Fecal Levels Research Articles

The role of fecal matrix metalloprotease-9 as a non-invasive marker in diagnosis and assessment of clinical activity in inflammatory bowel disease patients

BackgroundInflammatory bowel disease is characterized by chronic and relapsing inflammation of the gastrointestinal tract, including two prominent forms: Crohn’s disease and ulcerative colitis. Determining diagnostic biomarkers for predicting disease activity and treatment response remains a challenging aspect.Aim of the workThe purpose of our research was to compare fecal CP and fecal MMP-9, two non-invasive biomarkers for inflammatory bowel disease (IBD), and to find out how fecal MMP-9 levels relate to disease activity by looking at how they relate to clinical, endoscopic, and histologic scores of disease activity.Patients and methodsThis study was performed on 80 subjects divided into 3 groups: group A: 30 patients with Crohn’s disease evidenced by endoscopy ileocolonoscopy, upper GI endoscopy, and tissue biopsy (15 patients with active disease and 15 patients in remission). Group B: 30 patients with ulcerative colitis disease evidenced by colonoscopy and tissue biopsy (15 patients with active disease and 15 patients in remission). Group C: 20 age-matched and sex-matched healthy controls. All participants underwent a thorough history review, comprehensive physical examination, complete laboratory tests, and C-reactive protein measurements. A quantitative enzyme-linked immunosorbent assay was used to determine the levels of fecal matrix metalloproteinase MMP 9 for both the patients and the controls. Ulcerative colitis was evaluated using the Mayo score, Montreal classification, and the Riley histological score. Additionally, Crohn’s disease was assessed with the Crohn’s Disease Activity Index, the Simple Endoscopic Score for Crohn’s Disease, and the D’Haens histological score.ResultsComparing fecal MMP-9 with fecal calprotectin (FC), we found that fecal MMP-9 was superior to FC in differentiating active Crohn’s disease from inactive Crohn’s disease, although there was no significant difference between FC and MMP-9 (P-value = 0.561). However, in ulcerative colitis, FC was superior to MMP-9 in distinguishing active UC from inactive UC, but again, there was no significant difference between FC and MMP-9 (P-value = 0.0731).In both the ulcerative colitis and Crohn’s disease groups, fecal MMP-9 could discriminate between patients in remission and those with active disease. Fecal matrix metalloproteinase-9 (MMP-9) was discovered to be a significant marker for assessing the clinical activity of both Crohn’s disease (CD) and ulcerative colitis (UC), with an AUC of 0.998 for CD and 0.991 for UC. Fecal MMP-9 demonstrated great sensitivity (93.33%), specificity (100%), positive predictive value (PPV) of 100%, and negative predictive value (NPV) of 93.7% (with a P-value < 0.001) using cutoff values of > 0.34 ng/mL for CD and > 0.36 ng/mL for UC. There was a strong positive correlation between fecal MMP-9 and endoscopic and clinical scores of disease activity.ConclusionFecal MMP-9 has emerged as a promising biomarker for evaluating the clinical activity of both Crohn’s disease and ulcerative colitis. It demonstrated superior diagnostic performance compared to fecal calprotectin in distinguishing active from inactive disease, especially in Crohn’s disease. Although fecal calprotectin outperformed MMP-9 in identifying active ulcerative colitis, the differences between the two markers were not statistically significant, suggesting that they may complement each other in clinical practice. Furthermore, fecal MMP-9 is capable of assessing the activity of endoscopically visible inflammatory bowel disease (IBD), which could help reduce the need for invasive endoscopic procedures.

Sexual Dimorphism in the Immunometabolic Role of Gpr183 in Mice

Abstract Aim Excessive eating and intake of a Western diet negatively affect the intestinal immune system, resulting in compromised glucose homeostasis and lower gut bacterial diversity. The G protein-coupled receptor GPR183 regulates immune cell migration and intestinal immune response and has been associated with tuberculosis, type 1 diabetes, and inflammatory bowel diseases. We hypothesized that with these implications, GPR183 has an important immunometabolic role and investigated this using a global Gpr183 knock-out mouse model. Material and methods Wild-type (WT) and Gpr183 deficient (Gpr183-/-) mice were fed a high-fat, high-sucrose diet (HFSD) for 15 weeks. We investigated changes in weight, body composition, fecal IgA levels, fecal microbiome, and glucose tolerance before and after the diet. Macrophage infiltration into visceral fat was determined by flow cytometry, and hepatic gene expression was measured. Results and conclusions A sexual dimorphism was discovered, where female Gpr183-/- mice showed adverse metabolic outcomes compared to WT counterparts with inferior glucose tolerance, lower fecal IgA levels, and increased macrophage infiltration in visceral fat. In contrast, male Gpr183-/- mice had significantly lower fasting blood glucose after diet than male WT mice. Liver gene expression showed reduced inflammation and macrophage markers in Gpr183-/- livers, regardless of sex, while the pancreatic islet area did not differ between the groups. No conclusive differences were found after microbiome sequencing. To conclude, Gpr183 maintains metabolic homeostasis in female but not in male mice independent of diet. If confirmed in humans, future therapy targeting GPR183 should consider this sexual dimorphism.

