Studies on the mechanisms of Helicobacter pylori inhibition by Syzygium aromaticum aqueous extract.

BACKGROUND: The aqueous extract of the dried buds of Syzygium aromaticum (SAAE) have the potential to alleviate Helicobacter pylori infection, but the specific molecular mechanism has not been fully elucidated. PURPOSE: This study aimed to investigate the underlying mechanisms of SAAE on H. pylori pathogenicity. METHODS: The inhibitory kinetics and anti-H. pylori adhesive capacity assays were conducted to examine the effects of SAAE on the growth and adhesive capability of H. pylori. The H. pylori outer membrane vesicles (OMVs) were purified from the culture supernatant through high-speed centrifugation, filtration, and two rounds of ultracentrifugation. Their characteristics and protein composition were then identified using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and qualitative proteomics study. Subsequently, the effect of SAAE on the pathogenicity of H. pylori OMVs was investigated using the Griess reagent assay, enzyme-linked immunosorbent assay (ELISA), quantitative proteomics study, TEM, and western blotting assay. RESULTS: SAAE exhibited inhibitory effects on H. pylori growth and adhesion. The isolated H. pylori OMVs showed particle size of 27-242 nm and Zeta potential of -9.67 ± 0.53 mV. A total of 599 proteins were identified in the OMVs. Proteomics study indicated that the differential expressed proteins induced by OMVs with or without SAAE commonly enriched in P53 and autophagy pathways. Besides, SAAE counteracted the increased production of pro-inflammatory cytokines and attenuated the induction of cell autophagy caused by H. pylori OMVs. Furthermore, SAAE normalized the abnormal regulation of downstream targets (AIFM2 and IGFBP3) in the P53 signaling pathway caused by H. pylori OMVs. CONCLUSION: SAAE can inhibit the growth and adhesion of H. pylori, reduce the inflammation and autophagy induced by H. pylori OMVs, and combated the abnormal regulation of P53 signaling pathway caused by H. pylori OMVs. These findings may help elucidate the mechanisms through which SAAE reduces the pathogenicity of H. pylori.

1,3,6-Trigalloylglucose: A Novel Potent Anti-Helicobacter pylori Adhesion Agent Derived from Aqueous Extracts of Terminalia chebula Retz.

1,3,6-Trigalloylglucose is a natural compound that can be extracted from the aqueous extracts of ripe fruit of Terminalia chebula Retz, commonly known as "Haritaki". The potential anti-Helicobacter pylori (HP) activity of this compound has not been extensively studied or confirmed in scientific research. This compound was isolated using a semi-preparative liquid chromatography (LC) system and identified through Ultra-high-performance liquid chromatography-MS/MS (UPLC-MS/MS) and Nuclear Magnetic Resonance (NMR). Its role was evaluated using Minimum inhibitory concentration (MIC) assay and minimum bactericidal concentration (MBC) assay, scanning electron microscope (SEM), inhibiting kinetics curves, urea fast test, Cell Counting Kit-8 (CCK-8) assay, Western blot, and Griess Reagent System. Results showed that this compound effectively inhibits the growth of HP strain ATCC 700392, damages the HP structure, and suppresses the Cytotoxin-associated gene A (Cag A) protein, a crucial factor in HP infection. Importantly, it exhibits selective antimicrobial activity without impacting normal epithelial cells GES-1. In vitro studies have revealed that 1,3,6-Trigalloylglucose acts as an anti-adhesive agent, disrupting the adhesion of HP to host cells, a critical step in HP infection. These findings underscore the potential of 1,3,6-Trigalloylglucose as a targeted therapeutic agent against HP infections.

Overexpression of PvSTK1 gene from Switchgrass (Panicum virgatum L.) affects flowering time and development of floral organ in transgenic Arabidopsis thaliana.

