RESUMEN

Bacterial culturomics is a set of techniques to isolate and identify live bacteria from complex microbial ecosystems. Despite its potential to revolutionize microbiome research, bacterial culturomics has significant challenges when applied to human gut microbiome studies due to its labor-intensive nature. Therefore, we established a streamlined culturomics approach with minimal culture conditions for stool sample preincubation. We evaluated the suitability of non-selective medium candidates for maintaining microbial diversity during a 30-day incubation period based on 16S rRNA gene amplicon analysis. Subsequently, we applied four culture conditions (two preincubation media under an aerobic/anaerobic atmosphere) to isolate gut bacteria on a large scale from eight stool samples of healthy humans. We identified 8141 isolates, classified into 263 bacterial species, including 12 novel species candidates. Our analysis of cultivation efficiency revealed that seven days of aerobic and ten days of anaerobic incubation captured approximately 91% and 95% of the identified species within each condition, respectively, with a synergistic effect confirmed when selected preincubation media were combined. Moreover, our culturomics findings expanded the coverage of gut microbial diversity compared to 16S rRNA gene amplicon sequencing results. In conclusion, this study demonstrated the potential of a streamlined culturomics approach for the efficient isolation of gut bacteria from human stool samples. This approach might pave the way for the broader adoption of culturomics in human gut microbiome studies, ultimately leading to a more comprehensive understanding of this complex microbial ecosystem.

Asunto(s)

Bacterias , Heces , Microbioma Gastrointestinal , ARN Ribosómico 16S , Humanos , Microbioma Gastrointestinal/genética , ARN Ribosómico 16S/genética , Heces/microbiología , Bacterias/genética , Bacterias/clasificación , Bacterias/aislamiento & purificación , ADN Bacteriano/genética

RESUMEN

The gut microbiota plays a pivotal role in metabolic disorders, notably type 2 diabetes mellitus (T2DM). In this study, we investigated the synergistic potential of combining the effects of Bifidobacterium longum NBM7-1 (CKD1) with anti-diabetic medicines, LobeglitazoneⓇ (LO), SitagliptinⓇ (SI), and MetforminⓇ (Met), to alleviate hyperglycemia in a diabetic mouse model. CKD1 effectively mitigated insulin resistance, hepatic steatosis, and enhanced pancreatic ß-cell function, as well as fortifying gut-tight junction integrity. In the same way, SI-CKD1 and Met- CKD1 synergistically improved insulin sensitivity and prevented hepatic steatosis, as evidenced by the modulation of key genes associated with insulin signaling, ß-oxidation, gluconeogenesis, adipogenesis, and inflammation by qRT-PCR. The comprehensive impact on modulating gut microbiota composition was observed, particularly when combined with MetforminⓇ. This combination induced an increase in the abundance of Rikenellaceae and Alistipes related negatively to the T2DM incidence while reducing the causative species of Cryptosporangium, Staphylococcaceae, and Muribaculaceae. These alterations intervene in gut microbiota metabolites to modulate the level of butyrate, indole-3-acetic acid, propionate, and inflammatory cytokines and to activate the IL-22 pathway. However, it is meaningful that the combination of B. longum NBM7-1(CKD1) reduced the medicines' dose to the level of the maximal inhibitory concentrations (IC50). This study advances our understanding of the intricate relationship between gut microbiota and metabolic disorders. We expect this study to contribute to developing a prospective therapeutic strategy modulating the gut microbiota.