Oral supplementation of heat-killed Enterococcus faecalis strain EC-12 relieves gastrointestinal discomfort and alters the gut microecology in academically stressed students.

Stress significantly affects gastrointestinal and mental health, and the gut microbiota plays a pivotal role in this process. Enterococcus faecalis strain EC-12 (EC-12) is a lactic acid bacterium that has several health benefits. To investigate the impact of oral supplementation with heat-killed EC-12 on the discomfort caused by stress, a randomised, double-blind, placebo-controlled trial was conducted with students under academic stress taking EC-12 (n= 14) or a placebo (n= 13) daily for one week. Improvement in the students' symptoms was assessed using the visual analogue scale. Faecal microbiota was characterised by next-generation sequencing of 16S rRNA genes, and faecal metabolites and short-chain fatty acids were analysed using a GC-MS metabolomics approach. Significant improvements in abdominal pain and rumbling of the stomach were found in the EC-12 group compared to the placebo group, but no changes were observed in mental symptoms or salivary cortisol levels. The relative abundance of E. faecalis significantly increased in the EC-12 group after the trial; however, the composition and diversity of the gut microbiota did not change significantly. Functional analysis of the gut microbiota suggested that EC-12 intake alters specific metabolic pathways. Although the levels of faecal short-chain fatty acids did not change between the groups before and after the trial, EC-12 intake altered the composition of faecal metabolites, with a significant increase in tryptamine levels. The ratio of students with improved symptoms to those with increased tryptamine levels was calculated based on the number of students with elevated faecal tryptamine levels who showed symptomatic improvements. The ratio of improved rumbling stomach was higher than that of other types of digestive discomfort. These results suggest that oral supplementation with EC-12 has a potentially beneficial effect on stress-induced gastrointestinal discomfort, which may occur through alterations in gut microbiota composition and metabolism. This study was registered at the University Hospital Medical Information Network Center (UMIN) under the UMIN ID: UMIN000048184.

The effect of environmental and anthropogenic factors on the microbiome of the sponge, Halichondria panicea, at three coastal sites with different bathing water quality in North east England

The breadcrumb sponge, Halichondria panicea, is a cosmopolitan marine species. Life functions, such as feeding, metabolism and defence, are maintained through microbial symbiosis. As such, perturbations to the symbiotic balance can be expected to affect the health and survival of the sponge. Although generally tolerant of environmental variables, such as temperature, pH and salinity, responses to anthropogenic factors are poorly understood. In this study, the microbial community of the H. panicea was examined over the course of 1 year. Sponge and seawater samples were collected in January, April, July and October 2022, from three locations with different levels of bathing water quality, according to the UK’s Environment Agency. Samples were sequenced using the 16S ribosomal RNA (rRNA) gene, and amplicon sequence variants (ASVs) were inferred from the generated data. Differences in bacterial diversity and abundance among sponge samples from the three locations were examined. A correlation test was used to study the effect of physical and chemical environmental factors along with faecal indicator bacteria on the abundance of the top ten most abundant bacterial phyla. Environmental factors (determined from seawater physicochemical properties) and pollution (determined from trace metals, nutrients and faecal bacteria levels) were found to play an important role in shaping the microbial community of this sponge. The sponge microbiome showed a noticeable seasonal shift, with some species flourishing in January and others emerging in April, notably the faecal and coliform bacteria. Sponge microbiomes from sites with poor-quality bathing water were generally less diverse and had lower microbial abundance, resulting in a greater range of intra-species dispersion than those of sponges living in excellent–good quality waters.

Does gut microbiota dysbiosis impact the metabolic alterations of hydrogen sulfide and lanthionine in patients with chronic kidney disease?

BackgroundChronic Kidney Disease (CKD) is characterized by a methionine-related metabolic disorder involving reduced plasma levels of hydrogen sulfide (H2S) and increased lanthionine. The gut microbiota influences methionine metabolism, potentially impacting sulfur metabolite dysfunctions in CKD. We evaluated whether gut microbiota dysbiosis contributes to H2S and lanthionine metabolic alterations in CKD.MethodsThe gut microbiota of 88 CKD patients (non-dialysis, hemodialysis, and transplant patients) and 26 healthy controls were profiled using 16 S-amplicon sequencing. H2S and lanthionine concentrations were measured in serum and fecal samples using the methylene blue method and LC-MS/MS, respectively.ResultsThe CKD population exhibited a tenfold increase in serum lanthionine associated with kidney dysfunction. Despite lanthionine retention, hemodialysis and transplant patients had significantly lower serum H2S than healthy controls. Fecal H2S levels were not altered or related to bloodstream H2S concentrations. Conversely, fecal lanthionine was significantly increased in CKD compared to healthy controls and associated with kidney dysfunction. Microbiota composition varied among CKD groups and healthy controls, with the greatest dissimilarity observed between hemodialysis and transplant patients. Changes relative to the healthy group included uneven Ruminococcus gnavus distribution (higher in transplant patients and lower in non-dialysis CKD patients), reduced abundance of the short-chain fatty acid-producing bacteria Alistipes indistinctus and Coprococcus eutactus among transplant patients, and depleted Streptococcus salivarius in non-dialysis CKD patients. A higher abundance of Methanobrevibacter smithii, Christensenella minuta, and Negativibacillus massiliensis differentiated hemodialysis patients from controls. No correlation was found between differentially abundant species and the metabolic profile that could account for the H2S and lanthionine alterations observed.ConclusionsThe metabolic deregulation of H2S and lanthionine observed in the study was not associated with alterations in the gut microbiota composition in CKD patients. Further research on microbial sulfur pathways may provide a better understanding of the role of gut microbiota in maintaining H2S and lanthionine homeostasis.

