580 research outputs found
Development of a direct ESI-MS method for measuring the tannin precipitation effect of proline-rich peptides and in silico studies on the proline role in tannin-protein interactions
Tannins are a heterogeneous class of polyphenols that are present in several plants and foods. Their ability to interact and precipitate proline-rich proteins leads to different effects such as astringency or antidiarrheal activity. Thus, evaluation of the tannin content in plant extracts plays a key role in understanding their potential use as pharmaceuticals and nutraceuticals. Several methods have been proposed to study tannin-protein interactions but few of them are focused on quantification. The purpose of the present work is to set up a suitable and time efficient method able to quantify the extent of tannin protein precipitation. Bradykinin, chosen as a model, was incubated with increasing concentrations of 1,2,3,4,6-penta-O-galloyl-\u3b2-D-glucose and tannic acid selected as reference of tannic compounds. Bradykinin not precipitated was determined by a mass spectrometer TSQ Quantum Ultra Triple Quadrupole (direct infusion analysis). The results were expressed as PC 50 , which is the concentration able to precipitate 50% of the protein. The type of tannin-protein interaction was evaluated also after precipitate solubilisation. The involvement of proline residues in tannin-protein interactions was confirmed by repeating the experiment using a synthesized peptide (RR-9) characterized by the same bradykinin sequence, but having proline residues replaced by glycine residues: no interaction occurred between the peptide and the tannins. Moreover, modelling studies on PGG-BK and PGG-RR-9 were performed to deeply investigate the involvement of prolines: a balance of hydrophobic and H-bond contacts stabilizes the PGG-BK cluster and the proline residues exert a crucial role thus allowing the PGG molecules to elicit a sticking effect
Phenolic extracts from extra virgin olive oils inhibit dipeptidyl peptidase iv activity: In vitro, cellular, and in silico molecular modeling investigations
Two extra virgin olive oil (EVOO) phenolic extracts (BUO and OMN) modulate DPP-IV activity. The in vitro DPP-IV activity assay was performed at the concentrations of 1, 10, 100, 500, and 1000 μg/mL, showing a dose-dependent inhibition by 6.8 ± 1.9, 17.4 ± 6.1, 37.9 ± 2.4, 57.8 ± 2.9, and 81 ± 1.4% for BUO and by 5.4 ± 1.7, 8.9 ± 0.4, 28.4 ± 7.2, 52 ± 1.3, and 77.5 ± 3.5% for OMN. Moreover, both BUO and OMN reduced the DPP-IV activity expressed by Caco-2 cells by 2.9 ± 0.7, 44.4 ± 0.7, 61.2 ± 1.8, and 85 ± 4.2% and by 3 ± 1.9, 35 ± 9.4, 60 ± 7.2, and 82 ± 2.8%, respectively, at the same doses. The concentration of the most abundant and representative secoiridoids within both extracts was analyzed by nuclear magnetic resonance ((1)H-NMR). Oleuropein, oleacein, oleocanthal, hydroxytyrosol, and tyrosol, tested alone, reduced the DPP-IV activity, with IC(50) of 472.3 ± 21.7, 187 ± 11.4, 354.5 ± 12.7, 741.6 ± 35.7, and 1112 ± 55.6 µM, respectively. Finally, in silico molecular docking simulations permitted the study of the binding mode of these compounds
Design, synthesis and preliminary biological evaluation of 3-cyclopropyl-4-phenoxy-1H-pyrazole derivatives as small molecular ligands of RAGE
Receptor for advanced glycation end products (RAGE) is a multiligand receptor belonging to the immunoglobulin superfamily and plays a crucial role in the development of many human diseases such as neurodegenerative diseases, diabetes, cardiovascular diseases and cancer.1 RAGE is involved in a number of cell processes such as neuroinflammation, apoptosis, proliferation and autophagy, and therefore it is of considerable interest as a promising drug target for innovative therapeutic approaches. It consists of an extracellular region, a short hydrophobic transmembrane spanning region, and a highly charged amino acid cytoplasmatic tail. The extracellular region contains a signal peptide, followed by one N-terminal V-type immunoglobulin domain and two C-type (C1 and C2) immunoglobulin domains.2 RAGE is able to interact with a large number of pro-inflammatory and regulatory molecules, such as advanced glycation end-products (AGEs), quinolinic acid, beta amyloid (A\u3b2), high mobility group box 1 (HMGB1), S100/calgranulin family proteins.3,4 However, due to the structural heterogeneity of these endogenous ligands, little is known about the key pharmacophore elements for ligand-RAGE interaction and the specific mode of binding. On these grounds, we aimed at designing new small molecules able to bind the VC1 extracellular domains of RAGE, in order to clarify the structural features that account for RAGE affinity and activation, and to identify new drug-like compounds. Following a process of structural simplification of known pyrazole-5-carboxamide RAGE ligands,1 we planned a set of novel derivatives characterized by a variously functionalized 3-cyclopropyl-4-phenoxy-1H-pyrazole scaffold (Figure 1). The design and synthesis of the new putative RAGE ligands will be presented and discussed, together with the results of their in vitro screening by means of a surface plasmon resonance (SPR)-based assay to estimate their binding ability to the RAGE extracellular domain. References 1. Bongarzone S., Savickas V., Luzi F., Gee A. D. J. Med. Chem. 2017, 60, 7213-7232. 