The anti-hyperuricemic and gut microbiota regulatory effects of a novel purine assimilatory strain, Lactiplantibacillus plantarum X7022.

PURPOSE: Probiotics have been reported to effectively alleviate hyperuricemia and regulate the gut microbiota. The aim of this work was to study the in vivo anti-hyperuricemic properties and the mechanism of a novel strain, Lactiplantibacillus plantarum X7022. METHODS: Purine content and mRNA expression of purine assimilation related enzymes were determined by HPLC and qPCR, respectively. Hyperuricemic mice were induced by potassium oxonate and hypoxanthine. Uric acid (UA), blood urea nitrogen, creatinine and renal inflammation were examined by kits. The expression of renal UA transporters was subjected to western blotting. Kidney tissues were sectioned for histological analysis. The fecal short-chain fatty acids (SCFAs) were determined by HPLC, and gut microbiota was investigated using the 16S rDNA metagenomic sequencing. RESULTS: L. plantarum X7022 possesses a complete purine assimilation pathway and can exhaust xanthine, guanine, and adenine by 82.1%, 33.1%, and 12.6%, respectively. The strain exhibited gastrointestinal viability as 44% at the dose of 109 CFU/mL in mice. After four-week administration of the strain, a significant decrease of 35.5% in the serum UA level in hyperuricemic mice was achieved. The diminished contents of fecal propionate and butyrate were dramatically boosted. The treatment also alleviated renal inflammation and restored renal damage. The above physiological changes may due to the inhibited xanthine oxidase (XO) activity, as well as the expressional regulation of UA transporters (GLUT9, URAT1 and OAT1) to the normal level. Notably, gut microbiota dysbiosis in hyperuricemic mice was improved with the inflammation and hyperuricemia related flora depressed, and SCFAs production related flora promoted. CONCLUSION: The strain is a promising probiotic strain for ameliorating hyperuricemia.

Surface contacts strongly influence the elasticity and thermal conductivity of silica nanoparticle fibers.

Granular materials are often encountered in science and engineering disciplines, in which controlling the particle contacts is one of the critical issues for the design, engineering, and utilization of their desired properties. The achievable rapid fabrication of nanoparticles with tunable physical and chemical properties facilitates tailoring the macroscopic properties of particle assemblies through contacts at the nanoscale. Models have been developed to predict the mechanical properties of macroscopic granular materials; however, their predicted power in the case of nanoparticle assemblies is still uncertain. Here, we investigate the influence of nanocontacts on the elasticity and thermal conductivity of a granular fiber comprised of close-packed silica nanoparticles. A complete elastic moduli characterization was realized by non-contact and non-destructive Brillouin light spectroscopy, which also allowed resolving the stiffness of the constituent particles in situ. In the framework of effective medium models, the strong enhancement of the elastic moduli is attributed to the formation of adhesive nanocontacts with physical and/or chemical bondings. The nanoparticle contacts are also responsible for the increase in the fiber thermal conductivity that emphasizes the role of interface thermal resistance, which tends to be ignored in most porosity models. This insight into the fundamental understanding of structure-property relationships advances knowledge on the manipulation of granular systems at the nanoscale.

Selective Adsorption of C60 in the Supramolecular Nanopatterns of Donor-Acceptor Porphyrin Derivatives.

The nanostructure of active layers consisting of donor and acceptor molecules is responsible for the separation and transfer processes of charge carriers, which may result in different photoelectric conversion efficiencies of organic photovoltaic cells (OPVCs). Therefore, intensive study on the relationships among nanostructures, intermolecular interactions, and molecular chemical skeletons is necessary for preparing controlled nanostructures of active layers by designing photovoltaic molecules. In this research, the self-assembled nanopatterns of three (DPP-ZnP-E)2-based molecules on highly oriented pyrolytic graphite surface were probed by scanning tunneling microscopy and analyzed by density functional theory calculations. The results indicated that different bridges, diethynylene, diethynylene-dithienyl, and diethynylene-phenylene, in (DPP-ZnP-E)2-based molecules not only made a difference to intermolecular interactions and cooperated with molecule-substrate interactions, consequently affecting the packed nanopattern, but also influenced the adsorption of fullerene acceptors in the nanopatterns of (DPP-ZnP-E)2-based molecules. C60 molecules were found to be selectively adsorbed atop the dithienyl groups of (DPP-ZnP-E)2-2T donor molecules probably by S···π interactions compared with (DPP-ZnP-E)2 or (DPP-ZnP-E)2-Ph molecules. This study on the assembled nanopatterns of the three (DPP-ZnP-E)2-based molecules would be conductive to (DPP-ZnP-E)2-based optoelectronic materials design in OPVCs.