Asunto(s)

Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Microbioma Gastrointestinal , Resistencia a la Insulina , Metformina , Ratones , Animales , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Regulación hacia Arriba , Diabetes Mellitus Experimental/tratamiento farmacológico , Metformina/farmacología , Metformina/uso terapéutico

RESUMEN

Type 2 diabetes (T2D) is known as adult-onset diabetes, but recently, T2D has increased in the number of younger people, becoming a major clinical burden in human society. The objective of this study was to determine the effects of Bifidobacterium and Lactiplantibacillus strains derived from the feces of 20 healthy humans on T2D development and to understand the mechanism underlying any positive effects of probiotics. We found that Bifidobacterium longum NBM7-1 (Chong Kun Dang strain 1; CKD1) and Lactiplantibacillus rhamnosus NBM17-4 (Chong Kun Dang strain 2; CKD2) isolated from the feces of healthy Korean adults (n = 20) have anti-diabetic effects based on the insulin sensitivity. During the oral gavage for 8 weeks, T2D mice were supplemented with anti-diabetic drugs (1.0-10 mg/kg body weight) to four positive and negative control groups or four probiotics (200 uL; 1 × 109 CFU/mL) to groups separately or combined to the four treatment groups (n = 6 per group). While acknowledging the relatively small sample size, this study provides valuable insights into the potential benefits of B. longum NBM7-1 and L. rhamnosus NBM17-4 in mitigating T2D development. The animal gene expression was assessed using a qRT-PCR, and metabolic parameters were assessed using an ELISA assay. We demonstrated that B. longum NBM7-1 in the CKD1 group and L. rhamnosus NBM17-4 in the CKD2 group alleviate T2D development through the upregulation of IL-22, which enhances insulin sensitivity and pancreatic functions while reducing liver steatosis. These findings suggest that B. longum NBM7-1 and L. rhamnosus NBM17-4 could be the candidate probiotics for the therapeutic treatments of T2D patients as well as the prevention of type 2 diabetes.

RESUMEN

The human gut microbiota is a complex ecology comprising approximately 10 to 100 trillion microbial cells. Most of the bacteria detected by 16s rRNA sequencing have yet to be cultured, but intensive attempts to isolate the novel bacteria have improved our knowledge of the gut microbiome composition and its roles within human host. In our culturomics study, a novel gram-negative, motile, obligately anaerobic, rod-shaped bacteria, designated as strain ICN-92133T, was isolated from a fecal sample of a 26-year-old patient with Crohn's disease. Based on the 16s rRNA sequence of strain ICN-92133T, the phylogeny analysis placed the strain into the family Selenomonadaceae, showing 93.91% similarity with the closely related Massilibacillus massiliensis strain DSM 102838T. Strain ICN-92133T exhibited a genome size of 2,679,003 bp with a GC content of 35.5% which was predicted to contain 26 potential virulence factors and five antimicrobial resistance genes. In comparative genomic analysis, strain ICN-92133T showed digital DNA-DNA Hybridization and OrthoANI values lower than 21.9% and 71.9% with the closest type strains, respectively. In addition, comparing phenotypic, biochemical, and cellular fatty acids with those of closely related strains revealed the distinctiveness of strain ICN-92133T. Based on the taxonogenomic results, strain ICN-92133T is proposed as a novel species belonging to a new genus. Therefore, we suggest the name of the new genus Selenobaculum gen. nov. within the family Selenomonadaceae and strain ICN-92133T (= KCTC 25622T = JCM 36070T) as a type strain of new species Selenobaculum gbiensis sp. nov.

Asunto(s)

Enfermedad de Crohn , Microbioma Gastrointestinal , Humanos , Adulto , Microbioma Gastrointestinal/genética , ARN Ribosómico 16S/genética , Bacterias , Firmicutes , ADN

RESUMEN

The gut microbiome is critical in human health, and various dietary factors influence its composition and function. Among these factors, animal products, such as meat, dairy, and eggs, represent crucial sources of essential nutrients for the gut microbiome. However, the correlation and characteristics of livestock consumption with the gut microbiome remain poorly understood. This review aimed to delineate the distinct effects of meat, dairy, and egg products on gut microbiome composition and function. Based on the previous reports, the impact of red meat, white meat, and processed meat consumption on the gut microbiome differs from that of milk, yogurt, cheese, or egg products. In particular, we have focused on animal-originated proteins, a significant nutrient in each livestock product, and revealed that the major proteins in each food elicit diverse effects on the gut microbiome. Collectively, this review highlights the need for further insights into the interactions and mechanisms underlying the impact of animal products on the gut microbiome. A deeper understanding of these interactions would be beneficial in elucidating the development of dietary interventions to prevent and treat diseases linked to the gut microbiome.