Flowering means that the plant enters the reproductive growth stage, which is a crucial stage in the plant life cycle. Delaying flowering time to prolong vegetative growth is an important method to increase biomass yield and saccharification efficiency in switchgrass, It is of great significance to study the molecular mechanism of plant flowering and regulate the process of plant flowering in the process of biomass production. In this study, we identified 55 serine/threonine-protein kinase genes related to flower development from the switchgrass transcriptome database. Simultaneously, we cloned one of them, PvSTK1, whose expression level and differential fold were significantly higher than other members. PvSTK1 is located on chromosome 8N and its protein was in the cell membrane, cytoplasm, and nucleus. The spatio-temporal expression analysis of the PvSTK1 in switchgrass displayed that the PvSTK1 is crucial in vegetative period, however, not in the transition to reproductive period. Overexpression of PvSTK1 in Arabidopsis resulted in down-regulation of flower-promoting genes and up-regulation of flower-suppressing genes, thereby delaying flowering. In addition, PvSTK1 caused atrophy of the ovules of the florets at the base of the inflorescence, leading to sterility of the florets. The function of PvSTK1 is to inhibit the development of floral organs, and its overexpression can prolong its vegetative period. In the future, overexpression of the PvSTK1 gene in switchgrass will change the flowering time and increase yield and utilization efficiency of biomass.

Anthocyanin Biosynthesis and a Regulatory Network of Different-Colored Wheat Grains Revealed by Multiomics Analysis.

Colored wheat has always been a popular research area because of its high performance in the field and significant medical uses. Progress has been made mapping the genes of purple or blue grains; however, the reason why different grain colors form in wheat is not well understood. We created wheat lines with different grain colors (purple and blue) using the white grain cultivar Xiaoyan22 and located the candidate region related to the purple and blue grains in chromosome 2A, 2B, and 4D, 2A, respectively, by the bulked segregant RNA-seq. The transcriptomic and metabolomic analyses of the three grains at different developmental stages indicated that the upregulation of flavonoid 3'-hydroxylase/flavonoid 3',5'hydroxylase 2 and TaMYC1/TaMYC4 was important for the formation of purple/blue grains. The blue TaMYC4 had 16 nonsynonymous single nucleotide variants verified by Sanger sequencing and possessed a different splicing mode in the bHLH_MYC_N domain compared with the reference database. Targeted high-performance liquid chromatography-mass spectrometry/mass spectrometry analysis of anthocyanins found that the purple and blue grains contained more pelargonidin, cyanidin, and delphinidin, respectively. This study provides a comprehensive understanding of the different color formations of wheat grains and useful information about genetic improvements in wheat and other crops.

Identification of mcr-10 carried by self-transmissible plasmids and chromosome in Enterobacter roggenkampii strains isolated from hospital sewage water.

The recent emergence of plasmid-borne mobilized colistin resistance (mcr) genes largely challenges the clinical use of colistin. Monitoring the distribution of mcr genes in environment is important for aiding to develop effective control measures. In this study, we aimed to evaluate the occurrence of a recent reported mcr variant, mcr-10, in hospital sewage water. mcr-10 was identified in three Enterobacter roggenkampii strains with high-level colistin resistance (MIC ≥ 16 mg/L). The three strains were assigned to different sequence types suggesting a sporadic dissemination of mcr-10 in the sewage water. Pairwise comparisons of the predicted protein structures of ten mcr homologues revealed that MCR-10 shares a higher similarity with MCR-3, MCR-4, MCR-7, and MCR-9. Overexpression in Escherichia coli Top10 showed that the activity of mcr-10 against colistin is lower than that of mcr-9. mcr-10 expression can be specifically induced by colistin, and it was co-upregulated with phoPQ to mediate the high-level colistin resistance. The mcr-10 gene was detected on self-transmissible plasmids in two isolates and on the chromosome in the other one. Blasting in Genbank suggested that the two mcr-10-bearing plasmids (pECL981-1 and pECL983-1) were novel plasmids, and replicon typing showed that they belong to IncFIB-FII and IncFIB, respectively. Plasmid-curing assay evidence that pECL981-1 was lack of fitness cost for the host. Three novel types of the genetic context were found for the mcr-10 gene in the three isolates. The structure xerC-mcr10 was dominant in mcr-10-positive genomes (39/42) retrieved in Genbank, suggesting that xerC might be involved in the mobilization of mcr-10. To our knowledge, this is the first report of mcr-10-producing E. roggenkampii detected in hospital sewage water. Our study highlights that continuous monitoring of mcr genes in hospital sewage water is imperative for understanding and tackling the dissemination.