On-Surface Synthesis of Self-Assembled Monolayers of Benzothiazole Derivatives Studied by STM and XPS.

On-surface synthesis has gradually become a prevalent approach to constructing two-dimensional functional monolayers on various substrates. In the present work, the synthesis of self-assembled monolayers (SAMs) of benzothiazole derivatives was conducted at the liquid/solid interface for the first time. Two kinds of nanostructures were achieved on the highly oriented pyrolytic graphite (HOPG) surface via the condensation reaction between aromatic aldehyde derivatives and 2-aminothiophenol (ATP). The formation of thiazole-based self-assemblies was revealed by scanning tunneling microscopy (STM) and further confirmed by X-ray photoelectron spectroscopy (XPS). The successful synthesis of the benzothiazole derivatives not only extends the scope of on-surface reactions but also can be applied in designing multifunctional SAMs at the interface.

Efficient Synthesis of Ir-Polyoxometalate Cluster Using a Continuous Flow Apparatus and STM Investigation of Its Coassembly Behavior on HOPG Surface.

Taking advantage of a continuous-flow apparatus, the iridium(III)-containing polytungstate cluster K12Na2H2[Ir2Cl8P2W20O72]·37H2O (1) was obtained in a reasonable yield (13% based on IrCl3·H2O). Compound 1 was characterized by Fourier transform IR, UV-visible, (31)P NMR, electrospray ionization mass spectrometry (ESI-MS), and thermogravimetric analysis measurements. (31)P NMR, ESI-MS, and elemental analysis all indicated 1 was a new polytungstate cluster compared with the reported K14[(IrCl4)KP2W20O72] compound. Intriguingly, the successful isolation of 1 relied on the custom-built flow apparatus, demonstrating the uniqueness of continuous-flow chemistry to achieve crystalline materials. The catalytic properties of 1 were assessed by investigating the activity on catalyzing the electro-oxidation of ruthenium tris-2,2'-bipyridine [Ru(bpy)3](2+/3+). The voltammetric behavior suggested a coupled catalytic behavior between [Ru(bpy)3](3+/2+) and 1. Furthermore, on the highly oriented pyrolytic graphite surface, 1,3,5-tris(10-carboxydecyloxy) benzene (TCDB) was used as the two-dimensional host network to coassemble cluster 1; the surface morphology was observed by scanning tunneling microscope technique. "S"-shape of 1 was observed, indicating that the cluster could be accommodated in the cavity formed by two TCDB host molecules, leading to a TCDB/cluster binary structure.

Solution concentration controlled self-assembling structure with host-guest recognition at the liquid-solid interface.

In the present investigation, we reported the fabrication of a chicken-wire porous 2D network formed by triphenylene-2,6,10-tricarboxylic acid (H3TTCA) at the liquid-solid interface. When coronene (COR) molecules were added into the system, the H3TTCA honey-comb network was broken and the reconstructed structures of the H3TTCA/COR host-guest systems were subsequently formed. Scanning tunneling microscopic (STM) measurements and density function theory (DFT) calculations were utilized to reveal the structural variety in the co-assembly of H3TTCA/COR controlled by the solution concentration at 1-heptanoic acid/HOPG interface.

Development of a two-step process for production of 3-hydroxypropionic acid from glycerol using Klebsiella pneumoniae and Gluconobacter oxydans.

In this work, a two-step process was developed for the production of 3-hydroxypropionic acid from glycerol. In the first step, glycerol was converted to 1,3-propanediol by Klebsiella pneumonia. In the second step, the 1,3-propanediol was converted into 3-hydroxypropionic acid by Gluconobacter oxydans. In a 7.0 L bioreactor, the whole process took 54 h, consumed 480 g glycerol and produced 242 g 3-hydroxypropionic acid. The conversion rate of glycerol to 3-hydroxypropionic acid was 50.4 % (g g(-1)). The final concentration of 3-hydroxypropionic acid arrived 60.5 g L(-1). The process was effective for 3-HP production from glycerol and it might provide a new approach to the biosynthesis of 3-HP from a cheap starting material. Moreover, in this paper, it was first reported that the by-product of 3-hydroxypropionic acid production from 1,3-propandeiol was acrylic acid.

