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FDA-approved bone morphogenetic protein 2 (BMP2) has serious side effects due to the super high dose requirement. Heparin is one of the most well-studied sulfated polymers to stabilize BMP2 and improve its functionality. However, the clinical use of heparin is questionable because of its undesired anticoagulant activity. Recent studies suggest that poly(glutamic acid) (pGlu) has the potential to improve BMP2 bioactivity with less safety concerns; however, the knowledge on pGlu's contribution remains largely unknown. Therefore, we aimed to study the role of pGlu in BMP2-induced osteogenesis and its potential application in bone tissue engineering. Our data, for the first time, indicated that both low (L-pGlu) and high molecular weight pGlu (H-pGlu) were able to significantly improve the BMP2-induced early osteoblastic differentiation marker (ALP) in MC3T3-E1 preosteoblasts. Importantly, the matrix mineralization was more rapidly enhanced by H-pGlu compared to L-pGlu. Additionally, our data indicated that only α-H-pGlu could significantly improve BMP2's activity, whereas γ-H-pGlu failed to do so. Moreover, both gene expression and mineralization data demonstrated that α-H-pGlu enabled a single dose of BMP2 to induce a high level of osteoblastic differentiation without multiple doses of BMP2. To study the potential application of pGlu in tissue engineering, we incorporated the H-pGlu+BMP2 nanocomplexes into the collagen hydrogel with significantly elevated osteoblastic differentiation. Furthermore, H-pGlu-coated 3D porous gelatin and chitosan scaffolds significantly enhanced osteogenic differentiation through enabling sustained release of BMP2. Thus, our findings suggest that H-pGlu is a promising new alternative with great potential for bone tissue engineering applications.

Control of polymer molecular weight is critical for tailoring structure-function properties; however, traditional molecular weight characterization techniques have limited ability to determine the molecular weight of polymers in real time without sample removal from the reaction mixture, with spatial resolution, and of insoluble polymers. In this work, a fluorescence lifetime imaging microscopy (FLIM) method was developed that overcomes these limitations. The method is demonstrated with polynorbornene and polydicyclopentadiene, polymers derived from ruthenium-catalyzed ring-opening metathesis polymerization (ROMP). The polymer

To investigate the effects of salivary esterases on biostability of collagen treated by galloylated polyphenols.Human dentin was microtomed into 6-μm-thick films, which were demineralized and treated for 60 s using solutions containing 0.6% and 2% of one of the crosslinkers: tannic acid (TAC), epigallocatechin gallate (EGCG), epigallocatechin (EGC), and N-[3-dimethylaminopropyl]-N'-ethylcarbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS), and for 1 h using EDC/NHS. Half of the treated and untreated (control) films were subjected to human saliva incubation. Collagen biostability was assessed via exogenous protease biodegradation by weight loss and hydroxyproline release, and endogenous MMPs by in situ zymography. The degradation products of galloylated polyphenols (TAC and EGCG) by saliva were monitored using proton nuclear magnetic resonance (Collagen films treated with TAC and EGCG exhibited significantly improved biostability (p 0.05); however, the enhanced biostability was severely reduced after saliva incubation (p 0.001). For EDC/NHS treated collagen, saliva incubation showed negligible effect on the biostability.The galloyl group plays important role in collagen crosslinking, inducing higher biostability. However, galloylated polyphenols crosslinked on collagen are highly susceptible to metabolism of human saliva by salivary esterase, dramatically compromising the enhanced biostability.

Lysozyme (LYS) and hyaluronan with low (HA1: 3 kDa), medium (HA2: 120 kDa), and high (HA3: 1200 kDa) molecular weights were used to fabricate lysozyme-hyaluronan colloidal nanoparticles using a green self-assembly method. Fourier transform infrared spectroscopy indicated that hydrogen bonding, hydrophobic and electrostatic interactions promoted the formation of the colloidal nanoparticles. The hydrophobic area of prepared colloidal nanoparticles was quantified using a pyrene fluorescent probe, and the results showed that the LYS-HA3 nanoparticles had the strongest hydrophobic capacity. Furthermore, 5-fluorouracil (5-Fu) was used to evaluate encapsulation performance, demonstrating that the LYS-HA3 nanoparticles had the highest encapsulation ability (90 %). All prepared 5-Fu-loaded lysozyme-hyaluronan (5-Fu@LYS-HA) colloidal nanoparticles exhibited excellent long-term storage stability at 4 °C for 60 days. Cellular uptake and in vitro release results indicated that the LYS-HA2 nanoparticles exhibited the highest cellular uptake efficiency, and the LYS-HA3 nanoparticles had the best slow-release effect, while the release process was mainly controlled by the combination of Fickian diffusion and structural relaxation, respectively. This study demonstrates the influence of molecular weight on the conformational and structural properties of colloidal nanoparticles, which has implications for the design of insoluble drug self-assembly systems.