RESUMEN

Recently, several probiotic products have been developed; however, most probiotic applications focused on prokaryotic bacteria whereas eukaryotic probiotics have received little attention. Saccharomyces cerevisiae yeast strains are eukaryotes notable for their fermentation and functional food applications. The present study investigated the novel yeast strains isolated from Korean fermented beverages and examined their potential probiotic characteristics. We investigated seven strains among 100 isolates with probiotic characteristics further. The strains have capabilities such as auto-aggregation tendency, co-aggregation with a pathogen, hydrophobicity with n-hexadecane,1,1-diphenyl-2-picrylhydrazyl scavenging effect, survival in simulated gastrointestinal tract conditions and the adhesion ability of the strains to the Caco-2 cells. Furthermore, all the strains contained high cell wall glucan content, a polysaccharide with immunological effects. Internal transcribed spacer sequencing identified the Saccharomyces strains selected in the present study as probiotics. To examine the effects of alleviating inflammation in cells, nitric oxide generation in raw 264.7 cells with S. cerevisiae showed that S. cerevisiae GILA could be a potential probiotic strain able to alleviate inflammation. Three probiotics of S. cerevisiae GILA strains were chosen by in vivo screening with a dextran sulfate sodium-induced colitis murine model. In particular, GILA 118 down-regulates neutrophil-lymphocyte ratio and myeloperoxidase in mice treated with DSS. The expression levels of genes encoding tight junction proteins in the colon were upregulated, cytokine interleukin-10 was significantly increased, and tumor necrosis factor-α was reduced in the serum.

Asunto(s)

Colitis , Probióticos , Humanos , Animales , Ratones , Saccharomyces cerevisiae/metabolismo , Sulfato de Dextran/efectos adversos , Células CACO-2 , Modelos Animales de Enfermedad , Colitis/inducido químicamente , Inflamación , Probióticos/metabolismo

RESUMEN

We demonstrate the generation of high-energy femtosecond pulses by a spectrally combined four-diode-pumped Ti:sapphire laser with semi-conductor saturable absorber mirror mode locking. To achieve energy scaling, the laser cavity was extended using a design based on the Herriott multipass cell. The laser operates at a 17.5 MHz repetition rate and generates pulses with energies as high as 16.3 nJ and 80 fs in duration. The signal-to-noise ratio at the fundamental frequency showed an extinction ratio of >50 dB relative to the carrier. This compact single laser was applied to video-rate forward and backward coherent anti-Stokes Raman scattering spectro-microscopy with a pixel dwell time of 122 ns, which is the lowest dwell time ever achieved, to the best of our knowledge.

RESUMEN

Many probiotic species have been used as a fermentation starter for manufacturing functional food materials. We have isolated Bifidobacterium animalis subsp. lactis LDTM 8102 from the feces of infants as a novel strain for fermentation. While Glycine max has been known to display various bioactivities including anti-oxidant, anti-skin aging, and anti-cancer effects, the immune-modulatory effect of Glycine max has not been reported. In the current study, we have discovered that the extract of Glycine max fermented with B. animalis subsp. lactis LDTM 8102 (GFB 8102), could exert immuno-modulatory properties. GFB 8102 treatment increased the production of immune-stimulatory cytokines in RAW264.7 macrophages without any noticeable cytotoxicity. Analysis of the molecular mechanism revealed that GFB 8102 could upregulate MAPK2K and MAPK signaling pathways including ERK, p38, and JNK. GFB 8102 also increased the proliferation rate of splenocytes isolated from mice. In an animal study, administration of GFB 8102 partially recovered cyclophosphamide-mediated reduction in thymus and spleen weight. Moreover, splenocytes from the GFB 8102-treated group exhibited increased TNF-α, IL-6, and IL-1ß production. Based on these findings, GFB 8102 could be a promising functional food material for enhancing immune function.

