Quasi-spherical silver nanoparticles for human prolactin detection by surface-enhanced Raman spectroscopy.

Prolactin is a polypeptide hormone made of 199 amino acids; 50% of the amino acid chain forms helices, and the rest forms loops. This hormone is typically related to initiating and maintaining lactation, although it is also elevated in various pathological conditions. Serum prolactin levels of 2 to 18 ng ml-1 in men, up to 30 ng ml-1 in women, and 10 to 210 ng ml-1 in pregnant women are considered normal. Immunoassay techniques used for detection are susceptible to error in different clinical conditions. Surface-enhanced Raman spectroscopy (SERS) is a technique that allows for obtaining the protein spectrum in a simple, fast, and reproducible manner. Nonetheless, proper characterization of human prolactin's Raman/SERS spectrum at different concentrations has so far not been deeply discussed. This study aims to characterize the Raman spectrum of human prolactin at physiological concentrations using silver nanoparticles (AgNPs) as the SERS substrate. The Raman spectrum of prolactin at 20 ng ul-1 was acquired. Quasi-spherical AgNPs were obtained using chemical synthesis. For SERS characterization, decreasing dilutions of the protein were made by adding deionized water and then a 1 : 1 volume of the AgNPs colloid. For each mixture, the Raman spectrum was determined. The spectrum of prolactin by SERS was obtained with a concentration of up to 0.1 ng ml-1. It showed characteristic bands corresponding to the side chains of aromatic amino acids in the protein's primary structure and the alpha helices of the secondary structure of prolactin. In conclusion, using quasi-spherical silver nanoparticles as the SERS substrate, the Raman spectrum of human prolactin at physiological concentration was determined.

Phenolic compounds of Phellinus spp. with antibacterial and antiviral activities.

Phellinus Quél is one of the largest genera of Hymenochaetaceae; it comprises about 220 species widely distributed on Earth. Most Phellinus species are lignicolous mushrooms that accumulate bioactive compounds. This research studied the phenolic composition of Phellinus spp. and their relationship with antibacterial and antiviral capacity. Phenolics were extracted from Phellinus badius, P. fastuosus, and P. grenadensis; their antiviral and antibacterial activities were evaluated against Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica, and Escherichia coli O157: H7; and the bacteriophages MS2 and Φ- × 174. Gallic acid, chlorogenic acid, caffeic acid, epicatechin, ferulic acid, catechin, 1,3-dicaffeoylquinic acid, p-coumaric acid, and rutin were found in different proportions among Phellinus spp. Total phenolic content ranged from 96 to 209 mg GAE/g, and total flavonoids from 10 to 27 QE/g. The minimum inhibitory concentrations of P. badius, P. grenadensis, and P. fastuosus against E. coli O157: H7 were 13, 20, and 27 mg/mL, against S. enterica were 20, 30, and 15 mg/mL, and against L. monocytogenes were 10, 15, and 25 mg/mL, respectively. The phenolic content was better correlated with the antibacterial effect against E. coli O157: H7 and L. monocytogenes (r = 0.8-0.9), but not against S. enterica (r = 0.05). The antiviral activity of the extracts (0.9 mg/mL) was 29 to 41% against MS2 and 27 to 38% for Φ-X174 virus (r = 0.8-0.9). In silico analysis showed binding energy values of - 7.9 and - 4.8 kcal/mol between the identified phenolic compounds and the M and G proteins of each virus. The antibacterial and antiviral properties of Phellinus species were correlated with the phenolic content.

Computational and Experimental Analysis of Gold Nanorods in Terms of Their Morphology: Spectral Absorption and Local Field Enhancement.

A nanoparticle's shape and size determine its optical properties. Nanorods are nanoparticles that have double absorption bands associated to surface plasmon oscillations along their two main axes. In this work, we analize the optical response of gold nanorods with numerical simulations and spectral absorption measurements to evaluate their local field enhancement-which is key for surface-enhanced Raman spectroscopic (SERS) applications. Our experimental results are in good agreement with finite element method (FEM) simulations for the spectral optical absorption of the nanoparticles. We also observed a strong dependence of the optical properties of gold nanorods on their geometrical dimension and shape. Our numerical simulations helped us reveal the importance of the nanorods' morphology generated during the synthesis stage in the evaluation of absorption and local field enhancement. The application of these gold nanorods in surface-enhancement Raman spectroscopy is analyzed numerically, and results in a 5.8×104 amplification factor when comparing the values obtained for the nanorod deposited on a dielectric substrate compared to the nanorod immersed in water.

TNF-α detection using gold nanoparticles as a surface-enhanced Raman spectroscopy substrate.

