scholarly journals Delivery of DNA into Human Cells by Functionalized Lignin Nanoparticles

Materials ◽  
2022 ◽  
Vol 15 (1) ◽  
pp. 303
Author(s):  
Michael K. Riley ◽  
Wilfred Vermerris
Keyword(s):  
Light Scattering ◽  
Zeta Potential ◽  
Plant Cell Wall ◽  
Fluorescent Protein ◽  
Plant Biomass ◽  
Paper Industry ◽  
Chemical Properties ◽  
Dna Encoding ◽  
Mobility Shift ◽  
The Impact

Lignin is an aromatic plant cell wall polymer that is generated in large quantities as a low-value by-product by the pulp and paper industry and by biorefineries that produce renewable fuels and chemicals from plant biomass. Lignin structure varies among plant species and as a function of the method used for its extraction from plant biomass. We first explored the impact of this variation on the physico-chemical properties of lignin nanoparticles (LNPs) produced via a solvent exchange procedure and then examined whether LNPs produced from industrial sources of lignin could be used as delivery vehicles for DNA. Spherical LNPs were formed from birch and wheat BioLignin™ and from poplar thioglycolic acid lignin after dissolving the lignin in tetrahydrofuran (THF) and dialyzing it against water. Dynamic light scattering indicated that the diameter of these LNPs was dependent on the initial concentration of the lignin, while electrophoretic light scattering indicated that the LNPs had a negative zeta potential, which became less negative as the diameter increased. The dynamics of LNP formation as a function of the initial lignin concentration varied as a function of the source of the lignin, as did the absolute value of the zeta potential. After coating the LNPs with cationic poly-l-lysine, an electrophoretic mobility shift assay indicated that DNA could adsorb to LNPs. Upon transfection of human A549 lung carcinoma basal epithelial cells with functionalized LNPs carrying plasmid DNA encoding the enhanced green fluorescent protein (eGFP), green foci were observed under the microscope, and the presence of eGFP in the transfected cells was confirmed by ELISA. The low cytotoxicity of these LNPs and the ability to tailor diameter and zeta potential make these LNPs of interest for future gene therapy applications.

scholarly journals Purity and structural composition of lignin isolated from Miscanthus x giganteus by sub-critical water extraction with associated modifiers

2020 ◽  
Vol 1 (1) ◽  
pp. 1-12 ◽  
Keyword(s):  
Paper Industry ◽  
Chemical Properties ◽  
Value Added ◽  
Hydroxyl Groups ◽  
Physical And Chemical Properties ◽  
Miscanthus X Giganteus ◽  
Lignin Recovery ◽  
Physical And Chemical ◽  
The Impact ◽  
And Chemical Properties

Lignin is an industrial by-product produced from the pulping and paper industry, where the process generates lignin in the form of lignosulphonates. While there are many applications for lignin, there are all low value and attempts to add value to lignin are hindered by its complex physicochemical nature and the presence of sulphur. Adopting the biorefining concept, the study evaluates the impact of direct (DE) and sequential extraction (SE) of Miscanthus x giganteus using sub-critical water with associated modifiers; ethanol and carbon dioxide on the physical and chemical properties of the extracted lignin. Isolated lignins were characterised by a Fourier Transform Infrared Spectroscopy (FTIR). Although higher delignification was achieved by DE and SE about 81.5% and 58.0%, respectively, the lignin recovered from the SE process showed remarkably higher purity with 91.5%. Lignin recovery did not differ considerably for either processing method. FTIR revealed a qualitative reduction in the intensity of bonds corresponding to hydroxyl groups for the lignin derived from DE rather than SE processing routes. These indicated that the lignin derived from SE had potential for subsequent preparation in lignin value-added bio-based materials.

scholarly journals Characterisation of the Effect of the Spatial Organisation of Hemicellulases on the Hydrolysis of Plant Biomass Polymer

2020 ◽  
Vol 21 (12) ◽  
pp. 4360
Author(s):  
Thomas Enjalbert ◽  
Marion De La Mare ◽  
Pierre Roblin ◽  
Louise Badruna ◽  
Thierry Vernet ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Plant Biomass ◽  
Reducing Sugar ◽  
Intrinsic Activity ◽  
X Rays ◽  
Spatial Organisation ◽  
The Impact ◽  
Hydrolysis Of