Asunto(s)

Bifidobacterium animalis , Probióticos , Animales , Antioxidantes/metabolismo , Ciclofosfamida , Citocinas/metabolismo , Humanos , Inmunidad , Interleucina-6/metabolismo , Ratones , Extractos Vegetales/metabolismo , Glycine max/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

As the number of households that raise dogs and cats is increasing, there is growing interest in animal health. The gut plays an important role in animal health. In particular, the microbiome in the gut is known to affect both the absorption and metabolism of nutrients and the protective functions of the host. Using probiotics on pets has beneficial effects, such as modulating the immune system, helping to reduce stress, protecting against pathogenic bacteria and developing growth performance. The goals of this review are to summarize the relationship between probiotics/the gut microbiome and animal health, to feature technology used for identifying the diversity of microbiota composition of canine and feline microbiota, and to discuss recent reports on probiotics in canines and felines and the safety issues associated with probiotics and the gut microbiome in companion animals.

RESUMEN

Objective: The role of the gut microbiome in the onset and development of atopic dermatitis (AD) has been postulated. Thus, we investigated the gut microbial compositions in infants with and without AD and compared the gut bacterial flora of their mothers. Methods: The prospective and cross-sectional study participated in 44 pairs of mothers and children. We selected infants born via full-term normal vaginal delivery and no history of antibiotic or probiotic use and infection during the first three months of life. The 15 pairs, consisting of nine healthy infants and six AD infants, were included in this study. Fecal samples of mothers and infants were analyzed within 30 days of delivery and at 12 months, respectively. Microbes in the fecal samples of mothers and infants were subjected to analysis of 16S rRNA amplicon sequencing. Results: The abundance of specific taxonomic groups was notably different, but microbial diversity and phylogenetic distances were not significantly different in either maternal or infant groups according to the presence of infant AD. A total of 12 species were selected as differential species in infants with AD compared to healthy infants. Six species were significantly different in the mothers of infants with AD compared to the mothers of healthy infants. Akkermansia muciniphila was only detected in healthy infants and their mothers. Conclusion: The presence of Akkermansia muciniphila in mothers and children after vaginal delivery is associated with the onset and development of AD.

Asunto(s)

Dermatitis Atópica , Microbioma Gastrointestinal , Akkermansia , Niño , Estudios Transversales , Dermatitis Atópica/microbiología , Femenino , Microbioma Gastrointestinal/genética , Humanos , Lactante , Madres , Filogenia , Estudios Prospectivos , ARN Ribosómico 16S/genética

RESUMEN

The interest in skin microbiome differences by ethnicity, age, and gender is increasing. Compared to other ethnic groups, studies on the skin microbiome of Koreans remains insufficient; we investigated facial skin microbiome characteristics according to gender and age among Koreans. Fifty-one healthy participants were recruited, the facial skin characteristics of each donor were investigated, their skin bacterial DNA was isolated and metagenomic analysis was performed. The donors were divided into two groups for age and sex each to analyze their skin microbiomes. Moreover, we investigated the correlation between the skin microbiome and clinical characteristics. The alpha diversity of the skin microbiome was significantly higher in the elderly, and beta diversity was significantly different according to age. The comparative skin microbials showed that the genus Lawsonella was more abundant in the younger age group, and Enhydrobacter was predominant in the older age group. Staphylococcus and Corynebacterium were more abundant in males, while Lactobacillus was more abundant in females. Lawsonella had a negative correlation with skin moisture and brown spots. Staphylococcus and Corynebacterium both had negative correlations with the number of UV spots and positive correlations with transepidermal water loss (TEWL). Furthermore, Staphylococcus aureus had a negative correlation with skin moisture parameters.

