Asymptomatic Clostridium perfringens Inhabitation in Intestine Can Cause Inflammation, Apoptosis, and Disorders in Brain.

Clostridium perfringens (CP) is a foodborne pathogen. The bacterium can also inhabit human gut without symptoms of foodborne illness. However, the clinical symptoms of long-term inhabitation have not been known yet. Therefore, the objective of this study was to elucidate the relationship between intestinal CP and other internal organs. Phosphate-buffered saline (PBS) and CP were orally injected into 5-week-old (YOUNG) and 12-month-old C57BL6/J (ADULT) mice. Gene expression levels related to inflammation (tumor necrosis factor-α [TNF-α], interleukin [IL]-1ß, and IL-6) and oxidative stress (superoxide dismutase [SOD]1, SOD2, SOD3, glutathione reductase [GSR], glutathione peroxidase [GPx]3, and catalase [CAT]) responses were evaluated in the brain, small intestine, and liver. In addition, apoptosis-related (BCL2-associated X [BAX]1 and high-mobility group box-1 [HMGB1]) and brain disorder-related genes (CCAAT-enhancer-binding protein [C/EBP]-ß, C/EBPδ, C/EBP homologous protein [CHOP], and amyloid precursor protein [APP]) as brain damage markers were examined. The protein expressions in the brain were also measured. Gene expression levels of inflammation and oxidative stress responses were higher (p < 0.05) in brains of CP-YOUNG and CP-ADULT mice, compared with PBS-YOUNG and PBS-ADULT, and the gene expression levels were higher (p < 0.05) in brains of CP-ADULT mice than CP-YOUNG mice. Apoptosis-related (BAX1 and HMGB1) and brain disorder-related genes (C/EBPß, C/EBPδ, CHOP, and APP) were higher (p < 0.05) in brains of CP-challenged mice, compared with PBS-challenged mice. Even oxidative stress response (GPx and SOD2), cell damage-related (HMGB1), and ß-amyloid proteins were higher (p < 0.05) in brains of CP- than in PBS-challenged mice. C/EBP protein was higher (p < 0.05) in CP-YOUNG, compared with PBS-YOUNG mice. However, these clinical symptoms were not observed in small intestine and liver. These results indicate that although asymptomatic intestinal CP do not cause foodborne illness, their inhabitation may cause brain inflammation, oxidative stress, apoptosis, and cell damage, which may induce disorders, especially for the aged group.

Effects of mushroom extract on textural properties and muscle protein degradation of bovine longissimus dorsi muscle.

We investigated the effects of Sarcodon aspratus, Agaricus bisporus, and Lentinula edodes aqueous extracts on the tenderization of bovine longissimus dorsi muscle. Meat quality and muscle protein degradation were examined as well. Beef chunks were marinated in distilled water (control), 5% S. aspratus (SA), 5% A. bisporus (AB), or 5% L. edodes (LE) extracts. SA was shown to have a higher enzymatic activity (p < 0.001) and water-holding capacity than LE (p < 0.01). SA and AB extracts exhibited lower shear force values compared with the control (p < 0.05). SA, AB, and LE showed superior muscle proteolytic effects compared with the control. SA demonstrated the ability to degrade myosin heavy chains and actin, which was not observed after AB and LE extract treatments. This suggests that SA extract may affect tenderization. Taken together, our results show that aqueous extract of S. aspratus affects the tenderness of the bovine longissimus dorsi muscle.

Facile Lithium Densification Kinetics by Hyperporous/Hybrid Conductor for High-Energy-Density Lithium Metal Batteries.