Characterization, Semirational Design for pH Robustness, and the Application in Bioactive Peptide Production of a X-Prolyl Dipeptidyl Aminopeptidase from Lactococcus lactis MY-3.

PepXLcMY-3, an X-prolyl dipeptidyl aminopeptidase derived from Lactobacillus lactis MY-3, was screened and recombinantly expressed in Escherichia coli. The enzyme could exhibit about 40% activity within the pH range of 6.0-10. To further improve the pH robustness, site E396 located in the active pocket was discovered through alanine scanning. The mutant E396I displayed both developed activity and kcat/Km. The optimal pH of E396I shifted from 6.0 to 10 compared to WT, with the relative activity within the pH range of 6.0-10 significantly increased. The site K648 was then proposed by semirational design. The activity of mutant E396I/K648D reached 4.03 U/mg. The optimal pH was restored to 6.0, and the pH stability was further improved. E396I/K648D could totally hydrolyze ß-casomorphin 7 within 30 min. The hydrolysate showed 64.5% inhibition on angiotensin I converting enzyme, which was more efficient than those produced by E396I and WT, 23.2 and 44.7%, respectively.

[Advances of studies on culture and product functions of Euglena gracilis].

Euglena gracilis is a unicellular eukaryote between animal and plant cells, which is widely distributed in nature. E. gracilis has both plant and animal characteristics, and can grow photoautotrophically, heterotrophically and mixotrophically. E. gracilis also features on abundant and various cellular composition. Recently, extensive researches on unique cellular components of E. gracilis have revealed its application in the field of medicine, food, and feedstuff, in terms of improving immunity, fighting inflammation, and lowering uric acid levels. The application prospects of paramylon in biomedical area were also discovered. As food ingredients, food additives, feedstuffs and cosmetic ingredients, E. gracilis has been certified domestically and overseas. A series of products have been developed overseas, especially in Japan. However, the research and development of E. gracilis are still in its infancy in China, and there is huge space for development. At present, the research and potential application of cultivation and product functions of E. gracilis have been rarely reviewed. This review systematically examines both the domestic and abroad research of cultivation and production of E. gracilis, as well as the biological activity of E. gracilis powder and paramylon. The existing problems in the application, exploitation, and possible development direction of E. gracilis in the future are prospected. This review might be useful for establishing and optimizing large-scale and efficient heterotrophic technology, as well as developing related products of E. gracilis with specific functions.

Lactiplantibacillus plantarum X7022 Plays Roles on Aging Mice with Memory Impairment Induced by D-Galactose Through Restoring Neuronal Damage, Relieving Inflammation and Oxidative Stress.

In this study, Lactiplantibacillus plantarum X7022 was applied to ameliorate memory impairment of aging mice induced by D-galactose. The strain showed specific choloylglycine hydrolysis ability based on in vitro investigation. Morris water maze test showed L. plantarum X7022 administration improved learning ability and spatial memory of aging mice. The gavage of L. plantarum X7022 displayed a promising ability of relieving cerebral oxidative stress and hippocampal inflammatory condition according to the increased GSH level and SOD activity and decreased MDA level, as well as decreased TNF-α, IL-1ß, and IL-6 levels. The intervention with the strain could protect neuron by regulating cell apoptosis and AChE overexpression and inhibiting amyloid-ß deposition, as well as affect neuron functions by regulating CREB-BDNF signaling pathways and iNOS expression. Besides, the strain could improve fecal SCFA contents and increase the abundance of anti-inflammatory and antioxidant-related genera such as Lactobacillus, Akkermansia, and Adlercreutzia. These results suggest that L. plantarum X7022 could be a prospective therapeutic alternative for the improvement of memory impairment among the elderly.

Lactobacillus curieae sp. nov., isolated from stinky tofu brine.

