PGE₂-EP4 pathway is in charge of LF41-mediated attenuation of hepatic TNF-α expression.

<p><b>(A)</b> ELISA for PGE₂ secretion by the terminal ileum and total PGE₂ levels in the liver of mice (n = 8) orally treated either for 10 days with PBS, L-LF41, or H-LF41, or for 3 weeks with PBS or H-LF41. * P < 0.05 PBS. <b>(B)</b> ELISA for hepatic IL-10 protein concentration of mice (n = 8) fed either PBS or H-LF41 for 10 days. P > 0.05 compared to PBS. <b>(C)</b> q-PCR for hepatic <i>Cox1</i> or <i>Cox2</i> mRNA levels of mice (n = 8) fed either PBS or H-LF41 for 10 days. Results are expressed as fold change relative to PBS. P > 0.05 compared to PBS. <b>(D)</b> Western blot assay for representative hepatic COX-1 and COX-2 protein levels of mice (n = 4) orally treated with either PBS or H-LF41 for 10 days. Hepatic COX-2 protein levels from a mouse receiving 2 h of stimulation with LPS (0.5 mg/kg BW; single IP injection) were determined as a positive control (left lane). <b>(E)</b> Mice (H-LF41-treated groups: n = 10 per group; PBS-treated groups: n = 8 per group) were pretreated with 10 days of PBS or H-LF41, either alone or combined with administration of either a specific inhibitor for PGE₂ receptor EP-4, ONA-AE3-208 (I-EP4), or its vehicle (Vehicle). Hepatic <i>Tnf</i> mRNA levels were assayed by q-PCR 2 h after LPS treatment. Results are expressed as fold change relative to PBS+LPS.* P < 0.05; <b>&</b> P < 0.05 compared to H-LF41+LPS; <b>n.s.</b>, non-statistical difference. All values except that of Western blot are shown as mean ± SEM. Results are representative of 2 similar experiments.</p

Validation of q-PCR for quantitation of LF and effect of LF41 administration on LF-specific 16S rRNA levels in intestinal tissues.

<p><b>(A)</b> LF41, BC41, or LGG was cultured in MRS broth at 37°C overnight. An aliquot of culture from each culture was dilution-plated on MRS agar (to enumerate each strain). Total bacterial genomic DNA was isolated from an aliquot of each culture and analyzed by q-PCR using the primers specific to the 16S rRNA of either LF or LGG. Black and white triangles denote log numbers of 16S rRNA gene copies determined by LF- and LGG-specific q-PCR, respectively; black squares denote log numbers of bacteria determined by serial dilution. <b>(B)</b> MRS broth was co-inoculated with LGG and low, middle, or high dose of LF41, grown at 37°C overnight. Total bacterial genomic DNA was isolated from an aliquot of each sample and analyzed by q-PCR using the primers specific to 16S rRNA of <i>Lactobacillus</i>, LF, or LGG. The samples of a, b, and c denote the co-cultures of LGG with low, middle, and high dose of LF41, respectively; “R(LF)” and “R(LGG)” denote the ratios of the respective 16S rRNA gene copies determined by LF- and LGG-specific q-PCR to the gene copies by <i>Lactobacillus</i>-specific q-PCR. <b>(B)(C)(D)</b> Mice (n = 8) were orally inoculated either for 10 days with PBS, L-LF41, or H-LF41, or for 3 weeks with PBS or H-LF41, and LF-specific 16S rRNA gene levels in terminal ileum <b>(B)</b>, proximal colon <b>(C)</b>, and distal jejuna <b>(D)</b> determined by q-PCR. Results are expressed as log₁₀ of the 16S rRNA gene copies per mg of tissue samples. Values of are shown as mean ± SEM. * P < 0.05 compared to L-LF41 or H-LF41 (21 days); <b>+</b> P < 0.05 compared to H-LF41 (10 days); nd, not detected. Results are representative of 2 experiments with similar results.</p

LF41-mediated upregulation of PGE₂ is abrogated by COX-2 blockade, but facilitated by IL-10 blockade in a COX-2-dependent manner.

