Formation of Three-Dimensional Polysuccinimide Electrospun Fiber Meshes Induced by the Combination of CaCl2 and Humidity.

Even though electrospinning is getting more and more attention, the preparation of 3D nanofibrous meshes is still a big challenge that limits the application of electrospun materials, especially in tissue engineering. To overcome this problem, several solutions are introduced but most of them focus on the postprocessing of the electrospun meshes. This paper presents a straightforward novel method that utilizes the joint effect of the addition of CaCl2 and the relative environmental humidity (RH), which can induce the random 3D formation of polysuccinimide (PSI) electrospun fibers with different such as wrinkled or ribbon-like structures. Although the effect of humidity and inorganic salt additives on the micro and macrostructure of electrospun fibers is known, the connection between the two in this manner has never been presented. To investigate the effect, fibers with different PSI and CaCl2 concentrations at different humidity RH levels are prepared, and their microstructure is visualized with high-resolution scanning electron microscopy (SEM). To reveal the nature of the interaction between the polymer and the CaCl2, Fourier-transformed infrared (FTIR), X-ray diffraction (XRD), and thermogravimetry (TGA) measurements are carried out and 3D nanofibrous structures are obtained.

Structural and functional comparison of Saccharomonospora azurea strains in terms of primycin producing ability.

Emerging and re-emerging microbial pathogens, together with their rapid evolution and adaptation against antibiotics, highlight the importance not only of screening for new antimicrobial agents, but also for deepening knowledge about existing antibiotics. Primycin is a large 36-membered non-polyene macrolide lactone exclusively produced by Saccharomonospora azurea. This study provides information about strain dependent primycin production ability in conjunction with the structural, functional and comparative genomic examinations. Comparison of high- and low-primycin producer strains, transcriptomic analysis identified a total of 686 differentially expressed genes (DEGs), classified into diverse Cluster of Orthologous Groups. Among them, genes related to fatty acid synthesis, self-resistance, regulation of secondary metabolism and agmatinase encoding gene responsible for catalyze conversion between guanidino/amino forms of primycin were discussed. Based on in silico data mining methods, we were able to identify DEGs whose altered expression provide a good starting point for the optimization of fermentation processes, in order to perform targeted strain improvement and rational drug design.

Specific Condis Crystal-like Mesophase of Poly(butylene succinate-co-butylene adipate).

Understanding the properties of polymers, such as their crystallinity, is crucial for their material performance and predicting their behavior during and after use, especially in the case of environmentally friendly (bio)degradable polymers, enabling optimized design. In this work, for the first time, a pressure-induced condis crystal-like mesophase of poly(butylene succinate-co-butylene adipate) (PBSA) is presented. The phase behavior of pressed films obtained from commercial PBSA with 25% butylene adipate units is investigated at various processing temperatures from room temperature to 100 °C, pressed at a pressure of the press jaws and at 2-5 t for 1-5 min. The characterization and quantification evaluation of the condis crystal-like mesophase of pressed PBSA formed at temperatures above the glass transition is investigated by X-ray diffraction, polarized optical microscopy (POM), and differential scanning calorimetry (DSC) methods. Our results demonstrate that pressed PBSA films at 60 °C show a condis crystal-like mesophase, characterized by the presence of reflections at wide angles, birefringence by POM, as well as a higher melting point (endotherm) by DSC. The resulting oriented mesomorphic green polymer can, in a sustainable manner, expand further technological applications of (bio)degradable polymers, especially in the medical field, and open up opportunities for further research that could provide such polymers with tailored persistence and degradation, thus changing the shelf life.

Phytobiotic-Prebiotic Feed Additive Containing a Combination of Carob Pulp, Chicory, and Fenugreek Improve Growth Performance, Carcass Traits, and Fecal Microbiota of Fattening Pigs.

