Hydrolyzed fumonisins HFB1 and HFB2 are acylated in vitro and in vivo by ceramide synthase to form cytotoxic N-acyl-metabolites

Fumonisins B1 and B2 (FB1 and FB2) are the most abundant members of the fumonisins--mycotoxins that are produced by Fusarium verticillioides and are natural inhibitors of ceramide synthase. Their hydrolyzed forms, HFB1 and HFB2 (also called AP1 and AP2) are found in some foods, and they are not only inhibitors of ceramide synthase but also undergo acylation by this enzyme. This study characterized the conversion of HFB1 and HFB2 by ceramide synthase to their respective N-acylated metabolites using rat liver microsomes and palmitoyl-CoA or nervonoyl-CoA as cosubstrates, and examined animals that had been dosed with hydrolyzed fumonisins to ascertain if acylation occurs in vivo. Using an HPLC-MS/MS method that allowed the sensitive and selective detection of the acylation products, both HFB1 and HFB2 were found to be metabolized in vitro to nervonoyl- or palmitoyl-HFB1 and -HFB2 (i.e. C24:1-HFB1/2 and C16-HFB1/2, respectively). The apparent vmax was considerably higher for formation of C24:1HFB1 (157 pmol/min/mg protein) than for formation of C16HFB1 (8.7 pmol/min/mg protein). The acylation products also inhibited ceramide synthase and significantly reduced the number of viable cells in an in vitro [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] assay using a human colonic cell line (HT29). Furthermore, HPLC-MS/MS analysis of tissues from rats given intraperitoneal doses of HFB1 confirmed that formation of N-acyl-HFB1 occurs in vivo to produce metabolites with fatty acids of various chain lengths. The contribution of acylated HFB1 and HFB2 metabolites to fumonisin toxicity in vivo warrants further investigation.     

Assessment of green cleaning effectiveness on polychrome surfaces by MALDI‐TOF mass spectrometry and microscopic imaging

This article proposes an innovative methodology which employs nondestructive techniques to assess the effectiveness of new formulations based on ionic liquids, as alternative solvents for enzymes (proteases), for the removal of proteinaceous materials from painted surfaces during restoration treatments. Ionic liquids (ILs), also known as “designer” solvents, because of their peculiar properties which can be adjusted by selecting different cation‐anion combinations, are potentially green solvents due totheir low vapour pressure. In this study, two ionic liquids were selected: IL1 (1‐butyl‐3‐methylimidazolium tetrafluoroborate ([BMIM][BF₄])) and IL2 (1‐ethyl‐3‐methylimidazolium ethylsulphate ([EMIM][EtSO₄])). New formulations were prepared with these ILs and two different proteases (E): one acid (E1—pepsin) and one alkaline (E2—obtained from Aspergillus sojae). These formulations were tested on tempera and oil mock‐up samples, prepared in accordance with historically documented recipes, and covered with two different types of protein‐based varnishes (egg white and isinglass—fish glue). A noninvasive multiscale imaging methodology was applied before and after the treatment to evaluate the cleaning's effectiveness. Different microscopic techniques—optical microscopy (OM) with visible and fluorescent light, scanning electron microscopy (SEM) and atomic force microscopy (AFM)—together with Matrix‐Assisted Laser Desorption/Ionization—Time of Flight Mass Spectrometry (MALDI‐TOF MS) were applied on areas cleaned with the new formulations (IL + E) and reference areas cleaned only with the commercial enzyme formulations (gels). MALDI‐TOF proved particularly very useful for comparing the diversity and abundance of peptides released by using different enzymatic systems. Microsc. Res. Tech. 77:574–585, 2014. © 2014 Wiley Periodicals, Inc.     

Chemical,technological, and nutritional characteristics of two lines of “farro” (Triticum turgidum ssp. dicoccum)

In recent years, the renewed interest for foods with a natural image has increased the demand for dry pasta produced from “hulled” wheat such as the Triticum turgidum ssp. dicoccum, also known as “farro”. In order to contribute to the general knowledge, two lines of farro were considered in this study. To have a comparison, an old cultivar of Triticum turgidum ssp. durum (Senatore Cappelli) in addition to a commercial semolina were also examined. All semolina samples were used to produce pasta samples. Results showed some differences among pasta samples that seem to be due not to the presence of specific protein subunits but especially to the quantitative ratio between the different subunits. Results also reconfirmed the role played by the drying technology that is able to affect the sensory characteristics of pasta products.     