<p><b>(A)(B)</b> ELISA for PGE₂ secretion by the terminal ileum (A) and PGE₂ amount in the liver (B) of mice (PBS-treated groups: n = 5–6 per group; H-LF41-treated groups: n = 8 per group) treated with a combination of either PBS or H-LF41 for 10 days, either alone or in combination with the COX-2-specific inhibitor celecoxib, its vehicle (vehicle), IL-10-specific antibody (Anti-IL-10), its isotype control (Is-IL-10), or Anti-IL-10 together with either celecoxib or its vehicle. * P < 0.05; <b>&</b> P < 0.05 compared to H-LF41; <b> P > 0.05 compared to PBS; n.s., non-statistical difference. (C)(E) Western blot assay for representative protein levels of COX-2 in epithelial cells (ECs) of the terminal ileum and COX-2 and COX-1 in the liver from mice (n = 4) either given 10 days H-LF41 treatment together with IL-10 blockade (C), or treated with 10 days of PBS, killed-LF41, H-LF41, or killed-LF41 together with H-LF41 (killed-LF41+H-LF41) (E). (D) ELISA for hepatic PGE₂ amount in mice (n = 8) fed PBS or H-LF41 for 10 days or given a combination of 10 days gavage of killed-LF41 or killed-LF41+H-LF41,either singly or combined with COX-2 blockade. * P < 0.05; & P < 0.05 compared to H-LF41; </b> P > 0.05 compared to PBS. All values except that of Western blot are shown as mean ± SEM. Results are representative of 2 similar experiments.</p

Ten days of H-LF41 treatment significantly enhances ileal expression of COX-2 and IL-10.

<p><b>(A)</b> q-PCR for mRNA levels of several factors associated with innate and adaptive immune responses in the terminal ileum collected from mice (n = 10) fed either for 10 days with PBS, L-LF41, or H-LF41 (upper panel), or for 3 weeks with either PBS or H-LF41 (lower panel). Results are expressed as fold change relative to “PBS”. * P < 0.05 compared to PBS. <b>(B)</b> MPO expression in the terminal ileum from mice (n = 6) treated with either PBS or H-LF41 for 10 days. P > 0.05 compared to PBS. <b>(C)</b> Epithelial cells (ECs) from the terminal ileum and its underlying lamina propria cells (LPCs) were isolated from mice (n = 8) orally given10 days supplement of PBS or H-LF41. <i>Cox2</i> and <i>Il10</i> mRNA levels in these cells were determined by q-PCR. Results are expressed as fold change relative to PBS. * P < 0.05compared to PBS. <b>(D)</b> Western blot assay for representative COX-2 protein levels in ECs and LPCs of the terminal ileum of mice (n = 4) fed either PBS or H-LF41 for 10 days. “RI” denotes the mean relative luminous intensity of the targeted protein band, which is positively correlated with the real luminance; the RI in the control group is set at 1.00. All values except that of Western blot are shown as mean ± SEM. Results are representative of 2 similar experiments.</p

Orally-pretreated LF41 attenuates LPS-induced TNF-α expression and hepatic injury.

<p><b>(A)</b> C57BL/6 mice (n = 8) either untreated or treated with antibiotic formula (Ab) were given single IP injection with LPS (500 μg/kg body weight). Mice were killed 2 and 16 h after LPS treatment for determination of hepatic TNF-α gene levels (left panel) by q-PCR and serum ALT activity (right panel), respectively. Results in the left panel are expressed as fold change relative to LPS. P > 0.05 compared to LPS. <b>(B)(C)</b> Mice (LPS-treated groups: n = 8–10 per group; the remainder: n = 6 per group) were given daily IG inoculation either for 10 consecutive days ofL-LF41, H-LF41, killed-LF41, LGG, BC41, or PBS, or for 21 consecutive days of either PBS or H-LF41 (right panel), and then single IP injection with LPS or PBS. Hepatic <i>Tnf</i> mRNA levels by q-PCR (B) and serum ALT activity (C) were determined. Results of (B) are expressed as fold change relative to PBS+PBS. H-LF41+LPS denotes 10 days of oral challenge with H-LF41 and then LPS injection, and other similar abbreviations conform to the same rule. * P < 0.05 compared to PBS+LPS. <b>(D)</b> Mice (LPS-treated groups: n = 8 per group; the remainder: n = 6 per group) were treated for 10 days with either PBS or H-LF41 and then challenged with LPS. Mice were killed 2 h after LPS treatment to test hepatic and serum TNF-α protein levels by ELISA.* P < 0.05 compared to PBS+LPS. <b>(E) (F)</b> Mice(LPS-treated groups: n = 12–14 per group; the remainder: n = 6–7 per group) pretreated for 10 days with PBS or H-LF were challenged with PBS or LPS. 20 h after the challenge, the inflammatory foci in the liver were determined (E), and representative histological outcomes of liver tissue were shown (F). * P < 0.05 compared to PBS+LPS. a: PBS+PBS; b: H-LF41+PBS; c: PBS+LPS; d: LF41+LPS. Values are shown as mean ± SEM. Results of <b>(A)</b> are representative of 2 experiments with similar results, and the remainder 3 experiments with similar results.</p