The purpose of this study was to determine the effectiveness of a phytobiotic-prebiotic feed additive (PPFA, which contains a combination of chicory and extracts of carob pulp and fenugreek) in the diets of fattening pigs on growth indicators, carcass characteristics, and fecal microbiota. A total of 329 crossbred pigs were randomly divided into two dietary treatments, including a basal diet without additives as the control group and a basal diet supplemented with 1 kg/T PPFA as the trial group. The PPFA supplementation led to a significant increase in the body weight gain and average daily gain of the trial group compared to those of the control group after 70 days of feeding. Through the S-EUROP evaluation system, we also found that the fattening pigs fed PPFA significantly improved their carcass indicators. Furthermore, it was shown that PPFA regulated porcine intestinal microbiota, including promoting the growth of the beneficial commensal bacteria (i.e., Bifidobacterium and Lactobacillus) while inhibiting some potential pathogen bacteria (i.e., Bacteroidaceae and Campylobacteraceae). Our work revealed that the phytobiotic-prebiotic feed additive containing carob pulp, chicory, and fenugreek positively influences the intestinal microbiota, growth performance, and carcass traits in fattening swine.

Surface Affinity of Tetramethylammonium Iodide in Aqueous Solutions: A Combined Experimental and Computer Simulation Study.

The surface affinity of tetramethylammonium iodide (TMAI) in aqueous solutions is investigated by surface tension measurements and molecular dynamics computer simulations. Experiments, performed in the entire composition range of solubility using the pendant drop method with two different setups, clearly reveal that TMAI is a weakly capillary active salt. Computer simulations performed with the AMBER force field reproduce the experimental data very well, while two other major force fields (i.e., CHARMM and OPLS) can still reproduce the experimental trend qualitatively; however, even qualitative reproduction of the experimental trend requires scaling down the ion charges according to the Leontyev-Stuchebrukhov correction. On the other hand, the GROMOS force field fails in reproducing the experimentally confirmed capillary activity of TMAI. Molecular dynamics simulation results show that, among the two ions, iodide has a clearly larger surface affinity than tetramethylammonium (TMA+). Further, the adsorption of the I- anions is strictly limited to the first molecular layer beneath the liquid-vapor interface, which is followed by several layers of their depletion. On the other hand, the net negative charge of the surface layer, caused by the excess amount of I- with respect to TMA+, is compensated by a diffuse layer of adsorbed TMA+ cations, extending to or beyond the fourth molecular layer beneath the liquid surface.

Characterization of Electrospun Polysuccinimide-Dopamine Conjugates and Effect on Cell Viability and Uptake.

Biocompatible nanofibrous systems made by electrospinning have been studied widely for pharmaceutical applications since they have a high specific surface and the capability to make the entrapped drug molecule amorphous, which increases bioavailability. By covalently conjugating drugs onto polymers, the degradation of the drug as well as the fast clearance from the circulation can be avoided. Although covalent polymer-drug conjugates have a lot of advantages, there is a lack of research focusing on their nano-formulation by electrospinning. In this study, polysuccinimide (PSI) based electrospun fibrous meshes conjugated with dopamine (DA) are prepared. Fiber diameter, mechanical properties, dissolution kinetics and membrane permeability are thoroughly investigated, as these are crucial for drug delivery and implantation. Dopamine release kinetics prove the prolonged release that influenced the viability and morphology of periodontal ligament stem cells (PDLSCs) and SH-SY5Y cells. The presence of dopamine receptors on both cell types is also demonstrated and the uptake of the conjugates is measured. According to flow cytometry analysis, the conjugates are internalized by both cell types, which is influenced by the chemical structure and physical properties. In conclusion, electrospinning of PSI-DA conjugates alters release kinetics, meanwhile, conjugated dopamine can play a key role in cellular uptake.

Local Structure of DMF-Water Mixtures, as Seen from Computer Simulations and Voronoi Analysis.