Inhibitory effect of organic acids on arcobacters in culture and their use for control of Arcobacter butzleri on chicken skin

The inhibitory effects of 17 organic acids (C₂-C₁₆ fatty acids, sorbic, benzoic, phenylacetic, fumaric, succinic, lactic, malic and citric) on Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii were investigated by determining their IC₅₀ values, defined as the concentration of acid at which the target DNA sequence was expressed at 50% of the positive control level in cultures incubated at 30 °C for 24 h. DNA was analysed by real-time PCR. The Arcobacter strains tested were inhibited by all the organic acids, with the sensitivities in the order A. skirrowii > A. cryaerophilus > A. butzleri. Eight acids with IC₅₀ values of < 1 mg/mL against A. butzleri were tested for their effects on A. butzleri inoculated on chicken carcasses at a concentration of 5 log CFU/g of skin. Inoculated halved carcasses were immersed in solutions of the acids at 5 mg/mL for 1 min. Samples of skin were collected from carcass halves after storage at 4 °C for 0, 1, 2 or 3 days for enumeration of arcobacters on Muller-Hinton agar. All eight tested acids suppressed bacterial proliferation. The highest inhibitory activities were observed for benzoic, citric, malic and sorbic acids. Subsequent sensory analysis revealed benzoic acid to be the most suitable organic acid for chicken skin treatment.     

In-vitro cell exposure studies for the assessment of nanoparticle toxicity in the lung—A dialog between aerosol science and biology

The introduction of engineered nanostructured materials into a rapidly increasing number of industrial and consumer products will result in enhanced exposure to engineered nanoparticles. Workplace exposure has been identified as the most likely source of uncontrolled inhalation of engineered aerosolized nanoparticles, but release of engineered nanoparticles may occur at any stage of the lifecycle of (consumer) products. The dynamic development of nanomaterials with possibly unknown toxicological effects poses a challenge for the assessment of nanoparticle induced toxicity and safety.In this consensus document from a workshop on in-vitro cell systems for nanoparticle toxicity testing¹ an overview is given of the main issues concerning exposure to airborne nanoparticles, lung physiology, biological mechanisms of (adverse) action, in-vitro cell exposure systems, realistic tissue doses, risk assessment and social aspects of nanotechnology. The workshop participants recognized the large potential of in-vitro cell exposure systems for reliable, high-throughput screening of nanoparticle toxicity. For the investigation of lung toxicity, a strong preference was expressed for air–liquid interface (ALI) cell exposure systems (rather than submerged cell exposure systems) as they more closely resemble in-vivo conditions in the lungs and they allow for unaltered and dosimetrically accurate delivery of aerosolized nanoparticles to the cells. An important aspect, which is frequently overlooked, is the comparison of typically used in-vitro dose levels with realistic in-vivo nanoparticle doses in the lung. If we consider average ambient urban exposure and occupational exposure at 5 mg/m³ (maximum level allowed by Occupational Safety and Health Administration (OSHA)) as the boundaries of human exposure, the corresponding upper-limit range of nanoparticle flux delivered to the lung tissue is 3×10^?5–5×10^-3 μg/h/cm² of lung tissue and 2–300 particles/h/(epithelial) cell. This range can be easily matched and even exceeded by almost all currently available cell exposure systems.The consensus statement includes a set of recommendations for conducting in-vitro cell exposure studies with pulmonary cell systems and identifies urgent needs for future development. As these issues are crucial for the introduction of safe nanomaterials into the marketplace and the living environment, they deserve more attention and more interaction between biologists and aerosol scientists. The members of the workshop believe that further advances in in-vitro cell exposure studies would be greatly facilitated by a more active role of the aerosol scientists. The technical know-how for developing and running ALI in-vitro exposure systems is available in the aerosol community and at the same time biologists/toxicologists are required for proper assessment of the biological impact of nanoparticles.     