Background: TNF-α is a cytokine involved in inflammation. Surface-enhanced Raman spectroscopy (SERS) could be useful in its detection. Aim: Identify the TNF-α in an aqueous solution, using gold nanoparticles (AuNPs) as a SERS substrate. Materials & methods: Raman and SERS spectra were obtained from TNF-α samples, combined with AuNPs, with decreasing concentrations of TNF-α. The samples were analyzed using optical transmission spectroscopy, dynamic light scattering, and transmission electron microscopy. Results: Transmission electron microscopy/dynamic light scattering determined a change in the average diameter of the TNF-α/AuNPs (∼9.6 nm). Raman bands obtained were associated with aromatic amino acid side chains. We observe Raman signals for TNF-α concentrations as low as 0.125 pg/ml. Conclusion: TNF-α signal at physiological concentrations was determined with SERS.

Proteomic profiling of integral membrane proteins associated to pathogenicity in Vibrio parahaemolyticus strains.

Vibrio parahaemolyticus has been recognized as the causal agent of early mortality syndrome and is currently considered an emerging shrimp disease causing losses of millions in the aquaculture industry. Integral membrane proteins are widely recognized as pathogenicity factors involved in essential mechanisms for V. parahaemolyticus infection, which makes them attractive as therapeutic targets. However, their physico-chemical properties and weak expression has resulted in under-representation of these proteins in conventional bottom-up proteomics, making integral membrane proteomics a challenging task. Integral membrane proteins from a bacterial strain isolated from the hepatopancreases of white shrimp with early mortality syndrome and identified by 16S rRNA sequencing as V. parahaemolyticus and an ATCC strain that is pathogenic for humans were obtained by a sequential extraction method and subjected to relative quantification and identification by isobaric Tags for Relative and Absolute Quantitation. A homology database search resulted in identification of more than two hundred proteins, 35 of which are recognized as pathogenic factors showed statistically significant differential accumulation between the strains. These proteins are mainly associated with adherence, secretion systems, cell division, transport, lysogenization, movement and virulence. Identification of pathogenicity-related proteins in V. parahaemolyticus provides valuable information for developing strategies based on molecular mechanisms that inhibit these proteins, which may be useful therapeutic targets for assisting the shrimp and aquaculture industry.

Potential of medicinal plants as antimicrobial and antioxidant agents in food industry: a hypothesis.

Many food preservation strategies can be used for the control of microbial spoilage and oxidation; however, these quality problems are not yet controlled adequately. Although synthetic antimicrobial and antioxidant agents are approved in many countries, the use of natural safe and effective preservatives is a demand of food consumers and producers. This paper proposes medicinal plants, traditionally used to treat health disorders and prevent diseases, as a source of bioactive compounds having food additive properties. Medicinal plants are rich in terpenes and phenolic compounds that present antimicrobial and antioxidant properties; in addition, the literature revealed that these bioactive compounds extracted from other plants have been effective in food systems. In this context, the present hypothesis paper states that bioactive molecules extracted from medicinal plants can be used as antimicrobial and antioxidant additives in the food industry.

Polar fractionation affects the antioxidant properties of methanolic extracts from species of genus Phellinus quel. (higher Basidiomycetes).

The main objective of this study was to evaluate the antioxidant capacity of methanolic extracts from species of genus Phellinus: Ph. fastuosus, Ph. grenadensis, Ph. Merrillii, and Ph. Badius, in their respective polar fractions (aqueous) and nonpolar extracts (ethyl acetate), through tests of free-radical inactivation and hemolysis inhibition. The fungus species that gave the extract with the highest phenol content, total flavonoids, and antioxidant capacity [DPPH·, Trolox equivalents antioxidant capacity (TEAC), and hemolysis inhibition] was Ph. Merrillii, followed by Ph. fastuosus, Ph. Grenadensis, and Ph. Badius. The antioxidant capacities of the extracts, in descending order, were as follows: Ph. Merrillii (nonpolar), Ph. Fastuosus (nonpolar), Ph. Grenadensis (nonpolar), Ph. Fastuosus (polar), Ph. Merrillii (polar), Ph. Grenadensis (polar), Ph. Badius (nonpolar), and Ph. Badius (polar). Antioxidant capacity in the above Phellinus fungi species had EC50 values for DPPH inhibition of 0.45, 0.88, 1.31, 1.89, 2.14, 2.22, 3.42, and 6.00 mg/mL, respectively; TEAC values of 10400.29, 7635.53, 4855.05, 4415.39, 4041.68, 2989.2, 1937.7, and 842.42 µmol TE/g, respectively; and hemolysis inhibition values of 72.83, 66.95, 50.87, 50.28, 48.5, 42.82, 42.37, and 37.91%, respectively. In general, the fungus extract with the highest antioxidant capacity was the nonpolar fraction of Ph. Merrillii. The Phellinus species studied represent potential natural sources of bioactive compounds with antioxidant activity.