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5 pages, 547 KB  
Case Report
Recurrent Hepatic Encephalopathy After Abdominal Surgery in a Non-Cirrhotic Patient: A Case Report
by Sebastiano Ziola, Giuseppe Cittadini, Andrea Pasta, Sara Labanca, Giulia Pieri, Simona Marenco and Edoardo G. Giannini
Reports 2026, 9(2), 112; https://doi.org/10.3390/reports9020112 - 4 Apr 2026
Viewed by 337
Abstract
Background and Clinical Significance: Hepatic encephalopathy (HE) is a reversible brain dysfunction typically associated with cirrhosis and portal hypertension. In these patients, portosystemic shunts allow ammonia and other toxins to bypass hepatic metabolism, leading to neurological symptoms. However, HE can also occur in [...] Read more.
Background and Clinical Significance: Hepatic encephalopathy (HE) is a reversible brain dysfunction typically associated with cirrhosis and portal hypertension. In these patients, portosystemic shunts allow ammonia and other toxins to bypass hepatic metabolism, leading to neurological symptoms. However, HE can also occur in non-cirrhotic patients through congenital shunts or, less commonly, through iatrogenic shunts following abdominal trauma or surgery. This case is clinically significant as it illustrates a rare presentation of recurrent HE caused by a de novo portosystemic shunt following major abdominal surgery in a patient without underlying liver disease. Case Presentation: A 76-year-old male was admitted with confusion, lethargy, and flapping tremors. His medical history included a total pancreatectomy for pancreatic adenocarcinoma six months prior. Laboratory tests revealed hyperammonemia and altered liver enzymes likely related to ongoing chemotherapy, but no signs of hepatic insufficiency or cirrhosis. A review of recent CT imaging identified a new portosystemic shunt between the portal territory and the azygous vein that was absent prior to his pancreatectomy. This iatrogenic shunt likely formed via the re-vascularization of vestigial vessels following surgical de-vascularization. The patient was successfully managed with lactulose and rifaximin. At 3-month follow-up, no further HE episodes had occurred. Conclusions: This case highlights that HE should be considered in patients without cirrhosis presenting with altered mental status and hyperammonemia, especially following abdominal surgery. It underscores the importance of a multidisciplinary approach and meticulous re-evaluation of imaging to identify iatrogenic vascular shunts that may be amenable to medical or interventional management. Full article
(This article belongs to the Section Gastroenterology)
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16 pages, 1649 KB  
Article
Standardizing Intestinal Permeability Assessment: Optimization of Gluten Dose and Urine Collection Times for u-GIP and Lactulose:Mannitol Ratio in Healthy Volunteers
by Raquel Rodríguez-Ramírez, María Auxiliadora Fernández Peralbo, Ángel Cebolla and Carolina Sousa
Int. J. Mol. Sci. 2026, 27(5), 2286; https://doi.org/10.3390/ijms27052286 - 28 Feb 2026
Viewed by 410
Abstract
Urinary gluten immunogenic peptides (u-GIPs) have been proposed as a complementary marker to classical intestinal permeability tests based on lactulose, mannitol, and the lactulose:mannitol ratio (LMR). However, the effects of gluten dose, urine collection interval, and sampling strategy on their performance remain insufficiently [...] Read more.
Urinary gluten immunogenic peptides (u-GIPs) have been proposed as a complementary marker to classical intestinal permeability tests based on lactulose, mannitol, and the lactulose:mannitol ratio (LMR). However, the effects of gluten dose, urine collection interval, and sampling strategy on their performance remain insufficiently defined. This study evaluated these variables to support protocol standardization. Data from four standardized protocols including 46 healthy adults exposed to 0, 2, 4, or 10 g of gluten were analyzed. All participants ingested fixed doses of lactulose and mannitol. Urine was collected cumulatively (0–6 h and 0–15 h) or by individual voids. u-GIP levels were measured by lateral-flow immunoassay, and lactulose and mannitol by ion chromatography. u-GIP excretion showed a clear dose dependence. Lactulose excretion increased transiently only at the 10 g dose during the 0–6 h interval, while mannitol excretion and LMR were unaffected. The u-GIP excretion index showed linear proportionality at the 2 g and 4 g doses but exhibited saturation kinetics at the 10 g dose. The 4 g dose showed the lowest interindividual variability. Sampling strategies yielded equivalent results. A 4 g gluten challenge combined with a 6 h urine collection demonstrated effectiveness in healthy volunteers and may be suitable for clinical application. Further research involving larger cohorts of both healthy individuals and patients with intestinal hyperpermeability is required to validate this method. Full article
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18 pages, 491 KB  
Article
Relationship Between Diet Quality, Intestinal Permeability, and Gut Microbiota Features in Individuals with Obesity
by Sarah M. Eaton, Weiwen Chai, Olivia Moss, Edward C. Deehan, Victoria Texieira Reis, Ali Keshavarzian and Heather E. Rasmussen
Nutrients 2026, 18(5), 775; https://doi.org/10.3390/nu18050775 - 27 Feb 2026
Viewed by 662
Abstract
Background/Objectives: This study examined relationships between diet quality, as determined using three a priori-defined dietary patterns (Healthy Eating Index of 2010 dietary guidelines [HEI-2010], Mediterranean Dietary Pattern [MDP], and Dietary Approaches to Stop Hypertension [DASH]), intestinal permeability, and features of the gut microbiota [...] Read more.
