135 research outputs found
Gas Plasma Pre-Treatment Increases Antibiotic Sensitivity and Persister Eradication in Methicillin-Resistant Staphylococcus aureus
Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of serious nosocomial infections, and recurrent MRSA infections primarily result from the survival of persister cells after antibiotic treatment. Gas plasma, a novel source of ROS (reactive oxygen species) and RNS (reactive nitrogen species) generation, not only inactivates pathogenic microbes but also restore the sensitivity of MRSA to antibiotics. This study further found that sublethal treatment of MRSA with both plasma and plasma-activated saline increased the antibiotic sensitivity and promoted the eradication of persister cells by tetracycline, gentamycin, clindamycin, chloramphenicol, ciprofloxacin, rifampicin, and vancomycin. The short-lived ROS and RNS generated by plasma played a primary role in the process and induced the increase of many species of ROS and RNS in MRSA cells. Thus, our data indicated that the plasma treatment could promote the effects of many different classes of antibiotics and act as an antibiotic sensitizer for the treatment of antibiotic-resistant bacteria involved in infectious diseases
Tissue factor pathway inhibitor-2 was repressed by CpG hypermethylation through inhibition of KLF6 binding in highly invasive breast cancer cells
<p>Abstract</p> <p>Background</p> <p>Tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated Kunitz inhibitor that inhibits plasmin and trypsin-mediated activation of zymogen matrix metalloproteinases involved in tumor progression, invasion and metastasis. Here, we have investigated the mechanism of DNA methylation on the repression of TFPI-2 in breast cancer cell lines.</p> <p>Results</p> <p>We found that both protein and mRNA of TFPI-2 could not be detected in highly invasive breast cancer cell line MDA-MB-435. To further investigate the mechanism of TFPI-2 repression in breast cancer cells, 1.5 Kb TFPI-2 promoter was cloned, and several genetic variations were detected, but the promoter luciferase activities were not affected by the point mutation in the promoter region and the phenomena was further supported by deleted mutation. Scan mutation and informatics analysis identified a potential KLF6 binding site in TFPI-2 promoter. It was revealed, by bisulfite modified sequence, that the CpG island in TFPI-2 promoter region was hypermethylated in MDA-MB-435. Finally, using EMSA and ChIP assay, we demonstrated that the CpG methylation in the binding site of KLF-6 diminished the binding of KLF6 to TFPI-2 promoter.</p> <p>Conclusion</p> <p>In this study, we found that the CpG islands in TFPI-2 promoter was hypermethylated in highly invasive breast cancer cell line, and DNA methylation in the entire promoter region caused TFPI-2 repression by inducing inactive chromatin structure and decreasing KLF6 binding to its DNA binding sequence.</p
SacB-SacR Gene Cassette As the Negative Selection Marker to Suppress Agrobacterium Overgrowth in Agrobacterium-Mediated Plant Transformation
Agrobacterium overgrowth is a common problem in Agrobacterium-mediated plant transfor-mation. To suppress the Agrobacterium overgrowth, various antibiotics have been used during plant tissue culture steps. The antibiotics are expensive and may adversely affect plant cell differentiation and reduce plant transformation efficiency. The SacB-SacR proteins are toxic to most Agrobacterium tumefaciens strains when they are grown on culture medium supĀ¬plemented with sucrose. Therefore, SacB-SacR genes can be used as negative selection markers to suppress the overgrowth of Agrobacterium tumefaciens in the plant tissue culture process. We generated a mutant Agrobacterium tumefaciens strain GV2260 (recA-SacB/R) that has the SacB-SacR cassette inserted into the bacterial genome at the recA gene locus. The mutant Agrobacterium strain is sensitive to sucrose but maintains its ability to transform plant cells in both transient and stable transformation assays. We demonstrated that the mutant strain GV2260 (recA-SacB/R) can be inhibited by sucrose that reduces the overgrowth of Agrobacterium and therefore improves the plant transformation efficiency. We employed GV2260 (recA-SacB/R) to generate stable transgenic N. benthamiana plants expressing a CRISPR-Cas9 for knocking out a WRKY transcripĀ¬tion factor
Mechanism of Virus Inactivation by Cold Atmospheric-Pressure Plasma and Plasma-Activated Water
