Effect of Internalin InlJ of Listeria monocytogenes on Phage Sensitivity and Biofilm Formation

In order to explore the role and function of the inlJ gene of Listeria monocytogenes (Lm) in phage sensitivity and biofilm, the inlJ gene-deficient strain Lm NJ05-ΔinlJ was constructed by homologous recombination. The growth, adhesion and invasion characteristics of the defective strain were identified. The results showed that compared with the wide-type strain Lm NJ05, the adhesion and invasion of RAW264.7 cells by Lm NJ05-ΔinlJ were significantly reduced to 20.05% and 4.42%, respectively. The efficiency of plaque formation was enhanced by 2.72 folds and phage vB-LmoM-NJ05 had a stronger lytic activity on Lm NJ05-ΔinlJ. Phage vB-LmoM-NJ05 at titers of 105 and 108 PFU/mL could completely inhibit and remove the biofilm of Lm NJ05-ΔinlJ, respectively. Transcriptional analysis of biofilm formation-related genes showed that the transcriptional levels of the degU, agrA, agrD, luxS, yneA, recA and hpt genes were significantly decreased to nearly zero in the defective strain after interacting with phage vB-LmoM-NJ05. In conclusion, deletion of the inlJ gene can enhance the phage sensitivity of Lm, and down-regulate the ability of cell invasion and biofilm formation. Therefore, the inlJ gene not only regulate Lm but also affect its interaction with phage, which lays a foundation for the development and application of phage biocontrol

Circulating Monocytes Act as a Common Trigger for the Calcification Paradox of Osteoporosis and Carotid Atherosclerosis via TGFB1-SP1 and TNFSF10-NFKB1 Axis

BackgroundOsteoporosis often occurs with carotid atherosclerosis and causes contradictory calcification across tissue in the same patient, which is called the “calcification paradox”. Circulating monocytes may be responsible for this unbalanced ectopic calcification. Here, we aimed to show how CD14+ monocytes contribute to the pathophysiology of coexisting postmenopausal osteoporosis and carotid atherosclerosis.MethodsWe comprehensively analyzed osteoporosis data from the mRNA array dataset GSE56814 and the scRNA-seq dataset GSM4423510. Carotid atherosclerosis data were obtained from the GSE23746 mRNA dataset and GSM4705591 scRNA-seq dataset. First, osteoblast and vascular SMC lineages were annotated based on their functional expression using gene set enrichment analysis and AUCell scoring. Next, pseudotime analysis was applied to draw their differentiated trajectory and identify the key gene expression changes in crossroads. Then, ligand–receptor interactions between CD14+ monocytes and osteoblast and vascular smooth muscle cell (SMC) lineages were annotated with iTALK. Finally, we selected calcification paradox-related expression in circulating monocytes with LASSO analysis.ResultsFirst, we found a large proportion of delayed premature osteoblasts in osteoporosis and osteogenic SMCs in atherosclerosis. Second, CD14+ monocytes interacted with the intermediate cells of the premature osteoblast and osteogenic SMC lineage by delivering TGFB1 and TNFSF10. This interaction served as a trigger activating the transcription factors (TF) SP1 and NFKB1 to upregulate the inflammatory response and cell senescence and led to a retarded premature state in the osteoblast lineage and osteogenic transition in the SMC lineage. Then, 76.49% of common monocyte markers were upregulated in the circulating monocytes between the two diseases, which were related to chemotaxis and inflammatory responses. Finally, we identified 7 calcification paradox-related genes on circulating monocytes, which were upregulated in aging cells and downregulated in DNA repair cells, indicating that the aging monocytes contributed to the development of the two diseases.ConclusionsOur work provides a perspective for understanding the triggering roles of CD14+ monocytes in the development of the calcification paradox in osteoporosis- and atherosclerosis-related cells based on combined scRNA and mRNA data. This study provided us with an elucidation of the mechanisms underlying the calcification paradox and could help in developing preventive and therapeutic strategies