A lactic acid bacterium, strain CCTCC M 2011381(T), isolated from the brine of the traditional Chinese snack, stinky tofu, was studied to determine its taxonomic position. It was a Gram-stain-positive, non-motile, facultatively anaerobic rod-shaped bacterium that did not exhibit catalase activity. The DNA G+C content of the strain was 44.1 % and its peptidoglycan was characterized by the presence of meso-diaminopimelic acid. Levels of 16S rRNA gene sequence similarity between strain CCTCC M 2011381(T) and the most closely related species Lactobacillus senioris JCM 17472(T), Lactobacillus parafarraginis JCM 14109(T) and Lactobacillus diolivorans JCM 12183(T) were 96.5, 96.4 and 96.4 %, respectively. Combined with data from high-resolution genomic markers recA, rpoA and pheS, strain CCTCC M 2011381(T) was classified as representing a novel species. The strain could also be distinguished from other related species of the genus Lactobacillus by its physiological and biochemical characteristics. Based on the phylogenetic, physiological and biochemical data, it is proposed that the new isolate can be classified as representing a novel species of the genus Lactobacillus, for which the name Lactobacillus curieae sp. nov. (type strain CCTCC M 2011381(T) = S1L19(T) = JCM 18524(T)) is proposed.

Dynamics of an SEIR model with media coverage mediated nonlinear infectious force.

Media coverage can greatly impact the spread of infectious diseases. Taking into consideration the impacts of media coverage, we propose an SEIR model with a media coverage mediated nonlinear infection force. For this novel disease model, we identify the basic reproduction number using the next generation matrix method and establish the global threshold results: If the basic reproduction number $ \mathcal{R}_{0} < 1 $, then the disease-free equilibrium $ P_{0} $ is stable, and the disease dies out. If $ \mathcal{R}_{0} > 1 $, then the endemic equilibrium $ P^{*} $ is stable, and the disease persists. Sensitivity analysis indicates that the basic reproduction number $ \mathcal{R}_{0} $ is most sensitive to the population recruitment rate $ \Lambda $ and the disease transmission rate $ \beta _{1} $.

TRIM59 attenuates ox-LDL-induced endothelial cell inflammation, apoptosis, and monocyte adhesion through AnxA2.

Background: Atherosclerosis (AS), a chronic inflammatory vascular disease, is a cause of heart attack and ischemic stroke. Tripartite motif-containing protein 59 (TRIM59), a member of the tripartite motif family, has been reported to be involved in inflammatory diseases. This study was to investigate the role of TRIM59 in oxidized low-density lipoprotein (ox-LDL)-induced endothelial cells and examine the mechanism of TRIM59. Methods: To simulate a cellular model of AS in vitro, varying concentrations of ox-LDL (i.e., 20, 40, 60, 80, and 100 µg/mL) were used to treat the human umbilical vein endothelial cells (HUVECs) for 24 h. The messenger ribonucleic acid (RNA) and protein levels of TRIM59, lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), and annexin 2 (AnxA2) were examined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. The transfection efficacy of overexpression (Ov)-TRIM59 and small-interfering RNA-AnxA2 was examined by RT-qPCR and western blot. Cell counting kit-8 assays, lactate dehydrogenase (LDH) assays, enzyme-linked immunosorbent assays, and terminal-deoxynucleotidyl transferase mediated nick end labeling staining were used to examine viability, LDH expression, inflammation, and apoptosis in HUVECs. The protein levels of B-cell lymphoma 2, Bcl-2-associated X (BAX), cleaved caspase3, intercellular adhesion molecule 1, and vascular cell adhesion molecule 1 were assessed by western blot. Additionally, the adhesion of THP-1 to ox-LDL-induced HUVECs was detected using monocyte adhesion assays and the binding of TRIM59 and AnxA2 was verified by co-immunoprecipitation. Results: This study showed that TRIM59 expression was decreased in the ox-LDL-induced HUVECs while LOX-1 expression was increased. After transfection with Ov-TRIM59, TRIM59 in ox-LDL-induced HUVECs was increased, and TRIM59 overexpression alleviated the viability damage, inflammation, and apoptosis of the ox-LDL-induced HUVECs. In addition, THP-1 adhesion to the ox-LDL-induced HUVECs was also suppressed by TRIM59 overexpression. This study also showed that TRIM59 could bind to AnxA2 and promote AnxA2 expression in ox-LDL-stimulated HUVECs. Moreover, the rescue experiments revealed that TRIM59 suppressed the viability damage, inflammation, apoptosis, and monocyte adhesion of the ox-LDL-induced HUVECs via AnxA2. Conclusions: TRIM59 protected against ox-LDL-induced AS by binding to AnxA2.