ABSTRACT Viruses cause serious pathogenic contamination that severely affects the environment and human health. Cold atmospheric-pressure plasma efficiently inactivates pathogenic bacteria; however, the mechanism of virus inactivation by plasma is not fully understood. In this study, surface plasma in argon mixed with 1% air and plasma-activated water was used to treat water containing bacteriophages. Both agents efficiently inactivated bacteriophages T4, Ļ174, and MS2 in a time-dependent manner. Prolonged storage had marginal effects on the antiviral activity of plasma-activated water. DNA and protein analysis revealed that the reactive species generated by plasma damaged both nucleic acids and proteins, consistent with the morphological examination showing that plasma treatment caused the aggregation of bacteriophages. The inactivation of bacteriophages was alleviated by the singlet oxygen scavengers, demonstrating that singlet oxygen played a primary role in this process. Our findings provide a potentially effective disinfecting strategy to combat the environmental viruses using cold atmospheric-pressure plasma and plasma-activated water. IMPORTANCE Contamination with pathogenic and infectious viruses severely threatens human health and animal husbandry. Current methods for disinfection have different disadvantages, such as inconvenience and contamination of disinfection by-products (e.g., chlorine disinfection). In this study, atmospheric surface plasma in argon mixed with air and plasma-activated water was found to efficiently inactivate bacteriophages, and plasma-activated water still had strong antiviral activity after prolonged storage. Furthermore, it was shown that bacteriophage inactivation was associated with damage to nucleic acids and proteins by singlet oxygen. An understanding of the biological effects of plasma-based treatment is useful to inform the development of plasma into a novel disinfecting strategy with convenience and no by-product
Fabrication, microstructure, mechanical, and electrochemical properties of NiMnFeCu high entropy alloy from elemental powders
Transition metal based high entropy alloys (HEAs) are often used in electrocatalytic (water electrolysis) applications due to the synergistic effect operating among its constituent elements and unpaired electrons in d orbitals of the concerned metal. In this study, a low cost NiMnFeCu high entropy alloy was successfully synthesised using the combined techniques of mechanical milling (MA) and vacuum sintering. Xāray diffraction was used to analyse the phase composition, optical microscopy, and scanning electron microscopy were used to characterise the fabricated materialās microstructure and chemical homogeneity, thermal, and mechanical properties were tested using the differential scanning calorimetry method and a universal testing machine, respectively. Electrochemical workstation was used to carry out preliminary electrochemical studies such as linear sweep voltammetry (LSV), cyclic voltammetry (CV) and chronoamperometry. The results showed that the asā sintered NiMnFeCu HEA possessed a singleā phase FCC structure. The HEA NiMnFeCu sintered at 1050 Ā°C (S4) and 1000 Ā°C (S2) with a holding time of 2 h showed a yield strength of 516.3 MPa and 389.8 MPa, respectively, and the microāhardness values were measured to be 233.45 Ā± 9 HV and 198.7 Ā± 8 HV, respectively. Preliminary electrochemical studies proved that the alloy sintered at 1000 Ā°C (S2) with a holding time of 2 h exhibited excellent electrocatalytic properties with a measured overpotential of 322 mV at 10 mA cmā»Ā² at 100 cycles of CV and good stability for 10 h when compared to stateāofātheāart electrocatalytic materials IrOā and RuOā This suggested that the HEA NiMnFeCu fabricated under the condition S2 could potentially be used for industrialāscale water electrolysis as it possesses permissible mechanical and good electrochemical properties
Preparation of Colon-Targeted Acetylharpagide Tablets and its Release Properties in vivo and in vitro
Ethno Pharmacological Relevance: Acetylharpagide is a monomeric compound extracted from Ajuga decumbens, widely used for remedying infectious and inflammatory diseases in Southern China.Aim of the Study: The present study designed and investigated the formulation of colon-targeted acetylharpagide tablets according to the dual controlled release mechanisms of time-delay and pH-sensitivity.Materials and Methods: The core tablets of acetylharpagide were coated with the material used in time-delay systems such as ethyl cellulose and suitable channeling agent, followed by pH-dependent polymers, polyacrylic resin II and III in a combination of 1:4. Furthermore, the release and absorption performance of colon-targets tables were evaluated in vitro and in vivo. In the in vitro tests, the optimized formulation was not released in simulated gastric fluid in 2 h; the release was <5% at pH 6.8 simulated intestinal fluids for 4 h; the drug was completely released within 5 h at pH 7.6 simulated colon fluid. In the in vivo tests, pharmacokinetic characteristics of the colon-targeted tablets were investigated in dogs.Results: The results indicated that the acetylharpagide tablets with the technology of colon-targeting caused delayed Tmax, prolonged absorption time, lower Cmax, and AUCINF_obs. Meanwhile, the apparent volume of distribution (Vz_F_bs) of the colon-target tablets was higher than the reference.Conclusions: These results suggested that colon-targeted acetylharpagide tablets deliver the drug to the colon. The in vitro performance of colon-targeted acetylharpagide tablet was appropriately correlated with its performance in vivo
A transcriptomic analysis of Stylo [Stylosanthes guianensis (Aubl.) Sw.] provides novel insights into the basis of salinity tolerance