Synergism between enzymes is of crucial importance in cell metabolism. This synergism occurs often through a spatial organisation favouring proximity and substrate channelling. In this context, we developed a strategy for evaluating the impact of the geometry between two enzymes involved in nature in the recycling of the carbon derived from plant cell wall polymers. By using an innovative covalent association process using two protein fragments, Jo and In, we produced two bi-modular chimeric complexes connecting a xylanase and a xylosidase, involved in the deconstruction of xylose-based plant cell wall polymer. We first show that the intrinsic activity of the individual enzymes was preserved. Small Angle X-rays Scattering (SAXS) analysis of the complexes highlighted two different spatial organisations in solution, affecting both the distance between the enzymes (53 Å and 28 Å) and the distance between the catalytic pockets (94 Å and 75 Å). Reducing sugar and HPAEC-PAD analysis revealed different behaviour regarding the hydrolysis of Beechwood xylan. After 24 h of hydrolysis, one complex was able to release a higher amount of reducing sugar compare to the free enzymes (i.e., 15,640 and 14,549 µM of equivalent xylose, respectively). However, more interestingly, the two complexes were able to release variable percentages of xylooligosaccharides compared to the free enzymes. The structure of the complexes revealed some putative steric hindrance, which impacted both enzymatic efficiency and the product profile. This report shows that controlling the spatial geometry between two enzymes would help to better investigate synergism effect within complex multi-enzymatic machinery and control the final product.

scholarly journals The Impact of Filler Geometry on Polylactic Acid-Based Sustainable Polymer Composites

Molecules ◽  
2020 ◽  
Vol 26 (1) ◽  
pp. 149
Author(s):  
Karol Leluk ◽  
Stanisław Frąckowiak ◽  
Joanna Ludwiczak ◽  
Tomasz Rydzkowski ◽  
Vijay Kumar Thakur
Keyword(s):  
Polymer Composites ◽  
Polylactic Acid ◽  
Matrix Material ◽  
Chemical Properties ◽  
Precipitated Calcium Carbonate ◽  
Cellulose Fibres ◽  
Shape Factors ◽  
Pure Cellulose ◽  
Physico Chemical ◽  
The Impact

Recently, biocomposites have emerged as materials of great interest to the scientists and industry around the globe. Among various polymers, polylactic acid (PLA) is a popular matrix material with high potential for advanced applications. Various particulate materials and nanoparticles have been used as the filler in PLA based matrix. One of the extensively studied filler is cellulose. However, cellulose fibres, due to their hydrophilic nature, are difficult to blend with a hydrophobic polymer matrix. This leads to agglomeration and creates voids, reducing the mechanical strength of the resulting composite. Moreover, the role of the various forms of pure cellulose and its particle shape factors has not been analyzed in most of the current literature. Therefore, in this work, materials of various shapes and shape factors were selected as fillers for the production of polymer composites using Polylactic acid as a matrix to fill this knowledge gap. In particular, pure cellulose fibres (three types with different elongation coefficient) and two mineral nanocomponents: precipitated calcium carbonate and montmorillonite were used. The composites were prepared by a melt blending process using two different levels of fillers: 5% and 30%. Then, the analysis of their thermomechanical and physico-chemical properties was carried out. The obtained results were presented graphically and discussed in terms of their shape and degree of filling.

scholarly journals Silver Nanoparticles Stabilized with Phosphorus-Containing Heterocyclic Surfactants: Synthesis, Physico-Chemical Properties, and Biological Activity Determination

Nanomaterials ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1883
Author(s):  
Martin Pisárčik ◽  
Miloš Lukáč ◽  
Josef Jampílek ◽  
František Bilka ◽  
Andrea Bilková ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Biological Activity ◽  
Zeta Potential ◽  
Chain Length ◽  
Alkyl Chain ◽  
Chemical Properties ◽  
Alkyl Chain Length ◽  
Microbial Pathogens ◽  
Physico Chemical ◽  
Phosphorus Containing

