Glia-derived adenosine in the ventral hippocampus drives pain-related anxiodepression in a mouse model resembling trigeminal neuralgia.

Glial activation and dysregulation of adenosine triphosphate (ATP)/adenosine are involved in the neuropathology of several neuropsychiatric illnesses. The ventral hippocampus (vHPC) has attracted considerable attention in relation to its role in emotional regulation. However, it is not yet clear how vHPC glia and their derived adenosine regulate the anxiodepressive-like consequences of chronic pain. Here, we report that chronic cheek pain elevates vHPC extracellular ATP/adenosine in a mouse model resembling trigeminal neuralgia (rTN), which mediates pain-related anxiodepression, through a mechanism that involves synergistic effects of astrocytes and microglia. We found that rTN resulted in robust activation of astrocytes and microglia in the CA1 area of the vHPC (vCA1). Genetic or pharmacological inhibition of astrocytes and connexin 43, a hemichannel mainly distributed in astrocytes, completely attenuated rTN-induced extracellular ATP/adenosine elevation and anxiodepressive-like behaviors. Moreover, inhibiting microglia and CD39, an enzyme primarily expressed in microglia that degrades ATP into adenosine, significantly suppressed the increase in extracellular adenosine and anxiodepressive-like behaviors. Blockade of the adenosine A2A receptor (A2AR) alleviated rTN-induced anxiodepressive-like behaviors. Furthermore, interleukin (IL)-17A, a pro-inflammatory cytokine probably released by activated microglia, markedly increased intracellular calcium in vCA1 astrocytes and triggered ATP/adenosine release. The astrocytic metabolic inhibitor fluorocitrate and the CD39 inhibitor ARL 67156, attenuated IL-17A-induced increases in extracellular ATP and adenosine, respectively. In addition, astrocytes, microglia, CD39, and A2AR inhibitors all reversed rTN-induced hyperexcitability of pyramidal neurons in the vCA1. Taken together, these findings suggest that activation of astrocytes and microglia in the vCA1 increases extracellular adenosine, which leads to pain-related anxiodepression via A2AR activation. Approaches targeting astrocytes, microglia, and adenosine signaling may serve as novel therapies for pain-related anxiety and depression.

Reducing Dielectric Confinement Effect Enhances Carrier Separation in Two-dimensional Hybrid Perovskite Photocatalysts.

Two-dimensional organic-inorganic hybrid perovskites ( OIHPs) with alternating structure of the organic and inorganic layers have a natural quantum well structure. The difference of dielectric constants between organic and inorganic layers in this structure results in the enhancement of dielectric confinement effect, which exhibits a large exciton binding energy and hinders the separation of electron-hole pairs. Herein, a strategy to reduce the dielectric confinement effect by narrowing the dielectric difference between organic amine molecule and [PbBr6]4- octahedron is put forward. The Ethanolamine (EOA) contains hydroxyl groups, resulting in the positive and negative charge centers of O and H non-overlapping,which generated a larger polarity and dielectric constant. The reduced dielectric constant produces a smaller exciton binding energy (71.03 meV) of (C2H7NO)2PbBr4 ((EOA)2PbBr4) than (C8H11N)2PbBr4 ((PEA)2PbBr4 (156.07 meV), and promotes the dissociation of electrons and holes. The increasing of lifetime of photogenerated carrier in (EOA)2PbBr4 are proved by femtosecond transient absorption spectra. DFT calculations have also indicated that the small energy shift of the total density of states (DOS) between the C/H/N and the Pb/Br in (EOA)2PbBr4 favors the separation of electrons and holes. In addition, this work demonstrates the application of (PEA)2PbBr4 and (EOA)2PbBr4 in the field of photocatalytic CO2 reduction.

Hierarchical NiCo@NiOOH@CoMoO4 Core-Shell Heterostructure on Carbon Cloth for High-Performance Asymmetric Supercapacitors.