Background/Objectives: This study examined relationships between diet quality, as determined using three a priori-defined dietary patterns (Healthy Eating Index of 2010 dietary guidelines [HEI-2010], Mediterranean Dietary Pattern [MDP], and Dietary Approaches to Stop Hypertension [DASH]), intestinal permeability, and features of the gut microbiota in a diverse, obese sample. Methods: This was a post hoc, cross-sectional study including 103 healthy, obese individuals (43.8 ± 11.3 years, BMI: 37.5 ± 6.1 kg/m2, 64.1% African American). Dietary intake was assessed using the Vioscreen food frequency questionnaire. Intestinal permeability was assessed via urinary sugar excretion and microbiota features were characterized using 16S rRNA gene amplicon sequencing. Relationships between dietary pattern adherence, intestinal permeability, and gut microbiota were assessed using correlation coefficients and a general linear model. Results: Higher dietary pattern scores correlated with lower levels of intestinal permeability measures such as 24 h urinary sucralose (HEI-2010: r = −0.33, p = 0.002; MDP: r = −0.31, p = 0.004; DASH: r = −0.38, p < 0.0001) and 24 h sucralose-to-lactulose ratio (HEI-2010: r = −0.23, p = 0.03; MDP: r = −0.32, p = 0.003; DASH: r = −0.24, p = 0.03). Fruit intake consistently correlated with lower intestinal permeability measures (p < 0.05) across all three dietary patterns. Higher DASH scores correlated with lower Proteobacteria (r = −0.28, p = 0.004) and higher Verrucomicrobia (r = 0.30, p = 0.002) phylum abundance. Conclusions: The current results suggest a potential role for diet quality in promoting intestinal health. Full article
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12 pages, 968 KB  
Review
Possible Crosstalk Between Small Intestinal Bacterial Overgrowth (SIBO) and Atopic Manifestations—A Short Overview
by Michał Terlecki, Wiktoria Brzeczek, Martyna Kowalczyk, Emilia Kiełczyńska, Klaudia Kukla, Gabriela Osmulska and Krzysztof Gomułka
Int. J. Mol. Sci. 2026, 27(4), 1865; https://doi.org/10.3390/ijms27041865 - 15 Feb 2026
Viewed by 1335
Abstract
Small intestinal bacterial overgrowth (SIBO) is an increasingly recognized condition that influences immune responses. It may be linked to atopic disorders such as bronchial asthma (BA), food allergies (FA), chronic spontaneous urticaria (CSU), and mast cell activation syndrome (MCAS). The aim of our [...] Read more.
Small intestinal bacterial overgrowth (SIBO) is an increasingly recognized condition that influences immune responses. It may be linked to atopic disorders such as bronchial asthma (BA), food allergies (FA), chronic spontaneous urticaria (CSU), and mast cell activation syndrome (MCAS). The aim of our study was to perform a structured literature search to assess the possible correlation between SIBO and the presentation of atopic disorders. The prevalence of SIBO was highest in patients with BA (60–100%) and FA (50–87.5%), followed by MCAS (30.9%) and CSU (27.9%). The diagnosis of SIBO was based on lactulose or glucose breath tests. SIBO exacerbated symptoms of atopic diseases, and treating it within BA and MCAS improved the symptoms, in contrast to CSU. The present evidence suggests a possible crosslink between SIBO and atopic manifestations. Bacterial overgrowth appears to trigger the Th2 immune response via the mucosal pathway and low-grade endotoxemia. These result in the increased synthesis of interleukins involved in allergic reactions (IL-4, IL-5, IL-13). Further studies are essential to confirm the clinical significance of this association. The “gut–allergy axis” may offer new therapeutic options and possibly improve quality of life in patients with atopy. Full article
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14 pages, 24295 KB  
Article
Rational Engineering of Cellobiose 2-Epimerase Through Flexible Loop Modulation and Structure-Guided Sequence Alignment for Enhanced Lactulose Synthesis
by Xinyan Mao, Hongbin Zhang, Chao Hu, Chunhui Ma, Xueqin Hu and Jingwen Yang
Biomolecules 2026, 16(2), 206; https://doi.org/10.3390/biom16020206 - 28 Jan 2026
Viewed by 501
Abstract
Lactulose, a valuable functional disaccharide with pharmaceutical and food applications, is efficiently synthesized via enzymatic isomerization of lactose. This study developed an integrated strategy combining protein engineering of cellobiose 2-epimerase (CsCE) from Caldicellulosiruptor saccharolyticus and process optimization to enhance lactulose production. A dual-track [...] Read more.