Phosphorus-containing heterocyclic cationic surfactants alkyldimethylphenylphospholium bromides with the alkyl chain length 14 to 18 carbon atoms were used for the stabilization of silver nanodispersions. Zeta potential of silver nanodispersions ranges from +35 to +70 mV, which indicates the formation of stable silver nanoparticles (AgNPs). Long-chain heptadecyl and octadecyl homologs of the surfactants series provided the most intensive stabilizing effect to AgNPs, resulting in high positive zeta potential values and smaller diameter of AgNPs in the range 50–60 nm. A comparison with non-heterocyclic alkyltrimethylphosphonium surfactants of the same alkyl chain length showed better stability and more positive zeta potential values for silver nanodispersions stabilized with heterocyclic phospholium surfactants. Investigations of biological activity of phospholium-capped AgNPs are represented by the studies of antimicrobial activity and cytotoxicity. While cytotoxicity results revealed an increased level of HepG2 cell growth inhibition as compared with the cytotoxicity level of silver-free surfactant solutions, no enhanced antimicrobial action of phospholium-capped AgNPs against microbial pathogens was observed. The comparison of cytotoxicity of AgNPs stabilized with various non-heterocyclic ammonium and phosphonium surfactants shows that AgNPs capped with heterocyclic alkyldimethylphenylphospholium and non-heterocyclic triphenyl-substituted phosphonium surfactants have the highest cytotoxicity among silver nanodispersions stabilized by the series of ammonium and phosphonium surfactants.

scholarly journals Dye Separation and Antibacterial Activities of Polyaniline Thin Film-Coated Poly(phenyl sulfone) Membranes

Membranes ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 25
Author(s):  
Javed Alam ◽  
Arun Kumar Shukla ◽  
Mohammad Azam Ansari ◽  
Fekri Abdulraqeb Ahmed Ali ◽  
Mansour Alhoshan
Keyword(s):  
Zeta Potential ◽  
Surface Properties ◽  
Antibacterial Activities ◽  
Pani Film ◽  
Force Microscopy ◽  
Water Permeation ◽  
Membrane Pores ◽  
Membrane Performance ◽  
Phenyl Sulfone ◽  
The Impact

We fabricated a nanofiltration membrane consisting of a polyaniline (PANI) film on a polyphenylsulfone (PPSU) substrate membrane. The PANI film acted as a potent separation enhancer and antimicrobial coating. The membrane was analyzed via scanning electron microscopy and atomic force microscopy to examine its morphology, topography, contact angle, and zeta potential. We aimed to investigate the impact of the PANI film on the surface properties of the membrane. Membrane performance was then evaluated in terms of water permeation and rejection of methylene blue (MB), an organic dye. Coating the PPSU membrane with a PANI film imparted significant advantages, including finely tuned nanometer-scale membrane pores and tailored surface properties, including increased hydrophilicity and zeta potential. The PANI film also significantly enhanced separation of the MB dye. The PANI-coated membrane rejected over 90% of MB with little compromise in membrane permeability. The PANI film also enhanced the antimicrobial activity of the membrane. The bacteriostasis (BR) values of PANI-coated PPSU membranes after six and sixteen hours of incubation with Escherichia coli were 63.5% and 95.2%, respectively. The BR values of PANI-coated PPSU membranes after six and sixteen hours of incubation with Staphylococcus aureus were 70.6% and 88.0%, respectively.

scholarly journals Functionalization of Cellulose-Based Hydrogels with Bi-Functional Fusion Proteins Containing Carbohydrate-Binding Modules

Materials ◽  
2021 ◽  
Vol 14 (12) ◽  
pp. 3175
Author(s):  
Mariana Barbosa ◽  
Hélvio Simões ◽  
Duarte Miguel F. Prazeres
Keyword(s):  
Fusion Proteins ◽  
Fluorescent Protein ◽  
Protein A ◽  
Chemical Properties ◽  
Main Idea ◽  
Bioactive Molecules ◽  
Scaffolding Protein ◽  
Carbohydrate Binding ◽  
Biological Interactions ◽  
Carbohydrate Binding Modules

