Electron Paramagnetic Resonance Gives Evidence for the Presence of Type 1 Gonadotropin-Releasing Hormone Receptor (GnRH-R) in Subdomains of Lipid Rafts

This study investigated the effect of type 1 gonadotropin releasing hormone receptor (GnRH-R) localization within lipid rafts on the properties of plasma membrane (PM) nanodomain structure. Confocal microscopy revealed colocalization of PM-localized GnRH-R with GM₁-enriched raft-like PM subdomains. Electron paramagnetic resonance spectroscopy (EPR) of a membrane-partitioned spin probe was then used to study PM fluidity of immortalized pituitary gonadotrope cell line αT3-1 and HEK-293 cells stably expressing GnRH-R and compared it with their corresponding controls (αT4 and HEK-293 cells). Computer-assisted interpretation of EPR spectra revealed three modes of spin probe movement reflecting the properties of three types of PM nanodomains. Domains with an intermediate order parameter (domain 2) were the most affected by the presence of the GnRH-Rs, which increased PM ordering (order parameter (S)) and rotational mobility of PM lipids (decreased rotational correlation time (τc)). Depletion of cholesterol by methyl-β-cyclodextrin (methyl-β-CD) inhibited agonist-induced GnRH-R internalization and intracellular Ca²⁺ activity and resulted in an overall reduction in PM order; an observation further supported by molecular dynamics (MD) simulations of model membrane systems. This study provides evidence that GnRH-R PM localization may be related to a subdomain of lipid rafts that has lower PM ordering, suggesting lateral heterogeneity within lipid raft domains.     

A new generation of cyanide ion-selective membranes for flow injection application: Part III. A simple approach to the determination of toxic metal-cyanide complexes without preliminary separation

A new flow injection approach to total weak acid-dissociable (WAD) metal–cyanide complexes is proposed, which eliminates the need of a separation step (such as gas diffusion or pervaporation) prior to the detection. The cornerstone of the new methodology is based on the highly selective flow-injection potentiometric detection (FIPD) system that makes use of thin-layer electroplated silver chalcogenide ion-selective membranes of non-trivial composition and surface morphology: Ag_2 + δSe_1 − xTe_x and Ag_2 + δSe. An inherent feature of the FIP-detectors is their specific response to the sum of simple CN⁻ + Zn(CN)₄²⁻ + Cd(CN)₄²⁻. For total WAD cyanide determination, ligand exchange (LE) and a newly developed electrochemical pre-treatment procedure for release of the bound cyanide were used. The LE pre-treatment ensures complete recovery only when the sample does not contain Hg(CN)₄²⁻. This limitation is overcome by implementing electrochemical pre-treatment which liberates completely the bound WAD cyanide through cathodic reduction of the complexed metal ions. A complete recovery of toxic WAD cyanide is achieved in the concentration range from 156 μg L⁻¹ up to 13 mg L⁻¹. A three-step protocol for individual and group WAD cyanide speciation is proposed for the first time. The speciation protocol comprises three successive measurements: (i) of non-treated, (ii) LE-exchange pre-treated; (iii) electrochemically pre-treated sample. In the presence of all WAD complexes this procedure provides complete recovery of the total bound cyanide along with its quantitative differentiation into the following groups: (1) Hg(CN)₄²⁻; (2) CN⁻ + Cd(CN)₄²⁻ + Zn(CN)₄²⁻; (3) Cu(CN)₄³⁻ + Ni(CN)₄²⁻ + Ag(CN)₂⁻. The presence of a 100-fold excess in total of the following ions: CO₃²⁻, SCN⁻, NH₄⁺, SO₄²⁻ and Cl⁻ does not interferes. Thus the proposed approach offers a step ahead to meeting the ever increasing demand for cyanide-species-specific methods. The equipment simplicity makes the procedure a good candidate for implementing in portable devices for in-field cyanide monitoring.     

Chemical Structure and In Vitro Antitumor Activity of Rhamnolipids from Pseudomonas aeruginosa BN10

A newly isolated indigenous strain BN10 identified as Pseudomonas aeruginosa was found to produce glycolipid (i.e., rhamnolipid-type) biosurfactants. Two representative rhamnolipidic fractions, RL-1 and RL-2, were separated on silica gel columns and their chemical structure was elucidated by a combination of nuclear magnetic resonance and mass spectroscopy. Subsequently, their cytotoxic effect on cancer cell lines HL-60, BV-173, SKW-3, and JMSU-1 was investigated. RL-1 was superior in terms of potency, causing 50 % inhibition of cellular viability at lower concentrations, as compared to RL-2. Furthermore, the results from fluorescent staining analysis demonstrated that RL-1 inhibited proliferation of BV-173 pre-B human leukemia cells by induction of apoptotic cell death. These findings suggest that RL-1 could be of potential for application in biomedicine as a new and promising therapeutic agent.     

