Green Synthesis of Antibacterial CuO Nanoparticles Based on the Synergy Between Snail Mucus and Ascorbic Acid.

Many biologically active compounds have been identified in the mucus of the garden snail , which are effective in the treatment of several diseases such as cancer, ulcers, wounds, etc. The incorporation of these compounds into the green synthesis of copper nanoparticles (CuONPs-Muc) was demonstrated in our previous study. Based on the synergistic effect of two reducing agents- snail mucus and ascorbic acid (AsA)-on CuSO.

In silico and physico-chemical characterization of cluster formation dynamics in peptide solutions.

Although antimicrobial peptides are considered one of the most promising alternatives to conventional antibiotics given the alarming increase in bacterial multidrug resistance, many aspects of their mechanism of action remain unclear, in particular the emergence and role of collective phenomena such as the spontaneous formation of nano-sized unstructured objects (clusters) and their effects on the biodynamics. We study this process using two novel peptides from the mucus of the garden snail as an example to reveal its dynamics and bioactivity implications through coordinated in silico and in vitro techniques - molecular dynamics simulations, UV-Vis and fluorescence spectroscopy, and antibacterial activity tests against two representative bacterial strains - one gram-negative ( 3458) and one gram-positive (). The results obtained confirm the impact of the aggregation processes of the peptides on their biological activity and provide insight into possible synergies in their action.

Key Proteins in Rat Cerebral Cortex: Application of Extract as a Neuroprotective Agent in Alzheimer's Type Dementia.

Alzheimer's disease (AD) is the most widespread neurodegenerative disorder. Recently, it was found that mucus extract from has beneficial effects on memory and cognitive processes in a rat scopolamine model of AD. The present study elucidated the mechanisms of action of standardized mucus snail extract (SE) enriched with a fraction above 20 kDa on Alzheimer-type dementia in rats.

Antibacterial Properties of Peptide and Protein Fractions from Mucus.

The discovery and investigation of new natural compounds with antimicrobial activity are new potential strategies to reduce the spread of antimicrobial resistance. The presented study reveals, for the first time, the promising antibacterial potential of two fractions from mucus with an MW < 20 kDa and an MW > 20 kDa against five bacterial pathogens- 1085, 1897, 8691, 3915, and 8754. Using de novo sequencing, 16 novel peptides with potential antibacterial activity were identified in a fraction with an MW < 20 kDa.

Development of CuO Nanoparticles from the Mucus of Garden Snail as New Antimicrobial Agents.

Several biologically active compounds involved in the green synthesis of silver and gold nanoparticles have been isolated from snail mucus and characterized. This paper presents a successful method for the application of snail mucus from as a bioreducing agent of copper sulfate and as a biostabilizer of the copper oxide nanoparticles (CuONPs-Muc) obtained. The synthesis at room temperature and neutral pH yielded nanoparticles with a spherical shape and an average diameter of 150 nm.

Antibacterial Action of Protein Fraction Isolated from Hemolymph against NBIMCC 8785.

Natural products and especially those from marine organisms are being intensively explored as an alternative to synthetic antibiotics. However, the exact mechanisms of their action are not yet well understood. The molecular masses of components in the hemolymph fraction with MW 50-100 kDa from were determined using ImageQuant™ TL v8.

Structural and functional characterization of cold-active sialidase isolated from Antarctic fungus P29.

The fungal strain, P29, isolated from a soil sample taken from Terra Nova Bay, Antarctica, was found to be a good producer of sialidase (P29). The present study was focused on the purification and structural characterization of the enzyme. P29 enzyme was purified using a Q-Sepharose column and fast performance liquid chromatography separation on a Mono Q column.

Assessment of the and Antitumor Activity of Hemocyanins from , , and in a Graffi Myeloid Tumor Model.

Hemocyanins are oxygen-transporting glycoproteins in the hemolymph of some invertebrate species that attracted scientific interest as potential anticancer agents. The present study aims to assess the and anticancer activity of hemocyanins isolated from , , and in the Graffi myeloid tumor model. The antitumor activity of the hemocyanins was determined by a MTT test and cytomorphological analysis by fluorescent and transmission electron microscopy.

Antitumor Activity of Bioactive Compounds from against Human Breast Cell Lines.