Lithium metal anode (LMA) emerges as a promising candidate for lithium (Li)-based battery chemistries with high-energy-density. However, inhomogeneous charge distribution from the unbalanced ion/electron transport causes dendritic Li deposition, leading to "dead Li" and parasitic reactions, particularly at high Li utilization ratios (low negative/positive ratios in full cells). Herein, an innovative LMA structural model deploying a hyperporous/hybrid conductive architecture is proposed on single-walled carbon nanotube film (HCA/C), fabricated through a nonsolvent induced phase separation process. This design integrates ionic polymers with conductive carbon, offering a substantial improvement over traditional metal current collectors by reducing the weight of LMA and enabling high-energy-density batteries. The HCA/C promotes uniform lithium deposition even under rapid charging (up to 5 mA cm-2) owing to its efficient mixed ion/electron conduction pathways. Thus, the HCA/C demonstrates stable cycling for 200 cycles with a low negative/positive ratio of 1.0 when paired with a LiNi0.8Co0.1Mn0.1O2 cathode (areal capacity of 5.0 mAh cm-2). Furthermore, a stacked pouch-type full cell using HCA/C realizes a high energy density of 344 Wh kg-1 cell/951 Wh L-1 cell based on the total mass of the cell, exceeding previously reported pouch-type full cells. This work paves the way for LMA development in high-energy-density Li metal batteries.

Electrolyte Design for High-Voltage Lithium-Metal Batteries with Synthetic Sulfonamide-Based Solvent and Electrochemically Active Additives.

Considering practical viability, Li-metal battery electrolytes should be formulated by tuning solvent composition similar to electrolyte systems for Li-ion batteries to enable the facile salt-dissociation, ion-conduction, and introduction of sacrificial additives for building stable electrode-electrolyte interfaces. Although 1,2-dimethoxyethane with a high-donor number enables the implementation of ionic compounds as effective interface modifiers, its ubiquitous usage is limited by its low-oxidation durability and high-volatility. Regulation of the solvation structure and construction of well-structured interfacial layers ensure the potential strength of electrolytes in both Li-metal and LiNi0.8Co0.1Mn0.1O2 (NCM811). This study reports the build-up of multilayer solid-electrolyte interphase by utilizing different electron-accepting tendencies of lithium difluoro(bisoxalato) phosphate (LiDFBP), lithium nitrate, and synthetic 1-((trifluoromethyl)sulfonyl)piperidine. Furthermore, a well-structured cathode-electrolyte interface from LiDFBP effectively addresses the issues with NCM811. The developed electrolyte based on a framework of highly- and weakly-solvating solvents with interface modifiers enables the operation of Li|NCM811 cells with a high areal capacity cathode (4.3 mAh cm-2) at 4.4 V versus Li/Li+.

Compositionally Sequenced Interfacial Layers for High-Energy Li-Metal Batteries.

Electrolyte additives with multiple functions enable the interfacial engineering of Li-metal batteries (LMBs). Owing to their unique reduction behavior, additives exhibit a high potential for electrode surface modification that increases the reversibility of Li-metal anodes by enabling the development of a hierarchical solid electrolyte interphase (SEI). This study confirms that an adequately designed SEI facilitates the homogeneous supply of Li+, nonlocalized Li deposition, and low electrolyte degradation in LMBs while enduring the volume fluctuation of Li-metal anodes on cycling. An in-depth analysis of interfacial engineering mechanisms reveals that multilayered SEI structures comprising mechanically robust LiF-rich species, electron-rich P-O species, and elastic polymeric species enabled the stable charge and discharge of LMBs. The polymeric outer SEI layer in the as-fabricated multilayered SEI could accommodate the volume fluctuation of Li-metal anodes, significantly enhancing the cycling stability Li||LiNi0.8Co0.1Mn0.1O2 full cells with an electrolyte amount of 3.6 g Ah-1 and an areal capacity of 3.2 mAh cm-2. Therefore, this study confirms the ability of interfacial layers formed by electrolyte additives and fluorinated solvents to advance the performance of LMBs and can open new frontiers in the fabrication of high-performance LMBs through electrolyte-formulation engineering.

Liquid electrolyte chemistries for solid electrolyte interphase construction on silicon and lithium-metal anodes.

