Secondary Metabolism Rearrangements in Linum usitatissimum L. after Biostimulation of Roots with COS Oligosaccharides from Fungal Cell Wall

In vitro culture of flax (Linum usitatissimum L.) was exposed to chitosan oligosaccharides (COS) in order to investigate the effects on the growth and secondary metabolites content in roots and shoots. COS are fragments of chitosan released from the fungal cell wall during plant–pathogen interactions. They can be perceived by the plant as pathogen-associated signals, mediating local and systemic innate immune responses. In the present study, we report a novel COS oligosaccharide fraction with a degree of polymerization (DP) range of 2–10, which was produced from fungal chitosan by a thermal degradation method and purified by an alcohol-precipitation process. COS was dissolved in hydroponic medium at two different concentrations (250 and 500 mg/L) and applied to the roots of growing flax seedlings. Our observations indicated that the growth of roots and shoots decreased markedly in COS-treated flax seedlings compared to the control. In addition, the results of a metabolomics analysis showed that COS treatment induced the accumulation of (neo)lignans locally at roots, flavones luteolin C-glycosides, and chlorogenic acid in systemic responses in the shoots of flax seedlings. These phenolic compounds have been previously reported to exhibit a strong antioxidant and antimicrobial activities. COS oligosaccharides, under the conditions applied in this study (high dose treatment with a much longer exposure time), can be used to indirectly trigger metabolic response modifications in planta, especially secondary metabolism, because during fungal pathogen attack, COS oligosaccharides are among the signals exchanged between the pathogen and host plant.     

Near-infrared photosensitizer based on a cycloimide derivative of chlorin p6: 13,15-N-(3'-hydroxypropyl)cycloimide chlorin p6

The 13,15-N-(3'-hydroxypropylcycloimide) chlorin p6 (CIC), which absorbs at 711 nm, possesses considerable photoinduced cell-killing activity. It is 43-, 61- and 110-fold more active than chlorin p6, 3-formyl-3-devinyl chlorin p6 and Photogem, respectively, and has no cytotoxicity without irradiation as estimated on A549 human adenocarcinoma cells. To attain the highest intracellular penetration and activity the monomeric form of CIC should be stabilized. This stabilization in an aqueous environment can be achieved using 0.002-0.005% of Cremophor EL emulsion (polyoxyethylene derivative of hydrogenated castor oil). The intracellular accumulation of CIC occurs in cytoplasm in a monomeric form bound to cellular membranes. This form of the dye is characterized by a high quantum yield of singlet oxygen generation (0.66 +/- 0.02). Besides diffuse staining of intracellular membranous structures, CIC accumulates 3- to 4-fold more intensely in mitochondria and Golgi apparatus, thus indicating these organelles to be the initial targets of its photodynamic action. The incubation time providing 50% accumulation level of CIC in cells is 30 +/- 5 min. The time for 50% release of CIC from the cells is 60 +/- 10 min. A 10-fold decrease in CIC intracellular penetration at 22 degrees C proves that temperature-sensitive mechanisms of transport, rather than diffusion, are responsible for the dye uptake. The average cytoplasmic concentration of CIC was seven times the extracellular concentration in the 0.2-1.6 microM range, used for the photodynamic activity measurements. The concentration of CIC and the light dose that correspond to ca 50% level of phototoxicity induce predominantly an apoptotic-type of cell death, whereas the conditions providing 100% level of phototoxicity induced necrosis. The results obtained indicate that cycloimide derivatives of chlorin p6 may serve as a base for the development of an efficient near-IR photosensitizer.     

Chelation with metal is not essential for antitumor photodynamic activity of sulfonated phthalocyanines

It is generally assumed that a central metal is essential for the efficiency of phthalocyanines in photodynamic therapy (PDT) of cancer. Contrary to the set opinion, the results of the present study indicate that the metal-free sulfonated phthalocyanines (H2PcSn, where n is the number of sulfonate groups per molecule) possess a considerable photoactivity. The relative phototoxicities of H2PcS1.5, H2PcS2.4, H2PcS3.1 and H2PcS3.8 on HEp2 human epidermoid carcinoma cells were 3.3, 20, 3.3 and 1, respectively, thus demonstrating dependence of the activity on the sulfonation degree, known for metallo-PcSn. A significant delay in tumor growth and a decrease in tumor regrowth rate were observed in mice after PDT with H2PcS2.4. The antitumor effect declined in the order H2PcS2.4 > H2PcS3.1 > H2PcS1.5 and vanished for H2PcS3.8. We demonstrate here that the high photodynamic activity of H2PcS2.4 can be explained by its physicochemical properties in living cells and tissues. Thus, H2PcSn (n is about 2) can be considered as a new alternative in PDT of light-accessible neoplasms and further clinic-oriented studies are warranted.     