Lactulose, a valuable functional disaccharide with pharmaceutical and food applications, is efficiently synthesized via enzymatic isomerization of lactose. This study developed an integrated strategy combining protein engineering of cellobiose 2-epimerase (CsCE) from Caldicellulosiruptor saccharolyticus and process optimization to enhance lactulose production. A dual-track engineering approach—incorporating flexible loop modulation (residues 161–193) and structure-guided sequence alignment with N-acetyl-D-glucosamine-2-epimerase—enabled the creation of two superior mutants, R17Q/L184S and R17Q/S142T. The R17Q/L184S variant exhibited a 37% increase in crude enzyme activity, improved thermostability (half-life of 200 min at 80 °C), and enhanced substrate affinity (Km reduced by 23.2%). R17Q/S142T achieved a 21% higher specific activity (24.08 U/mg), the highest among all variants. Structural and molecular dynamics analyses revealed that L184S enriched hydrogen bonding and hydrophobic interactions, improving structural rigidity, while S142T introduced allosteric regulation that facilitated catalytic efficiency. Under optimized conditions (70 °C, pH 7.5, 40% lactose, 20 U/mL enzyme, 3 h), lactulose yield reached 75.6% with >95% purity. This work demonstrates the successful application of synergistic enzyme engineering and process intensification for high-efficiency lactulose biosynthesis, providing viable candidates and system solutions for industrial-scale production. Full article
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15 pages, 1037 KB  
Article
Diagnostic Evaluation of Small Intestinal Microbial Overgrowth: A Cross-Sectional Comparison of Glucose and Lactulose Breath Tests
by Giulia Scalese, Luca Spina, Lucia Gallucci, Alessandra Cesarini, Emanuela Ribichini, Maddalena Diofebi, Ivan Tattoli, Lucia Pallotta, Anna Citarella and Carola Severi
J. Clin. Med. 2025, 14(24), 8920; https://doi.org/10.3390/jcm14248920 - 17 Dec 2025
Viewed by 2125
Abstract
Background/Objectives: Small intestinal microbial overgrowth (SIMO), including both small intestinal bacterial overgrowth (SIBO) and intestinal methanogen overgrowth (IMO), is commonly diagnosed using non-invasive breath tests, whose diagnostic performance and criteria remain inconsistent. This study aimed to assess SIMO prevalence using lactulose (LBT) [...] Read more.
Background/Objectives: Small intestinal microbial overgrowth (SIMO), including both small intestinal bacterial overgrowth (SIBO) and intestinal methanogen overgrowth (IMO), is commonly diagnosed using non-invasive breath tests, whose diagnostic performance and criteria remain inconsistent. This study aimed to assess SIMO prevalence using lactulose (LBT) and glucose breath tests (GBT), compare their diagnostic yields for SIBO and IMO, analyze associated gas profiles, clinical features, risk factors, and evaluate the diagnostic accuracy of a simplified fasting methane criterion for IMO. Methods: Cross-sectional study conducted on 564 outpatients (75.7% female) with suspected SIMO. Patients underwent LBT (n = 275), GBT (n = 289), or both (n = 47). Results: SIMO was diagnosed in 26.8% of patients. LBT identified significantly more SIMO than GBT (37.5% vs. 16.6%, p < 0.01), particularly for SIBO (24.4% vs. 4.8%, p < 0.01), while IMO detection was comparable (9.8% vs. 10.7%). Mixed overgrowth (dual SIBO/IMO positivity) showed a borderline trend favoring LBT. Methane peaks occurred significantly earlier than hydrogen in both BTs. Clinical symptoms did not significantly differ between SIMO subtypes or between test-positive and test-negative groups. The simplified fasting methane criterion showed limited diagnostic accuracy for IMO making it inadequate as a standalone diagnostic tool, requiring further validation before clinical implementation. Conclusions: GBT is the more reliable test for SIMO diagnosis due to LBT’s lower specificity. Clinical symptoms alone were not predictive of SIMO subtypes, while the different gas profile suggests a distinct spatial distribution of microbial populations with a higher proximal concentration of methanogenic Archaea. Full article
(This article belongs to the Section Gastroenterology & Hepatopancreatobiliary Medicine)
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14 pages, 1601 KB  
Article
Regional Gastrointestinal Permeability Patterns in Juvenile Idiopathic Arthritis: A Window into Subclinical Inflammation and Microbiota-Driven Disease Mechanisms
by Francesco La Torre, Francesca Marasciulo, Giovanni La Grasta, Vanessa Nadia Dargenio, Stefania Paola Castellaneta, Silvia Amati, Violetta Mastrorilli, Antonella Sisto, Fabio Cardinale, Ruggiero Francavilla and Fernanda Cristofori
Children 2025, 12(12), 1663; https://doi.org/10.3390/children12121663 - 8 Dec 2025
Viewed by 425
Abstract
Objectives: To assess gastrointestinal permeability (GP) in children with Juvenile Idiopathic Arthritis (JIA) using a segment-specific sugar probe approach to assess gastric, small intestinal, and colonic permeability, and to determine whether GP alterations are associated with disease activity. Methods: This prospective study included [...] Read more.