The fabrication of low-cost, effective, and highly integrated nanostructured materials through simple and reproducible methods for high-energy-density supercapacitors is highly desirable. Herein, an activated carbon cloth (ACC) is designed as the functional scaffold for supercapacitors and treated hydrothermally to deposit NiCo nanoneedles working as internal core, followed by a dip-dry coating of NiOOH nanoflakes core-shell and uniform hydrothermal deposition of CoMoO4 nanosheets serving as an external shell. The structured core-shell heterostructure ACC@NiCo@NiOOH@CoMoO4 electrode resulted in exceptional specific areal capacitance of 2920 mF cm-2 and exceptional cycling stability for 10 000 cycles. Moreover, the fabricated electrode is developed into an asymmetric supercapacitor which demonstrates excellent areal capacitance, energy density, and power density within the broad potential window of 1.7 V with a cycling life of 92.4% after 10 000 charge-discharge cycles, which reflects excellent cycle life. The distinctive core-shell structure, highly conductive substrate, and synergetic effect of coated material results in more electrochemical active sites and flanges for effective electrons and ion transportation. This unique technique provides a new perspective for cost-efficient supercapacitor applications.

Huangqi Jianzhong Tang treats chronic atrophic gastritis rats by regulating intestinal flora and conjugated bile acid metabolism.

Huangqi Jianzhong Tang (HQJZ) is effective for treating chronic atrophic gastritis (CAG). The present study was carried out to reveal the mechanism of HQJZ in CAG rats. The metabolism and microbial composition of the cecal contents in CAG rats were analyzed through the integration of an untargeted metabolomic approach using ultra-high-performance liquid chromatography coupled with the quadrupole-time of flight mass spectrometry (UHPLC-QTOF-MS) and 16S rRNA gene sequencing, respectively. Finally, MetOrigin analyses were performed to explore the relationship between differential metabolites and intestinal flora. The results showed that HQJZ could significantly regulate metabolic disorders, especially conjugated acid metabolites. 16S rRNA gene sequencing analysis illustrated that HQJZ decreased the abundance of Acetobacter, Desulfovibrio, Escherichia, and Shigella. MetOrigin metabolite traceability analysis showed that the six bile acids associated with HQJZ efficacy included three bacteria-host cometabolites, which were involved in the primary bile acid biosynthesis pathway. Research presented here confirmed that conjugated bile acid metabolism was key to the treatment of CAG by HQJZ and correlates strongly with Bacteroides acidifaciens and Prevotella copri. These findings provide new insights into the mechanisms to explain the efficacy of HQJZ.

Clinical Efficiency of Fixed-Bearing Unicompartmental Knee Arthroplasty Versus Total Knee Arthroplasty For Lateral Compartment Knee Osteoarthritis and the Effect on Recovery of Motor Function.

Objective: To evaluate the clinical efficiency of fixed-bearing unicompartmental knee arthroplasty (UKA) versus total knee arthroplasty (TKA) for lateral compartment knee osteoarthritis and the effect on the recovery of motor function. Methods: A total of 54 patients who underwent surgery for lateral compartment knee osteoarthritis and satisfied the inclusion criteria from September 2018 to February 2021 at our hospital were recruited and assigned to receive either UKA (UKA group, n=30) or TKA (TKA group, n=24) via random number table method. Among them, the randomization was carried out using an online web-based randomization tool (freely available at http://www.randomizer.org/). Inclusion criteria: 1) patients with lateral compartment knee osteoarthritis diagnosed by clinically relevant tests; 2) patients with structural and functional integrity of the knee ligaments; 3) all with a single knee lesion. Outcome measures included operative time, the reduction ratio of Hb 1d postoperatively, visual analog scale (VAS) score 7d postoperatively, length of hospital stay, postoperative Keen society score (KSS), Oxford knee score (OKS), range of motion (ROM), forgotten joint score (FJS), motor function recovery, and adverse events. Results: All patients were followed up postoperatively for 12-33 (21.71±7.45) months. Patients in the UKA group showed significantly shorter operative indices, a lower reduction ratio of Hb 1d postoperatively, and VAS scores of 7d postoperatively (P < .05). At 1 month and 6 months postoperatively, UKA resulted in significantly better KSS scores, OKS scores, ROM, and motor function recovery versus TKA (P < .05), while the difference of the above indices did not come up to the statistical standard at 1 year postoperatively (P < .05). At 1 year postoperatively, patients receiving UKA were associated with significantly higher FJS scores versus those given TKA. No documented thrombosis, knee, or prosthesis-related adverse events were observed during hospitalization and follow-up. Conclusion: In comparison to TKA, UKA resulted in smaller surgical incisions, improved postoperative healing, and greater restoration of knee function. Both arthroplasties are successful in alleviating pain and increasing knee function, although they are less effective in recovering patients' motor capabilities.

