The Antibacterial Drug Candidate SBC3 is a Potent Inhibitor of Bacterial Thioredoxin Reductase

Antibiotic resistance is a growing problem for public health and associated with increasing economic costs and mortality rates. Silver and silver-related compounds have been used for centuries due to their antimicrobial properties. In this work, we show that 1,3-dibenzyl-4,5-diphenyl-imidazol-2-ylidene silver(I) acetate/NHC*-Ag-OAc (SBC3) is a reversible, high affinity inhibitor of E. coli thioredoxin reductase (TrxR; K_i=10.8±1.2 nM). Minimal inhibition concentration (MIC) tests with different E. coli and P. aeruginosa strains demonstrated that SBC3 can efficiently inhibit bacterial cell growth, especially in combination with established antibiotics like gentamicin. Our results show that SBC3 is a promising antibiotic drug candidate targeting bacterial TrxR.     

Chemical basis of anti-listerial effects of rosemary herb during stomaching with fresh-cut vegetables

Fresh rosemary herb has demonstrated much stronger anti-listerial effects when stomached with contaminated fresh-cut vegetables than those obtained by application of rosemary essential oils. Various types of rosemary extracts (hydrodistillates, CO₂ extracts, hexane/acetone extracts) were analysed by GC–MS and tested for anti-listerial effects in vitro. Principal component analysis showed that verbenone levels were most correlated with anti-listerial effects, followed by levels of camphor. Stomaching rosemary herb greatly increased headspace levels of verbenone and camphor (by up to 20 times) compared to intact or chopped fresh rosemary. It is speculated that by crushing the herb under water, stomaching released enhanced levels of these components as lipophilic nanosomes which rapidly migrated to lipophilic surfaces, including the membranes of Listeria cells.     

Effects of acid adaptation on the survival of Listeria monocytogenes on modified atmosphere packaged vegetables

The survival and growth of Listeria monocytogenes and L. innocua on ready-to-use (RTU) packaged vegetables (lettuce, swedes, dry coleslaw mix, bean-sprouts) were studied. The effects of acid adaptation of Listeria spp. on their survival during subsequent storage were also investigated. Listeria innocua behaviour was similar to that of L. monocytogenes on all vegetables examined. The survival and growth patterns of Listeria varied with the packaged product. Populations on packaged lettuce and swedes significantly increased ( P  < 0.05, by 1–1.5 log cycles) during a 14-day storage period. During the same period, Listeria counts gradually decreased (by 1–1.5 log cycles) on coleslaw mix. Acid adaptation enhanced survival of Listeria spp. during storage in packages of vegetables which had relatively high in-pack CO₂ levels (i.e. 25% in packaged coleslaw, bean-sprouts). It is concluded that adapting listerial cells to mildly acidic conditions rendered cultures more resistant to relatively high (25–30%) CO₂ atmospheres.     

Effects of the indigenous microflora of minimally processed lettuce on the survival and growth of Listeria innocua

Interactions between the natural background microflora of shredded lettuce and Listeria innocua (in lieu of Listeria monocytogenes ) were studied. The effect of increasing the initial size of indigenous populations (from 10³ to 10⁶–10⁷ CFU g⁻¹) was tested for its ability to reduce L. innocua growth on shredded lettuce. Co-culture experiments were performed in model media, where bacterial isolates from the indigenous microflora were tested for possible inhibitory effects. Varying the size of the indigenous populations had no effect on L. innocua survival or growth. However, interactions with individual species and mixed populations from lettuce did affect the survival and growth of L. innocua in model media. In general, mixed populations diminished L. innocua growth. In the undiluted lettuce medium, the various species tested individually either reduced or did not affect the growth of L. innocua . However, when the medium was diluted, some species extended the survival of L. innocua . Competition between the indigenous microflora and L. innocua resided mostly with the Enterobacter spp. and not with the pseudomonads. Enterobacter cloacae was particularly effective in reducing L. innocua growth. Lactic acid bacteria also reduced L. innocua growth in undiluted media. It is concluded that interactions with the natural background microflora may play an important role in determining the dynamics of Listeria populations on shredded lettuce.     

Isolation and pulsed-field gel electrophoresis typing of Listeria monocytogenes from modified atmosphere packaged fresh-cut vegetables collected in Ireland