Objectives: To assess gastrointestinal permeability (GP) in children with Juvenile Idiopathic Arthritis (JIA) using a segment-specific sugar probe approach to assess gastric, small intestinal, and colonic permeability, and to determine whether GP alterations are associated with disease activity. Methods: This prospective study included 30 children with JIA and 22 healthy controls who underwent a validated multi-sugar absorption test. Urinary excretion of sucrose, lactulose, mannitol, and sucralose was measured to evaluate gastric, small intestinal, and colonic permeability. All JIA patients had discontinued immunosuppressive therapy for at least three months before testing. None had a relapse of the disease. Disease activity was assessed using the Juvenile Arthritis Disease Activity Score (JADAS10). Comparisons were conducted between patients and controls and between remission and active disease groups. Results: None of the participants reported gastrointestinal manifestations. The lactulose/mannitol (LA/MA) ratio, a global index of small intestinal permeability, showed no significant difference between JIA patients and controls, suggesting preserved overall barrier function. However, urinary excretion of lactulose, mannitol, and sucralose was significantly higher in JIA patients, while sucrose excretion was significantly lower, indicating segment-specific alterations in small intestinal, colonic, and gastric permeability. These abnormalities were consistently present, even in patients in clinical remission. No statistically significant differences were observed between remission and active disease groups, though a trend toward increased permeability was noted in the latter. Conclusions: Children with JIA exhibit segmental GP alterations that persist independently of clinical disease activity. Despite the relatively small population, this exploratory study suggests subclinical mucosal dysfunction and the need for further investigation into how the gut–joint axis may be playing a role in JIA pathogenesis, including via intestinal microbiota. Full article
(This article belongs to the Special Issue Diagnosis, Treatment and Care of Pediatric Rheumatology: 2nd Edition)
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17 pages, 580 KB  
Article
Ten-Year Atherosclerotic Cardiovascular Disease Risk in Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD): Separate Analyses from Romanian and Italian Cohorts Integrating Metabolic, Hepatic, and Gut–Liver Axis Markers
by Naomi-Adina Ciurea, Cristina Monica Pantea, Paul Grama, Irina-Bianca Kosovski, Ilaria Farella, Simona Bataga, Agostino Di Ciaula and Piero Portincasa
J. Clin. Med. 2025, 14(23), 8361; https://doi.org/10.3390/jcm14238361 - 25 Nov 2025
Viewed by 1723
Abstract
Background/Objectives: Metabolic dysfunction–associated steatotic liver disease (MASLD) is closely linked to atherosclerotic cardiovascular disease (ASCVD), but the prognostic value of liver fibrosis and gut–liver axis alterations remains uncertain. Methods: We conducted a prospective, observational study in two tertiary centers (in Romania [...] Read more.
Background/Objectives: Metabolic dysfunction–associated steatotic liver disease (MASLD) is closely linked to atherosclerotic cardiovascular disease (ASCVD), but the prognostic value of liver fibrosis and gut–liver axis alterations remains uncertain. Methods: We conducted a prospective, observational study in two tertiary centers (in Romania and Italy) and compared the outcomes with different tests available for fibrosis (FibroTest in Romania or acoustic radiation force impulse (ARFI) elastography in Italy) and intestinal permeability (IP) (by fecal zonulin in Romania or lactulose/mannitol ratio in Italy). Liver steatosis was confirmed at ultrasonography. Analyses followed a within-cohort strategy. Ten-year ASCVD categories were summarized separately per cohort, and within-cohort associations with elevated ASCVD risk (≥7.5%) were explored using univariate logistic regression with age-adjusted two-parameter checks. A pooled robustness analysis (n = 132) was then performed using multivariable logistic regression models for intermediate–high ASCVD risk (≥7.5%), adjusted for age (per 5 years), waist circumference (per 5 cm), total cholesterol (per 10 mg/dL), diabetes, and hypertension. A higher threshold (≥20%) yielded the same qualitative interpretation. Results: ASCVD was computable for 52 Romanian (low 78.8%, borderline 5.8%, intermediate 7.7%, high 7.7%) and 80 Italian participants (low 80.0%, borderline 6.2%, intermediate 12.5%, high 1.2%). In both cohorts, age was associated with higher ASCVD. Fibrosis severity (FibroTest or ARFI) and IP (zonulin or LA/MA) showed no associations with ASCVD. In pooled adjusted models, neither significant fibrosis nor high intestinal permeability was independently associated with ASCVD, whereas age and cardiometabolic comorbidities remained the dominant correlates. Conclusions: Across both cohorts, 10-year ASCVD risk was mainly determined by age and major cardiometabolic comorbidities. Neither liver fibrosis nor intestinal permeability contributed additional prognostic value in this setting, regardless of the assessment method. These data support prioritizing aggressive metabolic risk management and call for harmonized, longitudinal studies to clarify gut–liver contributions to cardiovascular outcomes. Full article
(This article belongs to the Section Gastroenterology & Hepatopancreatobiliary Medicine)
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16 pages, 1968 KB  
Article
Effect of Different Prebiotic Saccharides on Listeria monocytogenes Adherence to Human Adenocarcinoma Caco-2 Cell Line
by Tereza Kodešová, Ivo Doskočil, Eva Vlková and Hana Šubrtová Salmonová
Curr. Issues Mol. Biol. 2025, 47(11), 891; https://doi.org/10.3390/cimb47110891 - 28 Oct 2025
Viewed by 670
Abstract
Listeria monocytogenes (LM) is one of the most emerging pathogens responsible for the serious foodborne disease listeriosis. The risk of disease outbreaks can be reduced by suppressing the adherence of LM to the intestinal epithelial cells. This effect can be achieved by prebiotic [...] Read more.
