Graphene oxide-octadecylsilane incorporated monolithic nano-columns with 50 μm id and 100 μm id for small molecule and protein separation by nano-liquid chromatography

In this study, graphene oxide-octadecylsilane incorporated monolithic nano-columns were developed for protein analysis by nano liquid chromatography (nano LC). The monolithic column with 100 μm id was first prepared by an in situ polymerization using ethylene dimethacrylate (EDMA), 3-chloro-2-hydroxypropylmethacrylate (HPMA-Cl), and methacryloyl graphene oxide nanoparticles (MGONPs). MGONPs were synthesized by the treatment of 3-(trimethoxysilyl)propylmethacrylate (TMSPM) and GO. Tetrahydrofuran (THF) and dodecanol were used as the porogenic solvent. The resulting column was functionalized by dimethyloctadecylch lorosilane (DODCS) for the enhancement of hydrophobicity. The functionalization greatly improved the baseline separation of hydrophobic compounds such as polyaromatic hydrocarbons (PAHs). The optimized monolith with respect to total polymerization mixture was characterized by using Fourier-transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) X-ray diffraction (XRD) and chromatographic analyses. The blank monoliths without functionalization exhibited poor separation while a good separation performance of MGONPs functionalized monoliths was achieved. The monolith with 100 μm id was evaluated in protein separation in nano LC using RNase A, Cytochrome C, Lysozyme, Trypsin, and Ca isozyme II as the test proteins. It was shown that protein separation mechanism was based on large π-system of GO and hydrophobicity of the monolithic structure. Theoretical plates number up to 57 600 plates were achieved. The nano-column with 50 μm id was also prepared using the same polymerization mixture under the same chemical conditions. These nano-columns were employed for protein separation by nano LC, and the dependence of both nano-column performance on the internal diameter was also discussed.     

Apis mellifera syriaca Venom: Evaluation of Its Anticoagulant Effect,Proteolytic Activity,and Cytotoxicity along with Its Two Main Compounds—MEL and PLA2—On HeLa Cancer Cells

Bee venom (BV) is one of the most remarkable natural products that has been a subject of studies since ancient times. Recent studies have shown that Apis mellifera syriaca venom possesses antibacterial as well as cytotoxic effects on cancer cell lines. The venom contains a variety of bioactive molecules—mainly melittin (MEL) and phospholipase A2 (PLA2), as well as other compounds that are not well characterized. In this work, we continue the biological characterization of A. mellifera syriaca venom by testing its anticoagulant effect on human plasma using the prothrombin time (PT) test, as well as assessing its proteolytic activity. In addition, the cytotoxicity of the crude venom—and of its two main components, MEL and PLA2—was tested on HeLa cancer cell lines for the first time. The results obtained showed the capacity of A. mellifera syriaca venom to increase clotting time, thereby proving its anticoagulant effect. Moreover, the venom did not demonstrate a significant proteolytic activity unless administrated at concentrations ≥ 5 mg/mL. Finally, we showed that crude A. mellifera syriaca venom, along with MEL, exhibit a strong in vitro cytotoxic effect on HeLa cancer cell lines, even at low concentrations. In summary, our findings could serve as a basis for the development of new natural-based drug candidates in the therapeutic field.     

Snake Venom Components: Tools and Cures to Target Cardiovascular Diseases

Cardiovascular diseases (CVDs) are considered as a major cause of death worldwide. Therefore, identifying and developing therapeutic strategies to treat and reduce the prevalence of CVDs is a major medical challenge. Several drugs used for the treatment of CVDs, such as captopril, emerged from natural products, namely snake venoms. These venoms are complex mixtures of bioactive molecules, which, among other physiological networks, target the cardiovascular system, leading to them being considered in the development and design of new drugs. In this review, we describe some snake venom molecules targeting the cardiovascular system such as phospholipase A2 (PLA2), natriuretic peptides (NPs), bradykinin-potentiating peptides (BPPs), cysteine-rich secretory proteins (CRISPs), disintegrins, fibrinolytic enzymes, and three-finger toxins (3FTXs). In addition, their molecular targets, and mechanisms of action—vasorelaxation, inhibition of platelet aggregation, cardioprotective activities—are discussed. The dissection of their biological effects at the molecular scale give insights for the development of future snake venom-derived drugs.     

High performance liquid chromatography of glycoconjugates

Stationary phases were prepared for the analytical and micropreparative high performance affinity chromatography of carbohydrates and glycoproteins by covalent attachment of lectins and certain biogenic amines to the surface of microparticulate, macroporous diol-silica. Columns packed with immobilized concanavalin A or wheat germ agglutinin afforded rapid separation of both glycoproteins and low molecular weight carbohydrates. The combined use of the two lectins in a mixed bed column or in two columns in series by sequential elution with the appropriate haptenic sugars facilitated the separation of mixtures having a broad range of complex carbohydrates. Chromatographic experiments for the study of the interaction of certain biogenic amines with immobilized glycoproteins used as the stationary phase led to the development of siliceous bonded phases with serotonin, tryptamine or phenylpropanolamine ligates. The sorbents thus obtained were mildly hydrophobic and cationic in contact with eluents of pH 8 or less. They were used for the separation of various carbohydrates; in addition they also proved effective for the separation of protein mixtures under conditions similar to those customarily employed in electrostatic or hydrophobic interaction chromatography.     