Next-generation battery development necessitates the coevolution of liquid electrolyte and electrode chemistries, as their erroneous combinations lead to battery failure. In this regard, priority should be given to the alleviation of the volumetric stress experienced by silicon and lithium-metal anodes during cycling and the mitigation of other problems hindering their commercialization. This review summarizes the advances in sacrificial compound-based volumetric stress-adaptable interfacial engineering, which has primarily driven the development of liquid electrolytes for high-performance lithium batteries. Besides, we discuss how the regulation of lithium-ion solvation structures helps expand the range of electrolyte formulations and thus enhance the quality of solid electrolyte interphases (SEIs), improve lithium-ion desolvation kinetics, and realize longer-lasting SEIs on high-capacity anodes. The presented insights are expected to inspire the design and synthesis of next-generation electrolyte materials and accelerate the development of advanced electrode materials for industrial battery applications.

Correction: Liquid electrolyte chemistries for solid electrolyte interphase construction on silicon and lithium-metal anodes.

[This corrects the article DOI: 10.1039/D3SC03514J.].

Complete genome sequencing of Lactobacillus acidophilus 30SC, isolated from swine intestine.

Lactobacillus acidophilus 30SC has been isolated from swine intestines and considered a probiotic strain for dairy products because of its ability to assimilate cholesterol and produce bacteriocins. Here, we report the complete genome sequence of Lactobacillus acidophilus 30SC (2,078,001 bp) exhibiting strong acid resistance and enhanced bile tolerance.

Radish sprout alleviates DSS-induced colitis via regulation of NF-kB signaling pathway and modifying gut microbiota.

In this study, we investigated the effects of radish sprout ethanol extract (RSE) on inflammatory responses in the macrophages and a mouse model of colitis. RSE administration was found to effectively inhibit the phosphorylation of IκB and, in turn, the production of pro-inflammatory enzymes and cytokines in lipopolysaccharide-stimulated macrophages. In dextran sulfate sodium (DSS)-colitis mice, RSE administration prevented body weight and colon length reduction, while decreasing inflammation and mucosal necrosis. The diversity of the fecal microbiota was significantly increased in the group treated with RSE. In addition, RSE administration decreased the relative abundance of the phylum Proteobacteria, which includes many pathogens, and increased the abundance of the genus Akkermansia. Beta diversity analyses showed that RSE administration restored the gut microbiota composition close to that of healthy mice. For the first time, we identified glycosides of sinapic acid as part of hydroxycinnamic acids in RSE with colitis-alleviating effects. Notably, 1,2-O-disinapoyl glucoside substantially decreased nitric oxide generation in LPS-stimulated macrophages.

Isolation of Lactococcus lactis ssp. cremoris LRCC5306 and Optimization of Diacetyl Production Conditions for Manufacturing Sour Cream.

The sensory properties and flavor of sour cream are important factors that influence consumer acceptability. The present study aimed to isolate lactic acid bacteria with excellent diacetyl production ability and to optimize the fermentation conditions for sour cream manufacture. Lactococcus lactis ssp. cremoris was isolated as a lactic acid bacterium derived from raw milk. This strain showed the greatest diacetyl production among other strains and was named LRCC5306. Various culture conditions were optimized to improve the diacetyl production of LRCC5306. The highest diacetyl production was found to be at 105.04±2.06 mg/L, when 0.2% citric acid and 0.001% Fe2+ were added and cultured at 20°C for 15 h. Based on the optimal cultivation conditions, sour cream was manufactured using LRCC5306, with a viable count of 1.04×108 CFU/g and a diacetyl concentration of 106.56±1.53 mg/g. The electronic tongue system was used to compare the sensory properties of the sour cream; the fermented product exhibited sweetness and saltiness which was similar to that of an imported commercial product, but with slightly reduced bitterness and a significantly greater degree of sour taste. Therefore, our study shows that if cream is fermented using the LRCC5306, it is possible to produce sour cream with greatly improved sensory attractiveness, resulting in increased acceptance by consumers. Since this sour cream has a higher viable count of lactic acid bacteria, it is also anticipated that it will have a better probiotic effect.

Isolation of the Cholesterol-Assimilating Strain Pediococcus acidilactici LRCC5307 and Production of Low-Cholesterol Butter.