The bZIP Transcription Factors in Current Jasmine Genomes: Identification, Characterization, Evolution and Expressions.

Jasmine, a recently domesticated shrub, is renowned for its use as a key ingredient in floral tea and its captivating fragrance, showcasing significant ornamental and economic value. When cultivated to subtropical zone, a significant abiotic stress adaptability occurs among different jasmine varieties, leading to huge flower production changes and plantlet survival. The bZIP transcription factors (TFs) are reported to play indispensable roles in abiotic stress tolerance. Here, we performed a genome-level comparison of bZIPs using three-type jasmine genomes. Based on their physicochemical properties, conserved motif analysis and phylogenetic analysis, about 63 bZIP genes were identified and clustered in jasmine genomes, noting a difference of one member compared to the other two types of jasmines. The HTbZIP genes were categorized into 12 subfamilies compared with A. thaliana. In cis-acting element analysis, all genes contained light-responsive elements. The abscisic acid response element (ABRE) was the most abundant in HTbZIP62 promoter, followed by HTbZIP33. Tissue-specific genes of the bZIPs may play a crucial role in regulating the development of jasmine organs and tissues, with HTbZIP36 showing the most significant expressions in roots. Combined with complicated protein interactions, HTbZIP62 and HTbZIP33 might play a crucial role in the ABA signaling pathway and stress tolerance. Combined with RT-qPCR analysis, SJbZIP37/57/62 were more sensitive to ABA response genes compared with other bZIPs in DJ amd HT genomes. Our findings provide a useful resource for further research on the regulation of key genes to improve abiotic stress tolerance in jasmine.

Heterogeneously integrated InGaN-based green microdisk light-emitters on Si (100).

Heterogeneous integration of nitrides on Si (100) is expected to open the door to the new possibilities for this material system in the fields of high-speed integrated photonics and information processing. In this work, GaN epitaxial layer grown on the patterned sapphire substrate is transferred onto Si (100) by a combination of wafer bonding, laser lift-off and chemical mechanical polishing (CMP) processes. The GaN epilayer transferred is uniformly thinned down to 800 nm with a root mean square surface roughness as low as 2.33 Å. The residual stress within the InGaN quantum wells transferred is mitigated by 79.4% after the CMP process. Accordingly, its emission wavelength exhibits a blue shift of 8.8 nm, revealing an alleviated quantum-confined Stark effect. Based on this platform, an array of microcavities with diverse geometrics and sizes are fabricated, by which optically-pumped green lasing at ∼505.8 nm is achieved with a linewidth of ∼0.48 nm from ∼12 µm microdisks. A spontaneous emission coupling factor of around 10-4 is roughly estimated based on the light output characteristics with increasing the pumping densities. Lasing behaviors beyond the threshold suggest that the microdisk suffers less thermal effects as compared to its undercut counterparts. The electrically-injected microdisks are also fabricated, with a turn-on voltage of ∼2.0 V and a leakage current as low as ∼2.4 pA at -5 V. Being compatible with traditional semiconductor processing techniques, this work provides a feasible solution to fabricate large-area heterogeneously integrated optoelectronic devices based on nitrides.

Relationships between social capital, patient empowerment, and self-management of patients undergoing hemodialysis: a cross-sectional study.

BACKGROUND: Hemodialysis is the most common treatment of end-stage renal disease. However, it is associated with a range of symptoms affecting patients' daily activities and quality of life. Effective self-management has proven crucial for the alleviation of symptoms. According to Social Cognitive Theory, social capital and patient empowerment may be important variables for predicting self-management. To date, few studies have explored the mechanisms underlying these results. The study aimed to verify whether patient empowerment mediated the effect of social capital on the self-management of hemodialysis patients. METHODS: The study was performed with 245 hemodialysis patients from January 2021 to April 2021 in Taiyuan, China. Demographic and clinical characteristics, social capital, patient empowerment, and self-management of patients undergoing hemodialysis were measured with a self-reported questionnaire. Descriptive statistics were used to summarize the participants' demographic and clinical characteristics, and bootstrapping tests were used to verify whether patient empowerment mediated the association of social capital with self-management in patients undergoing hemodialysis. RESULTS: Mediation analysis indicated that social capital and patient empowerment significantly predicted self-management. Patient empowerment partially mediated the relationship between social capital and self-management in hemodialysis patients. CONCLUSIONS: The results suggest that hemodialysis patients show relatively poor self-management and that patient empowerment mediates both social capital and self-management. Strategies to mobilize patients' social networks and help them identify and utilize effective social resources may provide useful information regarding the implementation of optimal health management for their disease.

