Nanoengineered biocomposite tricomponent polymer based matrices for bone tissue engineering

Nanoengineered biodegradable constructs based on synthetic and natural polymers enriched with hydroxyapatite (HA) nanoparticles have been found to mimic the extracellular matrix of bone tissue. The main objective of this study was to create biocomposite nanostructured scaffolds by incorporating collagen and HA nanoparticles into poly(L-lactic acid)-co-poly(?-caprolactone) by electrospinning. The fiber diameter of the composite PLCL/Col and PLCL/Col/HA fibers was smaller compared to PLCL. In vitro biocompatibility of the scaffolds studied using human fetal osteoblasts and EDX analysis showed high deposition of calcium on PLCL/Col/HA. The results shows that PLCL/Col/HA nanofibrous constructs have huge potential as substrates for bone regeneration.     

The role of ammonium citrate washing on the characteristics of mechanochemical–hydrothermal derived magnesium-containing apatites

The role of citrate washing on the physical and chemical characteristics of magnesium-substituted apatites (HAMgs) was performed. HAMgs were synthesized by a mechanochemical–hydrothermal route at room temperature in as little as 1 h, which is five times faster than our previous work. Magnesium-substituted apatites had concentrations as high as 17.6 wt% Mg with a corresponding specific surface area (SSA) of 216 m²/g. A systematic study was performed to examine the influence of increasing magnesium content on the physical and chemical characteristics of the reaction products. As the magnesium content increased from 0 to 17.6 wt%, magnesium-doped apatite crystallite size decreased from 12 to 8.8 nm. The Mg/(Mg + Ca) ratio in the product was enriched relative to that used for the reacting precursor solution. During mechanochemical–hydrothermal reaction, magnesium doped apatites co-crystallize with magnesium hydroxide. Citrate washing serves to remove the magnesium hydroxide phase. The concomitant increase in surface area results because of the removal of this phase. Possible mechanisms for magnesium hydroxide leaching are discussed to explain the measured trends.     

RAPD with microsatellite as a tool for differentiation of Candida genus yeasts isolated in brewing

Fifteen wild yeast strains were isolated in two factories of a lager brewing company in Poland. Their identification with API 32C system showed mainly the presence of Candida sake species (7/15). To differentiate the isolates, randomly amplified polymorphic DNA (RAPD) with (GTG)(5), (GAC)(5), (GACA)(4) microsatellite primers and M13 core sequence (5'-GAG GGT GGC GGT TCT-3') were chosen. The results of patterns similarity are presented as dendrograms for each RAPD analysis and for overall patterns. On the overall patterns, all isolates identified as C. sake, except Strain No. 1, were regrouped in one cluster. Collection strain C. sake CBS 617 was similar in 46% to the cluster with six isolates (Strain Nos. 3, 6, 8, 11, 13, 14). The second reference strain C. sake CBS 159 and the Strain No. 1 were regrouped with other Candida species (collection strains) showing, respectively, only 20% and 42% of similarity to other C. sake strains. The similarity based on the overall dendrogram between isolate Nos. 3, 6, 8, 11, 13, 14 and C. sake CBS 617 was 49%. Between those strains and other Candida, the similarity was only 37%.     

The role of vacancies and local distortions in the design of new phase-change materials

Phase-change materials are of tremendous technological importance ranging from optical data storage to electronic memories. Despite this interest, many fundamental properties of phase-change materials, such as the role of vacancies, remain poorly understood. 'GeSbTe'-based phase-change materials contain vacancy concentrations around 10% in their metastable crystalline structure. By using density-functional theory, the origin of these vacancies has been clarified and we show that the most stable crystalline phases with rocksalt-like structures are characterized by large vacancy concentrations and local distortions. The ease by which vacancies are formed is explained by the need to annihilate energetically unfavourable antibonding Ge-Te and Sb-Te interactions in the highest occupied bands. Understanding how the interplay between vacancies and local distortions lowers the total energy helps to design novel phase-change materials as evidenced by new experimental data.     

Nanostructuring with a crosslinkable discotic material

A high-yielding synthesis afforded a hexa-peri-hexabenzocoronene carrying acrylate units at the end of six attached alkyl spacers. The polymerization of these acrylate moieties could be initiated with thermal energy and through direct photoactivation without the addition of a photoinitiator. This allowed the organization of the liquid-crystalline material to be fixed in either the crystalline state or the mesophase, which preserved the organization in the respective phase. The use of a focused synchrotron beam permitted selected regions of a thin film to be rendered insoluble. After "developing" the film in this lithographic process by dissolving the soluble, unpolymerized material, defined nano-objects remained on the substrate. In addition, the pronounced aromatic pi stacking of the novel material allows an organization in mesoporous membranes that could be fixed by thermal crosslinking. After the removal of the inorganic template, mechanically stable nanotubes were obtained, which were characterized by different microscopy techniques.     