Synthesis and characterization of carboxymethylcelluloses from non-wood pulps II. Rheological behavior of CMC in aqueous solution

This paper concerns the rheological behavior of carboxymethylcelulloses(CMC) derived after one and two successive steps from different non-woodbleached cellulose pulps. CMC rheological characterization was achieved in0.1M NaCl solution, as a function of polymer concentration. Theevidence of a critical concentration (C* < 1 g/L) is discussedfromsteady shear and dynamic experiments. Rheological properties of the CMC werefound to depend on the cellulose source reactivity and on their degree ofsubstitution (DS). Higher molecular weight of initial cellulose was accompaniedby higher apparent intrinsic viscosity of the CMC produced. Depending on theCMCconcentration and on the degree of etherification, the system behaves as asolution or as a gel. In the case of abaca CMC sample, it is shown that afteronly one step of chemical modification and above a polymer concentration of20 g/L, the system behaves as a gel. The gel behavior was studied asafunction of temperature. In the temperature range from 25 to 45°C, the rheological behavior was found to remain almostconstant due to the existence of dispersed swollen aggregates. This unusualcharacteristic represents an advantage for applications such as oil recovery inthe petroleum industry, where viscosity of the recovered fluid should not diminishwith temperature.     

The use of sulfamic acid as a primary standard in nonaqueous titrimetry

A method has been developed for the standardization of a typical nonaqueous base lithium methoxide, with an inorganic primary standard, sulfamic acid. 'T'his acid was found to dissolve readily in four independant basic solvents, dimethyl formamide, ethylenediamine, n-butylamine and dimethyl sulfoxide. Sulfamic acid could be titrated in each of these solvents, by both visual and potentiometric means with lithium methoxide dissolved in benzene-methanol. No gels nor precipitates resulted in the course of the titrations and the accuracy of the method was comparable to that obtained using benzoic acid as the primary standard. Conductometric titrations were also performed using sulfamic acid dissolved in two systems, dimethyl formamide and glacial acetic acid. In the latter solvent it was possible to titrate sulfamic acid with perchloric acid in glacial acetic acid.     

Enhanced reactivity of an α-nucleophile in water-dimethyl sulfoxide mixtures. A transition state effect.

The reactivity of the anion of α,α,α-trifluoroacetophenone oxime towards p-nitrophenylacetate is considerably enhanced relative to that of the similarly basic p-chlorophenoxide anion on addition of Me₂ SO to aqueous solutions. This is shown to be due to an enhanced stabilization of the transition state for the oxime reaction.     

Molecular Weight Characterization and Structural Properties of Controlled Molecular Weight Dextrans Synthesized by Acceptor Reaction using Highly Purified Dextransucrase

ABSTRACT

Controlled molecular weight dextrans were synthesized using a highly purified dextransucrase from Leuconostoc mesenteroides NRRL B-512F in a multi-step process. Maltose was used as acceptor for the first reaction step. The purified product obtained at a given reaction step was used as acceptor for the next reaction step. Dextrans of molecular weights ranging from 1,000 to 100,000 were thus obtained with a good yield (80 %). After purification, the molecular weight distribution of the products was characterized using size exclusion chromatography coupled with low angle laser light scattering (LALLS). Polydispersity of the products was shown to be similar to that of commerical dextrans.

¹³C NMR spectra and enzymatic hydrolysis data are consistent with the fact that the enzymatically synthesized dextrans are essentially composed of α(1->6) linkages. It was observed that controlled molecular weight dextrans were less branched than commercial products obtained by acidic hydrolysis of high molecular weight dextrans.     

High quality dispersions of hexabenzocoronene in organic solvents

We have studied the exfoliation and dispersion of hexabenzocoronene (HBC) in 28 different solvents. We see a wide range of dispersed concentrations and aggregation states, all of which can be related to the solvent properties. To a first approximation, the dispersed concentration is maximized for solvents with Hildebrand solubility parameter close to 21 MPa(1/2), similar to graphitic materials such as nanotubes and graphene. We have also studied the concentration dependence of the absorbance and photoluminescence of HBC for both a good solvent, cyclohexyl pyrrolidone (CHP), and a poor solvent, tetrahydrofuran (THF). In both cases, we observe features that can be associated with either individual molecules or aggregates, allowing us to establish metrics both for aggregate and individual molecule content. While the aggregate content always increases with concentration, good solvents disperse individual molecules at relatively high concentrations while poor solvents display aggregation even at low concentrations. Using these metrics, we determine that large populations of individual molecules are present at low concentrations in certain solvents with Hildebrand solubility parameters close to 21 MPa(1/2). However, the aggregation state of HBC is considerably more sensitive to solvent Hildebrand parameter for halogenated solvents than for amide solvents. We find a combination of high overall concentrations and large populations of individual molecules in four solvents: cyclohexyl pyrrolidone, 1-chloronaphthalene, 1-bromonaphthalene, and 1,2,4-trichlorobenzene. Scanning tunnelling microscopy (STM) measurements show the formation of self-assembled monolayers at the interface between a HBC-solvent dispersion and a highly oriented pyrolytic graphite (HOPG) substrate. Similar structures were observed on ultrathin supports by aberration-corrected transmission electron microscopy (TEM). Also observed were graphitic objects of size ~1 nm consistent with monomers or aggregated stacks of very few monomers. We believe this is strong evidence of the presence of individual molecules in dispersions prepared with appropriate solvents.