Dissemination of a 'rare' extended-spectrum ß-lactamase gene blaSFO-1 mediated by epidemic clones of carbapenemase-producing Enterobacter hormaechei in China.

An increasing trend of the coexistence of a rare extended-spectrum ß-lactamase gene blaSFO-1 and carbapenemase genes in Enterobacteriaceae has recently been noted. This study aimed to determine the epidemiological and genetic characterisation of SFO-1-positive carbapenem-resistant Enterobacter cloacae complex (CREC) isolates. A total of 61 CREC clinical isolates were collected in the framework of a national surveillance for carbapenem-resistant Enterobacteriaceae during 2011-2015 in China. Seven SFO-1-positive CREC isolates (11.5%) were identified in four provinces, suggesting a wide dissemination of the blaSFO-1 gene among the CREC population in China. Five SFO-1-positive CREC isolates were further identified by screening 1625 genomes of E. cloacae complex strains retrieved from GenBank. The 12 SFO-1-positive CREC isolates were further identified as Enterobacter hormaechei, of which 10 belonged to epidemic clones (ST93, ST114 and ST418), indicating that these clones might largely contribute to the dissemination of blaSFO-1. Phylogenomics analysis further identified the occurrence of clonal dissemination in the community setting. The blaSFO-1-bearing plasmids were assigned to various incompatibility groups with highly diverse sizes (~104-370 kb), suggesting a wide vector range of blaSFO-1. Two types of genetic context, with and without insertion sequence IS26, were identified for the blaSFO-1 gene. The genetic context flanked by IS26 was more prevalent, thus largely facilitating the mobility of blaSFO-1. This study revealed that the blaSFO-1 gene is not as rare as previously found and that epidemic clones of CREC are responsible for its dissemination in China. These findings highlight the potential of wide dissemination of low-prevalence antimicrobial resistance genes.

In Vivo Imaging of the Coupling between Neuronal and CREB Activity in the Mouse Brain.

Sensory experiences cause long-term modifications of neuronal circuits by modulating activity-dependent transcription programs that are vital for regulation of long-term synaptic plasticity and memory. However, it has not been possible to precisely determine the interaction between neuronal activity patterns and transcription factor activity. Here we present a technique using two-photon fluorescence lifetime imaging (2pFLIM) with new FRET biosensors to chronically image in vivo signaling of CREB, an activity-dependent transcription factor important for synaptic plasticity, at single-cell resolution. Simultaneous imaging of the red-shifted CREB sensor and GCaMP permitted exploration of how experience shapes the interplay between CREB and neuronal activity in the neocortex of awake mice. Dark rearing increased the sensitivity of CREB activity to Ca2+ elevations and prolonged the duration of CREB activation to more than 24 h in the visual cortex. This technique will allow researchers to unravel the transcriptional dynamics underlying experience-dependent plasticity in the brain.

Electrospun chitosan/PVA/bioglass Nanofibrous membrane with spatially designed structure for accelerating chronic wound healing.

Cutaneous wounds, especially chronic wounds, remain clinical challenges, and this is partially due to the complex healing process composed of four overlapping but distinct stages including hemostasis, inflammation, proliferation and remodeling. Therefore, wound dressings with spatially designed structures which can temporally regulate certain bioactive components to function at specific healing stages might be able to accelerate the healing process. In this study, nanobioglass incorporated chitosan-PVA (polyvinyl alcohol) trilayer nanofibrous membrane (nBG-TFM) was fabricated via sequential electrospinning. This membrane exhibited excellent biocompatibility, antibacterial activity and regeneration promotion effect. Furthermore, spatially designed structure optimized functions of each component and provided more suitable microenvironment as compared with uniform membrane. Rat full-thickness skin defects model and mice diabetic chronic wound model showed that nBG-TFM could achieve significantly accelerated and enhanced healing, in terms of complete re-epithelialization, improved collagen alignment and formation of skin appendages. It was revealed that nBG-TFM functioned through upregulating growth factors including VEGF and TGF-ß. Meanwhile inflammatory cytokines such as TNF-α and IL-1ß were downregulated. The technology presented in this study shed new light on designing functional wound dressings which can promote healing of chronic wounds.