A Partial Order Approach to Branching Time Logic Model Checking

Partial order techniques enable reducing the size of the state space used for model checking, thus alleviating the “state space explosion” problem. These reductions are based on selecting a subset of the enabled operations from each program state. So far, these methods have been studied, implemented, and demonstrated for assertional languages that model the executions of a program as computation sequences, in particular the linear temporal logic. The present paper shows, for the first time, how this approach can be applied to languages that model the behavior of a program as a tree. We study here partial order reductions for branching temporal logics, e.g., the logics CTL and CTL* (with the next time operator removed) and process algebra logics such as Hennesy–Milner logic (withτactions). Conditions on the selection of subset of successors from each state during the state-space construction, which guarantee reduction that preserves CTL* properties, are given. The experimental results provided show that the reduction is substantial.     

From Primary MSC Culture of Adipose Tissue to Immortalized Cell Line Producing Cytokines for Potential Use in Regenerative Medicine Therapy or Immunotherapy

For twenty-five years, attempts have been made to use MSCs in the treatment of various diseases due to their regenerative and immunomodulatory properties. However, the results are not satisfactory. Assuming that MSCs can be replaced in some therapies by the active factors they produce, the immortalized MSCs line was established from human adipose tissue (HATMSC1) to produce conditioned media and test its regenerative potential in vitro in terms of possible clinical application. The production of biologically active factors by primary MSCs was lower compared to the HATMSC1 cell line and several factors were produced only by the cell line. It has been shown that an HATMSC1-conditioned medium increases the proliferation of various cell types, augments the adhesion of cells and improves endothelial cell function. It was found that hypoxia during culture resulted in an augmentation in the pro-angiogenic factors production, such as VEGF, IL-8, Angiogenin and MCP-1. The immunomodulatory factors caused an increase in the production of GM-CSF, IL-5, IL-6, MCP-1, RANTES and IL-8. These data suggest that these factors, produced under different culture conditions, could be used for different medical conditions, such as in regenerative medicine, when an increased concentration of pro-angiogenic factors may be beneficial, or in inflammatory diseases with conditioned media with a high concentration of immunomodulatory factors.     

Cationic Peptides and Their Cu(II) and Ni(II) Complexes: Coordination and Biological Characteristics

Antimicrobial peptides are a promising group of compounds used for the treatment of infections. In some cases, metal ions are essential to activate these molecules. Examples of metalloantibiotics are, for instance, bleomycin and dermcidin. This study is focused on three new pseudopeptides with potential biological activity. The coordination behavior of all ligands with Cu(II) and Ni(II) ions has been examined. Various analytical methods such as potentiometric titration, UV-Vis and CD spectroscopies, and mass spectrometry were used. All compounds are convenient chelators for metal ion-binding. Two of the ligands tested have histidine residues. Surprisingly, imidazole nitrogen is not involved in the coordination of the metal ion. The N-terminal amino group, Dab side chains, and amide nitrogen atoms of the peptide bonds coordinated Cu(II) and Ni(II) in all the complexes formed. The cytotoxicity of three pseudopeptides and their complexes was evaluated. Moreover, their other model allowed for assessing the attenuation of LPS-induced cytotoxicity and anti-inflammatory activities were also evaluated, the results of which revealed to be very promising.     

The Mutation in wbaP cps Gene Cluster Selected by Phage-Borne Depolymerase Abolishes Capsule Production and Diminishes the Virulence of Klebsiella pneumoniae

Klebsiella pneumoniae is considered one of the most critical multidrug-resistant pathogens and urgently requires new therapeutic strategies. Capsular polysaccharides (CPS), lipopolysaccharides (LPS), and exopolysaccharides (EPS) are the major virulence factors protecting K. pneumoniae against the immune response and thus may be targeted by phage-based therapeutics such as polysaccharides-degrading enzymes. Since the emergence of resistance to antibacterials is generally considered undesirable, in this study, the genetic and phenotypic characteristics of resistance to the phage-borne CPS-degrading depolymerase and its effect on K. pneumoniae virulence were investigated. The K63 serotype targeting depolymerase (KP36gp50) derived from Klebsiella siphovirus KP36 was used as the selective agent during the treatment of K. pneumoniae 486 biofilm. Genome-driven examination combined with the surface polysaccharide structural analysis of resistant mutant showed the point mutation and frameshift in the wbaP gene located within the cps gene cluster, resulting in the loss of the capsule. The sharp decline in the yield of CPS was accompanied by the production of a larger amount of smooth LPS. The modification of the surface polysaccharide layers did not affect bacterial fitness nor the insensitivity to serum complement; however, it made bacteria more prone to phagocytosis combined with the higher adherence and internalization to human lung epithelial cells. In that context, it was showed that the emerging resistance to the antivirulence agent (phage-borne capsule depolymerase) results in beneficial consequences, i.e., the sensitization to the innate immune response.