Asunto(s)

Bacterias/clasificación , Cara/microbiología , Voluntarios Sanos , Piel/microbiología , Adulto , Factores de Edad , Anciano , Pueblo Asiatico , Bacterias/genética , Código de Barras del ADN Taxonómico , Femenino , Humanos , Masculino , Metagenómica , Persona de Mediana Edad , Pérdida Insensible de Agua , Adulto Joven

RESUMEN

Yak-Kong is a type of black soybean that is colloquially referred to as the "medicinal bean" and it elicits several beneficial effects that are relevant to human health, including attenuating the formation of skin wrinkles. It has previously been shown that soybean extracts elicit additional bioactivity that is fermented by lactic acid bacteria. In this study of lactic acid bacteria strains that were isolated from the stools of breast-feeding infants (<100 days old), we selected Bifidobacterium animalis subsp. Lactis LDTM 8102 (LDTM 8102) as the lead strain for the fermentation of Yak-Kong. We investigated the effects of LDTM 8102-fermented Yak-Kong on solar-ultraviolet irradiation (sUV)-induced wrinkle formation. In HaCaT cells, the ethanol extract of LDTM 8102-fermented Yak-Kong (EFY) effectively reduced sUV-induced matrix metalloproteinase-1 (MMP-1) secretion. The effect of EFY was superior to that of unfermented (UFY)- and Lactis KCTC 5854 (another Bifidobacterium animalis species)-fermented Yak-Kong. Additionally, EFY reduced sUV-induced MMP-1 mRNA expression and promoter activity, as well as the transactivation of AP-1 and phosphorylation of ERK1/2 and JNK1/2. Furthermore, EFY alleviated sUV-induced MMP-1 secretion, the destruction of the epidermis, and degradation of collagen in a three-dimensional (3D) skin culture model. EFY had a higher total polyphenol content and anti-oxidative activity than UFY. Twelve metabolites were significantly (≥2-fold) increased in Yak-Kong extract after fermentation by LDTM 8102. Among them, the metabolites of major isoflavones, such as 6,7,4'-trihydroxyisoflavone (THIF), exerted the reducing effect of MMP-1, which indicated that the isoflavone metabolites contributed to the effect of EFY on MMP-1 expression as active compounds. These findings suggest that EFY is a potent natural material that can potentially prevent sUV-induced wrinkle formation.

RESUMEN

The role of the gut microbiota in the pathogenesis of inflammatory bowel disease (IBD) has been in focus for decades. Although metagenomic observations in patients/animal colitis models have been attempted, the microbiome results were still indefinite and broad taxonomic presumptions were made due to the cross-sectional studies. Herein, we conducted a longitudinal microbiome analysis in a dextran sulfate sodium (DSS)-induced colitis mouse model with a two-factor design based on serial DSS dose (0, 1, 2, and 3%) and duration for 12 days, and four mice from each group were sacrificed at two-day intervals. During the colitis development, a transition of the cecal microbial diversity from the normal state to dysbiosis and dynamic changes of the populations were observed. We identified genera that significantly induced or depleted depending on DSS exposure, and confirmed the correlations of the individual taxa to the colitis severity indicated by inflammatory biomarkers (intestinal bleeding and neutrophil-derived indicators). Of note, each taxonomic population showed its own susceptibility to the changing colitis status. Our findings suggest that an understanding of the individual susceptibility to colitis conditions may contribute to identifying the role of the gut microbes in the pathogenesis of IBD.

RESUMEN

Probiotics are often infused into functional foods or encapsulated in a supplement form to maintain a healthy balance between the gut microbiota and their host. Because there are milk-based functional foods such as yogurt and cheese on the market, it has been suggested that milk-based probiotics could be incorporated into skim milk proteins in a liquid capsule. Skim milk is mainly composed of casein and whey protein, which create a strong natural barrier and can be used to encapsulate probiotics. In this study, we compared the encapsulated probiotics prepared with milkbased concentrated cell mixtures using commercial probiotics. Probiotic capsules were emulsified with skim milk proteins using vegetable oil to form a double coating layer. The product was heatstable when tested using a rheometer. The survival rate of the milk-based probiotic cells in the lower gastric environment with bile was significantly higher than commercial probiotics. Thus, milkencapsulated probiotics exhibited greater efficacy in the host than other types of probiotics, suggesting that the former could be more viable with a longer shelf life under harsh conditions than other form of probiotics. Our findings suggested that, compared with other types of probiotics, milkbased probiotics may be a better choice for producers and consumers.