Molecular dynamics simulations of mixtures of N,N-dimethylformamide (DMF) with water of various compositions, covering the entire composition range, are performed on the canonical (N,V,T) ensemble. The local structure of the mixtures is analyzed in terms of radial distribution functions and the contributions of the first five neighbors to them, various order parameters of the water molecules around each other, and properties of the Voronoi polyhedra of the molecules. The analyses lead to the following main conclusions. The two molecules are mixing with each other even on the molecular scale; however, small self-aggregates of both components persist even at their small mole fraction values. In particular, water-water H-bonds exist in the entire composition range, while water clusters larger than 3 and 2 molecules disappear above the DMF mole fraction values of about 0.7 and 0.9, respectively. The O atoms of the DMF molecules can well replace water O atoms in the hydrogen-bonding network. Further, the H-bonding structure is enhanced by the presence of the hydrophobic CH3 groups of the DMF molecules. On the other hand, the H-bonding network of the molecules gradually breaks down upon the addition of DMF to the system due to the lack of H-donating groups of the DMF molecules. Finally, in neat DMF, the molecules form weak, CH-donated H-bonds with each other; however, these H-bonds disappear upon the addition of water due to the increasing competition with the considerably stronger OH-donated H-bonds DMF can form with the water molecules.

Alternative to ZnO to establish balanced intestinal microbiota for weaning piglets.

A wide range of phytobiotic feed additives are available on the market claiming to have beneficial effects on the growth of the host animal and to promote the development of a balanced microflora. The present study investigated the effects of the phytobiotic-prebiotic mixture of curcumin, wheat germ, and chicory on the growth performance and on the intestinal microflora composition of weaning piglets. Post weaning diarrhea causes significant losses for the producers, most commonly it is prevented by feeding high doses of zinc oxide (ZnO). The effect of a phytobiotic-prebiotic feed additive (1 kg T-1) was compared to a positive control (3.1 kg T-1 ZnO) and to a negative control (no feed supplement) in an in vivo animal trial. There was no significant difference in the final body weight and average daily gain of the trial and positive control groups, and both groups showed significantly (P<0.05) better results than the negative control. The feed conversion ratio of the phytobiotic-prebiotic supplemented group was significantly improved (P<0.05) compared to both controls. Both phytobiotic-prebiotic mixture and ZnO were able to significantly reduce (P<0.05) the amount of coliforms after weaning, even though ZnO reduced the amount of coliforms more efficiently than the trial feed additive, it also reduced the amount of potentially beneficial bacteria. Metagenomic data also corroborated the above conclusion. In the trial and positive control groups, the relative abundance of Enterobacteriaceae decreased by 85 and 88% between 3 weeks and 6 weeks of age, while in the negative control group a slight increase occurred. Lactobacillaceae were more abundant in the trial group (29.98%) than in the positive (8.67%) or in the negative (22.45%) control groups at 6 weeks of age. In summary, this study demonstrated that a phytobiotic-prebiotic feed additive may be a real alternative to ZnO for the prevention of post weaning diarrhea and promote the development of a balanced gut system.

Liposomal Formulation of Botanical Extracts may Enhance Yield Triggering PR Genes and Phenylpropanoid Pathway in Barley (Hordeum vulgare).

This work aimed to study the plant conditioning effect and mode of action of a plant-based biostimulant used in organic farming. This new generation plant biostimulant, named ELICE16INDURES®, is rich in plant bio-active ingredients containing eleven supercritical botanical extracts encapsulated in nano-scale liposomes. The dose-response (10 to 240 g ha-1) relationship was tested in a field population of autumn barley (Hordeum vulgare) test crop, and underlying molecular mechanisms were studied. Applying nanotechnology, cell-identical nanoparticles may help the better uptake and delivery of active ingredients increasing resilience, vitality, and crop yield. The amount of harvested crops showed a significant increase of 27.5% and 39.9% interconnected to higher normalized difference vegetation index (NDVI) of 20% and 25% after the treatment of low and high dosages (20 and 240 g ha-1), respectively. Illumina NextSeq 550 sequencing, gene expression profiling, and KEGG-pathway analysis of outstanding dosages indicated the upregulation of pathogenesis-related (PR) and other genes-associated with induced resistance-which showed dose dependency as well.

