Aqueous modification of chitosan with itaconic acid to produce strong oxygen barrier film

In this study, the chemical modification of chitosan using itaconic acid as a natural-based unsaturated dicarboxylic acid was investigated. In an aqueous environment, the amine group of chitosan reacts with itaconic acid to produce a chitosan derivative with pyrrolidone-4-carboxylic acid group. On the basis of the elemental analysis, 15 of the amine groups of chitosan reacted, thus creating modified chitosan with amine and carboxylic acid functionalities. Due to the presence of amine and carboxylic acid groups, the surface charge properties of the chitosan were notably altered after itaconic acid modification. In an aqueous solution, the modified chitosan exhibited zwitterionic properties, being cationic at low pH and turning anionic when the pH was increased over 6.5, whereas the original chitosan remained cationic until pH 9. Furthermore, it was demostrated that the modified chitosan was suitable for the preparation of a self-standing film with similarly high transparency but notably higher mechanical strength and oxygen barrier properties compared to a film made from the original chitosan. In addition, the thermal stability of the modified chitosan film was higher than that of the original chitosan film, and the modified chitosan exhibited flame-retardant properties

Molecular features of biguanides required for targeting of mitochondrial respiratory complex I and activation of AMP-kinase.

BACKGROUND: The biguanides are a family of drugs with diverse clinical applications. Metformin, a widely used anti-hyperglycemic biguanide, suppresses mitochondrial respiration by inhibiting respiratory complex I. Phenformin, a related anti-hyperglycemic biguanide, also inhibits respiration, but proguanil, which is widely used for the prevention of malaria, does not. The molecular structures of phenformin and proguanil are closely related and both inhibit isolated complex I. Proguanil does not inhibit respiration in cells and mitochondria because it is unable to access complex I. The molecular features that determine which biguanides accumulate in mitochondria, enabling them to inhibit complex I in vivo, are not known. RESULTS: Here, a family of seven biguanides are used to reveal the molecular features that determine why phenformin enters mitochondria and inhibits respiration whereas proguanil does not. All seven biguanides inhibit isolated complex I, but only four of them inhibit respiration in cells and mitochondria. Direct conjugation of a phenyl group and bis-substitution of the biguanide moiety prevent uptake into mitochondria, irrespective of the compound hydrophobicity. This high selectivity suggests that biguanide uptake into mitochondria is protein mediated, and is not by passive diffusion. Only those biguanides that enter mitochondria and inhibit complex I activate AMP kinase, strengthening links between complex I and the downstream effects of biguanide treatments. CONCLUSIONS: Biguanides inhibit mitochondrial complex I, but specific molecular features control the uptake of substituted biguanides into mitochondria, so only some biguanides inhibit mitochondrial respiration in vivo. Biguanides with restricted intracellular access may be used to determine physiologically relevant targets of biguanide action, and for the rational design of substituted biguanides for diverse clinical applications

Cytokines impact natural killer cell phenotype and functionality against glioblastoma in vitro

ObjectiveNatural killer (NK) cells are a part of the innate immune system and first-line defense against cancer. Since they possess natural mechanisms to recognize and kill tumor cells, NK cells are considered as a potential option for an off-the-shelf allogeneic cell-based immunotherapy. Here, our objective was to identify the optimal cytokine-based, feeder-free, activation and expansion protocol for cytotoxic NK cells against glioblastoma in vitro.MethodsNK cells were enriched from human peripheral blood and expanded for 16 days with different activation and cytokine combinations. The expansion conditions were evaluated based on NK cell viability, functionality, expansion rate and purity. The cytotoxicity and degranulation of the expanded NK cells were measured in vitro from co‑cultures with the glioma cell lines U‑87 MG, U‑87 MG EGFR vIII, LN-229, U-118 and DK-MG. The best expansion protocols were selected from ultimately 39 different conditions: three magnetic cell‑selection steps (Depletion of CD3+ cells, enrichment of CD56+ cells, and depletion of CD3+ cells followed by enrichment of CD56+ cells); four activation protocols (continuous, pre-activation, re-activation, and boost); and four cytokine combinations (IL-2/15, IL‑21/15, IL‑27/18/15 and IL-12/18/15).ResultsThe expansion rates varied between 2-50-fold, depending on the donor and the expansion conditions. The best expansion rate and purity were gained with sequential selection (Depletion of CD3+ cells and enrichment of CD56+ cells) from the starting material and pre-activation with IL‑12/18/15 cytokines, which are known to produce cytokine-induced memory-like NK cells. The cytotoxicity of these memory-like NK cells was enhanced with re-activation, diminishing the donor variation. The most cytotoxic NK cells were produced when cells were boosted at the end of the expansion with IL-12/18/15 or IL-21/15.ConclusionAccording to our findings the ex vivo proliferation capacity and functionality of NK cells is affected by multiple factors, such as the donor, composition of starting material, cytokine combination and the activation protocol. The cytokines modified the NK cells' phenotype and functionality, which was evident in their reactivity against the glioma cell lines. To our knowledge, this is the first comprehensive comparative study performed to this extent, and these findings could be used for upscaling clinical NK cell manufacturing