Listeria monocytogenes (LM) is one of the most emerging pathogens responsible for the serious foodborne disease listeriosis. The risk of disease outbreaks can be reduced by suppressing the adherence of LM to the intestinal epithelial cells. This effect can be achieved by prebiotic supplementation. The aim of this work was to determine the effect of prebiotics beta-(1,3)-D-glucan, inulin, fructooligosaccharides, galactooligosaccharides, lactulose, raffinose, stachyose, human milk oligosaccharides (HMOs), and 2’-fucosyllactose on the ability of LM to adhere to the human adenocarcinoma Caco-2 cell line. Despite strain-specific variability, a statistically significant reduction in LM adhesion to intestinal epithelial cells was observed in the presence of beta-(1,3)-D-glucan (~60% reduction), inulin (~46%), and HMOs (~44%). In contrast, the remaining tested prebiotics did not show a significant impact on LM adhesion. These findings highlight the potential of specific prebiotics, especially beta-glucans, to limit LM adherence, suggesting a protective effect for the host. Full article
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15 pages, 1244 KB  
Article
Lactulose, Rifaximin, and Survival in Hepatic Encephalopathy: A Cohort Study of 120 Patients
by Luana Alexandrescu, Ionut Tiberiu Tofolean, Cristina Aftenie, Daria Maria Alexandrescu, Doina Ecaterina Tofolean, Alina Doina Nicoara, Alina Mihaela Stanigut, Andreea Nelson Twakor, Elena Rusu, Eugen Dumitru, Andrei Dumitru, Cristina Tocia, Alexandra Herlo, Miruna Alexa Mihu, Ioana Popescu, Elena Dina and Bogdan Cimpineanu
J. Clin. Med. 2025, 14(20), 7331; https://doi.org/10.3390/jcm14207331 - 17 Oct 2025
Viewed by 3471
Abstract
Background: Hepatic encephalopathy (HE) is a severe neuropsychiatric complication of advanced liver disease, driven primarily by ammonia accumulation due to impaired hepatic detoxification and portosystemic shunting. Lactulose is a cornerstone therapy, while rifaximin serves as an effective adjunct for reducing recurrence and hospitalizations. [...] Read more.
Background: Hepatic encephalopathy (HE) is a severe neuropsychiatric complication of advanced liver disease, driven primarily by ammonia accumulation due to impaired hepatic detoxification and portosystemic shunting. Lactulose is a cornerstone therapy, while rifaximin serves as an effective adjunct for reducing recurrence and hospitalizations. Methods: We conducted a retrospective cohort study at Constanța Emergency County Hospital from January 2022 to March 2025, including 120 adult patients diagnosed with HE. Inclusion criteria were confirmed diagnosis of cirrhosis with clinical and/or biochemical evidence of HE. Patients with other primary neurological disorders or incomplete medical records were excluded. Data on demographics, comorbidities, laboratory results, and medications were collected. Statistical analyses were performed employing descriptive statistics, Friedman’s two-way ANOVA by ranks for medication use, and Cox proportional hazards regression to assess survival associations. Results: The mean age was 61.19 years, with high prevalence of anemia (mean hemoglobin: 9.35 g/dL) and thrombocytopenia (mean: 121.86 × 103/µL). Inflammatory markers were elevated (mean CRP: 36.95 mg/L; ESR: 55.83 mm/h), and INR averaged 1.86. Lactulose was administered to 63.3% of patients, rifaximin to 52.5%, with diuretics, pantoprazole, and albumin being frequently used. Friedman’s analysis ranked lactulose highest in usage frequency. Cox regression indicated no significant short-term survival difference associated with toxic encephalopathy or rifaximin use. Conclusion: In this cohort, lactulose remained the primary treatment for HE, often complemented by supportive pharmacotherapy. While rifaximin use was limited, the overall medication patterns reflected standard practice priorities in HE management. Full article
(This article belongs to the Section Gastroenterology & Hepatopancreatobiliary Medicine)
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19 pages, 5117 KB  
Article
Comparative Genomics and Functional Profiling Reveal Lineage-Specific Metabolic Adaptations in Globally Emerging Fluoroquinolone-Resistant Salmonella Kentucky ST198
by Juned Ahmed, Rachel C. Soltys, Smriti Shringi, Jean Guard, Bradd J. Haley and Devendra H. Shah
Genes 2025, 16(9), 1051; https://doi.org/10.3390/genes16091051 - 8 Sep 2025
Cited by 1 | Viewed by 1184
Abstract
Background: Salmonella Kentucky comprises two major lineages, ST152 and fluoroquinolone-resistant (FluR) ST198, which have diverged genotypically and phenotypically along distinct evolutionary and epidemiological trajectories. ST198 is linked to global human disease, while ST152 is primarily animal-associated in the U.S. We hypothesized [...] Read more.