Lipophilicity of some guaianolides isolated from two endemic subspecies of Amphoricarpos neumayeri (Asteraceae) from Montenegro

In this study 10 guaianolide-type sesquiterpene gamma-lactones named amphoricarpolides, isolated from the aerial parts of two endemic subspecies of Amphoricarpos neumayeri (ssp. neumayeri and ssp. murbeckii Bosnjak), were investigated by means of reversed-phase thin-layer chromatography. Methanol-water and tetrahydrofuran-water binary mixtures were used as mobile phase in order to determine lipophilicity parameters R (0) (M) and C(0). Some of the investigated compounds were screened for their cytotoxic activity against HeLa and B16 cells. Chromatographically obtained lipophilicity parameters were correlated with calculated logP values and IC(50) values. Principal component analysis identified the dominant pattern in the chromatographically obtained data.     

Ultra-thin SiO2 layers on TiO2: improved photocatalysis by enhancing products' desorption

A study on the photocatalytic degradation of contaminants (salicylic acid and stearic acid) on titanium dioxide films overcoated with a few monolayers of silica is presented herein. A rather uncommon situation was observed, where the presence of the thin silica layers decreased the degradation rate of stearic acid while increasing that of salicylic acid. The results were explained by addressing the effect in the presence of silica films on the desorption of the intermediate products formed in the degradation of salicylic acid. This finding, which may apply also to other aromatic compounds, may have implications on the design and operation of photocatalytic devices for indoor applications, since ultrathin layers of silica are known to be formed over time on the photocatalyst. An interesting effect of the thickness of the silica sub-nanometre layer on the degradation rate of salicylic acid was explained in terms of gradual changes in the isoelectric point. If optimized, this effect can be utilized to precisely control adsorption or desorption and accordingly to induce specificity in the photocatalytic degradation of contaminants. A methodology for preparing a molecularly imprinting photocatalyst with an inert ultrathin layer in between the imprinted sites was presented. It was found that overcoating the area in between the imprinted sites preserved the benevolent effect of imprinting. While at present the imprinting effect was moderate, there is a reason to believe that this effect can be improved considerably by controlling the type of inert overlayer.     

Acid—base equilibria in the mixed solvent 80% dimethyl sulfoxide—20% water : Determination of pK Values and Investigation of the Conditions for Titration of some Aromatic Carboxylic Acids and their Conjugated Bases

The acid—base properties of some aromatic carboxylic acids of zero charge type HA have been investigated in the mixed solvent, 80% DMSO—20% water, by means of potentiometric measurements with a glass electrode. The pK_a values of the following aromatic carboxylic acids have been determined: benzoic, o-chlorobenzoic,p-chlorobenzoic, m-bromobenzoic, 2,4-dichlorobenzoic, 2,5-dichlorobenzoic, 2,6-dichlorobenzoic, 2,3,5-triiodobenzoic, 3,5-dinitrobenzoic,p-methylbenzoic, m-aminobenzoic, o-hydroxybenzoic 3,4-dihydroxybenzoic, 3,4,5-trihydroxybenzoic, o-methoxybenzoic, 2,3-dimethoxybenzoic, 5-amino-2-hydroxybenzoic, pyridine-2-carboxylic, thiophene-2-carboxylic, 1-naphthylacetic, diphenylglycolic. The mixed solvent investigated offers better titration conditions for the determination of these acids than water and some other non-aqueous solvents.     

Flavonoid constituents and free radical scavenging activity of Alchemilla mollis

Antioxidant capacity of the methanolic extract of Alchemilla mollis was measured by its ability to scavenge the DPPH radical. The EtOAc fraction obtained after partition of the total extract was found to be the most active radical scavenger (IC50 9.8 +/- 1.8 microg/mL) and was subjected to fractionation by Sephadex LH-20 CC. Further purification by RP-18 CC led to the isolation of eight flavonoid glycosides: cis- and trans-tiliroside (1 and 2), rhodiolgin (3), hyperoside (4), isoquercitrin (5), miquelianin (6), sinocrassoside D2 (7), and gossypetin-3-O-beta-D-galactopyranosyl-7-O-alpha-L-rhamnopyranoside (8). It was found that 8 is a new compound and its antioxidant activity is also reported. Identification of the isolated compounds was carried out by spectroscopic and spectrometric analysis (1D and 2D NMR, UV and MS).     

Multifractal analysis of the UV/VIS spectra of malignant ascites: Confirmation of the diagnostic validity of a clinically evaluated spectral analysis

Multifractal (MF) approach was applied for the analysis of ultraviolet/visible (UV/VIS) spectra as an independent confirmation of the diagnostic efficacy of UV/VIS spectral analysis of intraperitoneal fluids, ascites, taken from patients with a known clinical diagnosis. Recently, it was reported that from UV/VIS spectra differentiation of malignant from benign ascites is possible. Here, it was shown that by using MF analysis of UV/VIS spectra, the objective classification of UV/VIS spectra is possible. The applicability of UV/VIS analysis and MF classification of spectra were evaluated on N=68 cases, of which M=64 and B=4 were clinically confirmed as malignant and benign, respectively. The overall diagnostic efficacy was 89.71% when using on-line analysis of UV/VIS spectra (61 out of 68 samples were positively recognized: 58 malignant and 3 benign), and even 95.59% by using off-line MF classsification (65 out of 68 samples were classified correctly: 63 malignant and 2 benign). It can be inferred that UV/VIS spectral analysis of ascites, combined with MF analysis, could be suggested as a successful and safe screening method in the evaluation of intraperitoneal fluids.