The objective of the present study was to evaluate the cholesterol-assimilation ability of lactic acid bacteria (LAB), which were isolated from kimchi, a Korean traditional fermented cabbage. The isolated strain, using modified MRS medium, showed 30.5% cholesterol assimilation activity and was named Pediococcus acidilactici LRCC5307. Types and concentrations of bile were investigated for their effects on increasing the cholesterol assimilation ability of the LRCC5307 strain, a 74.5% decrease in cholesterol was observed when 0.2% bile salts were added. In addition, the manufacture of low-cholesterol butter using LRCC5307 was examined. After fermentation, LRCC5307 with butter showed 8.74 Log CFU/g viable cells, pH 5.43, and a 11% decrease in cholesterol. These results suggest that LRCC5307 could help in the production of healthier butter by decreasing cholesterol and including living LAB.

Dietary Whey Protein Supplementation Increases Immunoglobulin G Production by Affecting Helper T Cell Populations after Antigen Exposure.

Whey protein is a by-product of cheese and casein manufacturing processes. It contains highly bioactive molecules, such as epidermal growth factor, colony-stimulating factor, transforming growth factor-α and -ß, insulin-like growth factor, and fibroblast growth factor. Effects of whey protein on immune responses after antigen (hemagglutinin peptide) injection were evaluated in rats. Experimental diets were formulated based on NIH-31M and supplemented with 1% amino acids mixture (CON) or 1% whey protein concentrate (WPC) to generate isocaloric and isonitrogenous diets. Rats were fed the experimental diets for two weeks and then exposed to antigen two times (Days 0 and 14). Blood was collected on Days 0, 7, 14, and 21 for hematological analysis. The WPC group showed decreased IgA and cytotoxic T cells before the antigen injection (Day 0) but increased IgG, IL-2, and IL-4 after antigen injection due to increased B cells and T cells. Helper T cells were increased at Days 14 and 21, but cytotoxic T cells were not affected by WPC. WPC may activate adaptive immunity (IgG) against antigen by modulating helper T cells. Bioactive molecules might contribute to the immune-enhancing effects of whey protein concentrate.

A Non-yeast Kefir-like Fermented Milk Development with Lactobacillus acidophilus KCNU and Lactobacillus brevis Bmb6.

The use of yeast assist kefir fermentation, but also can cause food spoilage if uncontrolled. Hence, in this study, the microbial composition of an existing commercial kefir starter was modified to produce a functional starter, where Lactobacillus acidophilus KCNU and Lactobacillus brevis Bmb6 were used to replace yeast in the original starter to produce non-yeast kefir-like fermented milk. The functional starter containing L. acidophilus KCNU and L. brevis Bmb6 demonstrated excellent stability with 1010 CFU/g of total viable cells throughout the 12 weeks low-temperature storage. The newly developed functional starter also displayed a similar fermentation efficacy as the yeast-containing control starter, by completing the milk fermentation within 12 h, with a comparable total number of viable cells (108 CFU/mL) in the final products, as in control. Sensory evaluation revealed that the functional starter-fermented milk highly resembled the flavor of the control kefir, with enhanced sourness. Furthermore, oral administration of functional starter-fermented milk significantly improved the disease activity index score by preventing drastic weight-loss and further deterioration of disease symptoms in DSS-induced mice. Altogether, L. acidophilus KCNU and L. brevis Bmb6 have successfully replaced yeast in a commercial starter pack to produce a kefir-like fermented milk beverage with additional health benefits. The outcome of this study provides an insight that the specific role of yeast in the fermentation process could be replaced with suitable probiotic candidates.

Association between the body weight of growing pigs and the functional capacity of their gut microbiota.

