенетические аспекты нарушений метаболома кишечника и эндотоксемии у лиц высокого кардио-метаболического риска

Aim. To study the relationship between the functional activity of the intestinal microbiota and endotoxemia with impaired gastrointestinal motility and excessive bacterial growth in patients with metabolic syndrome (MS). Materials and methods. 64 patients with MS were examined. Comprehensive examination included collection of complaints, anamnesis of the disease, questionnaires on actual nutrition, clinical examination, anthropometric study, assessment of indicators of carbohydrate and lipid metabolism, a marker of systemic inflammatory response of the body (CRP) in the blood. Determination of the blood endotoxin level, quantitative and qualitative composition of the parietal microbiota of the small intestine was carried out on an Agilent gas chromatograph with mass selective and flame ionization detectors (Agilent Technologies, USA). A chromatographic study of metabolites of microorganisms (SCFA) in coprofiltrate was carried out. The motor function of the gastrointestinal tract (GIT) was studied using peripheral electrogastroenterocolography using a Gastroscan-GEM device. The control group consisted of 30 apparently healthy individuals aged from 18 to 62 years. Statistical processing of the obtained results was carried out using the Statistica 6.1 software. Results. In 82.9% of the examined patients with MS, clinical signs of intestinal lesions were noted. The electrical activity of the MC departments in the postprandial period was low in 70% of patients. It was found that with MS with a decrease in the daily consumption of dietary fiber in the composition of the diet, the BMI value increases, the energy value of the diet and the use of simple carbohydrates increase. During the analysis of the main biochemical blood constants, it was possible to establish that MS is accompanied by a violation of carbohydrate and lipid metabolism. The CRP level was statistically significantly different from the CG in MS regardless of the degree of excess body weight, while a direct correlation of moderate strength was established between the BMI values and the CRP level. During the chromatographic study of metabolites of microorganisms in feces, it was possible to establish statistically significant differences in the profile and concentration of short-chain fatty acids (SCFA) of persons with MS, both in comparison with CG and in comparison with groups with different BMI values. According to the results of the study of the blood endotoxin level in patients with MS, its significant increase was revealed, more than 6 times, in comparison with the CG. The electrical activity of the colon on an empty stomach was unchanged, but decreased after a food load. Discoordination of motility is observed between the small intestine (SI) and the large intestine, aggravated after food stimulation. A significant decrease in the rhythm of contractions is observed at the frequencies of the jejunum, ileum, and colon both on an empty stomach and in the postprandial period, which indicates a weakening of propulsive bowel contractions in patients with MS. In patients with MS, excessive bacterial growth in MC is observed mainly due to opportunistic microbiota strains. Correlation analysis showed a moderate negative relationship between endotoxin levels and MC electrical activity, between endotoxin levels and the colon-to-ileum ratio. Conclusion. The study of the level of endotoxin, intestinal microbiota metabolites (SCFA), indicators of bacterial overgrowth, motor-evacuation functions of the gastrointestinal tract made it possible to identify important pathogenetic patterns concerning the contributing role of the latter in the development of endotoxemia and intestinal metabolic disorders in patients with MS.

Influence of endotoxemy on development of non-alcoholic fatty liver disease

Objective — to investigate endotoxin levels and to assess the relationship between endotoxemia and metabolic parameters and the degree of steatosis and fibrosis in patients with non‑alcoholic fatty liver disease (NAFLD) against the background of metabolic syndrome (MS).
 Materials and methods. The study included 68 patients with NAFLD with metabolic disorders and 20 people of the control group. Women prevailed in both investigational groups, the mean age was 49.34 ± 10.8 years. All patients underwent evaluation of their anthropometric parameters, investigation of body composition with determination of the percentage of visceral adipose tissue (VAT). To reveal VAT dysfunction, visceral adiposity index (VAI) was calculated with M. C. Amato method. Steatometry was performed by determining the wave attenuation coefficient and carrying out shear wave elastography. Endotoxin levels in blood serum were determined using a set of reagents «LAL Chromogenic Endpoint Assay» manufactured by «Hycult Biotech» (Netherlands).
 Results. Bacterial endotoxin levels in peripheral blood in the examined patients were almost three times higher in comparison with the control group (p < 0.05). Patients with overweight and obesity had significantly higher endotoxin levels compared with the control group (p < 0.05). The significant increase in endotoxin levels has been defined in obese NAFLD patients compared to the NAFLD patients with normal weight (p < 0.05). The direct correlation was found between endotoxin parameters and parameters of abdominal and visceral obesity. In the group of patients with high VAT activity, the maximal values of this indicator were observed, which significantly differed from the corresponding indicator in patients with low and medium VAI (p < 0.05). There was a direct correlation between moderate endotoxin and insulin resistance, a direct weak relationship between endotoxin and lipid profile parameters. In patients with grade 2 and 3 steatosis, significant differences in the level of endotoxemia were observed compared with the control group (p < 0.05) and patients with grade 1 steatosis (p < 0.05). At high grades of liver fibrosis (F3F4) there was a tendency to reduce endotoxemia compared with the low‑grade fibrosis (p < 0.05).
 Conclusions. The revealed relationships between endotoxemia level and metabolic factors of NAFLD development indicate the pathogenetic role of bacterial endotoxemia in triggering a cascade of metabolic reactions with the subsequent accumulation of lipids in the liver tissue and the progression of steatosis and fibrosis.