This study is the first report describing the promising antitumor activity of biologically active compounds isolated from the hemolymph of marine snail -a fraction with Mw between 50 and 100 kDa and two structural subunits (RvH1 and RvH2), tested on a panel of human breast cell lines-six lines of different molecular subtypes of breast cancer MDA-MB-231, MDA-MB-468, BT-474, BT-549, SK-BR-3, and MCF-7 and the non-cancerous MCF-10A. The fraction with Mw 50-100 kDa (HRv 50-100) showed good antitumor activity manifested by a significant decrease in cell viability, altered morphology, autophagy, and p53 activation in treated cancer cells. An apparent synergistic effect was observed for the combination of HRv 50-100 with cis-platin for all tested cell lines.

Selective Cytotoxicity and Changes in Protein Expression of T24 Bladder Carcinoma Permanent Cell Line after Treatment with Hemocyanins.

Background: Some molluscan hemocyanins (Hcs) have significant immunological and antitumor potential, enabling their application in oncology. The antitumor activity of Hcs from marine snails Rapana venosa (RvH), giant keyhole limpet Megathura crenulata (KLH) and garden snails Helix lucorum (HlH), as well as their different derivatives, were studied in vitro on a permanent T24 cell line of bladder cancer and normal urothelial cell line HL 10/29 compared to doxorubicin.

Methods: The antiproliferative activity of the tested Hcs was determined using the WST-1 assay and BrdU ELISA assay.

Snail Mucus Protective Effect on Ethanol-Induced Gastric Ulcers in Mice.

Nowadays, an increased interest in natural compounds with preventive or therapeutic potential for various diseases has been observed. Given the involvement of oxidative stress in the pathogenesis of gastric ulcer (GU) and the wide range of bioactive compounds isolated from snails, this study aimed to investigate the protective effect of (Müller, 1774) mucus on ethanol-induced GUs. Male albino mice were divided into Control, Ethanol, Mucus + Ethanol and Mucus + Omeprazole treated groups.

Beneficial Effects of Snail Helix aspersa Extract in an Experimental Model of Alzheimer's Type Dementia.

Background: Alzheimer's disease (AD) is a complex neurodegenerative disease with multifactorial etiology, unsatisfactory treatment, and a necessity for broad-spectrum active substances for cure. The mucus from Helix aspersa snail is a mixture of bioactive molecules with antimicrobial, anti-inflammatory, antioxidant, and anti-apoptotic effects. So far there are no data concerning the capacity of snail extract (SE) to affect neurodegenerative disorders.

Effect and Mechanisms of Antibacterial Peptide Fraction from Mucus of against NBIMCC 8785.

Peptides isolated from the mucus of Cornu aspersum could be prototypes for antibiotics against pathogenic bacteria. Information regarding the mechanisms, effective concentration, and methods of application is an important tool for therapeutic, financial, and ecological regulation and a holistic approach to medical treatment. A peptide fraction with MW < 10 kDa was analyzed by MALDI-TOF-TOF using Autoflex™ III.

Microbial diversity of garden snail mucus.

The search for new natural compounds for application in medicine and cosmetics is a trend in biotechnology. One of the sources of such active compounds is the snail mucus. Snail physiology and the biological activity of their fluids (especially the mucus) are still poorly studied.

Molecular Cloning, Structure and Phylogenetic Analysis of a Hemocyanin Subunit from the Black Sea Crustacean (Crustacea, Malacostraca).

Hemocyanins are copper-binding proteins that play a crucial role in the physiological processes in crustaceans. In this study, the cDNA encoding hemocyanin subunit 5 from the Black sea crab (EvHc5) was cloned using EST analysis, RT-PCR and rapid amplification of the cDNA ends (RACE) approach. The full-length cDNA of EvHc5 was 2254 bp, consisting of a 5' and 3' untranslated regions and an open reading frame of 2022 bp, encoding a protein consisting of 674 amino acid residues.

De Novo Structural Determination of the Oligosaccharide Structure of Hemocyanins from Molluscs.

A number of studies have shown that glycosylation of proteins plays diverse functions in the lives of organisms, has crucial biological and physiological roles in pathogen-host interactions, and is involved in a large number of biological events in the immune system, and in virus and bacteria recognition. The large amount of scientific interest in glycoproteins of molluscan hemocyanins is due not only to their complex quaternary structures, but also to the great diversity of their oligosaccharide structures with a high carbohydrate content (2-9%). This great variety is due to their specific monosaccharide composition and different side chain composition.

An H NMR- and MS-Based Study of Metabolites Profiling of Garden Snail Mucus.

