Morinaga (Japan)

ABSTRACTHyperuricemia is associated with various diseases, and xanthine oxidase (XO) is the rate‐limiting enzyme in uric acid (UA) production. A previous study reported that Leu‐Asp‐Gln‐Trp (LDQW) in whey protein hydrolysate (WPH) suppressed lipid droplet accumulation in differentiated 3T3‐L1 adipocyte‐like cells. However, our understanding of LDQW remains limited, further, its efficacy against hyperuricemia has not been elucidated. This study evaluated the XO inhibitory activity of LDQW, one of the bioactive peptides in WPH. In this study, UA produced by the reaction between XO and xanthine was determined using two methods: monitoring the absorbance at 290 nm using absorptiometry and detection using liquid chromatography with tandem mass spectrometry (LC–MS/MS) analysis. Allopurinol was used as the positive control, whereas tryptophan and Ala‐Leu‐Pro‐Met (ALPM) were used for comparison. Both absorptiometry and LC–MS/MS analyses demonstrated that LDQW significantly inhibited XO activity in a concentration‐dependent manner. The LC–MS/MS analysis results indicated that LDQW, tryptophan, and ALPM inhibition ratios at 20 mM were 58.0% ± 2.8%, 4.4% ± 3.7%, and 45.0% ± 1.0%, respectively. Moreover, it was suggested that Asp‐Gln‐Trp, a potential digestive peptide predicted by the enzymatic digestion of LDQW in silico, also possessed XO inhibitory activity comparable to that of LDQW in LC–MS/MS analysis. These findings suggest that LDQW is a promising bioactive peptide with potential ameliorative effects against hyperuricemia, similar to those of other XO inhibitory peptides.

Plasmacytoid dendritic cells (pDCs) express Toll-like receptor 7 (TLR7) in the endosomes, recognize viral single-stranded RNA (ssRNA), and produce significant amounts of interferon (IFN)-α. Bovine lactoferrin (LF) enhances the response of IFN regulatory factors followed by the activation of IFN-sensitive response elements located in the promoter regions of the IFN-α gene and IFN-stimulated genes in the TLR7 reporter THP-1 cells in the presence of R-848, a TLR7 agonist. In ex vivo experiments using human peripheral blood mononuclear cells, LF enhances IFN-α levels in the supernatant in the presence of R-848. Additionally, it increases the expression of IFN-α, human leukocyte antigen (HLA)-DR, and CD86 in pDCs; HLA-DR and CD86 in myeloid dendritic cells; CD69 in CD56 dim natural killer and T killer cells; and IFN-γ in T helper type 1 and B cells in the presence of R-848. The inhibition of phagocytosis or neutralization of nucleolin, a receptor of LF, suppresses LF incorporation into pDCs. These results suggest that pDCs incorporate LF through phagocytosis or nucleolin-mediated endocytosis, and LF enhances TLR7 response in the endosome and subsequent IFN signaling pathway and activates innate and adaptive immune cells. We anticipate that LF modulates antiviral immunity against environmental ssRNA viruses and contributes to homeostasis.

The INFOGEST method is a valuable tool for understanding and monitoring food digestion as an alternative to in vivo assays. However, few studies have compared animal and alternative protein sources in terms of protein quality using the INFOGEST method. This study aimed to evaluate the protein quality of milk-, plant-, and insect-based protein materials by in vitro protein digestibility and in vitro digestible indispensable amino acid score (DIAAS), following the INFOGEST method. Milk-based protein materials had the highest protein digestibility (86.1-90.8%), followed by soy (85.1%) and wheat (82.3%). These materials had significantly higher protein digestibility compared with zein (65.1%), cricket (63.6%), and mealworm (69.5%). Additionally, the mean values of in vitro DIAAS of milk-based protein materials (105.0-137.5) were higher than those of plant- and insect-based protein materials (1.9-91.0). Milk-based protein materials have higher protein quality than plant- and insect-based protein materials by the nutritional evaluation following the INFOGEST digestion method.

To elucidate the pathogenesis of postpancreatectomy diabetes mellitus (PPDM). Forty-eight patients without diabetes undergoing either pancreatoduodenectomy (PD) (n = 20) or distal pancreatectomy (DP) (n = 28) were included. A 75-g oral glucose tolerance test was performed every 6 months. Microbiome composition and short-chain fatty acids (SCFAs) in feces were examined before and 6 months after surgery. The association of histological characteristics of the resected pancreas with PPDM was examined. During follow-up (median 3.19 years), 2 of 20 PD patients and 16 of 28 DP patients developed PPDM. Proteobacteria relative abundance, plasma glucagon-like peptide 1 (GLP-1), and fecal butyrate levels increased only after PD. Postsurgical butyrate levels were correlated with postsurgical GLP-1 levels. With no significant difference in the volume of the resected pancreas between the surgical procedures, both β-cell and α-cell areas in the resected pancreas were significantly higher in DP patients than in PD patients. In DP patients, the progressors to diabetes showed preexisting insulin resistance compared with nonprogressors, and both increased α- and β-cell areas were predictors of PPDM. Furthermore, in DP patients, α-cell and β-cell areas were associated with ALDH1A3 expression in islets. We postulate that a greater removal of β-cells contributes to the development of PPDM after DP. Islet expansion along with preexisting insulin resistance is associated with high cellular plasticity, which may predict the development of PPDM after DP. In contrast, PD is associated with alterations of gut microbiome and increases in SCFA production and GLP-1 secretion, possibly protecting against PPDM development.

