Effects and mechanisms of Portulaca oleracea L. polysaccharides on the activation of dendritic cells derived from mice immunized with FMD vaccine

Background Our previous study has showed that Portulaca oleracea L. (POL-P), as an immunoenhancer, could increase the IgG and isotypes antibody titers in mice immunized with foot and mouth disease (FMD) vaccines. However, the structural features and the mechanism of action are still unclear. Enhancing antigen presentation is one of the main ways that immunoenhancer boost immune response. Dendritic cells (DCs) are the most potent antigen presenting cell (APC), which stimulate the initial T cells directly and initiate the specific immune responses. In addition to extracellular factors and intracellular genetic factors, epigenetics plays a major role in the regulation of DCs. In this study, we obtained POL-P, and structural features and monosaccharide composition were analyzed. We evaluated the effect of POL-P on functional maturation of DCs derived from mice immunized with foot and mouth disease (FMD) vaccine and explored the related mechanism responsible for immunoenhancer. The levels of protein and gene related to IL-12p35 and IL-12p40 were determined by western blot and chromatin immunoprecipitation (ChIP) assays. The expressions of TLR2, TLR4 receptors and the downstream molecules of MyD88 and NF-κB were examined using immunohistochemistry. Results The average molecular weight (Mw) of the POL-P was 4×104 Da. The monosaccharide composition of the POL-P was mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose with a relative mass of 1.2%, 13.2%, 33.5%, 1.2%, 3.3%, 32.2% and 15.4%, respectively. We concluded that co-administration of POL-P with the FMD vaccine could significantly promote DCs maturation of phenotype and the immune function. In addition, the acetylation level of histone H3 of IL-12 was closely connected with the immune activity of DCs. Moreover, POL-P induced immune response was related to up-regulating protein expression of TLR2, TLR4, MyD88 and NF-κB in DCs. Conclusions Our evidence suggested that POL-P could be a potential immunostimulant in the regulation of DCs maturation for FMD vaccine.

Comparative Investigation of Cellular Effects of Polyethylene Glycol (PEG) Derivatives

Nowadays, polyethylene glycols referred to as PEGs are widely used in cosmetics, consumer care products, and the pharmaceutical industry. Their advantageous properties such as chemical stability, low immunogenicity, and high tolerability explain why PEGs are applied in many fields of pharmaceutical formulations including parenteral, topical, ophthalmic, oral, and rectal preparations and also in modern drug delivery systems. Given their extensive use, they are considered a well-known group of chemicals. However, the number of large-scale comparative studies involving multiple PEGs of wide molecular weight range is low, as in most cases biological effects are estimated upon molecular weight. The aim of this publication was to study the action of PEGs on Caco-2 cells and G. mellonella larvae and to calculate the correlation of these effects with molecular weight and osmolality. Eleven PEGs of different molecular weight were used in our experiments: PEG 200, PEG 300, PEG 400, PEG 600, PEG 1000, PEG 1500, PEG 4000, PEG 8000, PEG 10,000, 12,000, and PEG 20,000. The investigated cellular effects included cytotoxicity (MTT and Neutral Red assays, flow cytometry with propidium iodide and annexin V) and autophagy. The osmolality of different molecular weight PEGs with various concentrations was measured by a vapor pressure osmometer OSMOMAT 070 and G. mellonella larvae were injected with the solutions of PEGs. Sorbitol was used as controls of the same osmolality. Statistical correlation was calculated to describe the average molecular weight dependence of the different measured effects. Osmolality, the cytotoxicity assays, flow cytometry data, and larvae mortality had significant correlation with the structure of the PEGs, while autophagosome formation and the proportion of early apoptotic cells showed no statistical correlation. Overall, it must be noted that PEGs must be tested individually for biological effects as not all effects can be estimated by the average molecular weight.

