Periwinkle (Littorina littorea) as a sentinel species: a field study integrating chemical and biological analyses

The gastropod Littorina littorea (common periwinkle) is an abundant and widespread North Atlantic species. The characteristic development of Intersex in L. littorea has been widely applied as a biomarker for tributyltin (TBT) contamination. Here, we assess the potential of L. littorea as a novel sentinel species for evaluating the sublethal effects in wild populations of widely distributed contaminants. We collected animals from six sites across the South coast of England. Tissue concentrations of polycyclic aromatic hydrocarbons (PAHs), organotin compounds (OTCs), polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs) were measured and compared with biomarkers of damage to DNA (Comet assay), lysosomal stability (NRR assay), and endocrine disruption (Intersex development). There was a strong correlation between DNA damage and PAH bioaccumulation (n=6, r=0.84, p<0.05), as well as that between Intersex development and OTC pollution (n=6, r=0.91, p<0.05). The relationship between PAH bioaccumulation and DNA strand breaks was nonlinear, highlighting the need to consider the role of adaptive mechanisms in the interpretation of field results. These results illustrate the potential use of periwinkles for monitoring a wide range of priority pollutants.     

Micro-kinetic model of fructo-oligosaccharide synthesis for prebiotic products

A new micro-kinetic model of the enzyme-catalyzed synthesis of fructo-oligosaccharides (FOS) was developed. A commercial enzyme mixture Pectinex® Ultra SP-L derived from Aspergillus aculeatus was used. A variety of initial enzyme concentrations (1–5 vol%) and sucrose concentrations (400–600 g/L) were experimentally investigated and included in kinetic modeling. Several variations of kinetic mechanisms and corresponding models have been examined. A hybrid genetic algorithm was used to predict the kinetic parameters simultaneously for all experimental data. The best fitting model has been adopted, and with an average error of 13.34%, it describes the experimental data very well. The influence of initial concentrations on the conversion of sucrose and production of FOS is being carefully investigated. It was shown that the initial sucrose concentration significantly affects the highest level of FOS concentration, but the enzyme concentration controls the time at which maximum is reached as well as the rate of FOS decomposition.     

Antioxidant effectiveness of coffee extracts and selected constituents in cell‐free systems and human colon cell lines

Scope: Epidemiological studies suggest that coffee can reduce the risk of degenerative diseases such as diabetes type 2, cardiovascular disease and cancer. These beneficial effects have partly been attributed to the antioxidant activity of coffee. We determined composition and antioxidant potential of differentially roasted coffee extracts and investigated the impact of selected original constituents and roast products. Methods and results: Parameters studied were direct antioxidant activity (trolox equivalent antioxidant capacity/oxygen radical absorbing capacity), cellular reactive oxygen species (ROS) level, DNA damage and protein expression of NAD(P)H: quinone oxidoreductase, γ‐glutamylcysteine ligase and glutathione reductase in HT‐29/Caco‐2 cells at 24‐h incubation. All extracts showed distinct direct antioxidant activity: medium roasts>light roast AB1 (caffeoylquinic acid (CQA)‐rich Arabica Brazil extract); dark roast AB2 (N‐methylpyridinium (NMP)‐rich Arabica Brazil extract), and diminished t‐butylhydroperoxide‐induced ROS level in HT‐29 cells (AB2>medium roasts>AB1). NAD(P)H:quinone oxidoreductase 1 expression and γ‐glutamylcysteine ligase expression were distinctly induced by AB1 and 5‐CQA, but not by AB2 and NMP. 5‐CQA and caffeic acid exhibited highest trolox equivalent antioxidant capacity/oxygen radical absorbing capacity values (5‐CQA: 1.3/3.5 mM and caffeic acid: 1.3/3.9 mM trolox); ROS level was distinctly diminished by 5‐CQA (≥3 μM), catechol (30 μM) and trigonelline (≥30 μM), whereas menadione‐induced DNA damage in Caco‐2 cells was reduced by NMP compounds (1–30 μM). Conclusion: The results emphasize that both original constituents and roast products contribute to the cellular antioxidant effectiveness of coffee.     

Mitochondrial Side Effects of Surgical Prophylactic Antibiotics Ceftriaxone and Rifaximin Lead to Bowel Mucosal Damage

Despite their clinical effectiveness, a growing body of evidence has shown that many classes of antibiotics lead to mitochondrial dysfunction. Ceftriaxone and Rifaximin are first choice perioperative antibiotics in gastrointestinal surgery targeting fundamental processes of intestinal bacteria; however, may also have negative consequences for the host cells. In this study, we investigated their direct effect on mitochondrial functions in vitro, together with their impact on ileum, colon and liver tissue. Additionally, their impact on the gastrointestinal microbiome was studied in vivo, in a rat model. Rifaximin significantly impaired the oxidative phosphorylation capacity (OxPhos) and leak respiration in the ileal mucosa, in line with increased oxidative tissue damage and histological changes following treatment. Ceftriaxone prophylaxis led to similar changes in the colon mucosa. The composition and diversity of bacterial communities differed extensively in response to antibiotic pre-treatment. However, the relative abundances of the toxin producing species were not increased. We have confirmed the harmful effects of prophylactic doses of Rifaximin and Ceftriaxone on the intestinal mucosa and that these effects were related to the mitochondrial dysfunction. These experiments raise awareness of mitochondrial side effects of these antibiotics that may be of clinical importance when evaluating their adverse effects on bowel mucosa.     

