Validation of an Isocratic HPLC Method for Simultaneous Estimation of Major Phytosterols in Prunus spinosa L. extracts.

This study aimed to develop a rapid method for separation of stigmasterol, campesterol and ß-sitosterol in Prunus spinosa L. (sloe) fruit extracts by High Performance Liquid Chromatography system. Samples were prepared by Soxhlet extraction method and separated on a high strength silica C18 column using acetonitrile-methanol mobile phase and Photodiode Array Detector. The optimized method resulted in a linear calibration curve ranging from 1.7 ng mL-1 to 130 ng mL-1 for all three phytosterols. Analyses of internal and external phytosterol standards showed good linearity (R2 of 0.998 to 0.999); LOD and LOQ were determined to be 2.33×10-7-2.18×10-4 and 7.07×10-7-6.60×10-4 mg mL-1, respectively. Repeatability and reproducibility precision analyses showed acceptable values of RSD %. ß-sitosterol was the predominant phytosterol (51.53-81.03 % of total) among all samples. Method validation parameters indicated that this analytical method can be applied for accurate and precise determination of campesterol, stigmasterol and ß-sitosterol, in selected extracts.

Phase stability of aqueous mixtures of bovine serum albumin with low molecular mass salts in presence of polyethylene glycol.

The stability of bovine serum albumin (BSA) solutions against phase separation caused by cooling the system is studied under the combined influence of added poly(ethylene glycol) (PEG) and alkali halide salts in water as solvent. The phase stability of the system depends on the concentration of the added PEG and its molecular mass, the concentration of the low molecular mass electrolyte and its nature, as also on the pH of the solution. More specifically, the addition of NaCl to the BSA-PEG mixture promotes phase separation at pH = 4.0, where BSA carries the net positive charge in aqueous solution, and it increases the stability of the solution at pH=4.6, i.e., near the isoionic point of the protein. Moreover, at pH = 4.6, the cloud-point temperature decreases in the order from NaF to NaI and from LiCl to CsCl. The order of the salts at pH = 4.0 is exactly reversed: LiCl and NaF show the weakest effect on the cloud-point temperature and the strongest decrease in stability is caused by RbCl and NaNO3. An attempt is made to correlate these observations with the free energies of hydration of the added salt ions and with the effect of adsorption of salt ions on the protein surface on the protein-protein interactions. Kosmotropic salt ions decrease the phase stability of BSA-PEG-salt solutions at pH < pI, while exactly the opposite is true at pH = pI.

Cytotoxic activities of Hypericum perforatum L. extracts against 2D and 3D cancer cell models.

Six extracts were obtained from plant species Hypericum perforatum L., collected at Samsun in Turkey. The aim of this study was to examine the mechanisms of the anticancer activity of these extracts. Methanol, ethyl-acetate and hexane were used as a solvents for extraction from both branch-body part of the plant (extracts 1, 2 and 3) and from plant flowers (extracts 4, 5 and 6). The cytotoxic effects of the extracts were determined against 2D and 3D cancer cell models. Cell cycle changes of treated HeLa cells were analyzed by flow cytometry. Measurements of gene and microRNA expression levels in treated HeLa cells were done by quantitative real time PCR. Five examined extracts (2-6) exerted selective concentration-dependent cytotoxic effects on HeLa, K562, and A549 cancer cells, while the extract 1 exhibited very weak cytotoxicity. The extract 6 showed the highest intensity of cytotoxic activity. All tested extracts (2-6) demonstrated the ability to induce apoptosis in HeLa cells through activation of caspase-3. These extracts remarkably decreased gene expression levels of MMP2, MMP9, TIMP3, and VEGFA in HeLa cells. Flower extracts might have stronger effects on miR128/193a-5p/335 level changes than branch-body extracts. Hypericum perforatum extracts exerted weaker cytotoxic effects on 3D HeLa spheroids when compared with their effects on 2D monolayer HeLa cells. Taken together, results of our research may suggest the promising anticancer properties of the Hypericum perforatum extracts.

Efficacy and safety of three plant extracts based formulations of vagitories in the treatment of vaginitis: a randomized controlled trial.

Aim There are more and more herbal preparations that are used for the purpose of treatment and improvement of the clinical manifestation of vaginitis not only by patients themselves, but also by healthcare professionals. Plant species, St. John's wort, chamomile, calendula, yarrow, shepherd's purse and tea tree oil are all well known for their anti-inflammatory, antimicrobial and wound healing activity. This paper presents the results of a clinical study in which three herbal formulations/vagitories, based on extracts of St. John's wort, chamomile, calendula, yarrow, shepherd's purse and tea tree oil, were investigated for their effectiveness on vaginitis. Methods This was a randomized controlled clinical study that included 210 women with diagnosed vaginitis. Patients were divided into two basic groups, women in reproductive period and postmenopausal period. Three subgroups including 30 patients each received one of the three vagitorie formulations for 5 days, after which the effects on subjective and objective symptoms were monitored. Results Three types of vagitories based on plant extracts had a positive effect in the treatment of vaginitis. Vagitories based on tea tree oil showed better efficiency compared to vagitories with St. John's wort and vagitories based on extracts of five plants. Women in postmenopausal group reported better tolerability of St. John's wort-based and five herbs-based vagitories compared to tea tree oil based vagitories. Conclusion Investigated vagitories showed a positive effect on both objective and subjective symptoms of vagitnis. No serious side effects were reported.