Association of plasma bone morphogenetic protein-4 levels with arterial stiffness in hypertensive patients.

BACKGROUND: Arterial stiffness interacts with hypertension, becoming an early marker of hypertension-mediated target organ damage. This study aimed to assess the association between plasma concentrations of bone morphogenetic protein-4 (BMP-4) and arterial stiffness during hypertension. METHODS: Using cardio-ankle vascular index (CAVI) to determine arterial stiffness status, 204 individuals with essential hypertension were classified into two groups, high CAVI (abnormal) group (n = 94) and low (normal) CAVI group (n = 110). Data were collected including clinical characteristics and laboratory measurements. Plasma levels of BMP-4 were tested by using ELISA analysis. RESULTS: Plasma levels of BMP-4 were substantially greater in high CAVI group than that in low CAVI group [38.51 (31.79-50.83) pg/mL vs. 31.15 (29.38-32.37) pg/mL; p < 0.001]. As shown by spearman correlation analysis, BMP-4 concentrations were correlated with CAVI values in hypertensive individuals (r = 0.406, p < 0.001). After adjustment for potential confounders, elevated BMP-4 levels were related with high CAVI (OR, 1.070; 95% CI, 1.003-1.108; p < 0.001). The best BMP-4 cutoff value for identifying high CAVI, as determined by ROC curve analysis, was 33.34 pg/mL (AUC, 0.751; 95% CI, 0.683-0.818; p < 0.001). CONCLUSION: Plasma levels of BMP-4 are increased in hypertensive individuals with high CAVI. Elevated BMP-4 levels are strongly correlated with higher CAVI values, implying a predictive value of BMP-4 in arterial stiffness during hypertension.

Urine Tenofovir Levels Measured Using a Novel Immunoassay Predict Human Immunodeficiency Virus Protection.

New tools are needed to support pre-exposure prophylaxis (PrEP) adherence for human immunodeficiency virus (HIV) prevention, including those that enable real-time feedback. In a large, completed PrEP trial, adequate urine tenofovir levels measured using a novel immunoassay predicted HIV protection and showed good sensitivity and specificity for detectable plasma tenofovir.

Competitive aptasensor for the ultrasensitive multiplexed detection of cancer biomarkers by fluorescent nanoparticle counting.

Cancer biomarker quantification in human serum is of great importance for accurate patient diagnosis and informed clinical management. To date, ultrasensitive multiplexed detection of proteins without amplification is still a major challenge. Herein, we proposed a competitive aptasensor strategy for ultrasensitive multiplexed cancer biomarker detection by fluorescent nanoparticle (FNP) counting. The sequences are designed such that the binding abilities of linker DNA (L-DNA) with DNA-functionalized FNPs (DNA-FNPs) and aptamer are comparable. As long as one target binds with one molecule of aptamer, a signalling FNP forms a sandwich-structured nanocomposite, which was subsequently observed and enumerated with a fluorescence microscope. This 1 : 1 target-to-signal FNP production assured an improved sensitivity, benefiting from the reasonably good brightness and photostability of FNPs. For both singleplexed and multiplexed detection, this proposed strategy achieved an approximately 1000-fold improved limit of detection than the conventional method with the detection volume of 3.2 µL. Notably, the results for carcinoembryonic antigen (CEA) detection obtained directly from 9 human serum samples (colorectal/lung/healthy individuals) were consistent with that obtained by ELISA, showing potential application in clinical diagnosis.

Global transcriptomic analysis of Lactobacillus plantarum CAUH2 in response to hydrogen peroxide stress.