Synthesis of High Molecular Weight Stereo-Di-Block Copolymers Driven by a Co-Initiator Free Catalyst

Stereo-diblock copolymers of high molecular weight polylactide (PLA) were synthetized by the one pot-sequential addition method assisted by a heteroscorpionate catalyst without the need of a co-initiator. The alkyl zinc organometallic heteroscorpionate derivative (Zn(Et)(κ3-bpzteH)] (bpzteH = 2,2-bis(3,5-dimethylpyrazol-1-yl)-1-para-tolylethoxide) proved to assist in the mechanism of reaction following a coordination-insertion process. Kinetic studies along with the linear correlation between monomer and number average molecular weight (Mn) conversion, and the narrow polydispersities supported the truly living polymerization character of the initiator, whereas matrix-assisted laser desorption/Ionization-time of flight (MALDI-TOF) studies showed a very low order of transesterification. The high stereo-control attained for the afforded high molecular weight derivatives was revealed by homonuclear decoupled 1H NMR spectra and polarimetry measurements. The nanostructure of the PLA derivatives was studied by both wide-angle X-ray scattering (WAXS) and differential scanning calorimetry (DSC) and the stereocomplex phase of the PLA stereo-diblock copolymers was successfully identified.

Evaluation of Radiation Resistance of Polystyrene Using Molecular Dynamics Simulation

The scission rates of polystyrene and fluorinated polystyrene irradiated in an irradiation facility with Co-60 γ-rays were determined using molecular dynamics simulation and gel permeation chromatography (GPC) molecular weight distributions. The prediction was based on the assumption that γ-ray energy is transferred to the initial velocity of the primary knock-on atom. We employed a molecular dynamics simulation procedure to compute the changes in bond length between the connections for selected values of the absorbed dose and compared the calculated values with measurements made on the irradiated samples. The samples were exposed to four different absorbed doses of 25, 50, 75, and 100 kGy. The scission process and scission ratio were simulated with LAMMPS with ReaxFF potential for each bond, and we compared the simulation results with the experimental data especially measuring average molecular weight to evaluate the effect of fluorination on radiation enhancement.

Morphology, thermal and mechanical properties of electrospun polyvinylidene/polyethylene glycol composite nanofibers as form-stabilized phase change materials

Polyvinylidene fluoride (PVDF)/polyethylene glycol (PEG) form-stabilized composite phase change nanofibers with various mass percentage of PEG1000 (with average molecular weight of 1000) loadings were fabricated by electrospinning technique, in which PEG was served as phase change material (PCM) and PVDF as the supporting matrix to afford a mechanically strong structure. Effect of PEG1000 content on nanofiber morphology, phase transition properties, thermal stability, thermal energy storage and release performance, and mechanical properties were studied experimentally. The results showed that the incorporation of PEG1000 improved the spinning solution viscosity and high mass fraction of PEG1000 in the nanofibers led to decreased fiber diameter and melting temperature, and higher latent heat of fusion. In addition, mechanical test revealed that the fracture elongation of the electrospun PVDF/PEG composite nanofibrous membranes were initially increased with unobvious tensile strength changes, while the value of the elongation would decrease, and the tensile strength increase as the mass ratio of PVDF/PEG lower to 1:1. Furthermore, the melting temperature and maximum latent heat for PVDF/PEG were determined from DSC measurement as 51.8 °C, and 73.3 J/g, when the mass radio of PVDF/PEG was 1:1 (w/w), which was suitable utilized for thermo-regulating textiles or in heat storage devices.

E-Beam Cross-Linking of Complex Hydrogels Formulation: The Influence of Poly(Ethylene Oxide) Concentration on the Hydrogel Properties