Effect of COX-2 or IL-10 blockade on TNF-α expression and intestinal permeability in LF41-fed mice.

<p><b>(A)(B)</b> Mice (PBS-treated groups: n = 5 per group; H-LF41-treated groups: n = 7 per group) were given 10 days treatment of PBS or H-LF41, either alone or in combination with blockade of EP4, COX-2, or IL-10, or co-blockade of TNF-α with COX-2 or IL-10, and then given IG inoculation with FITC-Dextran. Three hours later, the FITC-Dextran amount in the blood was determined. I-EP4, EP4-specific inhibitor; vehicle, the vehicle for celecoxib; celecoxib, COX-2-specific inhibitor; Anti-TNF, TNF-α-specific antibody; Is-TNF, the isotype control for Anti-TNF; Anti-IL-10, IL-10-specific antibody; Is-IL-10, the isotype control for Anti-IL-10.* P < 0.05; & P < 0.05compared to H-LF41; <b>n.s.</b>, non-statistical difference. <b>(C)</b> ELISA for TNF-α secretion by the terminal ileum collected from mice (PBS-treated groups: n = 5 per group; H-LF41-treated groups: n = 7 per group) fed for 10 days PBS or H-LF41, either singly or in combination with blockade of COX-2, IL-10, or EP4. * P < 0.05; & P < 0.05 compared toH-LF41. <b>(D)</b> q-PCR for <i>Tnf</i> mRNA levels in the epithelial cells (ECs) and lamina propria cells (LPCs) of the terminal ileum from mice (n = 7) treated for 10 days with PBS or H-LF41 in the presence of celecoxib administration. Results are expressed as fold change relative to PBS. * P < 0.05. <b>(E)</b> q-PCR for <i>Tnf</i> mRNA levels in the HMNCs isolated from mice (PBS-treated groups: n = 6 per group; H-LF41-treated groups: n = 8–10 per group) given 10 days treatment of PBS or H-LF41, either alone or together with blockade of EP4, COX-2, or IL-10, or with co-blockade of IL-10 and COX-2. Results are expressed as fold change relative to PBS. * P < 0.05; <b>&</b> P < 0.05 compared to H-LF41. Values are shown as mean ± SEM. Results are representative of 2 similar experiments.</p

Natural light-micro aerobic condition for PSB wastewater treatment: a flexible, simple, and effective resource recovery wastewater treatment process

<p>Photosynthetic bacteria (PSB) have two sets of metabolic pathways. They can degrade pollutants through light metabolic under light-anaerobic or oxygen metabolic pathways under dark-aerobic conditions. Both metabolisms function under natural light-microaerobic condition, which demands less energy input. This work investigated the characteristics of PSB wastewater treatment process under that condition. Results showed that PSB had very strong adaptability to chemical oxygen demand (COD) concentration; with F/M of 5.2–248.5 mg-COD/mg-biomass, the biomass increased three times and COD removal reached above 91.5%. PSB had both advantages of oxygen metabolism in COD removal and light metabolism in resource recovery under natural light-microaerobic condition. For pollutants’ degradation, COD, total organic carbon, nitrogen, and phosphorus removal reached 96.2%, 91.0%, 70.5%, and 92.7%, respectively. For resource recovery, 74.2% of C in wastewater was transformed into biomass. Especially, coexistence of light and oxygen promote N recovery ratio to 70.9%, higher than with the other two conditions. Further, 93.7% of N-removed was synthesized into biomass. Finally, CO₂ emission reduced by 62.6% compared with the traditional process. PSB wastewater treatment under this condition is energy-saving, highly effective, and environment friendly, and can achieve pollution control and resource recovery.</p