2. Hudson B. I., Carter A. M., Harja E., Kalea A. Z., Arriero M., Yang H., Grant P. J., Schmidt A. M. FASEB J. 2008, 22, 1572-1580. 3. Xue J., Rai V., Singer D., Chabierski S., Xie J., Reverdatto S., Burz D. S., Schmidt A. M., Hoffmann R., Shekhtman A. Structure 2011, 19, 722\u2013732. 4. Koch M., Chitayat S., Dattilo B. M., Schiefner A., Diez J., Chazin W. J., Fritz, G. Structure 2010, 18, 1342-1352
Data on thermal and hydrolytic stability of both domiphen bromide and para-bromodomiphen bromide
In this data article, HPLC analyses were applied to investigate the hydrolytic and thermal stability of domiphen bromide a FDA approved OTC ingredients endowed with antimicrobial activity. The data obtained by stressing domiphen bromide in acid, base and thermal conditions enlarge the research article published by Fumagalli et al. [2]. The chromatograms herein presented reveal that domiphen bromide is stable under acidic and thermal stress while the treatment with base yield to a by-product. The para-bromo derivative, p-bromodomiphen bromide shows the same behavior under the up mentioned stressed conditions
A simulation tool for MRPC telescopes of the EEE project
The Extreme Energy Events (EEE) Project is mainly devoted to the study of the
secondary cosmic ray radiation by using muon tracker telescopes made of three
Multigap Resistive Plate Chambers (MRPC) each. The experiment consists of a
telescope network mainly distributed across Italy, hosted in different building
structures pertaining to high schools, universities and research centers.
Therefore, the possibility to take into account the effects of these structures
on collected data is important for the large physics programme of the project.
A simulation tool, based on GEANT4 and using GEMC framework, has been
implemented to take into account the muon interaction with EEE telescopes and
to estimate the effects on data of the structures surrounding the experimental
apparata.A dedicated event generator producing realistic muon distributions,
detailed geometry and microscopic behavior of MRPCs have been included to
produce experimental-like data. The comparison between simulated and
experimental data, and the estimation of detector resolutions is here presented
and discussed
New Eco-gas mixtures for the Extreme Energy Events MRPCs: results and plans
The Extreme Energy Events observatory is an extended muon telescope array,
covering more than 10 degrees both in latitude and longitude. Its 59 muon
telescopes are equipped with tracking detectors based on Multigap Resistive
Plate Chamber technology with time resolution of the order of a few hundred
picoseconds. The recent restrictions on greenhouse gases demand studies for new
gas mixtures in compliance with the relative requirements. Tetrafluoropropene
is one of the candidates for tetrafluoroethane substitution, since it is
characterized by a Global Warming Power around 300 times lower than the gas
mixtures used up to now. Several mixtures have been tested, measuring
efficiency curves, charge distributions, streamer fractions and time
resolutions. Results are presented for the whole set of mixtures and operating
conditions, %. A set of tests on a real EEE telescope, with cosmic muons, are
being performed at the CERN-01 EEE telescope. The tests are focusing on
identifying a mixture with good performance at the low rates typical of an EEE
telescope.Comment: 8 pages, 6 figures, proceedings for the "XIV Workshop on Resistive
Plate Chambers and Related Detectors" (19-23 February 2018), Puerto Vallarta,
Jalisco State, Mexic
The Extreme Energy Events HECR array: status and perspectives
The Extreme Energy Events Project is a synchronous sparse array of 52
tracking detectors for studying High Energy Cosmic Rays (HECR) and Cosmic
Rays-related phenomena. The observatory is also meant to address Long Distance
Correlation (LDC) phenomena: the network is deployed over a broad area covering
10 degrees in latitude and 11 in longitude. An overview of a set of preliminary
results is given, extending from the study of local muon flux dependance on
solar activity to the investigation of the upward-going component of muon flux
traversing the EEE stations; from the search for anisotropies at the sub-TeV
scale to the hints for observations of km-scale Extensive Air Shower (EAS).Comment: XXV ECRS 2016 Proceedings - eConf C16-09-04.