Reduction of blood cholesterol and ischemic injury in the hypercholesteromic rabbits with modified resveratrol, longevinex. [corrected]

The present study examined the efficacy of using longevinex, a commercially available resveratrol formulation, to lower blood cholesterol in hypercholesteromic rabbits. New Zealand white rabbits were randomly divided into two groups (n = 6 per group), one group was given high cholesterol diet for 3 months while the other group fed regular diet served as control. The high cholesterol diet fed group was further subdivided into two groups (n = 6 per group), one group was given longevinex resveratrol while the other group given vehicle only served as control. Longevinex was given by gavaging up to a period of 6 months. Longevinex-treated rabbits exhibited lowering of plasma cholesterol level. Inhibition of arterial plaque formation was noticed even after 1 month. Longevinex-treated hearts demonstrated improved ventricular recovery when isolated working hearts were subjected to 30 min of ischemia followed by 2 h of reperfusion. Aortic flow and developed pressure during post-ischemic reperfusion period were significantly higher for the longevinex-treated hearts compared to those in control group of hearts. Myocardial infarct size was also lower in the treated group compared to that for the untreated group. These results indicate cholesterol-lowering ability of longevinex, which appears to reflect in its ability to protect the hypercholesteromic hearts from ischemic reperfusion injury.

Voronoi Polyhedra as a Tool for the Characterization of Inhomogeneous Distribution in 1-Butyl-3-methylimidazolium Cation-Based Ionic Liquids.

The inhomogeneity distribution in four imidazolium-based ionic liquids (ILs) containing the 1-butyl-3-methylimidazolium (C4mim) cation, coupled with tetrafluoroborate (BF4), hexafluorophosphate (PF6), bis(trifluoromethanesulfonyl)amide (TFSA), and trifluoromethanesulfonate (TfO) anions, was characterized using Voronoi polyhedra. For this purpose, molecular dynamic simulations have been performed on the isothermal-isobaric (NpT) ensemble. We checked the ability of the potential models to reproduce the experimental density, heat of vaporization, and transport properties (diffusion and viscosity) of these ionic liquids. The inhomogeneity distribution of ions around the ring, methyl, and butyl chain terminal hydrogen atoms of the C4mim cation was investigated by means of Voronoi polyhedra analysis. For this purpose, the position of the C4mim cation was described successively by the ring, methyl, and butyl chain terminal hydrogen atoms, while that of the anions was described by their F or O atom. We calculated the Voronoi polyhedra distributions of the volume, the density, and the asphericity parameters as well as that of the radius of the spherical intermolecular voids. We carried out the analysis in two steps. In the first step, both ions were taken into account. The calculated distributions gave information on the neighboring ions around a reference one. In the second step, to distinguish between like and oppositely charged ions and then to get information on the inhomogeneity distribution of the like ions, we repeated the same calculations on the same sample configurations and removed one of the ions and considered only the other one. Detailed analysis of these distributions has revealed that the ring hydrogen atoms are mainly solvated by the anions, while the methyl and butyl terminal H atoms are surrounded by like atoms. The extent of this inhomogeneity was assessed by calculating the cluster size distribution that shows that the dimers are the most abundant ones.

Free thiol groups on poly(aspartamide) based hydrogels facilitate tooth-derived progenitor cell proliferation and differentiation.

Cell-based tissue reconstruction is an important field of regenerative medicine. Stem and progenitor cells derived from tooth-associated tissues have strong regeneration potential. However, their in vivo application requires the development of novel scaffolds that will provide a suitable three-dimensional (3D) environment allowing not only the survival of the cells but eliciting their proliferation and differentiation. Our aim was to study the viability and differentiation capacity of periodontal ligament cells (PDLCs) cultured on recently developed biocompatible and biodegradable poly(aspartamide) (PASP)-based hydrogels. Viability and behavior of PDLCs were investigated on PASP-based hydrogels possessing different chemical, physical and mechanical properties. Based on our previous results, the effect of thiol group density in the polymer matrix on cell viability, morphology and differentiation ability is in the focus of our article. The chemical composition and 3D structures of the hydrogels were determined by FT Raman spectroscopy and Scanning Electron Microscopy. Morphology of the cells was examined by phase contrast microscopy. To visualize cell growth and migration patterns through the hydrogels, two-photon microscopy were utilized. Cell viability analysis was performed according to a standardized protocol using WST-1 reagent. PDLCs were able to attach and grow on PASP-based hydrogels. An increase in gel stiffness enhanced adhesion and proliferation of the cells. However, the highest population of viable cells was observed on the PASP gels containing free thiol groups. The presence of thiol groups does not only enhance viability but also facilitates the osteogenic direction of the differentiating cells. These cell-gel structures seem to be highly promising for cell-based tissue reconstruction purposes in the field of regenerative medicine.

