Reduction of Campylobacter jejuni by natural antimicrobials in chicken meat-related conditions

Nowadays it is essential to test new preservation and decontamination procedures using naturally occurring chemicals against important pathogenic bacteria in meat. We tested the antimicrobial effect of rosemary extracts and the bacteriocin nisin against Campylobacter jejuni at a low storage temperature (8 °C) with or without short-term pre-freezing. The antimicrobial effect of rosemary extract was four times greater in laboratory media than in chicken meat juice. Furthermore, low temperature storage conditions prolonged the survival of C. jejuni in chicken meat juice. Nisin, with an approximately 1.0 log reduction was neither effective alone nor in combination with the extract. Pre-freezing with plant extract addition proved to be effective treatment by more than 3.0 log reduction in 48 h. The results in chicken meat food model again showed the synergistic effect of freezing and plant extract antimicrobial activity. As the combination of pre-freezing and plant extract treatment reduced the cell number by more than 2.0 log reduction, studies should be conducted to further evaluate this promising treatment for Campylobacter reduction in the poultry meat supply.     

Cytotoxic and antimitotic effects of N-containing Monascus metabolites studied using immortalized human kidney epithelial cells

Recently the first Monascus metabolites with a pyridine ring were detected, the monascopyridines A and B. They are formally dehydrogenated derivatives of the red rice pigments rubropunctamine and monascorubramine. Because of their structural similarity, the toxicological effects of these secondary metabolites were studied using immortalized human kidney epithelial cells. The cytotoxicity was determined with the following different endpoint detection methods: metabolic activity, trypan blue exclusion, and electronic cell counting. The compounds led to EC(50) values between 11 and 31 micromol/L but the pigments caused a stronger reduction of the cell viability. Also, the apoptotic potential was examined by measuring caspase 3 activity and detecting apoptotic bodies, but none of the tested compounds induced apoptosis. All four substances caused a rise of the mitotic index to about 9% (100 micromol/L monascopyridine A and B) and 20% (25 micromol/L rubropunctamine and monascorubramine). The significant decrease of the ratio of cells in the ana- and telophase to cells in the prometa- and metaphase proved a stop of the mitosis at the meta- to anaphase control point. The compounds caused mitotic arrest and the formation of structural damages like c-mitosis through interaction with the mitotic spindle. These effects point to an aneuploidy inducing potential, which is linked to cancer formation.     

New Monascus metabolites: structure elucidation and toxicological properties studied with immortalized human kidney epithelial cells

Monascus sp. are used for the production of red yeast rice, which has been applied in Asian food and medicine for centuries. The fungi form several secondary metabolites, among others the monascopyridines A and B. We have now detected two new monascopyridines (named C and D) with similar UV absorption spectra and maxima at 306-307 nm in red rice fermented with M. purpureus. The new monascopyridines have the same chromophores as the already known and differ in a missing gamma-lactone ring only. Monascopyridine C has a hexanoyl side chain, whereas monascopyridine D is the higher homologue with an octanoyl side chain. The toxicological properties of monascopyridines were studied using immortalized human kidney epithelial cells, displaying cytotoxic effects in micromolar concentrations with median effective concentration values between 20.7 and 43.2 micromol/L, depending on the compound and method used. The monascopyridines C and D did not induce apoptosis. However, they caused a rise of the mitotic index from 3.21 +/- 0.27% (control) to maximum 8.14 +/- 0.89% (monascopyridine D) by an accumulation of cells in the metaphase with a simultaneous decrease of cells in the ana- and telophase. Monascopyridine-induced metaphase arrest is caused by a partial or complete loss of the spindle apparatus as indicated by the occurrence of abnormal metaphases and immunological staining of alpha-tubulin. These effects are indicating an aneuploidic potential and therefore the monascopyridines C and D might contribute to tumor formations.     

Correlation among sintering process,porosity, and creep deformation of refractory concrete

The aim of this paper is to establish the correlation among sintering process, porosity, and important thermo-mechanical property of refractory concrete, i.e., creep. Creep deformation was investigated according to the standard laboratory procedure applied at three temperatures: 1200, 1300, and 1400 °C. Corundum and bauxite-based refractory concretes were investigated. The concretes are varying in chemical and mineralogical composition. Both loss of strength and degradation of material occur when refractory concrete is subjected to increased temperature and compressive static load. Measuring of thermo-mechanical properties can indicate and monitor the changes within microstructure. Variation of refractory concrete microstructure, as a consequence of sintering process, during exposure to constant compressive load and constant elevated temperature during certain time-intervals was investigated using scanning electron microscope and Image Pro Plus program for image analysis. Obtained results of the investigation proved that creep can be useful method when type of refractory concrete is to be chosen for an application.     

Reversible acetonitrile-induced inactivation/activation of thermolysin

Thermolysin is catalytically inactive in mixtures of 10-15 % acetonitrile in aqueous buffer. Unexpectedly, dilution of the inactive enzyme with acetonitrile leads to complete recovery of the catalytic activity in a similar way to dilution with aqueous buffer. Circular dichroism and fluorescence studies of thermolysin in the same solvent mixtures reveal discontinuous changes in the overall secondary and tertiary protein structure that correlate well with the reversible differences in catalytic activity. The spectra on either side of the minimum activity point are different from each other, a fact indicating that the enzyme may be able to access two active conformations which are thermodynamically stable in different solvent environments.     

Quantitative pulsed CEST-MRI at a clinical 3T MRI system

Objectives

The goal of this study was to quantify CEST related parameters such as chemical exchange rate and fractional concentration of exchanging protons at a clinical 3T scanner. For this purpose, two CEST quantification approaches—the AREX metric (for ‘apparent exchange dependent relaxation’), and the AREX-based Ω-plot method were used. In addition, two different pulsed RF irradiation schemes, using Gaussian-shaped and spin-lock pulses, were compared.

Materials and methods

Numerical simulations as well as MRI measurements in phantoms were performed. For simulations, the Bloch–McConnell equations were solved using a two-pool exchange model. MR experiments were performed on a clinical 3T MRI scanner using a cylindrical phantom filled with creatine solution at different pH values and different concentrations.

Results

The validity of the Ω-plot method and the AREX approach using spin-lock preparation for determination of the quantitative CEST parameters was demonstrated. Especially promising results were achieved for the Ω-plot method when the spin-lock preparation was employed.

Conclusion

Pulsed CEST at 3T could be used to quantify parameters such as exchange rate constants and concentrations of protons exchanging with free water. In the future this technique might be used to estimate the exchange rates and concentrations of biochemical substances in human tissues in vivo.