Asunto(s)

Cápsulas/química , Proteínas de la Leche/química , Probióticos/química , Reología , Animales , Bilis , Caseínas , Queso , Alimentos Funcionales , Leche , Temperatura , Termotolerancia , Yogur/microbiología

RESUMEN

Lactic acid bacteria (LAB) play an important role in dairy fermentations, notably as cheese starter cultures. During the cheese production and ripening period, various enzymes from milk, rennet, starter cultures, and non-starter LABs are involved in flavor formation pathways, including glycolysis, proteolysis, and lipolysis. Among these three pathways, starter LABs are particularly related to amino acid degradation, presumably as the origins of major flavor compounds. Therefore, we used several enzymes as major criteria for the selection of starter bacteria with flavor-forming ability. Lactococcus lactis subsp. lactis LDTM6802 and Lactococcus lactis subsp. cremoris LDTM6803, isolated from Korean raw milk and cucumber kimchi, were confirmed by using multiplex PCR and characterized as starter bacteria. The combinations of starter bacteria were validated in a miniature Gouda-type cheese model. The flavor compounds of the tested miniature cheeses were analyzed and profiled by using an electronic nose. Compared to commercial industrial cheese starters, selected starter bacteria showed lower pH, and more variety in their flavor profile. These results demonstrated that LDTM6802 and LDTM6803 as starter bacteria have potent starter properties with a characteristic flavor-forming ability in cheese.

Asunto(s)

Queso/microbiología , Lactococcus lactis/metabolismo , Lactococcus/metabolismo , Gusto , Fermentación , Microbiología de Alimentos , Lactobacillales/metabolismo

RESUMEN

Strain CA7T, a Gram-stain-negative, non-motile, non-spore-forming, aerobic and rod-shaped bacterial strain, was isolated from raw cow's milk collected from a farm affiliated with Chung-Ang University, Anseong, Korea, and characterized by a polyphasic approach. Optimal growth of strain CA7T was observed on tryptic soy agar at 30 °C and pH 7.0 with 0 % NaCl. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CA7T belonged to the genus Chryseobacterium. The most closely related strains (16S rRNA gene sequence similarity indicated in parentheses), based on the phylogenetic analysis, were Chryseobacterium rhizosphaerae KCTC 22548T (98.08 %), Chryseobacterium nakagawai CCUG 60563T (98.61 %), Chryseobacterium jejuense KACC 12501T (97.85 %) and Chryseobacterium aurantiacum KCTC 62135T (97.78 %). Whole genome sequencing indicated that the genome size was 5 125 723 bp and had a DNA G+C content of 37.4 mol%. Average nucleotide identity values for strain CA7T with C. rhizosphaerae, C. nakagawai, C. jejuense, C. aurantiacum, and the type species of the genus Chryseobacterium, C. gleum, were 80.2, 79.8, 79.8, 79.6 and 80.4 %, respectively. The digital DNA-DNA hybridization values of CA7T compared to C. rhizosphaerae, C. nakagawai, C. jejuense, C. aurantiacum and C. gleum were 24.1, 23.9, 23.9, 23.7 and 24.3 %, respectively. The major fatty acids were iso-C15 : 0, summed feature 9 (iso-C17 : 1 ω9c and/or C16 : 0 10-methyl), iso-C17 : 0 3-OH and summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1 ω7c). Menaquinone-6 was the only respiratory quinone. The major polar lipid was phosphatidylethanolamine. Based on this polyphasic taxonomic study, strain CA7T represents a novel species of the genus Chryseobacterium for which the name Chryseobacterium vaccae sp. nov. is proposed. The type strain is CA7T (=KACC 21402T=JCM 33749T).