Allosteric inhibition of carnosinase (CN1) by inducing a conformational shift
In humans, low serum carnosinase (CN1) activity protects patients with type 2 diabetes from diabetic nephropathy. We now characterized the interaction of thiol-containing compounds with CN1 cysteine residue at position 102, which is important for CN1 activity. Reduced glutathione (GSH), N-acetylcysteine and cysteine (3.2 \uc2\ub1 0.4, 2.0 \uc2\ub1 0.3, 1.6 \uc2\ub1 0.2 \uc2\ub5mol/mg/h/mM; p <.05) lowered dose-dependently recombinant CN1 (rCN1) efficiency (5.2 \uc2\ub1 0.2 \uc2\ub5mol/mg/h/mM) and normalized increased CN1 activity renal tissue samples of diabetic mice. Inhibition was allosteric. Substitution of rCN1 cysteine residues at position 102 (Mut1C102S) and 229 (Mut2C229S) revealed that only cysteine-102 is influenced by cysteinylation. Molecular dynamic simulation confirmed a conformational rearrangement of negatively charged residues surrounding the zinc ions causing a partial shift of the carnosine ammonium head and resulting in a less effective pose of the substrate within the catalytic cavity and decreased activity. Cysteine-compounds influence the dynamic behaviour of CN1 and therefore present a promising option for the treatment of diabetes
Advanced lipoxidation end products (ALEs) as RAGE binders : Mass spectrometric and computational studies to explain the reasons why
Advanced Lipoxidation End-products (ALEs) are modified proteins that can act as pathogenic factors in several chronic diseases. Several molecular mechanisms have so far been considered to explain the damaging action of ALEs and among these a pathway involving the receptor for advanced glycation end products (RAGE) should be considered. The aim of the present work is to understand if ALEs formed from lipid peroxidation derived reactive carbonyl species (RCS) are able to act as RAGE binders and also to gain a deeper insight into the molecular mechanisms involved in the protein-protein engagement. ALEs were produced in vitro, by incubating human serum albumin (HSA) with 4-hydroxy-trans 12 2-nonenal (HNE), acrolein (ACR) and malondialdehyde (MDA). The identification of ALEs was performed by MS. ALEs were then subjected to the VC1 Pull-Down assay (VC1 is the ligand binding domain of RAGE) and the enrichment factor (the difference between the relative abundance in the enriched sample minus the amount in the untreated one) as an index of affinity, was determined. Computation studies were then carried out to explain the factors governing the affinity of the adducted moieties and the site of interaction on adducted HSA for VC1-binding. The in silico analyses revealed the key role played by those adducts which strongly reduce the basicity of the modified residues and thus occur at their neutral state at physiological conditions (e.g. the MDA adducts, dihydropyridine-Lysine (DHPK) and N-2-pyrimidyl-ornithine (NPO), and acrolein derivatives, N-(3-formyl-3,4-dehydro-piperidinyl) lysine, FDPK). These neutral adducts become unable to stabilize ion-pairs with the surrounding negative residues which thus can contact the RAGE positive residues. In conclusion, ALEs derived from lipid peroxidation-RCS are binders of RAGE and this affinity depends on the effect of the adduct moiety to reduce the basicity of the target amino acid and on the acid moieties surrounding the aminoacidic target
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