Tropical areas have a large distribution of saline soils and tidal flats with a high salinity level. Salinity stress is a key factor limiting the widespread use of tropical forage such as Stylosanthes guianensis (Aubl.) Sw. This study was designed to screen the salinity tolerance of 84 S. guianensis accessions; In a greenhouse experiment, plants were subjected to Hoagland solution or Hoagland solution with 200 mM NaCl for up to 15 days. Salinity tolerant accession CIAT11365 and salinity sensitive accession FM05-2 were obtained based on withered leaf rate (WLR). Further verification of salinity tolerance in CIAT11365 and FM05-2 with different salinity gradients showed that salinity stress increased WLR and decreased relative chlorophyll content (SPAD), maximum photochemical efficiency of photosystem II (Fv/Fm), and photosynthetic rate (Pn) in FM05-2, but CIAT11365 exhibited lower WLR and higher SPAD, Fv/Fm, and Pn. Leaf RNA-Seq revealed that Ca2+ signal transduction and Na+ transport ability, salinity tolerance-related transcription factors and antioxidant ability, an increase of auxin, and inhibition of cytokinin may play key roles in CIAT11365 response to salinity stress. The results of this study may contribute to our understanding of the molecular mechanism underlying the responses of S. guianensis to salinity stress and also provide important clues for further study and in-depth characterization of salinity resistance breeding candidate genes in S. guianensis
Widely Targeted Metabolomics Revealed the Dynamic Changes of Metabolites during the Formation of Goose Fatty Liver
To understand the composition and dynamic changes of metabolites during the formation of goose fatty liver, the metabolite profiles of goose liver at three overfeeding stages were analyzed using widely targeted metabolomics. Three 70-day-old Landes geese with similar body conditions from the same batch were selected randomly for slaughter at the early (day 7), middle (day 16) and late (day 25) overfeeding stages, separately. The tip of the larger liver lobe was collected for widely targeted metabolomic analysis. The results showed that: (1) a total of 1 153 metabolites belonging to 19 classes including amino acids, organic acids, nucleotides and lipids were detected in the liver of geese at the three overfeeding stages; (2) principal component analysis (PCA) showed significant differences in the metabolic profiles of goose liver at the three stages, and identified 142 and 92 differential metabolites at the early versus middle stage, and the middle versus late stage, respectively, the major ones being amino acids and their derivatives, as well as organic acids and their derivatives; and (3) Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis indicated that the pathways involved in fatty acid biosynthesis, vitamin B6 metabolism, linoleic acid metabolism, lysine degradation, arginine biosynthesis, arachidonic acid metabolism and amino acid biosynthesis changed significantly during the formation of goose fatty liver. This study found that most of the differential metabolites were involved in fatty acid synthesis during goose fatty liver formation. Moreover, the contents of transport-related metabolites showed a continuous increasing trend. Findings in this study will not only enrich the theoretical knowledge of poultry liver metabolism, but also provide a theoretical basis for the precise nutritional regulation and efficient production of high-quality goose fatty liver
Development of graphitic carbon nitride quantum dots-based oxygen self-sufficient platforms for enhanced corneal crosslinking
Keratoconus, a disorder characterized by corneal thinning and weakening, results in vision loss. Corneal crosslinking (CXL) can halt the progression of keratoconus. The development of accelerated corneal crosslinking (A-CXL) protocols to shorten the treatment time has been hampered by the rapid depletion of stromal oxygen when higher UVA intensities are used, resulting in a reduced cross-linking effect. It is therefore imperative to develop better methods to increase the oxygen concentration within the corneal stroma during the A-CXL process. Photocatalytic oxygen-generating nanomaterials are promising candidates to solve the hypoxia problem during A-CXL. Biocompatible graphitic carbon nitride (g-C3N4) quantum dots (QDs)-based oxygen self-sufficient platforms including g-C3N4 QDs and riboflavin/g-C3N4 QDs composites (RF@g-C3N4 QDs) have been developed in this study. Both display excellent photocatalytic oxygen generation ability, high reactive oxygen species (ROS) yield, and excellent biosafety. More importantly, the A-CXL effect of the g-C3N4 QDs or RF@g-C3N4 QDs composite on male New Zealand white rabbits is better than that of the riboflavin 5ā-phosphate sodium (RF) A-CXL protocol under the same conditions, indicating excellent strengthening of the cornea after A-CXL treatments. These lead us to suggest the potential application of g-C3N4 QDs in A-CXL for corneal ectasias and other corneal diseases
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