Extracellular proteins are important factors in host-microbe interactions; however, the specific factors that enable bifidobacterial adhesion and survival in the gastrointestinal (GI) tract are not fully characterized. Here, we discovered that Bifidobacterium longum NCC2705 cultured in bacterium-free supernatants of human fecal fermentation broth released a myriad of particles into the extracellular environment. The aim of this study was to characterize the physiological properties of these extracellular particles. The particles, approximately 50 to 80 nm in diameter, had high protein and double-stranded DNA contents, suggesting that they were extracellular vesicles (EVs). A proteomic analysis showed that the EVs primarily consisted of cytoplasmic proteins with crucial functions in essential cellular processes. We identified several mucin-binding proteins by performing a biomolecular interaction analysis of phosphoketolase, GroEL, elongation factor Tu (EF-Tu), phosphoglycerate kinase, transaldolase (Tal), and heat shock protein 20 (Hsp20). The recombinant GroEL and Tal proteins showed high binding affinities to mucin. Furthermore, the immobilization of these proteins on microbeads affected the permanence of the microbeads in the murine GI tract. These results suggest that bifidobacterial exposure conditions that mimic the intestine stimulate B. longum EV production. The resulting EVs exported several cytoplasmic proteins that may have promoted B. longum adhesion. This study improved our understanding of the Bifidobacterium colonization strategy in the intestinal microbiome.IMPORTANCEBifidobacterium is a natural inhabitant of the human gastrointestinal (GI) tract. Morphological observations revealed that extracellular appendages of bifidobacteria in complex microbial communities are important for understanding its adaptations to the GI tract environment. We identified dynamic extracellular vesicle (EV) production by Bifidobacterium longum in bacterium-free fecal fermentation broth that was strongly suggestive of differing bifidobacterial extracellular appendages in the GI tract. In addition, export of the adhesive moonlighting proteins mediated by EVs may promote bifidobacterial colonization. This study provides new insight into the roles of EVs in bifidobacterial colonization processes as these bacteria adapt to the GI environment.

本研究は，圧痕深度評価法の測定値で圧痕性浮腫の病態を識別するカットオフ値を得ることを目的とした。圧痕性浮腫ありと診断され，症状が安定している患者65名（男性39名・女性26名）と，地域の体力測定会に任意で参加した一般健常者35名（男11名，女性24名）を対象とした。被験者に足底を床面に接地した端座位にさせて，第3中足骨骨頭の足背部の圧痕深度を圧痕深度測定機器で左右測定した。統計解析は，圧痕性浮腫の有無を従属変数，圧痕深度値，性別，年齢，身長，体重，Body Mass Index（BMI）を独立変数とした多重ロジスティック回帰分析を行い，Receiver Operating Characteristic Curve（ROC 曲線）からカットオフ値を求め，感度・特異度を算出した。その結果，圧痕性浮腫の有無に影響する因子として圧痕深度値が抽出され，オッズ比は135．0であった。ROC 曲線から圧痕深度値のカットオフ値は2．25，感度は0．83，特異度は1．00の値が算出された。本研究より，圧痕深度評価法は，圧痕性浮腫の有無を識別可能で有用な評価法であり，その病態識別値は2．25mm であることが示唆された。

Background:Probiotics use has been associated with modulation of inflammation and considered as a possible intervention for CNS diseases such as mild cognitive impairment (MCI) and dementia.Objective:We aimed to test the effect of the probiotic strain, Bifidobacterium breve A1 (MCC1274), to restore cognition in a physically healthy, suspected MCI population.Methods:In this randomized, double-blind, placebo-controlled trial, 80 healthy older adults suffering from MCI were divided into two even groups to receive once daily either probiotic (B. breve A1, 2×1010 CFU) or placebo for 16 weeks using a computer-generated algorithm. Cognitive functions were assessed by the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) and the Japanese version of the MCI Screen (JMCIS) tests before and after the study as primary and secondary endpoints, respectively.Results:79 participants completed the study, and no adverse events were observed. RBANS total score was significantly improved in probiotic group compared with placebo (mean between-group difference 11.3 [95% CI 6.7 to 15.8]; p < 0.0001) after 16 weeks of consumption, in particular with significant improvement in domain scores of immediate memory, visuospatial/constructional, and delayed memory (p < 0.0001), in both intention-to-treat (ITT) analysis and per-protocol (PP) analysis. JMCIS score was also improved versus placebo in ITT analysis (p = 0.052) and PP analysis (p = 0.036).Conclusion:Study results indicate B. breve A1 is a safe and effective approach for improving memory functions of suspected MCI subjects.

The infant’s gut microbiome is generally rich in the Bifidobacterium genus. The mother’s milk contains natural prebiotics, called human milk oligosaccharides (HMOs), as the third most abundant solid component after lactose and lipids, and of the different gut microbes, infant gut-associated bifidobacteria are the most efficient in assimilating HMOs. Indeed, the fecal concentration of HMOs was found to be negatively correlated with the fecal abundance of Bifidobacterium in infants. Given these results, two HMO molecules, 2′-fucosyllactose and lacto-N-neotetraose, have recently been industrialized to fortify formula milk. As of now, however, our knowledge about the HMO consumption pathways in infant gut-associated bifidobacteria is still incomplete. The recent studies indicate that HMO assimilation abilities significantly vary among different Bifidobacterium species and strains. Therefore, to truly maximize the effects of prebiotic and probiotic supplementation in commercialized formula, we need to understand HMO consumption behaviors of bifidobacteria in more detail. In this review, we summarized how different Bifidobacterium species/strains are equipped with varied gene sets required for HMO assimilation. We then examined the correlation between the abundance of the HMO-related genes and bifidobacteria-rich microbiota formation in the infant gut through data mining analysis of a deposited fecal microbiome shotgun sequencing dataset. Finally, we shortly described future perspectives on HMO-related studies.