Use of green solvents as pre-treatment of dissolving pulp to decrease CS2 consumption from viscose production

Choline chloride-based deep eutectic solvents are widely used in biomass processing. In this work, four different green solvent mixtures were used as pre-treatment of acid sulphite dissolving pulp with the hypothesis of increasing the possibilities to produce viscose fibres and decreasing the use of the harmful and toxic carbon disulphide in the process. The experiments were performed at two different pulp to solvent mass ratios. Pulp quality parameters were also measured to determine the suitability of the pretreatment: a-cellulose, viscosity, lignin and pentosan content. In addition, X-ray diffraction analysis of pulps at the best solid to liquid ratio was performed to obtain the influence of the crystallinity index. Best results were obtained with the use of lactic acid, with reactivity values close to 94%, giving a reduction of CS2 usage of 15.83%. Furthermore, a linear relationship between the crystallinity index calculated by the XRD and reactivity with a regression factor of 0.87 was found

The role of factors promoting genetic diversity within social insect colonies

Abstract The evolution of sociality is often associated with close relatedness and genetic similarity of interacting individuals. However, colonies of advanced social insects (e.g. ants, bees and wasps) characterized by large colony size and division of tasks, are also shaped by acquisition of genetic diversity by polyandry, polygyny, recombination and even by hybridization. The balance between forces selecting for high relatedness on one hand and for improved colony performance though increased genetic diversity on the other hand forms an intriguing area of research. My study has produced the first genetic linkage maps for ants (Acromyrmex echinatior and Pogonomyrmex rugosus) and social wasps (Vespula vulgaris). Together with the findings of earlier honeybee research, it is shown that advanced eusocial insects have higher recombination rates than any other insect (or animal) studied so far. The estimates obtained here were 14 cM/Mb for P. rugosus, 9.7 cM/Mb for V. vulgaris and 6.2 cM/Mb for A. echinatior. Pogonomyrmex harvester ants have a genetic caste determination system in which workers arise from mating between two hybridizing lineages whereas sexuals are produced by within-lineage mating. I evaluated the origin of the lineages and the caste determination system by using 751 variable nuclear genetic markers. Fertile hybrids would lead to introgression, particularly in genomic regions characterized by a high recombination rate and lack of strongly selected loci. The hybridizing lineages (lineage pairs J1/J2 and H1/H2) showed many fixed differences. Nineteen of them were in the constructed linkage map, scattered in different linkage groups. The results suggest that there has been no recent introgression. As the hybrids are viable (as workers), caste differentiation can be affected by many loci scattered throughout the ant genome or by a small number of very strongly selected loci. Genetic diversity in colonies of the ant Formica cinerea is affected by varying levels of polygyny. I tested the hypotheses that the prevalence of endosymbiotic bacteria can vary in polygynous colonies but be either very low or very high in monogynous colonies. However, I found no association between the level of polygyny and endosymbiont prevalence. In addition to Wolbachia, I found two other endosymbiotic bacteria Cardinium and Candidatus Serratia symbiotica which have not been earlier reported from ants. Genetic diversity in insect colonies is affected by polyandry and polygyny. My results indicate that high a recombination rate is also an important factor influencing diversity. Genotypically diverse progenies can enhance colony success, e.g. through effects on division of labour or defence against pathogens. Recombination differs from the other factors in its effects on genetic relatedness among colony members

Cationization of lignocellulosic fibers with betaine in deep eutectic solvent:facile route to charge stabilized cellulose and wood nanofibers

Abstract In this study, a deep eutectic solvent (DES [based on triethylmethylammonium chloride (TEMA) and imidazole]) was used as a reaction medium for cationization of cellulose fibers with trimethylglycine (betaine) hydrochloride in the presence of p-Toluenesulfonyl (tosyl) chloride. Cellulose betaine ester with a cationic charge up to 1.95 mmol/g was obtained at mild reaction conditions (four hours at 80 °C). The reaction was further demonstrated in the fabrication of cationic cellulose nanofibers (CCNFs) by a mild mechanical disintegration of cationized cellulose. In addition to CCNFs, cationic wood nanofibers (CWNFs) were produced directly from groundwood pulp (GWP) with a high lignin content (27 w%). Individualized CCNFs and CWNFs had a fiber diameter of 4.7 ± 2.0 and 3.6 ± 1.3 nm, respectively, whereas some larger fiber aggregates (diameter below 200 nm) were also observed, especially in the case of CWNFs