Hideaki Takahashi

Gut microbiota dysbiosis is implicated in diverse intestinal and systemic disorders, and prebiotics offer a practical strategy to modify host–microbe interactions. This review evaluates 1-kestose as a candidate precision bioactive component by integrating its chemical structure, enzymatic production, gastrointestinal fate, GH32-dependent microbial utilization, human evidence, and qPCR-based response monitoring. Many commercial fructooligosaccharides contain molecules with different degrees of polymerization, complicating structure–function interpretation. In contrast, 1-kestose is a high-purity trisaccharide fructooligosaccharide and the shortest member of the inulin-type fructans. By comparing 1-kestose with long-chain inulin, we examine how fructan chain length may influence colonic fermentation kinetics, substrate availability, and tolerability. We then discuss the role of GH32 substrate specificity in the selective microbial utilization of 1-kestose and related fructooligosaccharides, particularly by bifidobacteria and representative butyrate producers. Next, we review the mechanistic rationale and preclinical evidence for co-administration of 1-kestose and long-chain inulin. Human intervention studies have evaluated 1-kestose across gastrointestinal, metabolic, immune-related, neonatal, oncological, and bowel-habit contexts, with emerging evidence of potential benefits. One healthy-adult trial has also evaluated co-administration with long-chain inulin, although direct comparative trials remain an important future priority. Finally, we propose a research framework that integrates high-purity 1-kestose, GH32-dependent microbial selectivity, and qPCR-based baseline stratification and response monitoring. Prospective, independently replicated trials are needed to establish clinical effectiveness and determine the value of biomarker-guided intervention and combination strategies.

Abstract Background Paraprobiotics, the non-viable microbial cells with health benefits, have gained interest as candidates for preventing food allergies owing to their safety and stability. Heat-killed Lactiplantibacillus plantarum FM8 (FM8) has been reported to induce interleukin (IL)-10 production in dendritic cells in vitro; however, its in vivo efficacy as a standalone intervention remains unexplored. We investigated the preventive effect of heat-killed FM8 in a murine model of ovalbumin (OVA)-induced food allergy. Methods BALB/c mice were assigned to control, OVA-induced allergy, or three FM8 treatment groups receiving FM8 at low (2 × 10⁹ CFU/day), medium (1 × 10¹⁰ CFU/day), or high (5 × 10¹⁰ CFU/day) doses. Food allergy was induced by repeated OVA sensitization and challenge. Allergy symptoms, OVA-specific immunoglobulin E (IgE), and IL-10 levels were measured. Tight junction gene expression, mucin production, gut microbiota composition, and short-chain fatty acids (SCFAs) were also analyzed. Results FM8 supplementation attenuated allergic responses in a dose-dependent manner. Allergy symptom scores were reduced in the High group, and the rectal temperature decline following OVA challenge was ameliorated in both the Med and High groups. The sensitization-induced increase in OVA-specific IgE was also attenuated in these groups. IL-4 expression in Peyer’s patches was reduced, whereas colonic IL-10 expression and serum IL-10 levels were increased in the High group. FM8 supplementation was further associated with increased ileal expression of tight junction-related genes ( Occludin , Claudin-1 and Zo-1 ) and Muc2 , along with a trend toward increased fecal mucin levels; however, no comparable changes in tight junction-related gene expression were observed in the colon. Although taxa with reported SCFA-producing potential were enriched in the High group, cecal acetate and propionate levels remained unchanged, while butyrate levels were decreased. Conclusions FM8 may prevent food allergy by promoting IL-10-associated immunoregulation and enhancing gut barrier-related responses, without a corresponding increase in cecal SCFA concentrations. The contribution of SCFA-related mechanisms remains to be clarified, and further studies are needed to determine clinically feasible dosing and efficacy in humans.