Gastrointestinal microbiota impact host's biological activities, including digestion of indigestible feed components, energy harvest, and immunity. In this study, fecal microbiota of high body weight (HW) and low body weight (LW) growing pigs at 103 days of age were compared. Principal coordinates analysis separated the HW and LW groups into two clusters, indicating their potential differences between microbial community composition. Although the abundances of two major phyla, Firmicutes and Bacteroidetes, did not significantly differ between the HW and LW groups, some genera showed significant differences. Among them, Peptococcus and Eubacterium exhibited strong positive correlations with body weight (BW) and average daily gain (ADG) (Rho > 0.40), whereas Treponema, Desulfovibrio, Parabacteroides, and Ruminococcaceae_unclassified exhibited strong negative correlations with BW and ADG (Rho < -0.40). Based on these results, the structure of intestinal microbiota may affect growth traits in pigs through host-microbe interactions. Further in-depth studies will provide insights into how best to reshape host-microbe interactions in pigs and other animals as well.

In Situ Interfacial Tuning To Obtain High-Performance Nickel-Rich Cathodes in Lithium Metal Batteries.

Nickel-rich layered oxides are currently considered the most practical candidates for realizing high-energy-density lithium metal batteries (LMBs) because of their relatively high capacities. However, undesired nickel-rich cathode-electrolyte interactions hinder their applicability. Here, we report a satisfactory combination of an antioxidant fluorinated ether solvent and an ionic additive that can form a stable, robust interfacial structure on the nickel-rich cathode in ether-based electrolytes. The fluorinated ether 1,1,2,2-tetrafluoroethyl-1H,1H,5H-octafluoropentyl ether (TFOFE) introduced as a cosolvent into ether-based electrolytes stabilizes the electrolytes against oxidation at the LiNi0.8Mn0.1Co0.1O2 (NCM811) cathode while simultaneously preserving the electrochemical performance of the Li metal anode. Lithium difluoro(bisoxalato)phosphate (LiDFBP) forms a uniform cathode-electrolyte interphase that limits the generation of microcracks inside secondary particles and undesired dissolution of transition metal ions such as nickel, cobalt, and manganese from the cathode into the electrolyte. Using TFOFE and LiDFBP in ether-based electrolytes provides an excellent capacity retention of 94.5% in a Li|NCM811 cell after 100 cycles and enables the delivery of significantly increased capacity at high charge and discharge rates by manipulating the interfaces of both electrodes. This research provides insights into advancing electrolyte technologies to resolve the interfacial instability of nickel-rich cathodes in LMBs.

Antipathogenic properties of green tea polyphenol epigallocatechin gallate at concentrations below the MIC against enterohemorrhagic Escherichia coli O157:H7.

The inhibitory effects of green tea polyphenol epigallocatechin gallate (EGCG) on virulence phenotypes and gene expression regulated by quorum sensing (QS) in Escherichia coli O157:H7 were demonstrated at concentrations of 1 to 100 microg/ml, which are lower than the MIC (539 +/- 22 microg/ml). At 25 microg/ml, the growth rate was not affected, but autoinducer 2 concentration, biofilm formation, and swarm motility decreased to 13.2, 11.8, and 50%, respectively. Survival at 5 days of nematodes (Caenorhabditis elegans) that were fed the pathogen without and with EGCG were 47.1 and 76%, respectively. Real-time PCR data indicated decreased transcriptional level in many quorum sensing-regulated virulence genes at 25 microg/ml. Our results suggest that EGCG at concentrations below itsMIC has significant antipathogenic effects against E. coli O157:H7.

The role of disulfide bond isomerase A (DsbA) of Escherichia coli O157:H7 in biofilm formation and virulence.

The role of periplasmic disulfide oxidoreductase DsbA in Shiga toxin-producing Escherichia coli O157:H7 (STEC) was investigated. Deletion of dsbA (DeltadsbA) significantly decreased cell motility and alkaline phosphatase activity in STEC. STEC DeltadsbA also showed greater sensitivity to menadione and under low pH conditions. Significant reductions in surface attachment to both biotic (HT-29 epithelial cells) and abiotic (polystyrene and polyvinyl chloride) surfaces were observed in STEC DeltadsbA. In addition, no biofilm formation was detected in STEC DeltadsbA compared to wild-type cells in glass capillary tubes under continuous flow-culture system conditions. In the nematode model Caenorhabditis elegans-killing assay, the deletion of dsbA in STEC resulted in attenuated virulence compared to wild-type cells. STEC DeltadsbA was also found to have a reduced ability to colonize the nematode gut. These results suggest that DsbA plays important roles in biofilm formation and virulence in STEC cells.