Anti-Hyperuricemic, Nephroprotective, and Gut Microbiota Regulative Effects of Separated Hydrolysate of α-Lactalbumin on Potassium Oxonate- and Hypoxanthine-Induced Hyperuricemic Mice.

SCOPE: This study aims to investigate the anti-hyperuricemic and nephroprotective effects and the potential mechanisms of the separated gastrointestinal hydrolysates of α-lactalbumin on hyperuricemic mice. METHODS AND RESULTS: The gastrointestinal hydrolysate of α-lactalbumin, the hydrolysate fraction with molecular weight (MW) < 3 kDa (LH-3k), and the fragments with smallest MW among LH-3K harvested through dextran gel chromatography (F5) are used. Hyperuricemia mice are induced via daily oral gavage of potassium oxonate and hypoxanthine. F5 displays the highest in vitro xanthine oxidase (XO) inhibition among all the fractions separated from LH-3k. Oral administration of F5 significantly reduces the levels of serum uric acid (UA), creatinine, and urea nitrogen. F5 treatment could ameliorate kidney injury through alleviating oxidative stress and inflammation. F5 alleviates hyperuricemia in mice by inhibiting hepatic XO activity and regulating the expression of renal urate transporters. Gut microbiota analysis illustrates that F5 administration increases the abundance of some SCFAs producers, and inhibits the growth of hyperuricemia and inflammation associated genera. LH-3k exhibits similar effects but does not show significance as those of the F5 fraction. CONCLUSION: The anti-hyperuricemia and nephroprotective functions of F5 are mediated by inhibiting hepatic XO activity, ameliorating oxidative stress and inflammation, regulating renal urate transporters, and modulating the gut microbiota in hyperuricemic mice.

[Cloning and expression of key genes of butanol synthetic pathway in Escherichia coli].

OBJECTIVE: We constructed a recombinant Escherichia coli strain for butanol production by cloning the cDNA sequence of the key butanol synthetic pathway genes from Clostridium acetobutylicum ATCC824. METHODS: We amplified the genes of thil, adhE2 and BCS operon by PCR with C. acetobutylicum ATCC824 genome as a template. We constructed the recombinant strain E. coli pBAT (BCS operon-adhE2-thil/pTrc99a/MG1655). We used 0.1 mmol/l Isopropyl beta-D-thiogalactopyranoside (IPTG) to induce the recombinant E. coli pBAT for 5 h for recombinant protein expression. We measured acetyl-CoA acetyltransferase (THL), beta-hydroxybutyryl-CoA dehydrogenase (HBD), 3-hydroxybutyryl-CoA dehydratase (CRT), butyryl-CoA dehydrogenase (BCD) and butyraldehyde dehydrogenase (BYDH)/butanol dehydrogenase (BDH) activities in E. coli MG1655 and E. coli pBAT. The fermentation of E. coli pBAT was done in flask in aerobic, micro-aerobic and anaerobic mode separately. RESULTS: In the recombinant E. coli pBAT, THL activity was 0.160 U/mg protein, about 30 times higher than that of E. coli MG1655. HBD activity was 5 times higher than that of E. coli MG1655. CRT activity was 1.53 U/mg protein whereas not detectable in E. coli MG1655. BCD activity was about 32 times higher than that of E. coli MG1655. In addition, the results show that n-butanol could be produced under anaerobic and micro-aerobic conditions. The maximum n-buntanol concentration of 84 mg/l was detected in cultivation broth. CONCLUSION: The key genes of butanol synthetic pathway were expressed in E. coli and the recombinant strains would offer an alternative strategy for butanol biosynthesis.

The effects of angiotensin I-converting enzyme inhibitory peptide VGINYW and the hydrolysate of α-lactalbumin on blood pressure, oxidative stress and gut microbiota of spontaneously hypertensive rats.

