Kinetics of Immunolatex Deposition at Abiotic Surfaces under Flow Conditions: Towards Quantitative Agglutination Assays

Physicochemical properties of immunolatex, prepared by incubation of negatively charged polystyrene microparticles with polyclonal rabbit IgGs, were determined by a variety of experimental techniques. These comprised dynamic light scattering (DLS), laser Doppler velocimetry (LDV) and atomic force microscopy (AFM). The particle diffusion coefficient, the hydrodynamic diameter, the electrophoretic mobility, the zeta potential and the suspension stability were determined as a function of pH for different ionic strengths. The deposition of the immunolatex on bare and polyallylamine (PAH) functionalized mica was investigated using the microfluidic oblique impinging-jet cell, with an in situ, real-time image analysis module. The particle deposition kinetics was acquired by a direct particle enumeration procedure. The measurements enabled us to determine the range of pH where the specific deposition of the immunolatex on these substrates was absent. We argue that the obtained results have practical significance for conducting efficient flow immunoassays governed by specific antigen/antibody interactions.     

Design,Synthesis and Biological Activity of New Amides Derived from 3-Benzhydryl and 3-sec-Butyl-2,5-dioxo-pyrrolidin-1-yl-acetic Acid

The aim of this study was to design and synthesize two new series of pyrrolidine-2,5-dione-acetamides with a benzhydryl or sec-butyl group at position 3 as potential anticonvulsants. Their anticonvulsant activity was evaluated in standard animal models of epilepsy: the maximal electroshock (MES), the 6 Hz, and the subcutaneous pentylenetetrazole (scPTZ) tests. The in vivo studies revealed the most potent anticonvulsant activity for 15 (3-(sec-butyl)-1-(2-(4-(3-trifluoromethylphenyl)piperazin-1-yl)-2-oxoethyl)pyrrolidine-2,5-dione), with ED₅₀ values of 80.38 mg/kg (MES) and 108.80 mg/kg (6 Hz). The plausible mechanism of action was assessed in in vitro binding assays, in which 15 interacted effectively with voltage-gated sodium (site 2) and L-type calcium channels at a concentration of 100 μM. Subsequently, the antinociceptive activity of compounds 7 and 15 was observed in the hot plate test of acute pain. Moreover, compounds 7 , 11 and 15 demonstrated an analgesic effect in the formalin test of tonic pain. The hepatotoxic properties of the most effective compounds ( 7 , 11 and 15 ) in HepG2 cells were also investigated.     

Adipocyte Fatty Acid-Binding Protein as a Novel Marker of Psoriasis and Clinical Response to Acitretin

Psoriasis is a systemic disease associated with metabolic syndrome and cardiometabolic diseases. Adipocyte fatty acid-binding protein (A-FABP, FABP4) is a relevant mediator of lipid metabolism and several comorbidities development. Aim of the study was to explore the possible role of FABP4 in psoriasis and assess its relationship with disease activity, inflammation or metabolic disturbances, and impact of systemic treatment. Fasting blood samples were obtained from 33 patients with active plaque-type psoriasis before and after 12 weeks of therapy and from 11 healthy volunteers. Serum FABP4 concentrations were analyzed by the enzyme-linked immunosorbent assay (ELISA) and statistically analyzed for their correlations with clinical outcomes and the treatment introduced. Serum FABP4 levels were significantly increased in psoriatics compared to controls (p = 0.03). No relationship between the protein and psoriasis severity expressed through psoriasis area and severity index (PASI) was noted (p = 0.57). FABP4 did not correlate with CRP (p = 0.41), lipid profile, and body mass index (BMI) nor the glucose level or liver enzyme activity. FABP4 significantly correlated with morphotic blood elements. After total therapy, FABP4 did not statistically change (p = 0.07), but significantly decreased after administering acitretin (p = 0.03). FABP4 is a potential marker of psoriasis and clinical outcome after therapy with acitretin. Adipocyte-type FABP may be related to hematological disorders or obesity-mediated comorbidities in psoriasis.     

Application of Polyethylene/Polypropylene Glycol Ethers of Fatty Alcohols for Micelle-Mediated Extraction of Calendula anthodium

Classical extraction methods used for the isolation of active substances from the plant materials are expensive, complicated, and often environmentally unfriendly. The ultrasonic assisted micelle-mediated extraction (UAMME) method seems to be an interesting alternative. In this study, Calendula anthodium UAMME extract composition and antioxidant properties were examined. Several nonionic surfactants, from the group of ethoxy/propoxylated fatty alcohols, were applied in the extraction process. The results were compared with water and water-ethanoic extracts of marigold flowers. Bioflavonoid compositions of the extracts were determined using ultra-performance liquid chromatography–diode-array detector–mass spectrometry (UPLC-DAD-MS). Their antioxidant properties were evaluated using free radical-scavenging activity (2,2-Diphenyl-1-picrylhydrazyl, DPPH) measurement and Follin's method. Additionally, the structure of the selected flavonoids solubilized in the micelle solution was theoretically simulated using molecular dynamics (MD). The results confirmed that the UAMME method might be an alternative method for the sourcing of the plant extracts rich in natural antioxidants. The results indicated that UAMME extracts showed 19–37% higher DPPH radical inhibition than aqueous extracts. Furthermore, the selection of a suitable surfactant may thus provide the expected composition of the extract. Due to the use as eluents in UAMME, safe raw materials, the obtained marigold flower extracts can be applied as components in the cosmetic and pharmaceutical products.     