In the present study, we report on the complex hydrogels formulations based on collagen-poly(vinyl pyrrolidone) (PVP)-poly(ethylene oxide) (PEO) cross-linked by e-beam irradiation in an aqueous polymeric solution, aiming to investigate the influence of different PEO concentrations on the hydrogel properties. The hydrogel networks’ structure and their composition were investigated using equilibrium swelling degree, complex rheological analysis, and FT-IR spectroscopy. Rheological analysis was performed to determine the elastic (G′) and viscous (G″) moduli, the average molecular weight between cross-linking points (Mc), cross-link density (Ve), and the mesh size (ξ). The effect of the PEO concentration on the properties of the hydrogel was investigated as well. Depending on the PEO concentration added in their composition, the hydrogels swelling degree depends on the absorbed dose, being lower at low PEO concentrations. All hydrogel formulations showed higher G′ values (9.8 kPa) compared to G″ values (0.2 kPa), which shows that the hydrogels have a predominantly elastic behavior. They presented stability greater than 72 h in physiological pH buffers and reached equilibrium after 25 h. The Mc parameter is strongly dependent on the PEO concentration and the absorbed dose for all hydrogel compositions. The cross-linking density increased with the absorbed dose.

Production and Properties of Microbial Polyhydroxyalkanoates Synthesized from Hydrolysates of Jerusalem Artichoke Tubers and Vegetative Biomass

One of the major challenges in PHA biotechnology is optimization of biotechnological processes of the entire synthesis, mainly by using new inexpensive carbon substrates. A promising substrate for PHA synthesis may be the sugars extracted from the Jerusalem artichoke. In the present study, hydrolysates of Jerusalem artichoke (JA) tubers and vegetative biomass were produced and used as carbon substrate for PHA synthesis. The hydrolysis procedure (the combination of aqueous extraction and acid hydrolysis, process temperature and duration) influenced the content of reducing substances (RS), monosaccharide contents, and the fructose/glucose ratio. All types of hydrolysates tested as substrates for cultivation of three strains—C. necator B-10646 and R. eutropha B 5786 and B 8562—were suitable for PHA synthesis, producing different biomass concentrations and polymer contents. The most productive process, conducted in 12-L fermenters, was achieved on hydrolysates of JA tubers (X = 66.9 g/L, 82% PHA) and vegetative biomass (55.1 g/L and 62% PHA) produced by aqueous extraction of sugars at 80 °C followed by acid hydrolysis at 60 °C, using the most productive strain, C. necator B-10646. The effects of JA hydrolysates on physicochemical properties of PHAs were studied for the first time. P(3HB) specimens synthesized from the JA hydrolysates, regardless of the source (tubers or vegetative biomass), hydrolysis conditions, and PHA producing strain employed, exhibited the 100–120 °C difference between the Tmelt and Tdegr, prevailing of the crystalline phase over the amorphous one (Cx between 69 and 75%), and variations in weight average molecular weight (409–480) kDa. Supplementation of the culture medium of C. necator B-10646 grown on JA hydrolysates with potassium valerate and ε-caprolactone resulted in the synthesis of P(3HB-co-3HV) and P(3HB-co-4HB) copolymers that had decreased degrees of crystallinity and molecular weights, which influenced the porosity and surface roughness of polymer films prepared from them. The study shows that JA hydrolysates used as carbon source enabled productive synthesis of PHAs, comparable to synthesis from pure sugars. The next step is to scale up PHA synthesis from JA hydrolysates and conduct the feasibility study. The present study contributes to the solution of the critical problem of PHA biotechnology—finding widely available and inexpensive substrates.

Investigations on the Influence of Collagen Type on Physicochemical Properties of PVP/PVA Composites Enriched with Hydroxyapatite Developed for Biomedical Applications