Oleic Acid Protects Endothelial Cells from Silica-Coated Superparamagnetic Iron Oxide Nanoparticles (SPIONs)-Induced Oxidative Stress and Cell Death

Superparamagnetic iron oxide nanoparticles (SPIONs) have great potential for use in medicine, but they may cause side effects due to oxidative stress. In our study, we investigated the effects of silica-coated SPIONs on endothelial cells and whether oleic acid (OA) can protect the cells from their harmful effects. We used viability assays, flow cytometry, infrared spectroscopy, fluorescence microscopy, and transmission electron microscopy. Our results show that silica-coated SPIONs are internalized by endothelial cells, where they increase the amount of reactive oxygen species (ROS) and cause cell death. Exposure to silica-coated SPIONs induced accumulation of lipid droplets (LD) that was not dependent on diacylglycerol acyltransferase (DGAT)-mediated LD biogenesis, suggesting that silica-coated SPIONs suppress LD degradation. Addition of exogenous OA promoted LD biogenesis and reduced SPION-dependent increases in oxidative stress and cell death. However, exogenous OA protected cells from SPION-induced cell damage even in the presence of DGAT inhibitors, implying that LDs are not required for the protective effect of exogenous OA. The molecular phenotype of the cells determined by Fourier transform infrared spectroscopy confirmed the destructive effect of silica-coated SPIONs and the ameliorative role of OA in the case of oxidative stress. Thus, exogenous OA protects endothelial cells from SPION-induced oxidative stress and cell death independent of its incorporation into triglycerides.     

Stereodivergent Biocatalytic Formal Reduction of α-Angelica Lactone to (R)- and (S)-γ-Valerolactone in a One-Pot Cascade

The formal asymmetric and stereodivergent enzymatic reduction of α-angelica lactone to both enantiomers of γ-valerolactone was achieved in a one-pot cascade by uniting the promiscuous stereoselective isomerization activity of Old Yellow Enzymes with their native reductase activity. In addition to running the cascade with one enzyme for each catalytic step, a bifunctional isomerase-reductase biocatalyst was designed by fusing two Old Yellow Enzymes, thereby generating an unprecedented case of an artificial enzyme catalyzing the reduction of nonactivated C=C bonds to access (R)-valerolactone in overall 41 % conversion and up to 91 % ee. The enzyme BfOYE4 could be used as single biocatalyst for both steps and delivered (S)-valerolactone in up to 84 % ee and 41 % overall conversion. The reducing equivalents were provided by a nicotinamide recycling system based on formate and formate dehydrogenase, added in a second step. This enzymatic system provides an asymmetric route to valuable chiral building blocks from an abundant bio-based chemical.     

Characterization of Competitive Inhibitors of Plasmodium falciparum cGMP-Dependent Protein Kinase**

Plasmodium falciparum cGMP-dependent protein kinase (PfPKG) is an enticing antimalarial drug target. Novel chemotypes are needed because existing inhibitors have safety issues that may prevent further development. This work demonstrates isoxazole-based compounds are potent ATP competitive inhibitors of PfPKG and discloses a new analogue in this series. Isoxazoles 3 and 5 had K_i values that are comparable to a known standard, 4-[2-(4-fluorophenyl)-5-(1-methylpiperidine-4-yl)-1H pyrrol-3-yl] pyridine. They also exhibited excellent selectivity for PfPKG over the human orthologue and the gatekeeper mutant T618Q PfPKG, which mimics the less accessible binding site of the human orthologue. The human orthologue's larger binding site volume is predicted to explain the selectivity of the inhibitors for the P. falciparum enzyme.     

Effect of grey component replacement processing and varnishing on the light fastness of colours on cardboard prints

The visual appearance of packaging is one of its most important features. Electromagnetic irradiation causes a change in colour and often leads to colour fading (ie, to diminished visual appearance of a printed product). This research study aimed to determine the combined effect of grey component replacement image processing and varnishing on the protection of prints' appearance when exposed to accelerated ageing. For that purpose, two test charts were printed in a lithographic press and coated with water-based and ultraviolet-curable varnish before exposing them to xenon light. The prepared prints were evaluated by determining the tone values and colour difference, as well as the rub and abrasion resistance. In addition, Fourier Transform–infrared-attenuated total reflectance spectra were recorded to detect possible chemical changes caused by the (AcA). The results showed that the investigated period of (AcA) caused significant differences in the tone values of yellow, while the other primary colours proved to be more resistant. The water-based varnish provided better protection than the ultraviolet-curable varnish. The grey component replacement image processing diminished the colour difference caused by (AcA) on all the prepared print samples, and was particularly detected in tertiary colours containing the mid-range tone values of yellow (34%-68%). Both varnishes improved the rub resistance of aged prints, but only ultraviolet-curable varnish improved the abrasion resistance of both the unaged and the aged prints. In conclusion, applying varnish to prints ensured enhanced rub and abrasion resistance and diminished the colour change caused by (AcA). Finally, improved resistance to colour change was achieved by processing images with the grey component replacement method.