Peptide-Based 2-Aminophenylamide Probes for Targeting Endogenous Class I Histone Deacetylase Complexes

Lysine deacetylases or histone deacetylases (HDACs) remove acetylation markers from numerous cellular proteins, thereby regulating their function and activity. Recently established peptide probes containing the HDAC-trapping amino acid α-aminosuberic acid ω-hydroxamate (AsuHd) have been used to investigate the compositions of HDAC complexes in a site-specific manner. Here we report the new HDAC-trapping amino acid 2-amino-8-[(2-aminophenyl)amino]-8-oxooctanoic acid (AsuApa) and the utility of AsuApa-containing probes for HDAC complex profiling on a proteome-wide scale. Unlike AsuHd-containing probes, AsuApa enriched only HDACs 1, 2, and 3 efficiently and was the most potent probe tested for capturing the last of these. These findings indicate that the inherent specificity of reported small-molecule pimelic diphenylamide HDAC inhibitors is preserved in AsuApa and that this HDAC-trapping amino acid represents a potent tool for investigating class I HDAC complexes.     

Effects of quinoa and amaranth on zinc,magnesium and calcium content in beer wort

Pseudocereals such as quinoa and amaranth can be used as partial replacements for malt in the production of new beers. Quinoa and amaranth are of interest due to their high levels of nutrients and micronutrients, including minerals which can significantly improve the performance of brewing yeast and the fermentation rate. In this study, we investigated whether the use of quinoa or amaranth as partial replacements for malt affected the concentration of ions such as zinc and magnesium in beer wort. The use of amaranth, and in particular quinoa, increased the content of both zinc and magnesium ions substantially, even when only 10% of the barley malt was replaced. With 10% quinoa, Zn²⁺ and Mg²⁺ content increased by 41% and 49%, respectively, while the ratio of Mg²⁺ to Ca²⁺ rose from 1.4:1 to 1.9:1. Use of unmalted quinoa and amaranth appears a good way of enriching wort with essential metal ions.     

Evaluation of the use of lactic acid bacteria and Thymus vulgaris essential oil on Suffolk and Ile de France lamb breed (MuscuIus gluteus) quality parameters

In the present study, Lactobacillus plantarum (LUHS135), Thymus vulgaris essential oil (0.1% v/v) and their combination were used to treat Suffolk and Ile de France lamb meat. Changes in the microbiological profile and physicochemical parameters of meat were evaluated after 24 h of treatment at 4 °C. Treatments resulted in significantly lower mould/yeast, total enterobacteria (by up to 45%) counts, water-holding capacity (by 21–63%) and cooking loss (by 17–35%) in all samples. Moisture content increased, on average, by 14.5% in the Suffolk meat. Polyunsaturated fatty acid content increased after all treatments. Treatments and breed significantly affected the content of malondialdehyde and some biogenic amines. Sensory analysis showed that raw lamb meat colour was more acceptable after all treatments, while the odour acceptability was ranked highest after combined treatment. These findings suggest that the applied treatments could be used to improve microbiological safety and some quality characteristics and to increase the PUFA content of lamb meat.     

Transient Excursions to Membrane Core as Determinants of Influenza Virus Fusion Peptide Activity

Fusion of viral and host cell membranes is a critical step in the life cycle of enveloped viruses. In the case of influenza virus, it is mediated by subunit 2 of hemagglutinin (HA) glycoprotein whose N-terminal fragments insert into the target membrane and initiate lipid exchange. These isolated fragments, known as fusion peptides (HAfp), already possess own fusogenic activity towards liposomes. Although they have long been studied with the hope to uncover the details of HA-mediated fusion, their actual mechanism of action remains elusive. Here, we use extensive molecular dynamics simulations combined with experimental studies of three HAfp variants to fully characterize their free energy landscape and interaction with lipid bilayer. In addition to customary assumed peptides localization at lipid–water interface, we characterize membrane-spanning configurations, which turn out to be metastable for active HAfps and unstable for the fusion inactive W14A mutant. We show that, while the degree of membrane perturbation by surface peptide configurations is relatively low and does not show any mutation-related differences, the effect of deeply inserted configurations is significant and correlates with insertion depth of the N-terminal amino group which is the highest for the wild type HAfp. Finally, we demonstrate the feasibility of spontaneous peptide transition to intramembrane location and the critical role of strictly conserved tryptofan residue 14 in this process.