VGINYW is a highly active angiotensin I-converting enzyme (ACE) inhibitory peptide discovered from α-lactalbumin by an in vitro-in silico high throughput screening strategy. The aim of this study was to evaluate the antihypertensive effect of the peptide and the α-lactalbumin hydrolysates under 3 kDa (LH-3k), and illustrate the possible mechanism in spontaneously hypertensive rats (SHRs). SHRs were administered with VGINYW and LH-3k at doses of 5 mg per kg BW and 100 mg per kg BW, respectively. VGINYW and LH-3k could markedly decrease the systolic blood pressure (SBP) of the SHRs, and the maximal drops of 21 mmHg (2 h after administration) and 17 mmHg (4 h after administration) were achieved during the 8 hour test, respectively. When the agents were given once per day for 4 weeks, they caused a long-term decrease of 16 mmHg of SBP. VGINYW and LH-3k control the blood pressure through regulating the renin-angiotensin system by inhibiting the ACE activity and diminishing the angiotensin II level, and further upregulating the expression levels of the angiotensin-converting enzyme 2 and angiotensin type 2 receptor, and downregulating the expression of the angiotensin type 1 receptor. VGINYW and LH-3k could notably ameliorate the oxidative stress in the SHR as well. It is more important that the gavage of VGINYW and LH-3k could alleviate hypertension-associated intestinal microbiota dysbiosis by recovering the diversity of the gut microbiota and altering the key floras which are short chain fatty acid producers. In conclusion, VGINYW and LH-3k are effective functional ingredients for blood pressure control.

Metabolism Characteristics of Lactic Acid Bacteria and the Expanding Applications in Food Industry.

Lactic acid bacteria are a kind of microorganisms that can ferment carbohydrates to produce lactic acid, and are currently widely used in the fermented food industry. In recent years, with the excellent role of lactic acid bacteria in the food industry and probiotic functions, their microbial metabolic characteristics have also attracted more attention. Lactic acid bacteria can decompose macromolecular substances in food, including degradation of indigestible polysaccharides and transformation of undesirable flavor substances. Meanwhile, they can also produce a variety of products including short-chain fatty acids, amines, bacteriocins, vitamins and exopolysaccharides during metabolism. Based on the above-mentioned metabolic characteristics, lactic acid bacteria have shown a variety of expanded applications in the food industry. On the one hand, they are used to improve the flavor of fermented foods, increase the nutrition of foods, reduce harmful substances, increase shelf life, and so on. On the other hand, they can be used as probiotics to promote health in the body. This article reviews and prospects the important metabolites in the expanded application of lactic acid bacteria from the perspective of bioengineering and biotechnology.

Guanosine ameliorates positive symptoms of schizophrenia via modulating 5-HT1A and 5-HT2A receptors.

Schizophrenia is a serious mental disorder characterized by hallucinations, delusions, and extremely disordered thinking and behavior. There are several hypotheses of pathogenesis in schizophrenia: dopaminergic, glutamatergic, or serotonergic hyperfunction. Guanosine reportedly protects the central nervous system by modulating the glutamatergic system. Thus, we assumed that guanosine may exert a positive effect on the pathophysiology of schizophrenia. Herein, we demonstrated that guanosine significantly reduced MK-801-induced hyperlocomotion and stereotyped behaviors, but showed no effect on hyperlocomotion induced by d-amphetamine, indicating that guanosine may directly affect the glutamatergic system. Guanosine dose-dependently reduced 5-HTP-induced wet dog shakes (WDS) and other serotonin syndromes (SS) behaviors, indicating that it might block serotonin 5-HT1A or 5-HT2A receptors. Finally, we confirm that that guanosine modulates serotonin 5-HT1A and 5-HT2A receptors and it might be anti-schizophrenic partly through pertussis toxin-sensitive Gi/o-coupled PI3K/Akt signaling. Collectively, this study provides possible compounds and mechanisms for therapeutic effects on schizophrenia.

Combination of a fluorescent dye and a Zn-S cluster and its biological application as a stain for bacteria.

An ionic-pair charge-transfer salt [C(15)H(16)N(3)](+)[Zn(8)S(SC(6)H(5))(15).H(2)O](-) (1) featuring a fluorescent dye and a wurtzite-like octanuclear Zn-S cluster shows high stability when staining bacteria Escherichia coli, Salmonella typhimurium, and Clostridium novyi NT.