The incidence of Listeria monocytogenes in modified atmosphere packaged fresh-cut fruits and vegetables from chill cabinets of a supermarket in Ireland was investigated over a 2-year period. Overall, 9.58% of fresh-cut produce was contaminated with Listeria spp. Various species of Listeria were isolated from samples, including L. monocytogenes, L. seeligeri, L. innocua, L. welshimeri, and L. ivanovii. No fruit samples contained detectable L. monocytogenes. Overall, a total of 21 L. monocytogenes isolates (2.9% of samples) were recovered from a range of products, including dry coleslaw mix (80% shredded cabbage and 20% shredded carrot), bean sprouts, and leafy vegetables such iceberg, romaine, and radicchio lettuce and mixed salad leaves (curly endive, escarole, and radicchio leaves). Dry coleslaw mix appeared to have the highest incidence of Listeria contamination (20%) compared with other products. Listeria contamination was more frequent (P < 0.05) during the summer and autumn months than during the winter and spring months. The 21 L. monocytogenes isolates were subsequently subtyped by genomic macrorestriction techniques using ApaI with pulsed-field gel electrophoresis (PFGE). PFGE of digested DNA produced bands of 79 to 518 kb. Four PFGE profiles were identified, and approximately 50% of the isolates were associated with profile 1. This study indicates that fresh-cut vegetables packaged under a modified atmosphere can support growth of numerous species of Listeria, including L. monocytogenes.     

Polyphenol Oxidase from Apple (Malus domestica Borkh. cv Bramley's Seedling): Purification Strategies and Characterization

ABSTRACT Polyphenol oxidase (PPO) was isolated from Bramley's Seedling apples with 75.7‐fold purification and 26.5% recovery by ammonium sulfate precipitation, phenyl sepharose chromatography, ion exchange chromatography, and hydroxyapatite chromatography. Molecular weight was estimated to be about 45 kDa by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS PAGE). Optimum PPO activity was at pH 6.5 and greater than 50% activity was retained during storage for 72 h at pH 5.5 to 6.5. Optimum temperature for activity was 30 °C and the enzyme had residual activity of greater than 50% during storage for 72 h at 20 °C to 30 °C and for 24 h at 40 °C to 50 °C. Of the substrates tested, activity was greatest with 4‐methylcatechol followed by catechol, pyrogallol, and (?)epicatechin. The most effective inhibitors tested were sodium metabisulfite and ascorbic acid.     

Quality of Shredded Carrots as Affected by Packaging Film and Storage Temperature

ABSTRACT: Different packaging films and storage temperatures were used to establish a range of equilibrium modified atmospheres for storage of shredded carrots. Quality and storage-life of the packaged shredded carrots were determined using sensory evaluation, microbial counts, and a range of physical tests. Minimal processing steps such as peeling and shredding caused physical damage, physiological stress, and enhanced microbial growth, leading to a reduced shelf life when compared to the whole vegetable. A P-plus microporous film (CO₂ permeability of 29 X 10³mL.m ^-2.d ^-1.atm ^-1) was the most suitable for the storage of shredded carrots. Findings indicated that deterioration in these products was triggered by the depletion of oxygen more than by the rise in carbon dioxide.     

Effects of argon enriched low-oxygen atmospheres and of high-oxygen atmospheres on the kinetics of polyphenoloxidase (PPO)

Abstract: The reported benefits of enrichment of air atmospheres with argon or oxygen for control of enzymatic browning were investigated by determining the effects of these atmospheres on PPO kinetics. Kinetics of purified apple PPO and a commercially available mushroom PPO were studied in an in vitro model system. Enrichment with argon produced greater inhibitory effects than the current industry practice of enrichment with nitrogen. Km^app values (mM) for apple PPO in 3%O₂/97%Ar, 3%O₂/97%N₂, and air, were 133, 87, and 48, respectively. The data indicate that inhibition by both gases is competitive, and also support the hypothesis that the greater inhibitory effect of argon was proportional to the size of the Van der Waals radius of argon against nitrogen (1.91Å against 1.54Å). Much smaller inhibitory effects were observed in the presence of 80% O₂ (Km^app 57mM), and the nature of this inhibition was less clear. The results suggest that the benefits of argon enrichment may be relatively small, and may require critical enzyme, substrate, and gas levels to be successful. However, these benefits may be exploitable commercially in some fresh‐cut products, and may allow less anoxic atmospheres to be used. Practical Application: Control of enzymatic browning without sulfites continues to be a challenge in some fresh‐cut products. While sporadic benefits of these atmospheres in control of enzymatic browning have been reported, results have been inconsistent in commercial practice. The results suggest that the benefits of argon enrichment may be relatively small, and may require critical enzyme, substrate, and gas levels to be successful. However, these benefits may be exploitable commercially in some fresh‐cut products, and allow less anoxic atmospheres to be used.     

Effects of Dietary Supplementation with Camelina Oil on Porcine Blood Lipids

ABSTRACT: The effects of dietary supplementation with camelina oil on porcine plasma fatty acid composition and on serum cholesterol/triglyceride concentrations were investigated and compared with the effects of fish oil. The diets consisted of a control diet and diets supplemented with 5% camelina oil, 10% camelina oil and 5% fish oil. The camelina and fish-oil diets increased plasma ω3 fatty acids and reduced plasma ω6 fatty acids. Plasma eicosapentaenoic acid (C20:5ω3, EPA) was increased more by the fish oils diet than the camelina-oil diets. Serum triglyceride levels were reduced by the camelina-oil diets.