Induction of serum amyloid A genes is associated with growth and apoptosis of HC11 mammary epithelial cells.

In this study, we examined the expression and functions of serum amyloid A (SAA) isoforms during apoptosis of HC11 mammary gland epithelial cells. Expression of SAA mRNAs and apoptosis were increased in HC11 cells by serum withdrawal and gradually decreased upon the addition of serum, or epidermal growth factor (EGF). TNFalpha treatment of HC11 cells also induced expression of SAA genes, and the effect on SAA1 and SAA2 expression was suppressed by treatment with MG132, and in cells transfected with a dominant negative mutant form of IkappaBalpha. Similar results were observed in response to interleukin-1 (IL-1), IL-6 and interferon gamma (IFNgamma). Furthermore, overexpression of the SAA1 and SAA2 isoforms suppressed growth and accelerated apoptosis of HC11 cells by increasing caspase 3/7 and caspase 8 activities, but the apoptotic effect of tumor necrosis factor alpha (TNFalpha) on HC11 cells was not enhanced. We found that expression of SAA1 and SAA2, but not SAA3, was regulated by an NFkappaB-dependent pathway, and that overexpression of SAA isoforms accelerated the apoptosis of HC11 cells.

WDNM1 is associated with differentiation and apoptosis of mammary epithelial cells.

In this study, we show that expression of the Westmead DMBA8 nonmetastatic cDNA 1 (WDNM1) gene was increased upon SFM and/or TNFalpha treatment, with a corresponding increase in apoptotic cells, and gradually decreased following re-stimulation with serum in HC11 mammary epithelial cells. TNFalpha induced WDNM1 expression showed the NFkappaB-dependent mechanism since it's expression was abrogated in IkappaBalphaM (super-repressor of NFkappaB)-transfected cells, but not those transfected with control vector. Furthermore, overexpression of WDNM1 suppressed growth and differentiation, and accelerated apoptosis of HC11 cells. Thus, our results demonstrate that WDNM1 gene expression, regulated by the TNFalpha-NFkappaB signal pathway, is associated with HC11 cell apoptosis.

Inhibition of Escherichia coli O157:H7 attachment by interactions between lactic acid bacteria and intestinal epithelial cells.

The intestinal epithelial cell (IEC) layer of the intestinal tract makes direct contact with a number of microbiota communities, including bacteria known to have deleterious health effects. IECs possess innate protective strategies against pathogenic challenge, which primarily involve the formation of a physicochemical barrier. Intestinal tract mucins are principal components of the mucus layer on epithelial surfaces, and perform a protective function against microbial damage. However, little is currently known regarding the interactions between probiotics/pathogens and epithelial cell mucins. The principal objective of this study was to determine the effects of Lactobacillus on the upregulation of MUC2 mucin and the subsequent inhibition of E. coli O157:H7 attachment to epithelial cells. In the current study, the attachment of E. coli O157:H7 to HT-29 intestinal epithelial cells was inhibited significantly by L. acidophilus A4 and its cell extracts. It is also important to note that the expression of MUC2 mucin was increased as the result of the addition of L. acidophilus A4 cell extracts (10.0 mg/ml), which also induced a significant reduction in the degree to which E. coli O157:H7 attached to epithelial cells. In addition, the mRNA levels of IL-8, IL-1beta, and TNF-alpha in HT-29 cells were significantly induced by treatment with L. acidophilus A4 extracts. These results indicate that MUC2 mucin and cytokines are important regulatory factors in the immune systems of the gut, and that selected lactobacilli may be able to induce the upregulation of MUC2 mucin and specific cytokines, thereby inhibiting the attachment of E. coli O157:H7.