Introduction. Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation. Methods. We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method. Results. The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, p<0.05 and 29%, p<0.01 measured by DNPH and FTIR-ATR methods, respectively). These results were consistent with the proteins phosphorylation analysis. Conclusion. The FTIR-ATR method may be applied to measure the level of plasma proteins peroxidation.
This paper presents the current state of the art of noninvasive glucose monitoring. In recent years, we can observe constant increase in the incidence of diabetes. About 40% of all performed blood tests apply to the glucose tests. Formerly, this lifestyle disease occurred mainly in rich countries, but now it is becoming more common in poorer countries. It is related to the increase in life expectancy, unhealthy diet, lack of exercise, and other factors. Untreated diabetes may cause many complications or even death. For this reason, daily control of glucose levels in people with this disorder is very important. Measurements with a traditional glucometer are connected with performing finger punctures several times a day, which is painful and uncomfortable for patients. Therefore, researches on other methods are ongoing. A method that would be fast, noninvasive and cheap could also enable testing the state of the entire population, which is necessary because of the number of people currently living with undiagnosed type 2 diabetes. Although the first glucometer was made in 1966, the first studies on glucose level measurement in tear film were documented as early as 1937. This shows how much a noninvasive method of diabetes control is needed. Since then, there have been more and more studies on alternative methods of glucose measurement, not only from tear fluid, but also from saliva, sweat, or transdermally.
CsgA is an aggregating protein from bacterial biofilms, representing a class of functional amyloids. Its amyloid propensity is defined by five fragments (R1–R5) of the sequence, representing non-perfect repeats. Gate-keeper amino acid residues, specific to each fragment, define the fragment’s propensity for self-aggregation and aggregating characteristics of the whole protein. We study the self-aggregation and secondary structures of the repeat fragments of Salmonella enterica and Escherichia coli and comparatively analyze their potential effects on these proteins in a bacterial biofilm. Using bioinformatics predictors, ATR-FTIR and FT-Raman spectroscopy techniques, circular dichroism, and transmission electron microscopy, we confirmed self-aggregation of R1, R3, R5 fragments, as previously reported for Escherichia coli, however, with different temporal characteristics for each species. We also observed aggregation propensities of R4 fragment of Salmonella enterica that is different than that of Escherichia coli. Our studies showed that amyloid structures of CsgA repeats are more easily formed and more durable in Salmonella enterica than those in Escherichia coli.
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