Nowadays, a great attention is directed into development of innovative multifunctional composites which may support bone tissue regeneration. This may be achieved by combining collagen and hydroxyapatite showing bioactivity, osteoconductivity and osteoinductivity with such biocompatible polymers as polyvinylpyrrolidone (PVP) and poly(vinyl alcohol) (PVA). Here PVA/PVP-based composites modified with hydroxyapatite (HAp, 10 wt.%) and collagen (30 wt.%) were obtained via UV radiation while two types of collagen were used (fish and bovine) and crosslinking agents differing in the average molecular weight. Next, their chemical structure was characterized using Fourier transform infrared (FT-IR) spectroscopy, roughness of their surfaces was determined using a stylus contact profilometer while their wettability was evaluated by a sessile drop method followed by the measurements of their surface free energy. Subsequently, swelling properties of composites were verified in simulated physiological liquids as well as the behavior of composites in these liquids by pH measurements. It was proved that collagen-modified composites showed higher swelling ability (even 25% more) compared to unmodified ones, surface roughness, biocompatibility towards simulated physiological liquids and hydrophilicity (contact angles lower than 90°). Considering physicochemical properties of developed materials and a possibility of the preparation of their various shapes and sizes, it may be concluded that developed materials showed great application potential for biomedical use, e.g., as materials filling bone defects supporting their treatments and promoting bone tissue regeneration due to the presence of hydroxyapatite with osteoinductive and osteoconductive properties.

A Novel Agarase, Gaa16B, Isolated from the Marine Bacterium Gilvimarinus agarilyticus JEA5, and the Moisturizing Effect of Its Partial Hydrolysis Products

We recently identified a β-agarase, Gaa16B, in the marine bacterium Gilvimarinus agarilyticus JEA5. Gaa16B, belonging to the glycoside hydrolase 16 family of β-agarases, shows less than 70.9% amino acid similarity with previously characterized agarases. Recombinant Gaa16B lacking the carbohydrate-binding region (rGaa16Bc) was overexpressed in Escherichia coli and purified. Activity assays revealed the optimal temperature and pH of rGaa16Bc to be 55 ∘C and pH 6–7, respectively, and the protein was highly stable at 55 ∘C for 90 min. Additionally, rGaa16Bc activity was strongly enhanced (2.3-fold) in the presence of 2.5 mM MnCl2. The Km and Vmax of rGaa16Bc for agarose were 6.4 mg/mL and 953 U/mg, respectively. Thin-layer chromatography analysis revealed that rGaa16Bc can hydrolyze agarose into neoagarotetraose and neoagarobiose. Partial hydrolysis products (PHPs) of rGaa16Bc had an average molecular weight of 88–102 kDa and exhibited > 60% hyaluronidase inhibition activity at a concentration of 1 mg/mL, whereas the completely hydrolyzed product (CHP) showed no hyaluronidase at the same concentration. The biochemical properties of Gaa16B suggest that it could be useful for producing functional neoagaro-oligosaccharides. Additionally, the PHP of rGaa16Bc may be useful in promoting its utilization, which is limited due to the gel strength of agar.

Effects of Radiation sterilization Dose on the Molecular Weight and Gelling Properties of Commercial Alginate Samples

To estimate the molecular weight (Mw) and gelling properties, a total of 26 alginate samples consisting of control (n = 13) and 15 kGy γ-irradiated (n = 13) samples were characterized through viscometric and gel permeation chromatography (GPC-MALLS) methods. Based on the observations, a remarkable decrease in the intrinsic viscosity of all samples of alginates was evident due to the effects of radiation, with a linear relationship between viscosity and concentration in 0.01 M NaCl solution. The correlation among the Mw, percentage mass recovery, radii of gyration (Rz/Rg), and percentage reduction of Mw assessed by GPC was significant. The Mw decreased dramatically (from 3.1 × 105 to 0.49 × 105 mole/g in sample no. 12) by the effect of radiation with momentous relation to the % reduction of the molecular weight. The highest molecular weight reduction (84%), which is the most sensitive to γ-radiation, and the average reduction rate was ≥50%. The mass recovery was 100% obtained from samples no. 1,3,4,5,7,12, and 13, while the rest of the samples’ recovery rate was significantly higher. The reduction rate of mass molecular weight (Mw) is higher than the average molecular weight (Mv), but they showed a sensitivity towards radiation, consequently their performance are different from each other. The stability test was performed as a critical behaviour in the control, recurrently same as in the irradiated samples. Thus, the sterilization dose of 15 kGy for the Mw distribution, and subsequently for the characterization, was significantly effective.