Background: Salmonella Kentucky comprises two major lineages, ST152 and fluoroquinolone-resistant (FluR) ST198, which have diverged genotypically and phenotypically along distinct evolutionary and epidemiological trajectories. ST198 is linked to global human disease, while ST152 is primarily animal-associated in the U.S. We hypothesized that lineage-specific metabolic adaptations contribute to their differing host associations and pathogenicity. Methods: We performed comparative metabolic profiling of ST198 (n = 3) and ST152 (n = 4) strains across 948 substrates and environmental conditions. Growth assays tested the ability of these lineages and other non-typhoidal Salmonella (NTS) serovars (n = 5) to utilize myo-inositol and lactulose as sole carbon sources. Comparative genomic analyses of 294 ST198, 173 ST152, and 1300 other NTS serovars identified nutrient utilization genes. Results: ST198 exhibited significantly higher respiratory activity and broader metabolic versatility across carbon, nitrogen/sulfur sources, and stress conditions. The canonical iol gene cluster for myo-inositol catabolism was conserved in ST198 but absent in ST152, which nonetheless showed weak growth on myo-inositol, suggesting an alternative metabolic pathway for myo-inositol may exist. We also report for the first time that, despite lineage-specific differences in metabolic efficiency, multiple NTS serovars, including S. Kentucky, can metabolize lactulose, a synthetic disaccharide traditionally associated with beneficial gut microbes. These results suggest the potential existence of a novel lactulose metabolic pathway in NTS. Conclusions: These findings highlight ST198’s metabolic adaptability and reveal novel metabolic capacities in NTS. A mechanistic understanding of nutrient utilization pathways, particularly of myo-inositol and lactulose, will provide novel insights into the mechanisms underlying nutrient metabolism that likely modulate the ecological success and pathogenic potential of NTS in human and animal hosts. Full article
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20 pages, 2357 KB  
Article
Betaine Supplementation Improves 60 km Cycling Time Trial Performance and One-Carbon Metabolism in Cyclists During Recovery
by David C. Nieman, Camila A. Sakaguchi, James C. Williams, Jackie Lawson and Kevin C. Lambirth
Nutrients 2025, 17(17), 2765; https://doi.org/10.3390/nu17172765 - 26 Aug 2025
Viewed by 4768
Abstract
Background/Objectives: This study examined the effects of 2 weeks of betaine versus placebo supplementation (3 g/d) on 60 km cycling performance, gut permeability, and shifts in plasma metabolites. Methods: Participants included 21 male and female non-elite cyclists. A randomized, placebo-controlled, double-blind, crossover design [...] Read more.
Background/Objectives: This study examined the effects of 2 weeks of betaine versus placebo supplementation (3 g/d) on 60 km cycling performance, gut permeability, and shifts in plasma metabolites. Methods: Participants included 21 male and female non-elite cyclists. A randomized, placebo-controlled, double-blind, crossover design was used with two 2-week supplementation periods and a 2-week washout period. Supplementation periods were followed by a 60 km cycling time trial. Six blood samples were collected before and after supplementation (overnight fasted state), and at 0 h, 1.5 h, 3 h, and 24 h post-exercise. Five-hour urine samples were collected pre-supplementation and post-60 km cycling after ingesting a sugar solution containing lactulose 5 g, 13C mannitol 100 mg, and 12C mannitol 1.9 g in 450 mL water. Other outcome measures included plasma intestinal fatty acid binding protein-1 (I-FABP), muscle damage biomarkers (serum creatine kinase, myoglobin), serum cortisol, complete blood cell counts, and shifts in plasma metabolites using untargeted metabolomics. Results: The time to complete the 60 km cycling bout differed significantly between the betaine and placebo trials (mean ± SE, 112.8 ± 2.3, 114.2 ± 2.6 min, respectively, (−1.41 ± 0.7 min) (effect size = 0.475, p = 0.042). No trial differences were found for I-FABP (interaction effect, p = 0.076), L:13CM (p = 0.559), the neutrophil/lymphocyte ratio (p = 0.171), serum cortisol (p = 0.982), serum myoglobin (p = 0.942), or serum creatine kinase (p = 0.694). Untargeted metabolomics showed that 214 metabolites exhibited significant trial treatment effects and 130 significant trial x time interaction effects. Betaine versus placebo supplementation was linked to significant increases in plasma betaine, dimethylglycine (DMG), sarcosine, methionine, S-adenosylhomocysteine (SAH), alpha-ketoglutaramate, and 5′methylthioadensone (MTA), and decreases in plasma carnitine and numerous acylcarnitines. Conclusions: Betaine supplementation modestly improved 60 km cycling performance but had no effect on gut permeability. The metabolomics data supported a strong influence of 2-week intake of betaine on the one-carbon metabolism pathway during the 24 h recovery period. Full article
(This article belongs to the Section Sports Nutrition)
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24 pages, 3673 KB  
Article
Multistrain Probiotics Plus Vitamin D Improve Gut Barrier Function and Gut Microbiota Composition in Irritable Bowel Syndrome Without Constipation: Results from a Double-Blind, Randomized, Placebo-Controlled Trial
by Lucrezia Laterza, Cesare Cremon, Gaetano Coppola, Carlo Romano Settanni, Rossella Maresca, Martina Strazzeri, Eleonora Durini, Valentina Petito, Franco Scaldaferri, Giorgio Gargari, Diego Mora, Elnaz Vojoudi Yazdi, Chiara Marangelo, Gianluca Ianiro, Lorenza Putignani, Maria Raffaella Barbaro, Giovanni Marasco, Giovanni Barbara and Antonio Gasbarrini
Nutrients 2025, 17(10), 1708; https://doi.org/10.3390/nu17101708 - 18 May 2025
Cited by 8 | Viewed by 7155
Abstract
Background: The disruption of the intestinal barrier and the imbalance of the gut microbiota (GM) seem to play a major role in the complex pathogenesis of irritable bowel syndrome (IBS). Specific microbial strains could improve the gut microenvironment, promoting anti-inflammatory pathways; similarly, vitamin [...] Read more.
