Investigation on the inactivation effect and mechanism of Listeria monocytogenes in fresh-cut cucumber during storage by ultrasound combined with sodium hypochlorite.

Fresh agricultural products are frequently contaminated with Listeria monocytogenes (L. monocytogenes), which threatens consumer health. The mechanism of the inhibitory effect of ultrasound and sodium hypochlorite (US-NaClO) on L. monocytogenes on fresh-cut cucumber remains poorly understood. Therefore, the bactericidal ability and mechanism of US-NaClO treatment on L. monocytogenes were studied on fresh-cut cucumber during storage using various approaches such as determination of intracellular material leakage, scanning electron microscopy, flow cytometry, and expression analysis of virulence genes. The results showed that the number of L. monocytogenes on fresh-cut cucumber was significantly reduced after ultrasound treatment for 5 min in combined with 75 ppm sodium hypochlorite treatment(P < 0.05). The US-NaClO treatment affected cell morphology, impaired cell membrane integrity, increased cell membrane permeability, and reduced the concentration of K+, inorganic phosphate, ATP, proteins, and DNA in bacterial cells, leading to the inactivation of microorganisms. In addition, the US-NaClO treatment downregulated expression of the virulence genes actA, hly, inlA, mpl, pclA, and plcB, thus decreasing the pathogenicity of bacteria. It can avoid contamination by pathogenic bacteria during the production of fresh-cut cucumber, while providing safety assurance for production.

Quantification of elongation stalls and impact on gene expression in yeast.

Ribosomal pauses are a critical part of cotranslational events including protein folding and localization. However, extended ribosome pauses can lead to ribosome collisions, resulting in the activation of ribosome rescue pathways and turnover of protein and mRNA. While this relationship has been known, there has been little exploration of how ribosomal stalls impact translation duration at a quantitative level. We have taken a method used to measure elongation time and adapted it for use in Saccharomyces cerevisiae to quantify the impact of elongation stalls. We find, in transcripts containing Arg CGA codon repeat-induced stalls, a Hel2-mediated dose-dependent decrease in protein expression and mRNA level and an elongation delay on the order of minutes. In transcripts that contain synonymous substitutions to nonoptimal Leu codons, there is a decrease in protein and mRNA levels, as well as similar elongation delay, but this occurs through a non-Hel2-mediated mechanism. Finally, we find that Dhh1 selectively increases protein expression, mRNA level, and elongation rate. This indicates that distinct poorly translated mRNAs will activate different rescue pathways despite similar elongation stall durations. Taken together, these results provide new quantitative mechanistic insight into the surveillance of translation and the roles of Hel2 and Dhh1 in mediating ribosome pausing events.

Infection behavior of Listeria monocytogenes on iceberg lettuce (Lactuca sativa var. capitata).

Iceberg lettuce among leafy vegetables is susceptible to contamination with foodborne pathogens, posing a risk of food microbial safety. Listeria monocytogenes (L. monocytogenes) is a highly lethal pathogen that can survive and proliferate on leafy vegetables. In this paper, the contamination stage, attachment site, internalization pathway, proliferation process, extracellular substance secretion and virulence factors expression of L. monocytogenes on iceberg lettuce were researched. Results showed that the contamination stage of L. monocytogenes on iceberg lettuce was 0-20 min, the proliferation stage was after 20 min. The attachment tissues were stomata and winkles. The internalization distance of L. monocytogenes in the midrib was farther than that in the leaf blade. They enhanced the movement ability of cells by up-regulating the expression of flaA and motA genes, and enhanced the adhesion ability of cells by up-regulating the expression of actA and inla genes, which was beneficial to the proliferation. During proliferation, cells gradually secreted extracellular substances to promote the biofilm formation on iceberg lettuce. The formation of biofilms experienced: individual bacteria, cell aggregation and biofilm maturation. Biofilms were more likely to form on the leaf blade of iceberg lettuce.

Quantification of elongation stalls and impact on gene expression in yeast.

Ribosomal pauses are a critical part of co-translational events including protein folding and localization. However, extended ribosome pauses can lead to ribosome collisions, resulting in the activation of ribosome rescue pathways and turnover of protein and mRNA. While this relationship has been known, the specific threshold between permissible pausing versus activation of rescue pathways has not been quantified. We have taken a method used to measure elongation time and adapted it for use in S. cerevisiae to quantify the impact of elongation stalls. We find, in transcripts containing Arg CGA codon repeat-induced stalls, a Hel2-mediated dose-dependent decrease in protein expression and mRNA level and an elongation delay on the order of minutes. In transcripts that contain synonymous substitutions to non-optimal Leu codons, there is a decrease in protein and mRNA levels, as well as similar elongation delay, but this occurs through a non-Hel2-mediated mechanism. Finally, we find that Dhh1 selectively increases protein expression, mRNA level, and elongation rate. This indicates that distinct poorly translated codons in an mRNA will activate different rescue pathways despite similar elongation stall durations. Taken together, these results provide new quantitative mechanistic insight into the surveillance of translation and the roles of Hel2 and Dhh1 in mediating ribosome pausing events.