Tryptophan Metabolites Improve Intestinal Mucosal Barrier via the Aryl Hydrocarbon Receptor-Interleukin-22 Pathway in Murine Dextran Sulfate Sodium-Induced Pouchitis.

Pouchitis is the commonest complication after ileal pouch-anal anastomosis for ulcerative colitis. The protective effect of tryptophan metabolites on the mucosal barrier may be an effective method for treating pouchitis. The role of tryptophan metabolites on pouchitis remained unclear. We aimed to establish a murine model of dextran sulfate sodium-induced pouchitis to examine the roles of tryptophan metabolites in its pathogenesis. This is a study combined clinical patient data and animal research. A total of 22 patients were enrolled: 5 with familial adenomatous polyposis after ileal pouch-anal anastomosis, eight ulcerative colitis patients after ileal pouch-anal anastomosis patients with pouchitis, and 9 ulcerative colitis patients after ileal pouch-anal anastomosis with normal pouch. The demographic data and fecal samples of patients were collected. Male C57BL/6 mice were purchased from a licensed breeder and underwent ileal pouch-anal anastomosis to establish murine model of pouch. The bloods, feces, tissues of mice were collected. This study was performed in an academic medical center in China. The demographic data of patients were observational collected. The mice underwent ileal pouch-anal anastomosis were divided into six groups: control group with chow diet, dextran sulfate sodium, 6-formylindolo[3,2-b] carbazole + dextran sulfate sodium, high tryptophan diet + dextran sulfate sodium, CH-223191 + dextran sulfate sodium, indole-3-carboxaldehyde + dextran sulfate sodium. Animals were sacrificed after dextran sulfate sodium for 7 days. Fecal tryptophan metabolite level and microbiome composition, the severity of pouchitis, intestinal mucosal barrier function, and activation of the aryl hydrocarbon receptor-interleukin 22 pathway were assessed. Patients with pouchitis had lower fecal microbial diversity and indole-3-acetic acid levels. In murine pouchitis model, high-tryptophan diet increased fecal levels of 3-indoleglyoxylic acid, indole-3-aldehyde, and indole. A high-tryptophan diet and intraperitoneal aryl hydrocarbon receptor ligand 6-formylindolo[3,2-b] carbazole injection alleviated pouchitis. Tryptophan metabolites improved pouch mucosal barriers. aryl hydrocarbon receptor inhibitors exacerbated experimental pouchitis and disrupted the mucosal barrier; however, the aryl hydrocarbon receptor ligand indole-3-carboxaldehyde reversed this effect. This study was limited by small human sample size and lacking an Aryl hydrocarbon receptor knockout mouse model. A high-tryptophan diet and aryl hydrocarbon receptor ligand alleviated dextran sulfate sodium-induced pouchitis in murine ileal pouch-anal anastomosis model, which might be through regulating epithelial tight junctions and promoting goblet-cell differentiation, as well as maintaining the integrity and function of the mucosal barrier. This study provides a rationale for the clinical application of Aryl hydrocarbon receptor ligands in the treatment of pouchitis. See Video Abstract.

Fecal glycoprotein 2 is a marker of gut microbiota dysbiosis and systemic inflammation

BackgroundAntimicrobial autoantigenic glycoprotein 2 (GP2) is an important component of the innate immune system which originates from the exocrine pancreas as well as from the small intestines. The relationship of GP2 with the intestinal microbiome as well as the systemic implications of increased fecal GP2 levels are, however, still unclear. Therefore, fecal samples from 2,812 individuals of the Study of Health in Pomerania (SHIP) were collected to determine GP2 levels (enzyme-linked immunosorbent assay) and gut microbiota profiles (16 S rRNA gene sequencing). These data were correlated and associated with highly standardised and comprehensive phenotypic data of the study participants.ResultsFecal GP2 levels were increased in individuals with higher body mass index and smokers, whereas lower levels were found in case of preserved exocrine pancreatic function, female sex or a healthier diet. Moreover, higher GP2 levels were associated with increased serum levels of high-sensitivity C-reactive protein, loss of gut microbial diversity and an increase of potentially detrimental bacteria (Streptococcus, Haemophilus, Clostridium XIVa, or Collinsella). At the same time, predicted microbial pathways for the biosynthesis of beneficial short-chain fatty acids or lactic acid were depleted in individuals with high fecal GP2. Of note, GP2 exhibited a stronger association to overall microbiome variation than calprotectin.ConclusionFecal GP2 is a biomarker of gut microbiota dysbiosis and associated with increased systemic inflammation. The intestines may be more important as origin for GP2 than pancreatic acinar cells. Future studies need to investigate the potential clinical value in disease specific patient cohorts.

RISE IN PLASMA BILE ACIDS FOLLOWING HYPOABSORPTIVE BARIATRIC SURGERIES PREDICT BENEFICIAL METABOLIC AND HOMEOSTATIC OUTCOMES IN MALE RATS.

