Viability study of clinical bacterial strains by capillary electrophoresis and flow cytometry approaches

One of the challenges medicine faces is the constantly growing resistance of pathogens to various classes of antibiotics. In this study, we investigated the use of capillary electrophoresis (CE) to characterize and assess the physiological states of three clinical bacterial strains—methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA), and Escherichia coli extended-spectrum β-lactamases (ESβL)—exposed to different antibiotics. All chosen bacteria are the leading causes of healthcare-associated and hospital-acquired invasive infections in adults. In the first part of the research, it was determined the optimal incubation time of the tested strains with antibiotics, represented as an optimal time of 24 h. In the second part, we have compared two approaches: flow cytometry (FC) as a standard method and CE as a proposed alternative approach. The viability of clinical strains treated with different class antibiotics calculated in CE measurements was strongly correlated (>0.83 for MSSA, >0.92 for ESβL and MRSA) with the viability obtained on the basis of FC measurements. As a result, CE has a chance to become a modern diagnostic method used in clinical practice. The CE cutoff was found to be 50%; above this value, the strain shows resistance to the action of the antibiotic.     

Pressurized planar electrochromatography as a supporting tool for qualitative toxicological chemical analysis with thin-layer chromatography and UV–Vis spectrometry

ABSTRACT

Pressurized planar electrochromatography (PPEC) was applied to support qualitative toxicological chemical analysis performed with thin-layer chromatography (TLC) and UV–Vis spectrometry. Based on retention/migration distance data of substances obtained in TLC and PPEC systems and database of their wavelength maxima of remission UV–Vis spectra as well, a combined fit factor was calculated for substance identification. The involvement of PPEC, TLC, and UV–Vis spectral data together in calculation of the combined fit factor lead to its lower values for substances, which were not identical with reference, in comparison with those when the combined fit factor was calculated using TLC and spectral data only. The results evidence that involvement of PPEC data in qualitative toxicological chemical analysis performed with TLC and UV–Vis spectrometry enhances reliability of it.     

A thermal,sorptive and spectral study of HDTMA-bentonite loaded with uranyl phosphate

The influence of phosphate ions on the thermal stability of complexes obtained by adsorption of uranium(VI) on organobentonite was determined. Organoclay samples were prepared by the reaction of hexadecyltrimethylammonium bromide with bentonite. The isotherms of sorption/desorption of U(VI) from aqueous solutions containing phosphate ions onto different forms of bentonite were measured using the batch method. The highest amount of uranium was absorbed on HDTMA-bentonite in the presence of phosphates. This may have been associated with the complexing of U(VI) ions by phosphate ions, which interacted with surfactant cations probably via electrostatic forces. A TG–DSC–MS study showed that the thermal decomposition of the surfactant sorbed on bentonite proceeded in two stages: at 200–400 and at 600–800 °C. The first stage involved defragmentation and oxidation of surfactant cations present in the interior and on the surface of the mineral. The second stage involved oxidation of charcoal and simultaneous dehydroxylation of the sorbent. The oxidation of surfactant cations and the dehydroxylation of the mineral were suppressed in the presence of phosphates.     

Application of ICP-MS,INAA and RNAA to the determination of some “difficult” elements in infant formulas

In this work a determination of selected elements in the infant formulas commercially available on the Polish market was done. 14 different materials (milk-based formulas and grain porridges) were analyzed. Both, inductively coupled plasma mass spectrometry (ICP-MS) and instrumental neutron activation analysis (INAA) were applied for the determination of As, Cr, Fe and Se, which are recognized as the problematic elements for ICP-MS. For As and Se, the radiochemical NAA was also used. The daily intake of Se and Fe in the age 0–6 months for non-breast fed infants was estimated and compared with present safety limits.

     

Stinging Nettle (Urtica dioica L.) as a Functional Component in Durum Wheat Pasta Production: Impact on Chemical Composition,In Vitro Glycemic Index,and Quality Properties

Stinging nettle (Urtica dioica L.) is a good source of biologically active compounds with proven beneficial health effects. This study aimed to investigate the effect of nettle herb supplementation on chemical composition, including the content of selected minerals and pigments, the in vitro glycemic response, and the cooking and sensory quality of extruded pasta. Tagliatelle-shaped pasta was produced under semi-technical scale by partial replacement of durum wheat semolina with 0, 1, 2, 3, 4, and 5% of lyophilized nettle. The partial substitution with freeze-dried nettle caused a statistically significant (p ≤ 0.05) increase in the content of minerals, especially calcium, iron, potassium, and magnesium in the products. The calcium content in the pasta fortified with 5%-addition of stinging nettle was 175.9 mg 100 g⁻¹ and this concentration was 5.8 times higher than in the control sample. At the same time, high content of chlorophylls and carotenoids (237.58 µg g⁻¹ and 13.35 µg g⁻¹, respectively) was noticed. Enriching pasta with a 0–5% addition of stinging nettle resulted in a statistically significant (p ≤ 0.05) increase in the content of the total dietary fiber (TDF) (from 5.1 g 100 g⁻¹ to 8.82 g 100 g⁻¹) and the insoluble dietary fiber (IDF) (from 2.29 g 100 g⁻¹ to 5.63 g 100 g⁻¹). The lowest hydrolysis index of starch (HI = 17.49%) and the lowest glycemic index (GI = 49.31%) were noted for the pasta enriched with 3% nettle.     