Beyond Yield Optimization: The Impact of Organosolv Process Parameters on Lignin Structure

When optimizing the process parameters of the acidic ethanolic organosolv process, the aim is usually to maximize the delignification and/or lignin purity. However, process parameters such as temperature, time, ethanol and catalyst concentration, respectively, can also be used to vary the structural properties of the obtained organosolv lignin, including the molecular weight and the ratio of aliphatic versus phenolic hydroxyl groups, among others. This review particularly focuses on these influencing factors and establishes a trend analysis between the variation of the process parameters and the effect on lignin structure. Especially when larger data sets are available, as for process temperature and time, correlations between the distribution of depolymerization and condensation reactions are found, which allow direct conclusions on the proportion of lignin's structural features, independent of the diversity of the biomass used. The newfound insights gained from this review can be used to tailor organosolv lignins isolated for a specific application.     

Solid–Water Interface Interaction of Selenium with Fe(II)-Bearing Minerals and Aqueous Fe(II) and S(-II) Ions in the Near-Field of the Radioactive Waste Disposal System

Selenium can be highly toxic in excess for both animals and humans. However, since its mobile forms can be easily adsorbed with ferric minerals, its mobility in the natural oxic environment is generally not an issue. Still, the removal and immobilization of the long-lived radioactive isotope ⁷⁹Se from the contaminated anoxic waters is currently a significant concern. ⁷⁹Se can be accessible in the case of radionuclidesˈ leaching from radioactive waste disposals, where anoxic conditions prevail and where ferrous ions and Fe(II)-bearing minerals predominate after corrosion processes (e.g., magnetite). Therefore, reductive and adsorptive immobilizations by Fe(II)-bearing minerals are the primary mechanisms for removing redox-sensitive selenium. Even though the information on the sorptive interactions of selenium and Fe(II)-bearing minerals seems to be well documented, this review focuses specifically on the state of the available information on the effects of the redox properties of Fe(II)-bearing solid phases (e.g., ferrous oxides, hydroxides, sulfides, and carbonates) on selenium speciation via redox transformation and co-occurring coprecipitation.     

Fertilization,but Not Post-Implantation Development,Can Occur in the Absence of Sperm and Oocyte Beta1 Integrin in Mice

Fertilization is a complex process that requires successive stages and culminates in the adhesion/fusion of gamete membranes. If the question of the involvement of oocyte integrins has been swept away by deletion experiments, that of the involvement of sperm integrins remains to be further characterized. In the present study, we addressed the question of the feasibility of sperm–oocyte adhesion/fusion and early implantation in the absence of sperm β1 integrin. Males and females with β1 integrin-depleted sperm and oocytes were mated, and fertilization outcome was monitored by a gestational ultrasound analysis. Results suggest that although the sperm β1 integrin participates in gamete adhesion/fusion, it is dispensable for fertilization in mice. However, sperm- and/or oocyte-originated integrin β1 is essential for post-implantation development. Redundancy phenomena could be at the origin of a compensatory expression or alternative dimerization pattern.     

GI,biliary tract and pancreas

Percutaneous stent-grafting is increasingly employed as a less invasive alternative to surgery for the treatment of infrarenal abdominal aortic aneurysms. It requires long-term imaging follow-up, to document the structural integrity of the device, to exculude perigraft channels and endograft leakages, as well as the shrinkage of the aneurysmal sac. The expectation of severe stent induced artifacts and safety concerns have prevented 3D MRA from being used. The purpose of this in vitro study was to investigate the imaging characteristics of a bifurcated stent graft with 3D MRA (3D Frourier transform fast spoiled GRE) at 1.5 T in comparison to those of CTA. Measurement of the stent wall thickness and luminal diameter were made on a agar gel embedded stent graft at five locations on both CTA and MRA images. The stent graft was depicted as a dark ring on MR images. Wall thickness measurments at the five locations of the stent graft overestimated the true stent thickness, while luminal diameters were slightly underestimated. Measurement differences between MR and CT were not statistically significant (P=0.67;P=0.85). Artifacts emanating from the platinum markers were considerably less severe on the MR-images. A wider area of signal loss was seen only at the insertion of the iliac stent leg into the aortic stent portion due to the overlap of two radio-opaque platinum markers. 3D MRA images should permit a comprehensive assessment of the arterial lumen, and of perivascular tissues.