The Impact of Alcohol-Induced Dysbiosis on Diseases and Disorders of the Central Nervous System.

The human digestive tract contains a diverse and abundant microbiota that is important for health. Excessive alcohol use can disrupt the balance of these microbes (known as dysbiosis), leading to elevated blood endotoxin levels and systemic inflammation. Using QIAGEN Ingenuity Pathway Analysis (IPA) bioinformatics tool, we have confirmed that peripheral endotoxin (lipopolysaccharide) mediates various cytokines to enhance the neuroinflammation signaling pathway. The literature has identified alcohol-mediated neuroinflammation as apossible risk factor for the onset and progression of neurodegenerative diseases, including Alzheimer's disease (AD) and Parkinson's disease (PD), and psychiatric disorders such as addiction to alcohol and other drugs. In this review, we discuss alcohol-use-induced dysbiosis in thegut and other body parts as a causal factor in the progression of Central Nervous System (CNS) diseases including neurodegenerative disease and possibly alcohol use disorder.

Endotoxins and Non-Alcoholic Fatty Liver Disease.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide. It occurs with a prevalence of up to 25%, of which 10–20% cases progress to nonalcoholic steatohepatitis (NASH), cirrhosis, and liver cancer. The histopathology of NASH is characterized by neutrophilic infiltration, and endotoxins from gram-negative rods have been postulated as a contributing factor. Elevations in endotoxin levels in the blood can be classified as intestinal and hepatic factors. In recent years, leaky gut syndrome, which is characterized by impaired intestinal barrier function, has become a significant issue. A leaky gut may prompt intestinal bacteria dysbiosis and increase the amount of endotoxin that enters the liver from the portal vein. These contribute to persistent chronic inflammation and progressive liver damage. In addition, hepatic factors suggest that liver damage can be induced by low-dose endotoxins, which does not occur in healthy individuals. In particular, increased expression of CD14, an endotoxin co-receptor in the liver, may result in leptin-induced endotoxin hyper-responsiveness in obese individuals. Thus, elevated blood endotoxin levels contribute to the progression of NASH. The current therapeutic targets for NASH treat steatosis and liver inflammation and fibrosis. While many clinical trials are underway, no studies have been performed on therapeutic agents that target the intestinal barrier. Recently, a randomized placebo-controlled trial examined the role of the intestinal barrier in patients with NAFLD. To our knowledge, this study was the first of its kind and study suggested that the intestinal barrier may be a novel target in the future treatment of NAFLD.

Modeling alcohol-associated liver disease in a human Liver-Chip.

SUMMARYAlcohol-associated liver disease (ALD) is a global health issue and leads to progressive liver injury, comorbidities, and increased mortality. Human-relevant preclinical models of ALD are urgently needed. Here, we leverage a triculture human Liver-Chip with biomimetic hepatic sinusoids and bile canaliculi to model ALD employing human-relevant blood alcohol concentrations (BACs) and multimodal profiling of clinically relevant endpoints. Our Liver-Chip recapitulates established ALD markers in response to 48 h of exposure to ethanol, including lipid accumulation and oxidative stress, in a concentration-dependent manner and supports the study of secondary insults, such as high blood endotoxin levels. We show that remodeling of the bile canalicular network can provide an in vitro quantitative readout of alcoholic liver toxicity. In summary, we report the development of a human ALD Liver-Chip as a powerful platform for modeling alcohol-induced liver injury with the potential for direct translation to clinical research and evaluation of patient-specific responses.

Outstanding Contributions of LAL Technology to Pharmaceutical and Medical Science: Review of Methods, Progress, Challenges, and Future Perspectives in Early Detection and Management of Bacterial Infections and Invasive Fungal Diseases.

The blue blood of the horseshoe crab is a natural, irreplaceable, and precious resource that is highly valued by the biomedical industry. The Limulus amebocyte lysate (LAL) obtained from horseshoe crab blood cells functions as a surprisingly sophisticated sensing system that allows for the extremely sensitive detection of bacterial and fungal cell-wall components. Notably, LAL tests have markedly contributed to the quality control of pharmaceutical drugs and medical devices as successful alternatives to the rabbit pyrogen test. Furthermore, LAL-based endotoxin and (1→3)-β-D-glucan (β-glucan) assay techniques are expected to have optimal use as effective biomarkers, serving as adjuncts in the diagnosis of bacterial sepsis and fungal infections. The innovative β-glucan assay has substantially contributed to the early diagnosis and management of invasive fungal diseases; however, the clinical significance of the endotoxin assay remains unclear and is challenging to elucidate. Many obstacles need to be overcome to enhance the analytical sensitivity and clinical performance of the LAL assay in detecting circulating levels of endotoxin in human blood. Additionally, there are complex interactions between endotoxin molecules and blood components that are attributable to the unique physicochemical properties of lipopolysaccharide (LPS). In this regard, while exploring the potential of new LPS-sensing technologies, a novel platform for the ultrasensitive detection of blood endotoxin will enable a reappraisal of the LAL assay for the highly sensitive and reliable detection of endotoxemia.