N-Trifluoroacetylated pyrazolines: Synthesis, characterization and antimicrobial studies.

A series of N-trifluoroacetyl-2-pyrazolines have been synthesized via cyclization of chalcones in the presence of trifluoroacetic acid and hydrazine as a base. The method used for the preparation of pyrazolines was found to be an efficient one as all of the compounds were obtained in good yield (up to 79%). Various spectroscopic techniques established the structures and additionally corroborated the compounds 2a and 2e by single crystal X-ray. Newly synthesized pyrazolines were investigated for their potential as antimicrobial agents. Compound 2a displayed promising antimicrobial activity against pathogenic Escherichia coli and Pseudomonas aeruginosa. Furthermore, the mechanism of the antimicrobial activity of 2a was demonstrated with the help of scanning electron microscopy (SEM), which revealed complete damage of the bacterial cell membrane, providing dead cell debris in the milieu. The minimum inhibitory concentration (MIC) observed was 79 and 90 µM against E. coli and P. aeruginosa, respectively. Hence, these compounds might be significantly useful in antimicrobial drug development.

Effects of temperature and polyols on the ciprofloxacin hydrochloride-mediated micellization of sodium dodecyl sulfate.

Herein, a conductivity method was engaged to explore the effects of a fluoroquinolone drug, namely ciprofloxacin hydrochloride (CFH)/CFH + polyols (organic compounds with multiple hydroxyl groups (glucose and fructose)), on the aggregation phenomenon of sodium dodecyl sulfate (SDS) at different temperatures (298.15-318.15 K) while maintaining a gap of 5 K. In this study, the critical micelle concentration (cmc) of the SDS/SDS + CFH mixture in water and polyols media was determined from plots of the specific conductivity versus the concentration of SDS to gain knowledge of the effects of CFH/CFH + polyols on the micelle formation behavior of SDS. The cmc value of the surfactant decreases in the presence of CFH in an aqueous medium; thus, CFH favors the micellization of SDS. The cmc values of SDS and the SDS + CFH mixture were enhanced in polyols media. The cmc values of SDS/SDS + CFH show a U-shaped behavior with temperature. The counterion dissociation (α) of the pure surfactant is higher in the presence of the drug and is further enhanced through an increase in the CFH concentration in water/polyols media. Different thermodynamic parameters, such as the Gibbs free energy of micellization , standard enthalpy , entropy , different transfer energies and enthalpy-entropy compensation parameters of micellization were determined and illustrated in detail to compare these parameters between the pure SDS and SDS + CFH mixture in polyols media. The negative values of for the SDS/SDS + CFH mixture in all cases indicate spontaneous micelle formation. The and values indicate the presence of both hydrophobic and electrostatic interactions amongst the studied components.

A Novel Acetylcholinesterase Biosensor: Core-Shell Magnetic Nanoparticles Incorporating a Conjugated Polymer for the Detection of Organophosphorus Pesticides.

To construct a sensing interface, in the present work, a conjugated polymer and core-shell magnetic nanoparticle containing biosensor was constructed for the pesticide analysis. The monomer 4,7-di(furan-2-yl)benzo[c][1,2,5]thiadiazole (FBThF) and core-shell magnetic nanoparticles were designed and synthesized for fabrication of the biosensing device. The magnetic nanoparticles were first treated with silica and then modified using carboxyl groups, which enabled binding of the biomolecules covalently. For the construction of the proposed sensor a two-step procedure was performed. First, the poly(FBThF) was electrochemically generated on the electrode surface. Then, carboxyl group modified magnetic nanoparticles (f-MNPs) and acetylcholinesterase (AChE), the model enzyme, were co-immobilized on the polymer-coated surface. Thereby, a robust and novel surface, conjugated polymer bearing magnetic nanoparticles with pendant carboxyl groups, was constructed, which was characterized using Fourier transform infrared spectrometer, cyclic voltammetry, scanning electron microscopy, and contact angle measurements. This novel architecture was then applied as an immobilization platform to detect pesticides. To the best of our knowledge, a sensor design that combines both conjugated polymer and magnetic nanoparticles was attempted for the first time, and this approach resulted in improved biosensor characteristics. Hence, this approach opens a new perspective in the field of enzyme immobilization and sensing applications. Paraoxon and trichlorfon were selected as the model toxicants. To obtain best biosensor performance, optimization studies were performed. Under optimized conditions, the biosensor in concern revealed a rapid response (5 s), a low detection limit (6.66 × 10(-3) mM), and high sensitivity (45.01 µA mM(-1) cm(-2)). The KM(app) value of poly(FBThF)/f-MNPs/AChE were determined as 0.73 mM. Furthermore, there was no considerable activity loss for 10 d for poly(FBThF)/f-MNPs/AChE biofilm.