Materials with novel and enhanced functionalities can be obtained by modifying cellulose with a range of biomolecules. This functionalization can deliver tailored cellulose-based materials with enhanced physical and chemical properties and control of biological interactions that match specific applications. One of the foundations for the success of such biomaterials is to efficiently control the capacity to combine relevant biomolecules into cellulose materials in such a way that the desired functionality is attained. In this context, our main goal was to develop bi-functional biomolecular constructs for the precise modification of cellulose hydrogels with bioactive molecules of interest. The main idea was to use biomolecular engineering techniques to generate and purify different recombinant fusions of carbohydrate binding modules (CBMs) with significant biological entities. Specifically, CBM-based fusions were designed to enable the bridging of proteins or oligonucleotides with cellulose hydrogels. The work focused on constructs that combine a family 3 CBM derived from the cellulosomal-scaffolding protein A from Clostridium thermocellum (CBM3) with the following: (i) an N-terminal green fluorescent protein (GFP) domain (GFP-CBM3); (ii) a double Z domain that recognizes IgG antibodies; and (iii) a C-terminal cysteine (CBM3C). The ability of the CBM fusions to bind and/or anchor their counterparts onto the surface of cellulose hydrogels was evaluated with pull-down assays. Capture of GFP-CBM3 by cellulose was first demonstrated qualitatively by fluorescence microscopy. The binding of the fusion proteins, the capture of antibodies (by ZZ-CBM3), and the grafting of an oligonucleotide (to CBM3C) were successfully demonstrated. The bioactive cellulose platform described here enables the precise anchoring of different biomolecules onto cellulose hydrogels and could contribute significatively to the development of advanced medical diagnostic sensors or specialized biomaterials, among others.

scholarly journals Assessment of Pellets from Three Forest Species: From Raw Material to End Use

Forests ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 447
Author(s):  
Miguel Alfonso Quiñones-Reveles ◽  
Víctor Manuel Ruiz-García ◽  
Sarai Ramos-Vargas ◽  
Benedicto Vargas-Larreta ◽  
Omar Masera-Cerutti ◽  
...  
Keyword(s):  
Chemical Properties ◽  
Ash Content ◽  
Hot Water ◽  
Raw Material ◽  
Chemical Components ◽  
Energy Yield ◽  
Cellulose Content ◽  
Fixed Carbon ◽  
End Use ◽  
The Impact

This study aimed to evaluate and compare the relationship between chemical properties, energy efficiency, and emissions of wood and pellets from madroño Arbutus xalapensis Kunth, tázcate Juniperus deppeana Steud, and encino colorado Quercus sideroxyla Humb. & Bonpl. in two gasifiers (top-lit-up-draft (T-LUD) and electricity generation wood camp stove (EGWCS)) in order to determine the reduction of footprint carbon. In accordance with conventional methodologies, we determined the extracts and chemical components (lignin, cellulose, holocellulose), and the immediate analyses were carried out (volatile materials, fixed carbon, ash content and microanalysis of said ash), as well as the evaluation of emission factors (total suspended particulate matter (PM2.5), CO, CO2, CH4, black carbon (BC), elemental carbon (EC), and organic carbon (OC)). The results were statistically analyzed to compare each variable among species and gasifiers. The raw material analyzed showed how the pH ranged from 5.01 to 5.57, and the ash content ranged between 0.39 and 0.53%. The content values of Cu, Zn, Fe, Mg, and Ca ranged from 0.08 to 0.22, 0.18 to 0.19, 0.38 to 0.84, 1.75 to 1.90, and 3.62 to 3.74 mg kg−1, respectively. The extractive ranges from cyclohexane were 2.48–4.79%, acetone 2.42–4.08%, methanol 3.17–7.99%, and hot water 2.12–4.83%. The range of lignin was 18.08–28.60%. The cellulose content ranged from 43.30 to 53.90%, and holocellulose from 53.50 to 64.02%. The volatile material range was 81.2–87.42%, while fixed carbon was 11.30–17.48%; the higher heating value (HHV) of raw material and pellets presented the ranges 17.68–20.21 and 19.72–21.81 MJ kg−1, respectively. Thermal efficiency showed statistically significant differences (p < 0.05) between pellets and gasifiers, with an average of 31% Tier 3 in ISO (International Organization for Standardization) for the T-LUD and 14% (ISO Tier 1) for EGWCS, with Arbutus xalapensis being the species with the highest energy yield. The use of improved combustion devices, as well as that of selected raw material species, can reduce the impact of global warming by up to 33% on a cooking task compared to the three-stone burner.

Can repeated soil amendment with biogas digestates increase soil suppressiveness toward non-specific soil-borne pathogens in agricultural lands?

