Carboxymethyl chitosan-N-alkylimine derivatives: Synthesis, characterization and use for preservation of symbiotic biofertilizer bacteria on chickpea seeds.

A series of carboxymethyl chitosan-N-alkylimine derivatives with side chain length of 4 to 10 carbons (CMCS-n, n = 4, 6, 8, 10) was prepared in a one-step solvent-free synthesis using Schiff base chemistry. The modified polysaccharides were characterized by their spectral, thermal and physical properties. The prepared polymers demonstrated an ability to spontaneous self-assembly with a clear correlation between critical aggregation concentration and the chain length of the alkyl substituent. N-alkylimine-CMCS derivatives were found to deliver hydrophobic (curcumin) and hydrophilic (ascorbic acid) active agents in unfavorable environments of water and oil, respectively. Then, N-alkylimine-CMCS derivatives were used as a platform for the delivery of symbiotic gram-positive bacteria Bacillus subtilis CJ onto chickpea seeds. These bacteria demonstrated a significantly higher survival rate (106 CFU/mL) in dried CMCS-6 derivative film than in other films tested. The seeds treated with N-alkylimine-CMCS coatings that contained B. subtilis CJ demonstrated up to 100-fold increase of this bacterial population on the seedlings in comparison to the pristine CMCS.

Carboxymethyl chitosan-quercetin conjugate: A sustainable one-step synthesis and use for food preservation.

Bioactive polysaccharide, carboxymethyl chitosan-quercetin (CMCS-q) was prepared by a one-step reaction utilizing Schiff base chemistry. Notably, the presented conjugation method involves neither radical reactions nor auxiliary coupling agents. Physicochemical properties and bioactivity of the modified polymer were studied and compared to those of the pristine carboxymethyl chitosan, CMCS. The modified CMCS-q demonstrated antioxidant activity by TEAC assay and antifungal activity by inhibiting spore germination of plant pathogen Botrytis cynerea. Then, CMCS-q was applied as an active coating on fresh-cut apples. The treatment resulted in enhanced firmness, inhibited browning and improved microbiological quality of the food product. The presented conjugation method allows retaining antimicrobial and antioxidant activity of quercetin moiety in the modified biopolymer. This method can be further used as a platform for binding ketone/aldehyde-containing polyphenols and other natural compounds to form various bioactive polymers.

A Novel Platform for Root Protection Applies New Root-Coating Technologies to Mitigate Soil-Borne Tomato Brown Rugose Fruit Virus Disease.

Tomato brown rugose fruit virus (ToBRFV) is a soil-borne virus showing a low percentage of ca. 3% soil-mediated infection when the soil contains root debris from a previous 30-50 day growth cycle of ToBRFV-infected tomato plants. We designed stringent conditions of soil-mediated ToBRFV infection by increasing the length of the pre-growth cycle to 90-120 days, adding a ToBRFV inoculum as well as truncating seedling roots, which increased seedling susceptibility to ToBRFV infection. These rigorous conditions were employed to challenge the efficiency of four innovative root-coating technologies in mitigating soil-mediated ToBRFV infection while avoiding any phytotoxic effect. We tested four different formulations, which were prepared with or without the addition of various virus disinfectants. We found that under conditions of 100% soil-mediated ToBRFV infection of uncoated positive control plants, root-coating with formulations based on methylcellulose (MC), polyvinyl alcohol (PVA), silica Pickering emulsion and super-absorbent polymer (SAP) that were prepared with the disinfectant chlorinated-trisodium phosphate (Cl-TSP) showed low percentages of soil-mediated ToBRFV infection of 0%, 4.3%, 5.5% and 0%, respectively. These formulations had no adverse effect on plant growth parameters when compared to negative control plants grown under non ToBRFV inoculation conditions.

The polyaminosaccharide-based buffers as a new type of zwitterionic buffering macromolecules for biochemical applications.

A new type of biocompatible buffers based on zwitterionic polyaminosaccharides is reported. The carboxy- and amino-groups containing carboxymethyl chitosan (CM-CS) was synthesized and reacted with hydrochloric/acetic acid resulting in CM-CS-HCl and CM-CS-HAc buffers with buffering capacity of 20.6 and 15.2 mM/pH. The new buffers were comprehensively characterized for their physicochemical properties and checked on enzymatic reactions of acetylcholinesterase (AChE) and alkaline phosphatase (ALP). Their performance was compared to the phosphate and Tris buffers. The chloride-free, CM-CS-HAc demonstrated excellent buffering activity with Michaelis constants of 0.50 and 1.00 mM and maximum reaction rates of 5.62 and 2.26 µmol/min/mL for AChE and ALP reactions, respectively. Toxicity studies on stress-sensitive bioreporter bacteria verified nontoxicity of CM-CS-HAc. Zwitterionic polyaminosaccharides overcome drawbacks of monomeric buffers, such as interference with enzyme active sites, cell membrane injury and purification difficulties. Therefore, they may become the next generation of effective buffers for biological and biochemical applications.

Highly Selective and Potent Ectonucleotide Pyrophosphatase-1 (NPP1) Inhibitors Based on Uridine 5'-Pα,α-Dithiophosphate Analogues.

Ectonucleotide pyrophosphatase/phosphodiesterase-1 (NPP1) hydrolyzes phosphodiester bonds of nucleotides such as ATP, resulting mainly in the formation of AMP and pyrophosphate. NPP1 activity plays a deleterious function in calcified aortic valve disease and calcium pyrophosphate deposition disease. Thus, inhibitors of NPP1 represent a medical need. We developed novel NPP1 inhibitors based on uridine 5'-Pα,α-dithiophosphate analogues, 9-12. All these analogues potently inhibited hNPP1 (80-100% inhibition) at 100 µM, with no, or minimal, inhibition of NPP3 and other ectonucleotidases (NTPDase1,2,3,8). These compounds showed nearly no activity at uracil-nucleotide sensitive P2Y2,4,6-receptors and thus represent highly selective NPP1 inhibitors. The most promising inhibitor was diuridine 5'-Pα,α,5″-Pα,α-tetrathiotetraphosphate, 12, exhibiting Ki of 27 nM. Analogues 9-12 proved to be highly stable to air oxidation and to acidic and basic pH. Docking simulations suggested that the enhanced NPP1 inhibitory activity and selectivity of analogue 12 could be attributed to the simultaneous occupancy of two sites (the AMP site and an alternative site) of NPP1 by this compound.