Schisandra rubriflora Fruit and Leaves as Promising New Materials of High Biological Potential: Lignan Profiling and Effect-Directed Analysis

The effect-directed detection (EDD) of Schisandra rubriflora fruit and leaves extracts was performed to assess their pharmacological properties. The EDD comprised TLC—direct bioautography against Bacillus subtilis, a DPPH assay, as well as α-glucosidase, lipase, tyrosinase, and acetylcholinesterase (AChE) inhibition assays. The leaf extracts showed stronger antioxidant activity than the fruit extract as well as inhibition of tyrosinase and lipase. The fruit extract was found to be extremely active against B. subtilis and to inhibit α-glucosidase and AChE slightly more than the leaf extracts. UHPLC–MS/MS analysis was carried out for the bioactive fractions and pointed to the possible anti-dementia properties of the dibenzocyclooctadiene lignans found in the upper TLC fractions. Gomisin N (518 mg/100 g DW), schisanhenol (454 mg/100 g DW), gomisin G (197 mg/100 g DW), schisandrin A (167 mg/100 g DW), and gomisin O (150 mg/100 g DW) were the quantitatively dominant compounds in the fruit extract. In total, twenty-one lignans were found in the bioactive fractions.     

Phytochemical Profile and Biological Activity of the Ethanolic Extract from the Aerial Part of Crocus alatavicus Regel & Semen Growing Wildly in Southern Kazakhstan

The composition of the ethanolic extract from the aerial parts of Crocus alatavicus Regel & Semen from southern Kazakhstan spontaneous flora was analyzed together with the determination of its antibacterial, antifungal, antiviral and anticancer activity. The phytochemical profile analysis by high-performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (HPLC/ESI-QTOF-MS) revealed the presence of multiple kaempferol derivatives. High-performance reverse-phase liquid chromatography combined with a photodiode-array detection (RP-HPLC/PDA) found that kaempferol 3-O-dihexoside and kaempferol 3-O-acyltetrahexoside accounted for 70.5% of the kaempferol derivatives. The minimum inhibitory concentration (MIC) values of the extract for all the tested reference microorganisms were high, reaching 10 mg/mL for yeasts and 20 mg/mL for bacteria. In contrast, antiviral activity was observed at 2 mg/mL, resulting in the inhibition of the HSV-1-induced cytopathic effect and the reduction in virus infectious titer by 1.96 log, as well as the viral load by 0.85 log. Among the tested prostate cancer cell lines, significant cytotoxic activity of the extract was noted only on the LNCaP cell line, with an IC₅₀ value of 1.95 mg/mL. The LNCaP cell line treated with 2 mg/mL of the extract showed a noticeably reduced number of spindle-shaped cells with longer cellular projections, a significant increase in the peak corresponding to the population of apoptotic cells in the sub-G1 phase and a decreased intracellular glutathione (GSH) level, suggesting the prooxidative properties of the extract. The obtained data provide novel information about the flavonoids present in the aerial part of C. alatavicus and suggest its potential application as a source of the compounds active against HSV-1 and metastatic, androgen-sensitive prostate cancer.     

NHC copper complexes functionalized with sulfoxide and sulfone moieties

A series of N‐heterocyclic copper carbene complexes bearing sulfoxide and sulfone moieties have been prepared. In case of new copper compounds with sulfone ligand, the solid‐state structures were determined using X‐Ray crystallography. Obtained complexes were investigated as catalysts in such transformations as: 1,3‐dipolar cycloaddition of alkynes and azides (CuAAC), A³ coupling reaction and β‐hydroboration and compared with standard copper catalytic systems.     

Status of the hard X‐ray microprobe beamline ID22 of the European Synchrotron Radiation Facility

The ESRF synchrotron beamline ID22, dedicated to hard X‐ray microanalysis and consisting of the combination of X‐ray fluorescence, X‐ray absorption spectroscopy, diffraction and 2D/3D X‐ray imaging techniques, is one of the most versatile instruments in hard X‐ray microscopy science. This paper describes the present beamline characteristics, recent technical developments, as well as a few scientific examples from recent years of the beamline operation. The upgrade plans to adapt the beamline to the growing needs of the user community are briefly discussed.