Exploring the Link Between Hepatic Perfusion and Endotoxemia in Hemodialysis

IntroductionThe liver receives gut-derived endotoxin via the portal vein, clearing it before it enters systemic circulation. Hemodialysis negatively impacts the perfusion and function of multiple organs systems. Dialysate cooling reduces hemodialysis-induced circulatory stress and protects organs from ischemic injury. This study examined how hemodialysis disrupts liver hemodynamics and function, its effect on endotoxemia, and the potential protective effect of dialysate cooling.MethodsFifteen patients were randomized to receive either standard (36.5°C dialysate temperature) or cooled (35.0°C) hemodialysis first in a two-visit crossover trial. We applied computed tomography (CT) liver perfusion imaging to patients before, 3 hours into and after each hemodialysis session. We measured hepatic perfusion and perfusion heterogeneity. Hepatic function was measured by indocyanine green (ICG) clearance. Endotoxin levels in blood throughout dialysis were also measured.ResultsDuring hemodialysis, overall liver perfusion did not significantly change, but portal vein perfusion trended towards increasing (P = 0.14) and perfusion heterogeneity significantly increased (P = 0.038). In addition, ICG clearance decreased significantly during hemodialysis (P = 0.016), and endotoxin levels trended towards increasing during hemodialysis (P = 0.15) and increased significantly after hemodialysis (P = 0.037). Applying dialysate cooling trended towards abrogating these changes but did not reach statistical significance compared to standard hemodialysis.ConclusionHemodialysis redistributes liver perfusion, attenuates hepatic function, and results in endotoxemia. Higher endotoxin levels in end-stage renal disease (ESRD) patients may result from the combination of decreased hepatic clearance function and increasing fraction of liver perfusion coming from toxin-laden portal vein during hemodialysis. The protective potential of dialysate cooling should be explored further in future research studies.

Consumption of Dietary Fiber from Different Sources during Pregnancy Alters Sow Gut Microbiota and Improves Performance and Reduces Inflammation in Sows and Piglets.

In pregnant and lactating sows, metabolism and immunity undergo drastic changes, which can lead to constipation, abortion, and intrauterine growth restriction (IUGR) and reduce production performance. Dietary fiber can regulate animal gut microbiota, alleviate inflammatory responses, and improve performance. Here, 48 sows (Large × Landrace) were randomly allocated to groups including, control, and with alfalfa meal (AM), beet pulp, and soybean skin dietary supplementation for 60 days of gestation. The AM diet decreased IUGR, increased food intake during lactation, and promoted the reproductive performance and physical condition of sows. Further, the AM diet significantly reduced markers of intestinal permeability (reactive oxygen species and endotoxin) in sow serum, and of systemic inflammation (interleukin-6 [IL-6] and tumor necrosis factor alpha) in sow feces and serum, as well as piglet serum, while it increased the anti-inflammatory marker, IL-10, in sow serum and feces. The AM diet also significantly affected gut microbiota by increasing the relative abundance of proinflammatory bacteria, while decreasing anti-inflammatory bacteria. Moreover, the total short-chain fatty acid (SCFA) content was higher in feces from sows fed an AM diet, with butyric acid content significantly higher during lactation, than in controls. Sow performance was correlated with intestinal permeability, inflammation, and gut microbiota, which were also vertically transmitted to piglets. Our results are significant for guiding feed management in the pig breeding industry. Further, the "sows to piglets" model provides a reference for the effect of dietary fiber on the gastrointestinal function of human mothers and infants.IMPORTANCE Although the direct effects of dietary fiber on gut microbiota composition have been studied extensively, systematic evaluation of different fiber sources on gut health and inflammatory responses of sows and their offspring has rarely been conducted. Excessive reactive oxygen species produced by overactive metabolic processes during late pregnancy and lactation of sows leads to increased endotoxin levels, disordered gut microbiota, decreased SCFA production, and secretion of proinflammatory factors, which in turn causes local inflammation of the gut, potential damage of the gut microbial barrier, increased gut permeability, increased blood endotoxin levels (resulting in systemic inflammation), and ultimately decreased sow and piglet performance. Our results showed that supplementation of the diet with alfalfa meal in mid and late pregnancy can reverse this process. Our findings lay a foundation for improving the gut health of sows and piglets and provide insights into the study of the gastrointestinal tract function in human mothers and infants.