An integrated production-distribution model for a deteriorating inventory item

We develop an integrated production-distribution model for a deteriorating item in a two-echelon supply chain. The supplier's production batch size is restricted to an integer multiple of the discrete delivery lot quantity to the buyer. Exact cost functions for the supplier, the buyer and the entire supply chain are developed. These lead to the determination of individual optimal policies, as well as the optimal policy for the overall, integrated supply chain. We outline a procedure for determining the optimal supply chain decisions with the objective of minimizing the total system cost. Our approach is illustrated through a numerical example.Integrated supply chain Deterioration Inventory JIT

Identification of Ramie Genes in Response to Pratylenchus coffeae Infection Challenge by Digital Gene Expression Analysis

Root lesion disease, caused by Pratylenchus coffeae, seriously impairs the growth and yield of ramie, an important natural fiber crop. The ramie defense mechanism against P. coffeae infection is poorly understood, which hinders efforts to improve resistance via breeding programs. In this study, the transcriptome of the resistant ramie cultivar Qingdaye was characterized using Illumina sequence technology. About 46.3 million clean pair end (PE) reads were generated and assembled into 40,826 unigenes with a mean length of 830 bp. Digital gene expression (DGE) analysis was performed on both the control roots (CK) and P. coffeae-challenged roots (CH), and the differentially expressed genes (DEGs) were identified. Approximately 10.16 and 8.07 million cDNA reads in the CK and CH cDNA libraries were sequenced, respectively. A total of 137 genes exhibited different transcript abundances between the two libraries. Among them, the expressions of 117 and 20 DEGs were up- and down-regulated in P. coffeae-challenged ramie, respectively. The expression patterns of 15 candidate genes determined by qRT-PCR confirmed the results of DGE analysis. Time-course expression profiles of eight defense-related genes in susceptible and resistant ramie cultivars were different after P. coffeae inoculation. The differential expression of protease inhibitors, pathogenesis-related proteins (PRs), and transcription factors in resistant and susceptible ramie during P. coffeae infection indicated that cystatin likely plays an important role in nematode resistance

Wellbore Fracture Mode and Fracture Pressure Drilled in Depleted Reservoir

Drilling fluid loss in depleted reservoir has been an import issue faced by further tapping the potential of old oil fields. Accurate evaluation of the fracture pressure is the foundation to avoid mud loss. Traditional views suggest that tensile failure is the only fracture mode and the fracture pressure should be determined by a tensile failure criterion, which are not suitable for wells drilled in the depleted reservoir. In this paper, the analysis focuses on the fracture mode and fracture pressure in depleted reservoir, and case studies show that three fracture modes may first occur, and the fracture mode will be changed with reservoir depletion which highly depends on reservoir depletion degree, well azimuth and deviation angle, and the in situ stress state; different failure criteria at different stages of reservoir depletion should be selected to accurately evaluate the fracture pressure. For the vertical well, fracture pressure is no longer a single linear reduction with reservoir depletion; instead, a three-step and two-step reduction may appear, and for the directional well, the fracture pressure is not always decreased; the other patterns such as increase and first increase then decrease may also appear for the wells drilled in reverse and strike fault stress regimes

Counterion-Induced Nanosheet-to-Nanofilament Transition of Lyotropic Bent-Core Liquid Crystals

The smart flexibility of phase transitions in liquid crystals (LCs) makes them suitable for various applications and is an important research field in contemporary science, engineering, and technology. Unlike most reports focused on bent-core LCs in the thermotropic situation, in our present study, we designed and synthesized a fully rigid bent-core molecule with the sulfonic acid group replacing conventional flexible chains. A rich variety of counterion-induced supramolecular LC phase behaviors have been systematically investigated. It was found that the smectic phase with nanosheets tends to transform to the hexagonal phase with nanofilaments when the protons of the sulfonic acid group are partially replaced by alkali metal ions. The experimental results show that the nanoaggregate and phase transition are controlled by the displacing ratio of alkali metal ions rather than the molecular concentration. Another interesting feature is that the achiral bent-core molecules self-assemble into columns by helical stacking and present macroscopic chirality, indicating that spontaneous chiral symmetry breaking occurs in the columnar phase. The fully rigid bent-core molecules reveal surprisingly hierarchical molecular self-assemblies with the smectic-to-hexagonal phase transition, which was not previously observed in supramolecular complexes. The findings will provide new possibilities for applications in LC-based photonic devices, biosystem switches, and supramolecular actuators

Molecular Cloning, Recombinant Expression and Antifungal Activity of BnCPI, a Cystatin in Ramie (Boehmeria nivea L.)