To overcome the deleterious effects of hydrogen peroxide, Lactobacillus plantarum elicits an adaptive response to oxidative stress. In this study, global transcriptomic analysis revealed that L. plantarum CAUH2 expanded its carbon source utilizing profile and enhanced glycolysis to produce more ATP to confront with H2O2 stress. Some antioxidant enzymes including NADH peroxidase, thioredoxin reductase and glutathione peroxidase were 6.11, 36.76 and 6.23-fold up-regulated at transcription level for H2O2 scavenging. Meanwhile, free ferrous iron (Fe2+) was maintained at low concentrations in the cytoplasm, which could limit Fenton reaction and reduce the production of hydroxyl radicals. To repair DNA lesion caused by H2O2, both base excision repair system and recombinational DNA repair pathway were employed by L. plantarum CAUH2. In addition, the expression of methionine sulfoxide reductases and thioredoxin were up-regulated to repair oxidized proteins. It is noteworthy that some transcriptional regulators (Spx, CcpA and MarR1) were predicted to participate in the adaptive response to H2O2 stress, suggesting that L. plantarum CAUH2 utilized a wide array of sensors to monitor oxidative stress and modulated the transcriptional regulation network under H2O2 stress. These findings provide novel insight into the protective mechanisms developed by L. plantarum to cope with oxidative stress.

The MarR Family Regulator BmrR Is Involved in Bile Tolerance of Bifidobacterium longum BBMN68 via Controlling the Expression of an ABC Transporter.

In order to colonize the human gastrointestinal tract and exert their beneficial effects, bifidobacteria must effectively cope with toxic bile salts in the intestine; however, the molecular mechanism underlying bile tolerance is poorly understood. In this study, heterologous expression of a MarR family transcriptional regulator, BmrR, significantly reduced the ox bile resistance of Lactococcus lactis NZ9000, suggesting that BmrR might play a role in the bile stress response. In silico analysis combined with reverse transcription-PCR assays demonstrated that bmrR was cotranscribed with bmrA and bmrB, which encoded multidrug resistance (MDR) ABC transporters. Promoter prediction and electrophoretic mobility shift assays revealed that BmrR could autoregulate the bmrRAB operon by binding to the bmr box (ATTGTTG-6nt-CAACAAT) in the promoter region. Moreover, heterologous expression of bmrA and bmrB in L. lactis yielded 20.77-fold higher tolerance to 0.10% ox bile, compared to the wild-type strain. In addition, ox bile could disrupt the DNA binding activity of BmrR as a ligand. Taken together, our findings indicate that the bmrRAB operon is autoregulated by the transcriptional regulator BmrR and ox bile serves as an inducer to activate the bile efflux transporter BmrAB in response to bile stress in Bifidobacterium longum BBMN68.IMPORTANCE Bifidobacteria are natural inhabitants of the human intestinal tract. Some bifidobacterial strains are used as probiotics in fermented dairy production because of their health-promoting effects. Following consumption, bifidobacteria colonize the lower intestinal tract, where the concentrations of bile salts remain nearly 0.05% to 2.0%. Bile salts, as detergent-like antimicrobial compounds, can cause cellular membrane disruption, protein misfolding, and DNA damage. Therefore, tolerance to physiological bile stress is indeed essential for bifidobacteria to survive and to exert probiotic effects in the gastrointestinal tract. In B. longum BBMN68, the MarR-type regulator BmrR was involved in the bile stress response by autoregulating the bmrRAB operon, and ox bile as an inducer could increase the expression of the BmrAB transporter to enhance the bile tolerance of BBMN68. Our study represents a functional analysis of the bmrRAB operon in the bile stress response, which will provide new insights into bile tolerance mechanisms in Bifidobacterium and other bacteria.

Thromboembolic risks associated with paroxysmal and persistent atrial fibrillation in Asian patients: a report from the Chinese atrial fibrillation registry.

BACKGROUND: Several studies have reported on atrial fibrillation (AF) outcomes, including thromboembolism in patients with paroxysmal and non-paroxysmal AF; however the findings still remain controversial on whether risks differ between these clinical subtypes and limited data are available in Asian cohorts. METHODS: We compared the risk of thromboembolism between paroxysmal and persistent AF patients, in a large contemporary Chinese cohort study. A total of 8529 non-valvular atrial fibrillation (NVAF) patients from the Chinese Atrial Fibrillation Registry (CAFR) study were enrolled. The study subjects were divided into two groups: paroxysmal AF (PaAF, defined as AF lasting within 7 days, n = 4642) and persistent AF (PeAF, lasting over 7 days, n = 3887) groups. RESULTS: In non-anticoagulated patients, PeAF group demonstrated a higher risk of stroke, all-cause death, cardiac/ non-cardiac death and composition of stroke/ transient ischemic attack (TIA)/peripheral thromboembolism (PT)/all-cause death, compared to the PaAF group. No significant difference was found in anticoagulated subjects. On multivariate analysis in non-anticoagulated patients, age ≥ 75 years (P = 0.046) and prior stroke/TIA/PT (P = 0.018) but not AF type (P = 0.63) were significantly associated with the risk of stroke/TIA/PT events. CONCLUSIONS: Stroke, all-cause death and cardiac/non-cardiac death in Chinese NVAF population was increased in non-anticoagulated PeAF patients compared with PaAF group, but same between anticoagulated PeAF and PaAF patients. After adjustment, AF type was not an independent predictor of thromboembolism in NVAF patients. CLINICAL TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR-OCH-13003729. Registered 22 October 2013.