The effects of three hypoabsorptive bariatric surgeries, Roux-en-Y gastric bypass (RYGB), biliopancreatic diversion with duodenal switch (BPD-DS) and single anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S), on bile acids (BAs) were assessed including the changes in BA plasma levels associated with the metabolic and homeostatic effects of the surgeries. Male Wistar rats, etheir fed high- (HF) or a low-fat (LF) diets, were divided into seven groups: RYGB HF, BPD-DS HF, SADI-S HF, sleeve-gastrectomy (SG) HF, sham-operation (SHAM) HF, SHAM LF and SHAM HF-pair-weighed to BPD-DS (SHAM HF-PW). The rats were treated 56 days. The results demonstrate the ability of RYGB, BPD-DS and SADI-S to raise plasma levels of BAs, whose elevations, most notably those of the secondary BAs (deoxycholic acid, ursodooxycholic acid and lithocholic acid) associated negatively with body weight gain, fat gain and fasting insulin levels and positively with plasma peptide tyrosine-tyrosine (PYY). Plasma BAs also correlated positively with the fecal levels of Clostridium, Sutterella and Enterobacteriaceae and negatively with Clostridiales_f_g_2, Christensenellaceae, Ruminococcaceae_g_2, Oscillibacter and Oscillospira. Additionally, they associated positively with the short-chain fatty acid (SCFA) levels of propionate, butyrate, isobutyrate, valerate and isovalerate. Altogether, the present study emphasizes the ability of RYGB, BPD-DS and SADI-S to induce circulating BA elevations that predict the beneficial consequences of those hypoabsorptive bariatric surgeries on energy and glucose homeostasis and circulating levels of PYY. The present results also reveal close associations between plasma BAs and SCFAs, whose variations following hypoabsorptive surgeries are also linked to significant fat losses and metabolic health improvements.

Metabolomic profiling reveals the step-wise alteration of bile acid metabolism in patients with diabetic kidney disease

BackgroundDiabetic kidney disease (DKD) is the major complication of diabetes concomitant with gut dysbiosis and glycometabolic disorder, which are strongly associated with bile acid (BA) metabolism. Yet studies investigating the BA metabolism involving in DKD pathogenesis are limited. This study aimed to explore the metabolomic profiling of BAs in DKD and analyze its association with DKD progression.MethodsAn ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was established to quantify BAs in the plasma, fecal and urine samples of patients with DKD or T2DM and healthy individuals (n = 30 for each group). The key BAs associated with DKD were identified by orthogonal partial least-squares discriminant analysis (OPLS-DA) and receiver-operating characteristic (ROC) curve. Polynomial regression and Pearson’s correlation analyses were performed to assess the correlation between the key BAs and the clinical indicators reflecting DKD progression.ResultsMetabolomic profiling of 50 kinds of BAs presented the markedly step-wise alterations of BAs in plasma and feces as well as the little in urine of patients with DKD. Eight kinds of BAs in the plasma, eight kinds in the feces and three kinds in the urine were abnormally expressed, accompanying with the increased conjugated/unconjugated ratios of cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid and hyocholic acid in the plasma, and of cholic acid, chenodeoxycholic acid and lithocholic acid in the feces. Moreover, the increased plasma level of glycochenodeoxycholic acid, and the increased fecal levels of glycolithocholic acid, 7-ketodeoxycholic acid and chenodeoxycholic acid-3-β-D-glucuronide are strongly correlated with the clinical indicators reflecting DKD progression, including eGFR, 24 h urinary protein and 24 h urinary microalbumin.ConclusionsOur study for the first time disclosed the specific alterations of BA metabolism reflecting the step-wise progression of DKD, providing the basis for early identification and therapeutical strategies for DKD.

Fecal zonulin as a prognostic marker of atopic march in children with food allergy

Introduction. The onset of allergic diseases most often occurs in early childhood with the onset of food allergies, which can subsequently lead to the implementation of the atopic march. Increased intestinal permeability with high production of zonulin, the main moderator of intestinal tight junctions, can be an important link in the development of comorbid allergic diseases.Material and methods. In order to study the significance of fecal zonulin as a marker for predicting the atopic march in children with food allergy, a cross-sectional retrospective study was conducted on 73 children aged 5 years who were diagnosed with food allergy (FA) to cow’s milk proteins in the first year of life. In all children, when the diagnosis was made in the first year of life, the content of zonulin in feces was determined using the ELISA method.Results. As a result of dynamic observation, all children with food allergy were divided into 2 groups: the first group consisted of children with food allergy who developed allergic rhinitis and/or bronchial asthma within 5 years (group I, n = 39), group 2 consisted of 34 children with food allergy who did not implement the atopic march within 5 years of observation (group II, n = 34). Our study showed statistically significant differences in the fecal zonulin level in the first year of life: group I Me = 2.39 ng/ml (Q1-Q3: 1.78–2.65 ng/ml), group II Me = 1.85 ng/ml (Q1-Q3: 0.49–0.91 ng/ml), p = 0.034. Strong direct correlations were found (Spearman correlation coefficient S = 0.681 (p &lt; 0.05)) between the zonulin level in feces at the onset of the disease and the development of allergic rhinitis and/or bronchial asthma up to 5 years of age, the data were confirmed by comparing the areas under the curves during ROC analysis, AUC in the study of fecal zonulin as a prognostic marker of the risk of atopic march in children is 0.887, the optimal threshold (cutoff point) is 1.94 ng/ml.Conclusions. Fecal zonulin level in children with food allergy can be an effective prognostic marker of atopic march development, its values in feces above 1.94 ng/ml allow us to predict with a high degree of probability the risk of atopic march development in children with food allergy to cow’s milk proteins within 5 years

Intestinal inflammation and permeability in patients recovered from SARS-CoV-2 infection.