Background: The disruption of the intestinal barrier and the imbalance of the gut microbiota (GM) seem to play a major role in the complex pathogenesis of irritable bowel syndrome (IBS). Specific microbial strains could improve the gut microenvironment, promoting anti-inflammatory pathways; similarly, vitamin D supplementation could play a role in enhancing the barrier integrity and modulating the immune response in the gut. This study aims to evaluate the efficacy of a new multistrain probiotic, combined with vitamin D, in improving gut barrier function in IBS without constipation. Methods: In this phase IIb double-blind randomized placebo-controlled, parallel-group, multicenter, clinical trial, 35 patients were treated for 12 weeks with OttaBac®, a high concentration multistrain probiotic plus cholecalciferol, or placebo and were followed up until week 16. Symptoms, quality of life, intestinal permeability, fecal biomarkers, and microbiota composition were evaluated at 0, 12, and 16 weeks. Results: Mean zonulin values showed a significant progressive reduction in the active group (−10.2 ng/mL at week 12, p = 0.0375; −19.5 ng/mL at week 16, p = 0.0002), with a significant difference between groups at week 16 in the per-protocol population (−19.01, p = 0.0053). The active group showed a more stable trend toward improvement in stool frequency and consistency at both week 12 and 16, with a significant improvement compared to the baseline and to the placebo group (−23.2, p = 0.0265, and 5.57 vs. −23.2, p = 0.0492, respectively). No differences were found in regards to the lactulose/mannitol ratio, Irritable Bowel Syndrome Severity Scoring System (IBS-SSS) and Short Form Health Survey (SF-36) total scores, plasmalemmal vesicle associated protein-1 (PV-1), and citrulline levels. In the active group, Bifidobacterium animalis subsp. lactis and Streptococcus thermophilus levels were increased (p < 0.05), while those for Lachnospira were decreased (p < 0.05), and significant changes in Actinobacteria and Proteobacteria were observed (p < 0.05). Lactate (p < 0.01) and acetate (p < 0.05) levels increased post-treatment. Correlation analysis pointed out a significant association between the microbial biomarkers and the symptoms (p < 0.05). Conclusions: Probiotic plus vitamin D could improve IBS-associated symptoms through gut microbiota modulation and gut barrier enhancement, with persistent benefits after treatment discontinuation. Full article
(This article belongs to the Section Prebiotics, Probiotics and Postbiotics)
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18 pages, 2566 KB  
Article
Selective Influence of Hemp Fiber Ingestion on Post-Exercise Gut Permeability: A Metabolomics-Based Analysis
by David C. Nieman, Camila A. Sakaguchi, James C. Williams, Wimal Pathmasiri, Blake R. Rushing, Susan McRitchie and Susan J. Sumner
Nutrients 2025, 17(8), 1384; https://doi.org/10.3390/nu17081384 - 19 Apr 2025
Cited by 1 | Viewed by 2137
Abstract
Objectives: This study investigated the effects of 2-week ingestion of hemp fiber (high and low doses) versus placebo bars on gut permeability and plasma metabolite shifts during recovery from 2.25 h intensive cycling. Hemp hull powder is a rich source of two bioactive [...] Read more.