New insights on low-temperature storage regulating garlic greening and the accumulation of pigment precursors via glutathione metabolism and energy cycles.

To explore regulation mechanism of temperature on garlic greening and pigment precursors' accumulation, greening capacities, pigment precursors and critical metabolites, enzyme and genes involved in glutathione and NADPH metabolism of garlic stored at five temperatures (4, 8, 16, 24 and 30 ℃) were analyzed. Results showed that garlic pre-stored at 4, 8 and 16 ℃ were more likely to green than ones at 24 and 30 ℃ after pickling. After 25 days, more S-1-propenyl-l-cysteine sulfoxide (1-PeCSO) were detected in garlic stored at 4, 8 and 16 ℃ (753.60, 921.85 and 756.75 mAU, respectively) than that at 24 and 30 ℃ (394.35 and 290.70 mAU). Pigment precursors' accumulation in garlic was mainly realized by glutathione and NADPH metabolism under low-temperature storage, through enhancements of activities or expressions for GR (GSR), GST (GST), γ-GT (GGT1, GGT2), 6PGDH (PGD) and ICDHc (IDH1). This study enriched the mechanism of garlic greening.

Effect of Ultrasound Combinated with Sodium Hypochlorite Treatment on Microbial Inhibition and Quality of Fresh-Cut Cucumber.

The influence of ultrasound combined with sodium hypochlorite (US-NaClO) treatment on microorganisms and quality of fresh-cut cucumber during storage were investigated. Ultrasound (400 W, 40 kHz, US: 5, 10 and 15 min) and sodium hypochlorite (NaClO: 50, 75, 100 ppm) were used to treat fresh-cut cucumber in a single or combined treatment and stored at 4 °C for 8 days and analyzed for texture, color and flavor. The results showed that US-NaClO treatment had a synergistic effect on the inhibition of microorganisms during storage. It could significantly reduce (p < 0.05) the number of microorganisms by 1.73 to 2.17 log CFU/g. In addition, US-NaClO treatment reduced the accumulation of malondialdehyde (MDA) during storage (4.42 nmol/g) and water mobility, and maintained the integrity of the cell membrane, delayed the increase of weight loss (3.21%), reduced water loss, thus slowing down the decline of firmness (9.20%) of fresh-cut cucumber during storage. The degradation of chlorophyll (6.41%) was reduced to maintain the color of freshly cut cucumbers. At the same time, US-NaClO could maintain the content of aldehydes, the main aromatic substance of cucumber, and reduced the content of alcohols and ketones during storage. Combined with the electronic nose results, it could maintain the cucumber flavor at the end of the storage period and reduce the odor produced by microorganisms. Overall, US-NaClO was helpful to inhibit the growth of microorganisms during storage, improve the quality of fresh-cut cucumber.

Rvb1/Rvb2 proteins couple transcription and translation during glucose starvation.

During times of unpredictable stress, organisms must adapt their gene expression to maximize survival. Along with changes in transcription, one conserved means of gene regulation during conditions that quickly repress translation is the formation of cytoplasmic phase-separated mRNP granules such as P-bodies and stress granules. Previously, we identified that distinct steps in gene expression can be coupled during glucose starvation as promoter sequences in the nucleus are able to direct the subcellular localization and translatability of mRNAs in the cytosol. Here, we report that Rvb1 and Rvb2, conserved ATPase proteins implicated as protein assembly chaperones and chromatin remodelers, were enriched at the promoters and mRNAs of genes involved in alternative glucose metabolism pathways that we previously found to be transcriptionally upregulated but translationally downregulated during glucose starvation in yeast. Engineered Rvb1/Rvb2-binding on mRNAs was sufficient to sequester mRNAs into mRNP granules and repress their translation. Additionally, this Rvb tethering to the mRNA drove further transcriptional upregulation of the target genes. Further, we found that depletion of Rvb2 caused decreased alternative glucose metabolism gene mRNA induction, but upregulation of protein synthesis during glucose starvation. Overall, our results point to Rvb1/Rvb2 coupling transcription, mRNA granular localization, and translatability of mRNAs during glucose starvation. This Rvb-mediated rapid gene regulation could potentially serve as an efficient recovery plan for cells after stress removal.

Centrifugal micropipette-tip with pressure signal readout for portable quantitative detection of myoglobin.

This paper describes a centrifugal micropipette-tip method for ELISA sample processing combined with a pressure meter for portable quantitative detection of acute myocardial infarction (AMI) biomarker myoglobin (Myo). The method enables sensitive and reliable quantification of Myo in 35 minutes. With the advantages of simplicity, speed, user friendliness, low cost, and small sample consumption, centrifugal micropipette-tip ELISA shows great potential for quantitative POC diagnostics for AMI.