Different works suggest a close link between Long COVID Gastrointestinal (GI) manifestations and the post-infection disorders of gut-brain interaction (PI-DGBIs). However, the actual mechanisms underlying long-term GI sequelae are still not clear. Our study was aimed to assess both intestinal inflammation and permeability among subjects recovered from SARS-CoV-2 infection and their eventual correlation with long-term GI sequelae. Eighty-six subjects attending the Post-COVID Service and recovered from SARS-CoV-2 infection for six months, were investigated for Long COVID manifestations. Those subjects complaining of long-term GI symptoms were further evaluated by Rome IV questionnaire to assess PI-DGBIs. Intestinal inflammation (by fecal calprotectin, FC), and permeability (by serum and fecal levels of zonulin) were evaluated in all subjects. The Hospital Anxiety and Depression Scale (HADS) and The Gastrointestinal Quality of Life Index (GIQLI) questionnaires were further provided to all participants. Thirty-seven subjects (43%) complained of long-term GI symptoms, while 49 subjects (57%) did not. Thirty-three subjects fulfilled Rome IV criteria for PI-DGBIs. FC values resulted higher in those subjects who did not complain GI symptoms (p=0.03), although remaining quite close to the normal range. No significant differences were shown regarding the assessment of intestinal permeability. By GIQLI, long-term gastrointestinal sequelae were inversely correlated with quality of life (p=0.009). Long COVID GI complaints unlikely recognize underlying local inflammatory mechanisms. Since the healthcare, economic, and social burden of post-COVID DGBIs, a deeper understanding of this emerging condition should be encouraged to improve management of the affected subjects.

Stress levels, hematological condition, and productivity of plasma-producing horses used for snake antivenom manufacture: A comparison of two industrial bleeding methods

The immunization and industrial bleeding of horses are essential stages for producing snake antivenoms. In Costa Rica, the traditional method involves stimulating the antibody response of horses by periodically injecting venoms, collecting hyperimmune plasma over three consecutive bleeding days, and repeating this process every eight weeks. While this method does not cause major physical or hematological issues in horses, the associated stress has not been evaluated. We compared this traditional method with an alternative method that involves injecting venoms, collecting hyperimmune plasma in a single bleeding day, and repeating the process every two weeks. We assessed stress (via serum and fecal cortisol levels and an ethological study), hematological parameters (hematocrit and hemoglobin concentration), and plasma productivity over eight months. Serum cortisol levels remained within the normal range for both methods throughout the immunization/bleeding cycle. However, serum and fecal cortisol levels were significantly higher in horses subjected to the traditional method compared to those in the alternative method. Neither method caused significant hematological alterations. Notably, the alternative method yielded a higher volume of plasma. We concluded that adopting the alternative method ensures horse welfare while improving industrial bleeding productivity. This approach may reduce costs and improve the availability of this essential treatment for vulnerable populations.

Gut-lung microbiota dynamics in mice exposed to Nanoplastics

Concern has grown over potential health effects of micro- and nanoplastics (M/NPs) exposure. There is significant interest in understanding their impact on animal and human microbiota due to its crucial role in preserving health, as research in this area is rapidly advancing. We conducted a sub-chronic exposure study involving 12 male mice, divided into two groups: a control group (n = 6) and a PET-NPs exposure group (n = 6). PET-NPs, administered by oral gavage at a dose of 0.5 mg/day in 0.1 ml/mice, were given daily for 28 days. Microbiota analyses were performed on lung, colon, oral cavity, and stool samples using 16S rRNA sequencing. Additionally, fecal short and medium-chain fatty acids were analyzed by GC/MS. No significant changes were observed in the fecal and oral microbiome of the treated mice, nor in the fecal fatty acid levels. However, there were prominent alterations in the colon, characterized by increased abundance of Gram-negative bacteria belonging to Veillonella and Prevotella genera, and of amino acid metabolism pathways, coupled with a decrease in Lactobacillus. PET-NPs ingestion caused unexpected alterations in the lung microbiome with an increase in the Pseudomonas and changes in microbial energy metabolism and nitrogen utilization. This study provides insights into the differential impact of PET-NPs exposure on various microbiome niches.