Asunto(s)

Chryseobacterium/clasificación , Leche/microbiología , Filogenia , Animales , Técnicas de Tipificación Bacteriana , Composición de Base , Bovinos , Chryseobacterium/aislamiento & purificación , ADN Bacteriano/genética , Ácidos Grasos/química , Hibridación de Ácido Nucleico , Fosfatidiletanolaminas/química , ARN Ribosómico 16S/genética , República de Corea , Análisis de Secuencia de ADN , Vitamina K 2/análogos & derivados , Vitamina K 2/química

RESUMEN

Inflammatory bowel disease (IBD) is a group of conditions involving chronic relapsing-remitting inflammation of the gastrointestinal tract with an unknown etiology. Although the cause-effect relationship between gut microbiota and IBD has not been clearly established, emerging evidence from experimental models supports the idea that gut microbes play a fundamental role in the pathogenesis of IBD. As microbiome-based therapeutics for IBD, the beneficial effects of probiotics have been found in animal colitis models and IBD patients. In this study, based on the dextran sulfate sodium (DSS)-induced colitis mouse model, we investigated Lactobacillus rhamnosus strain LDTM 7511 originating from Korean infant feces as a putative probiotic strain for IBD. The strain LDTM 7511 not only alleviated the release of inflammatory mediators, but also induced the transition of gut microbiota from dysbiotic conditions, exhibiting the opposite pattern in the abundance of DSS colitis-associated bacterial taxa to the DSS group. Our findings suggest that the strain LDTM 7511 has the potential to be used as a probiotic treatment for IBD patients in comparison to L. rhamnosus GG (ATCC 53103), which has been frequently used for IBD studies.

RESUMEN

Removal of sugar moieties from ginsenosides has been proposed to increase their biological effects in various disease models. In order to identify strains that can increase aglycone contents, we performed a screening using bacteria isolated from the feces of infants focusing on acid tolerance and ß-glucosidase activity. We isolated 565 bacteria and selected Bifidobacterium animalis subsp. lactis LT 19-2 (LT 19-2), which exhibited the highest ß-glucosidase activity with strong acid tolerance. As red ginseng (RG) has been known to exert immunomodulatory functions, we fermented RG using LT 19-2 (FRG) and investigated whether this could alter the aglycone profile of ginsenosides and improve its immunomodulatory effect. FRG increased macrophage activity more potently compared to RG, demonstrated by higher TNF-α and IL-6 production. More importantly, the FRG treatment stimulated the proliferation of mouse splenocytes and increased TNF-α levels in bone marrow-derived macrophages, confirming that the enhanced immunomodulatory function can be recapitulated in primary immune cells. Examination of the molecular mechanism revealed that F-RG could induce phosphorylations of ERK, p38, JNK, and NF-κB. Analysis of the ginsenoside composition showed a decrease in Rb1, Re, Rc, and Rb3, accompanied by an increase in Rd, Rh1, F2, and Rg3, the corresponding aglycone metabolites, in FRG compared to RG. Collectively, LT 19-2 maybe used as a probiotic strain to improve the bioactivity of functional foods through modifying the aglycone/glycoside profile.

Asunto(s)

Proteínas Bacterianas/metabolismo , Bifidobacterium animalis/enzimología , Fermentación , Ginsenósidos/farmacología , Factores Inmunológicos/farmacología , Macrófagos/efectos de los fármacos , Panax/microbiología , Probióticos/farmacología , beta-Glucosidasa/metabolismo , Animales , Bifidobacterium animalis/aislamiento & purificación , Heces/microbiología , Femenino , Ginsenósidos/metabolismo , Humanos , Factores Inmunológicos/metabolismo , Lactante , Recién Nacido , Interleucina-6/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Proteínas Quinasas Activadas por Mitógenos/metabolismo , FN-kappa B/metabolismo , Panax/metabolismo , Fosforilación , Probióticos/metabolismo , Células RAW 264.7 , Transducción de Señal , Factor de Necrosis Tumoral alfa/metabolismo