Photochromism of dihydroindolizines part X. Photo‐responsive self‐assembling organogelators based on photochromic dihydroindolizines and 11‐aminoundecanoic acid (AUDA)

Multi‐addressable photophysical properties of new synthesized photochromic materials based on photochromic dihydroindolizine system (DHI) covalently linked to N‐acyl‐11 aminoundecanoic acid (AUDA) or to its sodium salt or to its ester, through an amidic or urethane linkage have been studied. The DHI skeleton in these compounds is substituted in both the fluorene part (region A) or in the heterocyclic base (region B) with the gelling moieties. These molecules have been designed to respond to their environment. Interestingly, they are shown to act as efficient gelators for polar organic fluids, water and obviously they exhibit a thermosensitive answer as low molecular mass organogelators. In these fluids, the aggregative properties are totally suppressed upon conversion to neutral carboxylic species. The gels of these carboxylate sodium salts are shown to be markedly affected by light irradiation. Supramolecular gelating assemblies can be disrupted by the photoinduced ring opening of the DHI subunit, so that the macroscopic flowing property is recovered. Upon a further thermal treatment, the system is reversibly converted back to the supramolecular network. Controlled gelation could be achieved using temperature, light, or acidity as external stimuli. These new synthesized photochromic gels with their multi‐addressable properties will find their applications as super photoresponsive materials. Developing and tuning of the photophysical properties of the synthesized compounds by the amide and urethane substituents in the 4‐position of the fluorene and pyridazine regions have been achieved. The absorption maxima (λ_max) and the half‐lives (t_1/2) of the colored betaines were detected in all cases using UV/VIS spectrophotometric measurements. Irradiation of DHI 12‐20 in CH₂Cl₂ or in acetonitrile solutions at ambient temperature with polychromatic light leads to the formation of red to red‐violet colored betaines 12 ′ ‐20 ′. The kinetics of the bleaching process of betaines 12 ′ ‐20 ′ to DHIs 12‐20 were found to take place in the second range (96‐218 s) and fit well the first order thermal back reaction. Some of these DHIs showed a photostability higher than that of the standard one. These interesting photophysical properties will help this family of compounds to find useful applications. Copyright © 2008 John Wiley & Sons, Ltd.     

A novel, neutral hydroxylated octadecyl acrylate monolith with fast electroosmotic flow velocity and its application to the separation of various solutes including peptides and proteins in the absence of electrostatic interactions

A neutral hydroxylated octadecyl monolith (ODM-OH) for reversed-phase capillary electrochromatography has been developed. The ODM-OH was prepared by the in situ polymerization of octadecyl acrylate and pentaerythritol triacrylate (PETA) in a ternary porogenic solvent. Pentaerythritol triacrylate possesses a hydroxyl functional group, which imparts the monolith with a hydrophilic group, thus the acronym ODM-OH. The ODM-OH column exhibited cathodal EOF over a wide range of pH and ACN concentration in the mobile phase despite the fact that it was devoid of any fixed charges. This ODM-OH monolith exhibited stronger EOF than its counterpart the ODM made from the in situ polymerization of octadecyl acrylate and trimethylolpropane trimethacrylate. Similar to ODM, it is believed that the EOF was due to the adsorption of ions from the mobile phase onto the surface of the monolith thus imparting the neutral monolithic column the zeta potential necessary to support the EOF. The higher EOF exhibited by ODM-OH was due to the presence of polar OH groups on its surface, which would favor stronger adsorption of ions from the mobile phase. The wide applications of the neutral ODM-OH column were demonstrated in the separation of a wide range of small and large solutes. As a typical result, the ODM-OH was able to separate proteins quite rapidly yielding 200,000 plates/m.     

Global $$L^q$$-Gevrey Functions and Their Applications

The goal of this paper is to introduce a class of \(C^\infty \) functions derivatives of which satisfy quantitative size estimates. The estimates, called global \(L^q\) Gevrey estimates, first arose in the work of Boggess and Raich ( J. Fourier Anal. Appl. 19:180–224, 2013) when they investigated how to capture a particular type of exponential decay through estimates on the Fourier transform. In the present work, we refine the notion of global \(L^q\)-Gevrey functions and include a discussion of the function theory as well as the relationship to Gevrey classes and known function spaces. In addition, we present explicit examples of global \(L^q\)-Gevrey functions and ways to generate new global \(L^q\)-Gevrey functions from old ones. We conclude with three applications: The first is solving a Carleman-type problem for constructing functions derivatives of which are a given sequence of global \(L^q\)-Gevrey functions. The other two applications concern extensions of a given global \(L^q\)-Gevrey function: the first is constructing an almost analytic extension, and the second is building an approximate solution to a first-order complex vector field coefficients of which are global \(L^q\)-Gevrey functions.