Integrating fecal metabolomics and gut microbiome to reveal the mechanism of Schisandra chinensis-Acorus tatarinowii Schott treatment in Alzheimer’s disease rats

BackgroundSchisandra chinensis (Turcz.) Baill (Schisandraceae) and Acorus tatarinowii Schott (Araceae Juss) (Sc-At) are two traditional Chinese medicinal herbs that are widely used in the treatment of neurological disorders such as insomnia. In our previous study, we found that Sc-At could improve the therapeutic efficacy of Alzheimer’s disease by affecting aromatase activity and estrogen levels, among other pathways. Material and methodsIn this study, first, the ameliorative effect of Sc-At on cognitive dysfunction in Alzheimer’s disease model rats was verified by Y-maze test together with Elisa assay. Second, fecal untargeted metabolomics analysis was performed using a UPLC-Q-TOF/MS-based metabolomics approach. 16S rDNA sequencing was utilized to screen the differential flora between groups, and linear discriminantan alysis effect size (LEfSe) was used to find the target flora. Finally, Spearman correlation analysis was combined to find the relationship between differential flora and differential metabolites in feces. Results(1) The results of Y-maze test and Elisa assay indicated that Sc-At could improve the cognitive dysfunction of AD model rats. (2) Metabolomics results showed that fecal metabolite levels were significantly different from those of rats in the blank group, and 18 potential biomarkers in feces were screened, mainly affecting linoleic acid metabolism, steroid hormone biosynthesis, sphingomyelin metabolism, riboflavin metabolism, etc. The 16S rDNA results showed that the abundance and diversity of intestinal flora in AD rats were destroyed, and the Sc-At treatment was able to reverse these changes. (3) Spearman’s correlation analysis showed a significant correlation between differential metabolites in feces and intestinal flora, further suggesting that Sc-At treats Alzheimer’s disease through the gut-brain axis. ConclusionsIn this study, we explored the mechanism of Sc-At in the treatment of Alzheimer’s disease by integrating fecal untargeted metabolomics and 16S rDNA gene sequencing, and the results showed that Sc-At exerts therapeutic effects on Alzheimer’s disease by improving the intestinal flora and related metabolic pathways.

Evaluation of the Antibacterial Effects of the Punica granatum Peels Extracts Against some Pathogenic Bacteria: An In vivo and In vitro Study

Background: This study aimed to evaluate the antibacterial effects of Punica granatum extract on such pathogenic bacteria as Staphylococcus aureus and Escherichia coli. Materials and methods: The samples from 130 patients with skin infections in Baghdad, Iraq, aged between 15 and 60 over years were collected for this study. The study collected. Each isolate was positively identified using morphological, cultural, and biochemical assays as detailed in the reference. The P. granatum peels were air-dried and powdered. Then 25g were extracted using 500 mL of water and ethanol on Soxhlet equipment for 72 hours. The extracts were then cooled, filtered, and concentrated at 40oC to get the crude extract; it was kept at four degrees centigrade in dark vials until use. The extracts were tested for the presence of alkaloids, tannins, flavonoids, glycosides, as well as steroidal terpenes. The efficacy of antimicrobial effects was calculated using well-diffusion techniques on Muller Hinton Agar (MHA). The plates were injected with a standardized suspension of the test isolates against McFarland tube 0.5. Five wells, each measuring five millimeters in diameter, were evenly spaced out using a sterile standard core borer. The well bottoms were sealed with sterile molten nutritional agar to prevent the extract from leaking out from beneath the agar. The aqueous and ethanolic crude extracts dissolved in DMSO served as positive controls, while sterile water and 10% DMSO served as negative controls. Each extract was diluted to a final concentration of 50, 100, or 200mg/ml, and 25 ml was added to the appropriate well on the infected plate. The plates were then incubated for 24 hours at 37 degrees Celsius. A millimeter-calibrated ruler was used to measure the size of the resultant inhibitory zones. The zone of inhibition of the test microorganisms at that dose was calculated as the mean of three measurements. Results: Clinical isolates of E. coli and S. aureus were inhibited by pomegranate extracts at a concentration of 200mg/ml compared to other concentrations, and this extract concentration showed a non-significant difference with chloramphenicol (P&lt;0.01). The study revealed that pomegranate peel extract significantly reduced E. coli levels in feces and increased survival rates in rats. On the first day, E. coli concentrations were much higher in the control group (G2) compared to the treatment group (G3). By day 6, all rats in the control group had died, while all rats in the treatment group survived. Pomegranate peel extract shows notable antibacterial properties, impacting bacterial membrane permeability and cell survival. The variation in extract composition affects its efficacy. Conclusion, Pomegranate peel extract significantly reduced E. coli levels and improved survival rates in rats. On day 6, all rats in the control group died, while all in the treatment group survived. The extract's antibacterial effects and impact on bacterial membranes highlight its potential as a therapeutic agent.

