Authors:Kinga Horváth, Zs. Seregély, I. Dalmadi, Éva Andrássy and J. Farkas
The utility of chemosensor array (EN) signals of head-space volatiles of aerobically stored pork cutlets as a non-invasive technique for monitoring their microbiological load was studied during storage at 4, 8 and 12 °C, respectively. The bacteriological quality of the meat samples was determined by standard total aerobic plate counts (TAPC) and colony count of selectively estimated
(PS) spp., the predominant aerobic spoilage bacteria. Statistical analysis of the electronic nose measurements were principal component analysis (PCA), and canonical discriminant analysis (CDA). Partial least squares (PLS) regression was used to model correlation between microbial loads and EN signal responses, the degree of bacteriological spoilage, independently of the temperature of the refrigerated storage. Sensor selection techniques were applied to reduce the dimensionality and more robust calibration models were computed by determining few individual sensors having the smallest cross correlations and highest correlations with the reference data. Correlations between the predicted and “real” values were given on cross-validated data from both data reduced models and for full calibrations using the 23 sensor elements. At the same time, sensorial quality of the raw cutlets was noted subjectively on faultiness of the odour and colour, and drip formation of the samples. These preliminary studies indicated that the electronic nose technique has a potential to detect bacteriological spoilage earlier or at the same time as olfactory quality deterioration.
Authors:J. Farkas, Éva Andrássy, L. Mészáros and Anna Simon
Suspensions of a bioluminescent (luxAB) transformant of Listeria monocytogenes in pH 7.0 phosphate buffer were pressurised and the effect of the pressure treatment was monitored by plate counting. When the bacteria were suspended in NaCl- and nisin-free buffer the number of colony forming units (CFU) decreased by 3 and 6 log cycles after 300 MPA for 10 and 30 min, respectively. Supplementing the plating medium with 5% NaCl did not influence the colony forming capacity of non-pressurised cells, however, CFU of residual populations after respective treatments of 300 MPa for 10 and 30 min were reduced by a further 2 and 3.5 log cycles in case of salt containing plates. Nisin-addition to the plating medium caused less than one log unit decrease in the CFU of the non-pressurised population. However, the CFU of 10 min-pressurised sample was 4 log cycles less in the nisin-containing plates than in the nisin-free ones, whereas no colonies were formed in the nisin-containing plates even when 1 ml was inoculated from the originally 1010 CFU/ml population after 300 MPa for 30 min. The luciferase activities (bioluminescence intensities) decreased concomitant with the reduction of the viable cell counts, however, they were approx. 0.6-0.8 log units less in the presence of 5% NaCl in the pressurised suspension than those expected from the previously established linear correlation between the logarithmic light outputs and the logarithmic viable cell counts.
Authors:Franciska Könczöl, Nelli Farkas, Tímea Dergez, J. Belágyi and D. Lőrinczy
DSC and EPR experiments were performed on human erythrocyte membranes and DPPC vesicles in order to study the effect of the
anaesthetic drug tetracaine on structure and dynamics of the lipid region. Experiments using spin label technique showed that
tetracaine induced fluidity changes of the lipid region in the environment of the fatty acid probe molecules incorporated
into the membranes in the vicinity of the lipid-water interface. Similarly to EPR observations, DSC measurements reported
decrease of the main melting and the pretransition temperature in comparison to control DPPC vesicles, which is the sign of
destabilisation of the structure in the head group region of the lipids. Similar effect was observed in the case of erythrocytes
where the protein conformation was also controlled in the presence of drug. A separated membrane melting with well distinguished
membrane protein phase transition was found that was affected significantly by tetracaine. These results suggest that tetracaine
is able to modify not only the internal dynamics of erythrocyte membranes and produce destabilisation of the lipid structure,
but the protein system as well. These might lead to further damage of the biological functions.
Authors:T. Dergez, F. Könczöl, N. Farkas, J. Belágyi and D. Lőrinczy
Summary The heat capacity of contractile proteins actin and myosin was studied in psoas muscle of rabbit in strongly and weakly binding state of myosin to actin as a function of temperature by DSC. Deconvolution of the unfolding scans makes possible to characterize the structural domains of the macromolecules. We tried to approach the unfolding process in different intermediate state of ATP hydrolysis. The thermal transitions were calorimetrically irreversible, therefore the two-state irreversible model that describes fairly well the denaturation of different proteins was used for evaluation of the denaturation processes in muscle fibers in strongly (rigor, ADP) and weakly binding states (ATP·Vi, ADP·AlF4) of myosin to actin. Deconvolution resulted in four transitions, the first three transition temperatures were almost independent of the intermediate states of muscle, the last transition temperature was shifted to higher temperature, when the buffer solution was manipulated. The mean values in strongly binding states were Tm1=52.9±0.7°C, Tm2=57.9±0.7°C, Tm3=63.7±1.0°C and Tm4=67.8±0.7°C, but the last transition increased to higher temperature depending on the Pi analogue.