Comparative analysis of the complete mitochondrial genomes of Aspergillus niger mtDNA type 1a and Aspergillus tubingensis mtDNA type 2b.

To understand the differences in the organization of mitochondrial genomes of the very closely related Aspergillus niger and Aspergillus tubingensis species, we determined the complete genome sequence of the 1a mtDNA type of A. niger and 2b mtDNA type of A. tubingensis and now we provide a comparative analysis of the two mtDNAs. We found that (1) the organization (gene content and order) of the two genomes is almost identical and (2) the size difference between them is principally attributed to the different intron content of their cox1, atp9 and ndh4L genes.

Interpretation of intraspecific variability in mtDNAs of Aspergillus niger strains and rearrangement of their mtDNAs following mitochondrial transmissions.

Physical and functional maps of mitochondrial DNAs of Aspergillus niger strains representing different mitochondrial DNA RFLP patterns were constructed and compared. In spite of the high similarity in the organisation of mitochondrial DNAs among examined strains, differences could be easily recognised by applying molecular markers, such as the different intron content of the cox1 genes, the sequence of the intergenic regions between the Met- and His-tRNA genes and downstream of the tRNA-Gly gene. Intraspecific mitochondrial transfers between the heterokaryon incompatible mitochondrial oligomycin-resistant A. niger strain, as the donor, and other A. niger-sensitive strains bearing different RFLP patterns resulted in oligomycin-resistant progeny possessing either rearranged or unchanged donor mitochondrial DNA and recipient nuclei. Since the intergenic marker sequences of mitochondrial DNAs turned out to be identical in the donor and the progeny, it can be assumed that the oligomycin-resistant progeny inherit the mitochondrial DNA of the donor strain; this may either remain unchanged or may be modified by a mobile intron of the cox1 gene of the recipient mitochondria.

Mitochondrial DNA organisation of the mtDNA type 2b of Aspergillus tubingensis compared to the Aspergillus niger mtDNA type 1a.

A physical and functional map of Aspergillus tubingensis mtDNA type 2b was constructed and compared to Aspergillus niger typeI a mtDNA. The gene content and order, as well as the patterns of restriction sites, were similar. Two unidentified ORFs and several repeat elements were found in the region between the ndhl and ndh4 genes on both mtDNAs. The sizes of the A. niger and A. tubingensis mtDNAs were 31.23 and 33.06 kb. respectively, the difference was principally attributed to the altered intron content of their coxl genes.

Interpretation of mtDNA RFLP variability among Aspergillus tubingensis isolates.

Aspergillus tubingensis isolates collected from distant geographic areas were earlier classified into six groups on the basis of the mtDNA RFLP variability they exhibited (mtDNA types 2a-2f). In the present work, we investigated the reason for the intraspecific mtDNA variability and we describe here how this fungus, with a relatively small mitochondrial genome, can display intraspecific polymorphism due to intron acquisition and also sporadic point mutations affecting the recognition motifs of the restriction enzymes employed in the RFLP analysis. Three different LAGLI-DADG type group I introns were identified in the cox1 gene amongst the six mtDNA RFLP types. MtDNAs of types 2b and 2d contain all of the three introns, mtDNA of type 2f carries only one, and the other mtDNA types contain two introns each. Comparative analysis showed that the first and second introns of mtDNAs of types 2b and 2d are well distributed among fungi, indicating their active horizontal transfer capacity. The third intron occurs rarely among fungi and is restricted to a limited number of fungal species, namely to A. tubingensis and the yeast Candida stellata. It is interesting that this intron is present in a small mitochondrial genome such as that of A. tubingensis and, considering its rarity, its presence amongst black Aspergillus isolates is recommended to be considered as a tool to establish taxonomical unit(s) or to track down evolutionary divergence of closely related taxonomical units.