ZmSTK1 and ZmSTK2, encoding receptor-like cytoplasmic kinase, are involved in maize pollen development with additive effect.

Pollen germination and pollen tube growth are important physiological processes of sexual reproduction of plants and also are involved in signal transduction. Our previous study reveals that ZmSTK1 and ZmSTK2 are two receptor-like cytoplasmic kinases (RLCK) homologs in Zea mays as members of receptor-like protein kinase (RLK) subfamily, sharing 86% identity at the amino acid level. Here, we report that ZmSTK1 and ZmSTK2, expressed at late stages of pollen development, regulate maize pollen development with additive effect. ZmSTK1 or ZmSTK2 mutation exhibited severe pollen transmission deficiency, which thus influenced pollen fertility. Moreover, the kinase domains of ZmSTKs were cross-interacted with C-terminus of enolases detected by co-immunoprecipitation (Co-IP) and yeast two-hybrid system (Y2H), respectively. Further, the detective ZmSTK1 or ZmSTK2 was associated with decreased activity of enolases and also reduced downstream metabolite contents, which enolases are involved in glycolytic pathway, such as phosphoenolpyruvate (PEP), pyruvate, ADP/ATP, starch, glucose, sucrose and fructose. This study reveals that ZmSTK1 and ZmSTK2 regulate maize pollen development and indirectly participate in glycolytic pathway.

Isolation and characterization of a novel pollen-specific promoter in maize (Zea mays L.).

ZmSTK2_USP, located on the long arm of chromosome 4, belongs to the serine/threonine kinase gene in maize. The sequence analysis of 2100 bp upstream from the start codon ATG has shown that it contains cis-element motifs and two types of anther/pollen-specific promoter elements (GTGA and AGAAA), suggesting that it is the pollen-specific promoter. To investigate the function of ZmSTK2_USP promoter, the GUS gene fusion system was employed. In proZmSTK2_USP-GUS genetically modified plants, GUS activity was detected in mature pollen grains and pollen tubes but not found in other floral and vegetative tissues. These results show that proZmSTK2_USP is the pollen-specific promoter and drives pollen-specific activity during the middle stage of pollen development until pollen maturation.

Preparation and characterization of monodisperse molecularly imprinted polymers for the recognition and enrichment of oleanolic acid.

Monodisperse molecularly imprinted polymers for oleanolic acid were successfully prepared by a precipitation polymerization method using oleanolic acid as a template, methacrylic acid as a functional monomer, and divinylbenzene/ethylene glycol dimethacrylate as a crosslinker in a mixture of acetonitrile and ethanol (3:1, v/v). The imprinted polymers and nonimprinted polymers were characterized by using scanning electron microscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. The resulting imprinted polymers had average diameters of 3.15 µm and monodispersity values of 1.024. The results clearly demonstrate that use of ethanol as a cosolvent is indeed exceedingly effective in promoting the dissolution of oleanolic acid and in obtaining uniform microspheres. Molecular recognition properties and binding capability to oleanolic acid were evaluated by adsorption testing, which indicated that the imprinted polymers displayed optimal binding performance with a maximum adsorption capacity of 17.3 mg/g and a binding saturation time of 80 min. Meanwhile, the produced imprinted polymers exhibited higher selectivity to oleanolic acid than that for ursolic acid and rhein. Herein, the studies can provide theoretical and experimental references for the oleanolic acid molecular imprinted system.

Integrated transcriptomic and proteomic analysis of the bile stress response in a centenarian-originated probiotic Bifidobacterium longum BBMN68.