Abstract The gut microbiome is a potential source of non-invasive cancer biomarkers. We evaluated six fecal microbial markers and developed targeted qPCR-based logistic models for colorectal cancer (CRC) and pancreatic cancer (PC). Using LASSO with the 1-standard-error rule, four markers were selected for CRC (afb, nan, fsr, and 5ar) and three for PC (but, fsr, and saa). In post-selection leave-one-out cross-validation of fixed model structures, the CRC and PC models yielded AUCs of 0.824 and 0.780, respectively. Fixed-model application yielded AUCs of 0.716 for colorectal adenoma and 0.540 for the pancreatic high-risk group. In an exploratory Early PC versus high-risk comparison, the fecal qPCR score showed a higher AUC point estimate than CA19-9, while the difference was not statistically significant. Overall, the disease-specific model performance and fixed-model behavior across clinically related groups support further evaluation of model-derived cancer probability scores as exploratory cancer-assessment tools.

Background: Agaro-oligosaccharides (AOS) have been shown to modulate the gut microbiota in in vitro and animal studies; however, human evidence remains scarce. Methods: Herein, we conducted a four-week open-label, single-arm, non-randomized pilot trial in 18 healthy Japanese adults to examine the association of AOS intake at 200 mg/day with gut microbiota composition and bowel condition. Fecal samples collected before and after the intervention were analyzed using QIIME2-based 16S rRNA sequencing, and bowel condition was assessed with the Bristol Stool Form Scale. This study was registered in the UMIN Clinical Trials Registry (UMIN000056992). Results: AOS intake was not associated with significant changes in bowel condition. Gut microbiota analysis showed no significant alterations in overall community structure but revealed taxon-specific trends in the relative abundance of several bacterial taxa. Notably, nominal changes were observed in the abundance of the Ruminococcus gnavus group and Bacteroides uniformis after the intervention. In addition, quantitative PCR analysis showed an increase in 3,6-anhydro-L-galactose cycloisomerase (ACI) gene abundance after the intervention. Conclusions: These findings suggest that, in this exploratory pilot study, AOS intake was associated with a taxon-specific pattern in the gut microbiota. Further randomized controlled studies are needed to clarify the microbiota-related effects of AOS in humans.

Aims: To evaluate the diagnostic potential of previously reported (5ar, nan) and novel microbial gene markers in fecal and salivary microbiomes for the non-invasive detection of colorectal cancer (CRC) progression. Study Design: Cross-sectional study. Place and Duration of Study: Department of Gastroenterology and Hepatology, Fujita Health University, Toyoake, Japan, between June 2024 and July 2025. Methodology: Fecal samples were collected from healthy controls (n = 65) and patients with colorectal adenoma (n = 38) or carcinoma (n = 27). Salivary samples were collected from healthy controls (n = 34) and the same patient cohorts. Microbial gene abundances were quantified via quantitative PCR, and microbiome composition was assessed using 16S rRNA gene sequencing. Group comparisons used the Kruskal-Wallis and Dunn’s tests. Beta diversity differences were evaluated by PERMANOVA. Results: In fecal samples, the abundances of 5ar, nan, and the Fusicatenibacter saccharivorans 16S rRNA gene (fsr) showed a stepwise decline from the adenoma stage onward, with significantly lower levels in carcinoma versus controls (5ar, P = 0.0023; nan, P = 0.0012; fsr, P = 0.0004). Conversely, in saliva, the Fusobacterium periodonticum tyrosine phenol lyase gene (tpl) was significantly reduced only in the carcinoma group (P = 0.0180). Fecal beta diversity differed significantly between controls and both colorectal neoplasia groups (both P ≤ 0.001), whereas salivary beta diversity differed only between controls and carcinoma patients (P = 0.014). Conclusion: Fecal and salivary microbial gene markers may serve as non-invasive, rapid, and cost-effective biomarkers for colorectal neoplasia. Fecal markers captured microbiota disruption at the adenoma stage (representing the early phase of CRC progression), whereas salivary markers predominantly reflected alterations specific to carcinoma (representing the later phase). Integrating these multi-niche biomarkers provides a promising clinical platform for early detection, monitoring of precursor lesions, and postoperative surveillance; however, validation in larger, independent cohorts is warranted.