2020 ◽  
pp. 1-12
Author(s):  
L. M. Manici ◽  
F. Caputo ◽  
G. A. Cappelli ◽  
E. Ceotto
Keyword(s):  
Plant Growth ◽  
Biomass Production ◽  
Soil Amendment ◽  
Plant Biomass ◽  
Amended Soils ◽  
Soil Suppressiveness ◽  
Soil Microbial ◽  
Soil Borne Pathogens ◽  
The Impact ◽  
Plant Biomass Production

Abstract Soil suppressiveness which is the natural ability of soil to support optimal plant growth and health is the resultant of multiple soil microbial components; which implies many difficulties when estimating this soil condition. Microbial benefits for plant health from repeated digestate applications were assessed in three experimental sites surrounding anaerobic biogas plants in an intensively cultivated area of northern Italy. A 2-yr trial was performed in 2017 and 2018 by performing an in-pot plant growth assay, using soil samples taken from two fields for each experimental site, of which one had been repeatedly amended with anaerobic biogas digestate and the other had not. These fields were similar in management and crop sequences (maize was the recurrent crop) for the last 10 yr. Plant growth response in the bioassay was expressed as plant biomass production, root colonization frequency by soil-borne fungi were estimated to evaluate the impact of soil-borne pathogens on plant growth, abundance of Pseudomonas and actinomycetes populations in rhizosphere were estimated as beneficial soil microbial indicators. Repeated soil amendment with digestate increased significantly soil capacity to support plant biomass production as compared to unamended control in both the years. Findings supported evidence that this increase was principally attributable to a higher natural ability of digestate-amended soils to reduce root infection by saprophytic soil-borne pathogens whose inoculum was increased by the recurrent maize cultivation. Pseudomonas and actinomycetes were always more abundant in digestate-amended soils suggesting that both these large bacterial groups were involved in the increase of their natural capacity to control soil-borne pathogens (soil suppressiveness).

scholarly journals Phosphorylation within Intrinsic Disordered Region Discriminates Histone Variant macroH2A1 Splicing Isoforms—macroH2A1.1 and macroH2A1.2

Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 659
Author(s):  
Sebastiano Giallongo ◽  
Oriana Lo Re ◽  
Gabriela Lochmanová ◽  
Luca Parca ◽  
Francesco Petrizzelli ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Regulatory Element ◽  
Histone Variants ◽  
Histone H2a ◽  
Human Hepatoma Cells ◽  
Nucleosome Core ◽  
Intrinsically Disordered ◽  
Splicing Isoforms ◽  
The Impact

Background: Gene expression in eukaryotic cells can be governed by histone variants, which replace replication-coupled histones, conferring unique chromatin properties. MacroH2A1 is a histone H2A variant containing a domain highly similar to H2A and a large non-histone (macro) domain. MacroH2A1, in turn, is present in two alternatively exon-spliced isoforms: macroH2A1.1 and macroH2A1.2, which regulate cell plasticity and proliferation in a remarkably distinct manner. The N-terminal and the C-terminal tails of H2A histones stem from the nucleosome core structure and can be target sites for several post-translational modifications (PTMs). MacroH2A1.1 and macroH2A1.2 isoforms differ only in a few amino acids and their ability to bind NAD-derived metabolites, a property allegedly conferring their different functions in vivo. Some of the modifications on the macroH2A1 variant have been identified, such as phosphorylation (T129, S138) and methylation (K18, K123, K239). However, no study to our knowledge has analyzed extensively, and in parallel, the PTM pattern of macroH2A1.1 and macroH2A1.2 in the same experimental setting, which could facilitate the understanding of their distinct biological functions in health and disease. Methods: We used a mass spectrometry-based approach to identify the sites for phosphorylation, acetylation, and methylation in green fluorescent protein (GFP)-tagged macroH2A1.1 and macroH2A1.2 expressed in human hepatoma cells. The impact of selected PTMs on macroH2A1.1 and macroH2A1.2 structure and function are demonstrated using computational analyses. Results: We identified K7 as a new acetylation site in both macroH2A1 isoforms. Quantitative comparison of histone marks between the two isoforms revealed significant differences in the levels of phosphorylated T129 and S170. Our computational analysis provided evidence that the phosphorylation status in the intrinsically disordered linker region in macroH2A1 isoforms might represent a key regulatory element contributing to their distinct biological responses. Conclusions: Taken together, our results report different PTMs on the two macroH2A1 splicing isoforms as responsible for their distinct features and distribution in the cell.

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