Bifidobacteria are natural inhabitants of the human gastrointestinal tract and well known for their health-promoting effects. Tolerance to bile stress is crucial for bifidobacteria to survive in the colon and to exert their beneficial actions. In this work, RNA-Seq transcriptomic analysis complemented with proteomic analysis was used to investigate the cellular response to bile in Bifidobacterium longum BBMN68. The transcript levels of 236 genes were significantly changed (≥ threefold, p < 0.001) and 44 proteins were differentially abundant (≥1.6-fold, p < 0.01) in B. longum BBMN68 when exposed to 0.75 g l(-1) ox-bile. The hemolysin-like protein and bile efflux systems were significantly over produced, which might prevent bile adsorption and exclude bile, respectively. The cell membrane composition was modified probably by an increase of cyclopropane fatty acid and a decrease of transmembrane proteins, resulting in a cell membrane more impermeable to bile salts. Our hypothesis was later confirmed by surface hydrophobicity assay. The transcription of genes related to xylose utilization and bifid shunt were up-regulated, which increased the production of ATP and reducing equivalents to cope with bile-induced damages in a xylan-rich colon environment. Bile salts signal the B. longum BBMN68 to gut entrance and enhance the expression of esterase and sortase associated with adhesion and colonization in intestinal tract, which was supported by a fivefold increased adhesion ability to HT-29 cells by BBMN68 upon bile exposure. Notably, bacterial one-hybrid and EMSA assay revealed that the two-component system senX3-regX3 controlled the expression of pstS in bifidobacteria and the role of this target gene in bile resistance was further verified by heterologous expression in Lactococcus lactis. Taken altogether, this study established a model for global response mechanisms in B. longum to bile.

Microcin V Production in Lactobacillus plantarum LB-B1 Using Heterologous Leader Peptide from Pediocin PA-1.

Lactobacillus strains producing bacteriocins have attracted highly attention as probiotic cultures in animal nutrition since the use of antibiotics was forbidden in the livestock industry. Lactobacillus plantarum LB-B1 isolated from the fermented dairy product can produce pediocin PA-1, which has a strong inhibition of Listeria but hardly any influence on Gram-negative spoilage agents. In this work, L. plantarum LB-B1 was selected as the host to express microcin V using the leader peptide of pediocin PA-1. Well-diffusion assay combined with Tricine-SDS-polyacrylamide gel showed that microcin V could be successfully expressed and secreted in L. plantarum LB-B1. Meanwhile, the production of microcin V did not affect the secretion of pediocin PA-1. It is worthwhile noted that the supernatant from L. plantarum 8148-ColV had a more effective inhibition of Listeria than that from the control strain L. plantarum 8148. Furthermore, this supernatant also unexpectedly produced antibacterial activity against Staphylococcus aureus. Taken altogether, these results suggested that pediocin PA-1 and microcin V in the supernatant could generate synergistic effect, which not only enhanced the antibacterial ability but also expanded the antibacterial spectrum. Therefore, the recombinant strain has a great potential application as a probiotic to reduce the level of enteric pathogens in livestock industry.

Functional role of oppA encoding an oligopeptide-binding protein from Lactobacillus salivarius Ren in bile tolerance.

Lactobacillus salivarius is a member of the indigenous microbiota of the human gastrointestinal tract (GIT), and some L. salivarius strains are considered as probiotics. Bile tolerance is a crucial property for probiotic bacteria to survive the transit through the GIT and exert their beneficial effects. In this work, the functional role of oppA encoding an oligopeptide transporter substrate-binding protein from L. salivarius Ren in bile salt tolerance was investigated. In silico analysis revealed that the oppA gene encodes a 61.7-kDa cell surface-anchored hydrophilic protein with a canonical lipoprotein signal peptide. Homologous overexpression of OppA was shown to confer 20-fold higher tolerance to 0.5 % oxgall in L. salivarius Ren. Furthermore, the recombinant strain exhibited 1.8-fold and 3.6-fold higher survival when exposed to the sublethal concentration of sodium taurocholate and sodium taurodeoxycholate, respectively, while no significant change was observed when exposed to sodium glycocholate and sodium glycodeoxycholate (GDCA). Our results indicate that OppA confers specific resistance to taurine-conjugated bile salts in L. salivarius Ren. In addition, the OppA overexpression strain also showed significant increased resistance to heat and salt stresses, suggesting the protective role of OppA against multiple stresses in L. salivarius Ren.