Modulation of fecal microbiota and reductions in fecal antibiotic resistance genes (ARGs) driven by Weissella-fermented feed in growing pigs

Probiotics-induced feed fermentation can improve the composition of microbiota, leading to benefits in pig production. However, the influence of probiotics-driven feed fermentation on pollution reduction is limited. This study aimed to analyze the impact of Weissella-based feed fermentation on the chemical characteristics, changes in microbial abundance, and antibiotic resistance genes (ARGs). Moreover, the possible mechanism and the association among them was also analyzed. First, pigs reared on fermented feed exhibited improved growth performance. The fermentation group showed a significant reduction in emissions of total phosphorus (TP), total carbon (TC), organic matter (OM), copper (Cu), and zinc (Zn) levels in feces compared to the control group. The fermentation group also showed a significant decrease in the ARGs, especially for the tetX, tetW, tetQ, tetL, tetO, tet32, tet44, ermG, ermF, CfxA2, CfxA3, aph3-III, aadA, and ant9-I, compared to the control group. The primary functional microbiota, characterized by increased levels of Bifidobacterium, Megasphaera, and Mitsuokella, and decreased levels of Methanosphaera, and Ruminiclostridium, displayed both negative and positive correlations with ARGs, TC, TP, OM, Cu, and Zn. Furthermore, a significant association was observed between the alterations in microbiota and ARGs and the lactic acid concentration in the fermented feed. The molecular docking results showed a good fit between lactate dehydrogenase and three antibiotics, particularly tetracycline. In conclusion, these results offer novel targets and strategies to address environmental pollutants associated with pig farming.

Zonulin — effective biomarker for the diagnosis of intestinal lesions in patients with nonalcoholic fatty liver disease in COVID‑19

The SARS‑CoV‑2 virus mainly attacks the respiratory system, but can also affect several organs, in particular, causing gastrointestinal symptoms such as vomiting, diarrhea or abdominal pain in the early stages of the disease. Intestinal dysfunction leads to a change in intestinal microorganisms and an increase in the content of inflammatory cytokines. Liver dysfunction is also relatively common in patients with coronavirus disease 2019 (COVID19). Objective — to investigate the efficacy of zonulin in determining intestinal damage in patients with nonalcoholic fatty liver disease with COVID‑19. Materials and methods. The study included 76 patients with non‑alcoholic fatty liver disease (NAFLD) who had complaints indicating intestinal disorders during the outpatient stage of follow‑up after ­COVID‑19. From the anamnesis, it was found that all examined patients had a confirmed diagnosis of infection with SARS‑CoV‑2 virus. All patients had a mostly mild course of COVID‑19, which did not require hospitalization. Patients with NAFLD were divided into two groups depending on defecation violation: group I — 36 patients who, after an acute respiratory infection caused by the SARS‑CoV‑2 virus, developed periodic diarrhea (on average, in (38.7±5.2) days after a negative PCR result for SARS‑CoV‑2 virus); group II — 40 patients with constipation, which began to bother on average in (86.9±7.4) days after a negative PCR result. The control group included 20 practically healthy individuals. The level of zonulin was determined in blood serum and faeces by enzyme‑linked immunosorbent assay. Patients underwent faecal culture to assess the quantitative and qualitative composition of the colon microflora. Results. During the anthropometric examination, overweight and first‑degree obesity were detected more frequently in patients of both groups: overweight was diagnosed most frequently in patients of group I (38.9%) and first‑degree obesity in patients of group II (35.0%). The microbiological examination of faeces in both groups of patients revealed a decrease in the number of bifidus and lactobacilli, as well as Enterococcus and E. coli with normal enzymatic properties. In group I patients, Citrobacter and Staphylococcus were statistically significantly (p &lt;0.05) more often detected in increased concentrations in faeces, whereas in group II patients Enterobacter, Klebsiella and Clostridium were detected (p &lt;0.05). Significant changes were found in the analysis of zonulin levels in both serum and faeces in patients with NAFLD after COVID‑19. Serum zonulin levels in patients of group I increased 6.5‑fold, and in patients of group II—8.6‑fold (p &lt;0.001). An increase in the concentration of zonulin in the faeces was also detected — by 7.5 and 9.7 times, respectively (p &lt;0.001). A correlation was established between the level of zonulin in both blood serum and faeces and a decrease in the number of bifidus and lactobacilli in faeces in patients of both groups. Also, a strong correlation was found between the number of Enterobacter, Klebsiella and Clostridium and a medium intensity correlation between the number of Staphylococcus in the faeces in patients of group ІІ. In patients of group І, a correlation was found between the number of Staphylococcus and Citrobacter. The level of zonulin (both in blood serum and faeces) directly depended on changes in body mass index in the patients we examined, namely, overweight in patients of group І and obesity of the first degree in patients of group ІІ. Conclusions. In patients with NAFLD after COVID19, changes in the quantitative and qualitative composition of the colon microflora were found in the intestinal lesions, including a decrease in the number of bifidobacteria and lactobacilli, as well as an increase in the content of Staphylococcus and Citrobacter in the faeces in patients with predominant diarrhoea and Enterobacter, Klebsiella and Clostridium in patients with predominant constipation. An increase in the level of zonulin in the blood serum and faeces in patients with NAFLD after ­COVID‑19 was detected. A more pronounced deviation from the norm was found in patients of group ІІ (with complaints of constipation). A direct correlation was established between the level of zonulin in the blood serum and faeces and changes in the quantitative and qualitative composition of the colon microflora, as well as an increase in body mass index in patients with NAFLD after ­COVID‑19.