Phytocystatins play multiple roles in plant growth, development and resistance to pests and other environmental stresses. A ramie (Boehmeria nivea L.) phytocystatin gene, designated as BnCPI, was isolated from a ramie cDNA library and its full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence (691 bp) consisted of a 303 bp open reading frame (ORF) encoding a protein of 100 amino acids with deduced molecular mass of 11.06 kDa and a theoretical isoelectric point (pI) of 6.0. The alignment of genome DNA (accession no. MF153097) and cDNA sequences of BnCPI showed that an intron (~104 bp) exists in the coding region. The BnCPI protein contains most of the highly conserved blocks including Gly5-Gly6 at the N-terminal, the reactive site motif QxVxG (Q49V50V51S52G53), the L79-W80 block and the [LVI]-[AGT]-[RKE]-[FY]-[AS]-[VI]-x-[EDQV]-[HYFQ]-N (L22G23R24 F25A26V27 D28D29H30 N31) block that is common among plant cystatins. BLAST analysis indicated that BnCPI is similar to cystatins from Glycine max (77%), Glycine soja (76%), Hevea brasiliensis (75%) and Ricinus communis (75%). The BnCPI was subcloned into expression vector pSmart-I and then overexpressed in Escherichia coli BL21 (DE3) as a His-tagged recombinant protein. The purified reBnCPI has a molecular mass of 11.4 kDa determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE). Purified reBnCPI can efficiently inhibit the protease activity of papain and ficin toward BANA (Nα-benzoyl-L-arginine-2-naphthyamide), as well as the mycelium growth of some important plant pathogenic fungi. The data further contribute to our understanding of the molecular functions of BnCPI

Deletion of the sex-determining gene SXI1α enhances the spread of mitochondrial introns in Cryptococcus neoformans

Abstract Background Homing endonuclease genes (HEGs) are widely distributed genetic elements in the mitochondrial genomes of a diversity of eukaryotes. Due to their ability to self-propagate within and between genomes, these elements can spread rapidly in populations. Whether and how such elements are controlled in genomes remains largely unknown. Results Here we report that the HEG-containing introns in the mitochondrial COX1 gene in Cryptococcus neoformans are mobile and that their spread in sexual crosses is influenced by mating type (MAT) α-specific homeodomain gene SXI1α. C. neoformans has two mating types, MAT a and MATα. In typical crosses between strains of the two mating types, only a small portion ( 95%) of the diploid fusants inheriting the HEG-containing introns from the MATα parent, a frequency significantly higher than those of intronless mitochondrial genes. Conclusions Our results suggest that SXI1α not only determines uniparental mitochondrial inheritance but also inhibits the spread of HEG-containing introns in the mitochondrial genome in C. neoformans

Development Strategy of Microbial Safety Industry

Microbial safety is crucial for China’s social and economic development in the new era, and it is an important support for national food security, public health, and social stability. In this article, we analyze the development needs and current status of the microbial safety industry considering the policy environment in China. Subsequently, we clarify the domestic and global development patterns of the industry and summarize the challenges faced by the innovation and application of pathogenic microorganism detection and control technologies in China. Furthermore, we propose a development plan for building China’s microbial safety industry. To explore a high-quality development path, China should build innovation platforms and a large scientific database regarding microbial safety, enhance personnel development for this industry, encourage leading enterprises to develop according to standards and regulations, and strengthen major system innovation and engineering technology research in the microbial safety field