RESUMEN

Soy isoflavones, particularly genistein, have been shown to exhibit anti-obesity effects. When compared with the isoflavones genistin, daidzin, coumestrol, genistein, daidzein, 6-o-dihydroxyisoflavone, equol, 3'-o-dihydroxyisoflavone, and 8-o-dihydroxyisoflavone, a remarkably higher inhibitory effect on lipid accumulation was observed for orobol treatment during adipogenesis in 3T3-L1 cells. To identify the cellular target of orobol, its pharmacological effect on 395 human kinases was analyzed. Of the 395 kinases, orobol showed the lowest half maximal inhibitory concentration (IC50) for Casein Kinase 1 epsilon (CK1ε), and bound to this target in an ATP-competitive manner. A computer modeling study revealed that orobol may potentially dock with the ATP-binding site of CK1ε via several hydrogen bonds and van der Waals interactions. The phosphorylation of eukaryotic translation initiation factor 4E-binding protein 1, a substrate of CK1ε, was inhibited by orobol in isobutylmethylxanthine, dexamethasone and insulin (MDI)-induced 3T3-L1 cells. It was also found that orobol attenuates high fat diet-induced weight gain and lipid accumulation without affecting food intake in C57BL/6J mice. These findings underline orobol's potential for development as a novel agent for the prevention and treatment of obesity.

Asunto(s)

Fármacos Antiobesidad/uso terapéutico , Caseína Cinasa 1 épsilon/efectos de los fármacos , Flavonoides/farmacología , Obesidad/tratamiento farmacológico , Células 3T3-L1 , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Adipogénesis/efectos de los fármacos , Animales , Fármacos Antiobesidad/farmacología , Proteínas de Ciclo Celular/metabolismo , Dexametasona/farmacología , Dieta Alta en Grasa , Genisteína/química , Humanos , Insulina/farmacología , Metabolismo de los Lípidos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Obesidad/enzimología , Fosforilación , Aumento de Peso/efectos de los fármacos , Xantinas/farmacología

RESUMEN

Probiotics show low cell viability after oral administration because they have difficulty surviving in the stomach due to low pH and enzymes. For the oral delivery of probiotics, developing a formula that protects the probiotic bacteria from gastric acidity while providing living cells is mandatory. In this study, we developed tablets using a new pH-sensitive phthalyl inulin (PI) to protect probiotics from gastric conditions and investigated the effects of different compression forces on cell survival. We made three different tablets under different compression forces and measured survivability, disintegration time, and kinetics in simulated gastric-intestinal fluid. During tableting, there were no significant differences in probiotic viability among the different compression forces although disintegration time was affected by the compression force. A higher compression force resulted in higher viability in simulated gastric fluid. The swelling degree of the PI tablets in simulated intestinal fluid was higher than that of the tablets in simulated gastric fluid due to the pH sensitivity of the PI. The probiotic viability formulated in the tablets was also higher in acidic gastric conditions than that for probiotics in solution. Rapid release of the probiotics from the tablet occurred in the simulated intestinal fluid due to the pH sensitivity. After 6 months of refrigeration, the viability of the PI probiotics was kept. Overall, this is the first study to show the pH-sensitive properties of PI and one that may be useful for oral delivery of the probiotics.

Asunto(s)

Inulina/administración & dosificación , Inulina/química , Probióticos/administración & dosificación , Probióticos/química , Administración Oral , Fuerza Compresiva , Composición de Medicamentos , Estabilidad de Medicamentos , Almacenaje de Medicamentos , Jugo Gástrico/química , Concentración de Iones de Hidrógeno , Secreciones Intestinales/química , Inulina/farmacocinética , Viabilidad Microbiana , Probióticos/farmacocinética , Comprimidos/administración & dosificación , Comprimidos/química , Comprimidos/farmacocinética