Three- and Eight-Fold Interpenetrated ThSi₂ Metal–Organic Frameworks Fine-Tuned by the Length of Ligand

Two new interpenetrated ThSi₂ networks, {[Ag₄(bipy)₄(ox)]·2OH·16H₂O}_<i>n</i> (<b>1</b>) and {[Ag₂(dpb)₂(ox)]·10H₂O}_<i>n</i> (<b>2</b>) (bipy = 4,4′-bipyridine, dpb = 1,4-di­(pyridin-4-yl)­benzene and Na₂ox = sodium oxalate), were constructed from bidentate pyridyl-based organic tectons incorporating ox auxiliary ligand. Interestingly, both <b>1</b> and <b>2</b> are 3D frameworks with the same ThSi₂ topology but with substantial changes in the interpenetration degrees, which are well controlled by employing the pyridyl-based ligands with different lengths. The thermal stabilities and photoluminescence behaviors of them were also discussed

Three- and Eight-Fold Interpenetrated ThSi₂ Metal–Organic Frameworks Fine-Tuned by the Length of Ligand

Two new interpenetrated ThSi₂ networks, {[Ag₄(bipy)₄(ox)]·2OH·16H₂O}_<i>n</i> (<b>1</b>) and {[Ag₂(dpb)₂(ox)]·10H₂O}_<i>n</i> (<b>2</b>) (bipy = 4,4′-bipyridine, dpb = 1,4-di­(pyridin-4-yl)­benzene and Na₂ox = sodium oxalate), were constructed from bidentate pyridyl-based organic tectons incorporating ox auxiliary ligand. Interestingly, both <b>1</b> and <b>2</b> are 3D frameworks with the same ThSi₂ topology but with substantial changes in the interpenetration degrees, which are well controlled by employing the pyridyl-based ligands with different lengths. The thermal stabilities and photoluminescence behaviors of them were also discussed

Data_Sheet_1_Alterations in the intestinal microbiome and metabolic profile of patients with cirrhosis supplemented with lactulose, Clostridium butyricum, and Bifidobacterium longum infantis: a randomized placebo-controlled trial.xlsx

BackgroundLiver cirrhosis is commonly accompanied by intestinal dysbiosis and metabolic defects. Many clinical trials have shown microbiota-targeting strategies represent promising interventions for managing cirrhosis and its complications. However, the influences of the intestinal metagenomes and metabolic profiles of patients have not been fully elucidated.MethodsWe administered lactulose, Clostridium butyricum, and Bifidobacterium longum infantis as a synbiotic and used shotgun metagenomics and non-targeted metabolomics to characterize the results.ResultsPatients treated with the synbiotic for 12 weeks had lower dysbiosis index (DI) scores than placebo-treated patients and patients at baseline (NIP group). We identified 48 bacterial taxa enriched in the various groups, 66 differentially expressed genes, 18 differentially expressed virulence factor genes, 10 differentially expressed carbohydrate-active enzyme genes, and 173 metabolites present at differing concentrations in the Synbiotic versus Placebo group, and the Synbiotic versus NIP group. And Bifidobacteria species, especially B. longum, showed positive associations with many differentially expressed genes in synbiotic-treated patients. Metabolites pathway enrichment analysis showed that synbiotic significantly affected purine metabolism and aminoacyl-tRNA biosynthesis. And the purine metabolism and aminoacyl-tRNA biosynthesis were no longer significant differences in the Synbiotic group versus the healthy controls group. In conclusion, although littles influence on clinical parameters in the early intervention, the synbiotic showed a potential benefit to patients by ameliorating intestinal dysbiosis and metabolic defects; and the DI of intestinal microbiota is useful for the evaluation of the effect of clinical microbiota-targeting strategies on cirrhotic patients.Clinical Trial Registrationhttps://www.clinicaltrials.gov, identifiers NCT05687409.</p