Authors:D. Lőrinczy, N. Hartvig, N. Farkas and J. Belagyi
Differential scanning calorimetry and electron paramagnetic resonance experiments were performed on glycerinated muscle fibres
to study the effect of the binding of nucleotides (ADP and AMP⋅PNP) to myosin. The thermal unfolding of muscle fibres showed
three discrete domain regions with thermal stabilities of 52.2, 58.8 and 67.8°C. AMP⋅PNP markedly affected the transitions,
implying the strong interaction between AMP⋅PNP and catalytic domain, and partial dissociation of heads from actin. ADP produced
only small changes in transition temperatures.
Spectrum deconvolution performed on isothiocyanate-labelled fibres in AMP⋅PNP-state resulted in two populations; 50% of labels
was highly ordered with respect to fibre axis, whereas the other 50% of labels was randomly oriented. The conformation of
the myosin heads which showed high degree of order were in the strongly binding ADP-state, the heads being attached to actin
differ from those of heads in rigor. The results support the suggestion that the attached heads in strongly binding state
to actin might have different local conformations.
Authors:D. Lőrinczy, F. Könczöl, L. Farkas, J. Belagyi and C. Schick
Electron paramagnetic resonance (EPR, ST-EPR) and differential scanning calorimetry(DSC) were used in conventional and temperature
modulated mode to study internal motions and energetics of myosin in skeletal muscle fibres in different states of the actomyosin
ATPase cycle. Psoas muscle fibres from rabbit were spin-labelled with an isothiocyanate-based probe molecule at the reactive
sulfhydryl site (Cys-707) of the catalytic domain of myosin. In the presence of nucleotides (ATP, ADP, AMP⋅PNP) and ATP or
ADP plus orthovanadate, the conventional EPR spectra showed changes in the ordering of the probe molecules in fibres. In MgADP
state a new distribution appeared; ATP plus orthovanadate increased the orientational disorder of myosin heads, a random population
of spin labels was superimposed on the ADP-like spectrum.
In the complex DSC pattern, higher transition referred to the head region of myosin. The enthalpy of the thermal unfolding
depended on the nucleotides, the conversion from a strongly attached state of myosin to actin to a weakly binding state was
accompanied with an increase of the transition temperature which was due to the change of the affinity of nucleotide binding
to myosin. This was more pronounced in TMDSC mode, indicating that the strong-binding state and rigor state differ energetically
from each other. The different transition temperatures indicated alterations in the internal microstructure of myosin head
region The monoton decreasing TMDSC heat capacities show that Cp of biological samples should not be temperature independent.
Authors:M. Kiss, F. Könczöl, N. Farkas, D. Lőrinczy and J. Belagyi
The effect of free radicals obtained in hydroxyl and cerium(IV)-nitrilotriacetic acid free radical generating systems on contractile
proteins (actin, myosin and their complexes in glycerinated muscle fibres) was studied using differential scanning calorimetry
and spin trapping electron paramagnetic resonance technique. The analysis of spectra showed that selective attack of thiol
groups – Cys-257 and Cys-374 residues of actin, and among others Cys-707 residue of myosin – and random attack of sidechains
of the main proteins of muscle tissue produced structural and functional changes, which affected the ATP hydrolysis cycle
and very likely the dynamics of actin. The melting curves obtained on protein systems support the view that global conformational
changes accompany the local damage of free radicals.
Authors:D. Hovorka, Z. Farkaš, J. Spišiak, J. Krištín, L. Števula, I. Túnyi and A. Kaplíková
In the paper we present results of multi-analytical study of pottery fragments archaeologically ranked from the Older Linear till the Middle Danube Tumulus Cultures, e.g. originated in the time-span of approximately 4700 years. In the studied set of pottery fragments we didn’t observed substantial differences of the raw materials used and temperatures of firing/annealing comparing studied set through the whole mentioned time-span didn’t surpass 650 °C in any of artefacts studied. The most realistic is to rank temperatures of firing/annealing close to the 600 °C. Oxidizing/reducing conditions during firing/annealing changed. Above statemets are based on the application of the following laboratory methods: stereoscopic observation of natural splitting planes, thin sections studies under polarizing microscope, scanning electron microscope, X-ray diffraction studies, organic matter determination (its quantity as well as quality) and the archaeopal aeomagnetic study.
Authors:Z. Farkas, J. Márki-Zay, Judit Kucsera, Cs. Vágvölgyi, W. Golubev and Ilona Pfeiffer
Wickerhamomyces anomalus VKM Y-159 strain produces two types of toxin designated as WAKT a and WAKT b, encoded by chromosomal genes. The WAKT a toxin is heat-labile, pronase sensitive acting in pH range 3–4 affecting on several yeasts including pathogenic Candida species while the WAKT b toxin is protease- and thermo-resistant, acting in pH range 3–7 on two species, Candida alai and Candida norvegica. The rapid decrease of the number of viable cells after toxin treatment demonstrates that both toxins have cytocidic effect.