Objectives: This study investigated the effects of 2-week ingestion of hemp fiber (high and low doses) versus placebo bars on gut permeability and plasma metabolite shifts during recovery from 2.25 h intensive cycling. Hemp hull powder is a rich source of two bioactive compounds, N-trans-caffeoyl tyramine (NCT) and N-trans-feruloyl tyramine (NFT), with potential gut health benefits. Methods: The study participants included 23 male and female cyclists. A three-arm randomized, placebo-controlled, double-blind, crossover design was used with two 2-week supplementation periods and 2-week washout periods. Supplement bars provided 20, 5, or 0 g/d of hemp hull powder. Participants engaged in an intensive 2.25 h cycling bout at the end of each of the three supplementation periods. Five blood samples were collected before and after supplementation (overnight fasted state), and at 0 h-, 1.5 h-, and 3 h-post-exercise. Five-hour urine samples were collected pre-supplementation and post-2.25 h cycling after ingesting a sugar solution containing 5 g of lactulose, 100 mg of 13C mannitol, and 1.9 g of mannitol in 450 mL of water. An increase in the post-exercise lactulose/13C mannitol ratio (L:13CM) was used as the primary indicator of altered gut permeability. Other outcome measures included muscle damage biomarkers (serum creatine kinase, myoglobin), serum cortisol, complete blood cell counts, and shifts in plasma metabolites using untargeted metabolomics. Results: No trial differences were found for L:13CM, cortisol, blood cell counts, and muscle damage biomarkers. Orthogonal partial least-squares discriminant analysis (OPLSDA) showed distinct trial differences when comparing high- and low-dose hemp fiber compared to placebo supplementation (R2Y = 0.987 and 0.995, respectively). Variable Importance in Projection (VIP) scores identified several relevant metabolites, including 3-hydroxy-4-methoxybenzoic acid (VIP = 1.9), serotonin (VIP = 1.5), 5-hydroxytryptophan (VIP = 1.4), and 4-methoxycinnamic acid (VIP = 1.4). Mummichog analysis showed significant effects of hemp fiber intake on multiple metabolic pathways, including alpha-linolenic acid, porphyrin, sphingolipid, arginine and proline, tryptophan, and primary bile acid metabolism. Conclusions: Hemp fiber intake during a 2-week supplementation period did not have a significant effect on post-exercise gut permeability in cyclists (2.25 h cycling bout) using urine sugar data. On the contrary, untargeted metabolomics showed that the combination of consuming nutrient-rich hemp fiber bars and exercising for 135 min increased levels of beneficial metabolites, including those derived from the gut in healthy cyclists. Full article
(This article belongs to the Special Issue Sports Nutrition: Current and Novel Insigh (2nd Edition))
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19 pages, 4101 KB  
Article
L-Arabinose Alleviates Functional Constipation in Mice by Regulating Gut Microbiota and Metabolites
by Ke Liu, Huixuan Dong, Xinran Li, Chaoqun Hu, Fengya Cui, Shiji Li, Xiaolin Zhang, Yushan Du, Penghui Yang, Wenna Ji, Wenjie Sui and Jing Meng
Foods 2025, 14(5), 900; https://doi.org/10.3390/foods14050900 - 6 Mar 2025
Cited by 4 | Viewed by 3011
Abstract
Functional constipation ranks among the most common disorders impacting human health, which is manifested by difficulty in defecation and a complex etiology. L-Arabinose, a pentose found naturally in fruit rinds and cereal husks, has been reported to regulate glycolipid metabolism, improve glucose homeostasis, [...] Read more.
Functional constipation ranks among the most common disorders impacting human health, which is manifested by difficulty in defecation and a complex etiology. L-Arabinose, a pentose found naturally in fruit rinds and cereal husks, has been reported to regulate glycolipid metabolism, improve glucose homeostasis, and exhibit anti-inflammatory effects. However, the effect and precise mechanism of L-Arabinose on functional constipation remain unclear. In this study, the effect of L-Arabinose in alleviating functional constipation induced by diphenoxylate was evaluated. The model group consisted of functional constipation mice that did not receive any intervention. The positive drug group was treated with 2.0 g/kg lactulose, while the intervention group was given 0.5 g/kg, 0.75 g/kg, 1.0 g/kg, and 2.0 g/kg L-Arabinose, respectively. The data suggested that 20 days of L-Arabinose intervention could shorten the first black stool defecation time, increase fecal water content, and enhance the rate of small intestinal propulsion in mice with functional constipation induced by diphenoxylate. Additionally, L-Arabinose reversed the protein expression of functional constipation-related intestinal factors in the colon, characterized by a decrease in the expression of water channel proteins AQP3 and AQP4, as well as an increase in the expression of tight-junction proteins ZO-1, Claudin-1 and Occludin. Furthermore, L-Arabinose modulated the levels of hormones (MTL, Gas) and neurotransmitters (5-HT, VIP) related to the digestive systems of mice with constipation, resulting in elevated levels of 5-HT, MTL, and Gas and decreasing levels of VIP. Histopathological analysis also revealed that L-Arabinose intervention improved the intestinal inflammatory response. Furthermore, 16S rRNA sequencing and metabolomics of the intestinal microbiota demonstrated that L-Arabinose treatment improved both the intestinal microbiota composition and the metabolite levels. This study suggests that L-Arabinose can serve as a potential functional ingredient to promote intestinal health, enhance gastrointestinal motility and barrier function, regulate osmotic pressure, restore neurotransmitter levels, and effectively relieve functional constipation. Full article
(This article belongs to the Section Nutraceuticals, Functional Foods, and Novel Foods)
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