Gut microbiota metabolite trimethylamine N-oxide promoted NAFLD progression by exacerbating intestinal barrier disruption and intrahepatic cellular imbalance

Non-alcoholic fatty liver disease (NAFLD) has become the most common chronic liver disease worldwide, with the gut microbiota and its metabolites are important regulators of its progression. Trimethylamine N-oxide (TMAO), a metabolite of the gut microbiota, has been closely associated with various metabolic diseases, but its relationship with NAFLD remains to be elucidated. In this study, we found that fecal TMAO levels correlated with NAFLD severity. Moreover, TMAO promoted lipid deposition in HepG2 fatty liver cells and exacerbated hepatic steatosis in NAFLD rats. In the colon, TMAO undermined the structure and function of the intestinal barrier at various levels, further activated the TLR4/MyD88/NF-κB pathway, and inhibited the WNT/β-catenin pathway. In the liver, TMAO induced endothelial dysfunction with capillarization of liver sinusoidal endothelial cells, while modulating macrophage polarization. In conclusion, our study suggests that gut microbiota metabolite TMAO promotes NAFLD progression by impairing the gut and liver and that targeting TMAO could be an alternative therapeutic strategy for NAFLD.

Unique sterol metabolite shifts in inflammatory bowel disease and primary sclerosing cholangitis

Inflammatory bowel disease (IBD) triggers chronic intestinal inflammation and is linked to primary sclerosing cholangitis (PSC). Cholesterol homeostasis, tightly regulated under normal conditions, becomes disrupted in both inflammation and chronic liver disease. We analyzed fecal and serum levels of cholesterol synthesis precursors, oxysterols, and phytosterols in 87 patients with IBD (81 for serum analysis) including patients with Crohn's disease (CD) and ulcerative colitis (UC), 11 patients with PSC, 21 patients with PSC-IBD (18 for serum analysis), and 16 healthy controls (17 for serum analysis). Cholesterol was analysed by flow injection analysis on a high-resolution hybrid quadrupole-Orbitrap mass spectrometer and further serum sterols and all fecal sterols were analysed by a gas chromatograph mass spectrometer. Serum levels of lanosterol, 7-dehydrocholesterol, 7-beta-hydroxycholesterol, 27-hydroxycholesterol, and the plant sterols campesterol, stigmasterol, and sitosterol were similar across control and patient groups. Notably, serum lathosterol was elevated in CD patients compared to those with UC, PSC, PSC-IBD, and healthy controls. All other serum and fecal sterols showed no differences between CD and UC. Cholesterol synthesis precursors in serum, serum cholesterol levels, and both serum and fecal plant sterol levels decreased with increasing IBD severity. Consequently, serum cholesterol, campesterol, sitosterol, and fecal 5-beta sitostanol and 5-alpha sitostanol were negatively correlated with C-reactive protein and fecal calprotectin. The conversion of cholesterol to coprostanol in feces was impaired in IBD, PSC, and PSC-IBD, independent of bowel inflammation severity or liver disease extent. Patients with PSC, and to a lesser extent PSC-IBD, had elevated serum plant sterol levels, positively correlating with liver disease markers. In conclusion, in patients with IBD, cholesterol biosynthetic precursors, serum cholesterol levels, and fecal plant sterols decrease with intestinal inflammation. An inverse association of serum plant sterols with intestinal inflammation was observed in patients with IBD and a direct association of serum phytosterols with liver injury in patients with PSC. The conversion of fecal cholesterol to coprostanol was impaired in all patient cohorts. IBD and PSC alter serum sterol levels differently, whereas changes in fecal sterols are not disease specific and are moderate.

The extracellular vesicle of depressive patient-derived Escherichia fergusonii induces vagus nerve-mediated neuroinflammation in mice

BackgroundGut microbiota dysbiosis is closely associated with psychiatric disorders such as depression and anxiety (DA). In our preliminary study, fecal microbiota transplantation from volunteers with psychological stress and subclinical symptoms of depression (Vsd) induced DA-like behaviors in mice. Escherichia fergusonii (Esf) was found to be more abundant in the feces of Vsd compared to healthy volunteers. Therefore, we investigated the effect of Esf on DA-like behavior and neuroinflammation in mice with and without celiac vagotomy.Methods and resultsOrally gavaged Esf increased DA-like behaviors, tumor necrosis factor (TNF)-α, and toll-like receptor-4 (TLR4) expression, and NF-κB+Iba1+ and lipopolysaccharide (LPS)+Iba1+ cell populations, while decreasing serotonin, 5-HT1A receptor, and brain-derived neurotrophic factor (BDNF) expression in the hippocampus and prefrontal cortex. However, celiac vagotomy attenuated Esf-induced DA-like behavior and neuroinflammation. Orally gavaged extracellular vesicle (EV) from Vsd feces (vfEV) or Esf culture (esEV) induced DA-like behavior and inflammation in hippocampus, prefrontal cortex and colon. However, celiac vagotomy attenuated vfEV- or esEV-induced DA-like behaviors and inflammation in the brain alone, while vfEV- or esEV-induced blood LPS and TNF-α levels, colonic TNF-α expression and NF-κB-positive cell number, and fecal LPS level were not. Although orally gavaged fluorescence isothiocyanate-labeled esEV was translocated into the blood and hippocampus, celiac vagotomy decreased its translocation into the hippocampus alone.ConclusionsesEVs may be translocated into the brain via the vagus nerve and bloodstream, subsequently inducing TNF-α expression and suppressing serotonin, its receptor, and BDNF expression through the activation of TLR4-mediated NF-κB signaling, thereby contributing to DA pathogenesis.