Metabolic profiling based on H nuclear magnetic resonance (NMR) spectroscopy was applied with the aim to investigate the functional role of the metabolites in lyophilized mucus from the garden snail . Twenty metabolites were unambiguously identified by H, 1D TOCSY, 2D J-resolved, 2D COSY, and 2D HSQC NMR spectra with water suppression. The metabolic profiles of two fractions with low molecular weight (Mw < 1 kDa and Mw < 3 kDa) are very similar.

Antimicrobial Activities of Different Fractions from Mucus of the Garden Snail .

Natural products have long played a major role in medicine and science. The garden snail is a rich source of biologically active natural substances that might be an important source for new drugs to treat human disease. Based on our previous studies, nine fractions containing compounds with Mw <3 kDa; <10 kDa; <20 kDa; >20 kDa; >30 kDa>50 kDa and between 3 and 5 kDa; 5 and 10 kDa; and 10 and 30 kDa were purified from the mucus of and analyzed by tandem mass spectrometry (MALDI-TOF/TOF).

Hemocyanins from and Snails Exhibit in Vitro Antitumor Effects in Human Colorectal Adenocarcinoma.

Hemocyanins are oxygen-transporting glycoproteins in the hemolymph of arthropods and mollusks that attract scientific interest with their diverse biological activities and potential applications in pharmacy and medicine. The aim of the present study was to assess the in vitro antitumor activity of hemocyanins isolated from marine snail (RvH) and garden snails (HlH) and (HaH), as well the mucus of snails, in the HT-29 human colorectal carcinoma cell line. The effects of the hemocyanins on the cell viability and proliferation were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the alterations in the tumor cell morphology were examined by fluorescent and transmission electron microscopy.

Structural and conformational stability of hemocyanin from the garden snail Cornu aspersum.

Various aspects of biomedical applications of molluscan hemocyanins, associated with their immunogenic properties and antitumor activity, promoted us to perform structural studies on these glycoproteins. The stability and reassociation behavior of native Cornu aspersum hemocyanin (CaH) are studied in the presence of different concentrations of Ca2+ and Mg2+ ions and pH values using electron microscopy. Higher concentrations of those ions led to a more rapid reassociation of CaH, resulting in stable multidecamers with different lengths.

Antimicrobial Activity of Molluscan Hemocyanins from Helix and Rapana Snails.

For the first time the antimicrobial activities of hemocyanins from the molluscs Rapana venosa (RvH) and Helix aspersa (HaH) have been tested. From the hemolymph of the garden snail H. aspersa one structural subunit (βc-HaH ) and eight functional units (FUs, βc-HaH-a to βc-HaH-h) were isolated, and their N-terminal sequences and molecular weights, ranging between 45 and 65 kDa, determined.

Structure and Characterization of Eriphia verrucosa Hemocyanin.

Arthropod hemocyanins (Hcs) are a family of large extracellular oxygen-transporting proteins with high molecular mass and hexameric or multi-hexameric molecular assembly. This study reports for the first time the isolation and characterization of the structure of an arthropod hemocyanin from crab Eriphia verrucosa (EvH) living in the Black Sea. Its oligomeric quaternary structure is based on different arrangements of a basic 6 × 75 kDa hexameric unit, and four of them (EvH1, EvH2, EvH3, and EvH4) were identified using ion-exchange chromatography.

Synthesis of pyrazolo[4,3-e][1,2,4]triazine sulfonamides, novel Sildenafil analogs with tyrosinase inhibitory activity.

Tyrosinase is a multifunctional, glycosylated and copper-containing oxidase which catalyzes the first two steps in mammalian melanogenesis and is responsible for enzymatic browning reactions in damaged fruits during post-harvest handling and processing. Neither hyperpigmentation in human skin nor enzymatic browning in fruits are desirable. These phenomena have encouraged researchers to seek new potent tyrosinase inhibitors for use in foods and cosmetics.

Positions of the glycans in molluscan hemocyanin, determined by fluorescence spectroscopy.

Molluscan hemocyanins are glycoproteins with different quaternary and carbohydrate structures. It was suggested that the carbohydrate chains of some Hcs are involved in their antiviral and antitumor effect, as well in the organization of the quaternary structure of the molecules. Using a well-known complex for saccharide sensing, positions and access to the carbohydrate chains in the native hemocyanins from Rapana venosa (RvH) and Helix lucorum (HlH) and also their structural subunits (RvH1, RvH2 and βcHlH) and functional units (FUs) were analysed by fluorescence spectroscopy and circular dichroism.