The structural characteristics and immunoregulatory activities of neutral heteropolysaccharide (AVRP-N) separated from the roots of Apocynum venetum L. were extensively investigated. The results showed that the weight average molecular mass (Mw) of AVRP-N was 6.430 × 10

Homogeneous low molecular weight dextran can be used to improve microcirculation and expand blood volume. However, the synthesis and separation of low molecular weight dextran are chemically difficult and environmentally unfriendly. Here, a one-step strategy for the synthesis of homogeneous low molecular weight dextran was developed. Dextransucrase and dextranase were fused by the addition of different length linker peptides. An artificial bifunctional enzyme was created to directly convert sucrose into low molecular weight dextran (13,050 Da), and the related substrate channel mechanism was found. The substrate channel adaptability was studied by changing the length of the linker and its corresponding product behavior. Compared with the mixture of two free enzymes, the residence lag time demonstrates the degree of substrate channelization of a series of fusion enzymes. And found that the highest channelization degree is not equal to produce homogenous dextran. Whereas a fusion enzyme with the appropriate linker (the one with the best substrate channel adaptation) will produce dextran with a homogeneous molecular weight. By studying the temperature dynamics of the fusion enzyme to adjust the two-stage catalytic efficiency of the fusion enzyme, we have increased the yield of low molecular weight homogeneous dextran (Yield of 62 %).

Changes in the antioxidant protection system of Larix sibirica Ledeb at different pollution levels caused by emissions from a large aluminum smelter (BrAS) have been studied. We revealed that the content of peroxide (H

Acetaminophen is widely used to treat mild to moderate pain and to reduce fever. Under the worldwide COVID-19 pandemic, this over-the-counter pain reliever and fever reducer has been drastically consumed, which makes it even more abundant than ever in municipal wastewater and drinking water sources. Chlorine is the most widely used oxidant in drinking water disinfection, and chlorination generally causes the degradation of organic compounds, including acetaminophen. In this study, a new reaction pathway in the chlorination of acetaminophen, i.e., oxidative coupling reactions via acetaminophen radicals, was investigated both experimentally and computationally. Using an ultraperformance liquid chromatograph coupled to an electrospray ionization-triple quadrupole mass spectrometer, we detected over 20 polymeric products in chlorinated acetaminophen samples, some of which have structures similar to the legacy pollutants "polychlorinated biphenyls". Both C-C and C-O bonding products were found, and the corresponding bonding processes and kinetics were revealed by quantum chemical calculations. Based on the product confirmation and intrinsic reaction coordinate computations, a pathway for the formation of the polymeric products in the chlorination of acetaminophen was proposed. This study suggests that chlorination may cause not only degradation but also upgradation of a phenolic compound or contaminant.

We experimentally determined the characteristics and Langmuir parameters of the binding of calcium ions to different polycarboxylates. By using potentiometric titrations and isothermal titration calorimetry, the effects of side chain chemistry, pH value, and chain length were systematically investigated using the linear polymers poly(aspartic acid), poly(glutamic acid), and poly(acrylic acid). We demonstrate that for polymers with high polymerization degrees, the binding process is governed by higher-order effects, such as the change of apparent p

Structurally controlled high-molecular-weight (HMW) polystyrenes (PSts) and block copolymers consisting of HMW PSt segments were successfully synthesized by emulsion organotellurium-mediated radical polymerization (TERP). The hydrophilicity of the organotellurium group of TERP chain transfer agents (CTAs) was important for success, and CTAs

The transition toward a circular plastic economy and sustainable chemical industries requires the development and implementationofgreen methodsoffree radicalpolymerization.Inthis study, thehigh-speedradicalpolymerizationofN,N-diethylacrylamidewas achievedinhigh temperaturewater(150 degrees C, 0.5 MPa) without using potentially contaminating organic solvents. Precipitationpolymerizationwas completed within 2 min. The molecular weightofthe resulting polymer (27.7 x 103; dispersityof1.78) was comparable to thatofpolymersobtainedbytypicalpolymerizationat 70 degrees C and significantly exceeding thoseofpolymersobtainedbypolymerizationinhigh-temperatureorganic solvents. This implies that the main factor governing thepolymerizationrate is a kinetic effect due to hydrogen bonding between the carbonyl groups andwatermolecules; under these conditions, disproportionation termination was suppressed. Thepolymerizationprocess developed herein usinghigh-temperaturewateroffers the benefitsofa short reaction time and a low environmental impact, thus holding great promise for industrial applications., Article, Industrial & Engineering Chemistry Research 61(46) : 17012-17016(2022)

Sensitive quantification of methoxy poly(ethylene glycol) (mPEG)-conjugated therapeutics for pharmacokinetic determination is critical for mPEGylated drug development. However, sensitive measurement of low-molecular-weight (lmw) mPEG compounds remains challenging due to epitope competition between backbone-specific anti-PEG antibodies. Here, we engineered a high-affinity methoxy-specific anti-mPEG antibody for sensitive quantification of free mPEG molecules and mPEGylated therapeutics. The affinity-enhanced h15-2Y antibody variant shows a 10.3-fold increase in mPEG-binding activity compared to parental h15-2b. h15-2Y-based sandwich ELISA can effectively quantify lmw mPEG

Although exopolysaccharide (EPS) has been applied to various fields, EPS for UVR-mediated oxidative stress repair still needs further exploration. In this study, a novel EPS was isolated from the fermentation medium of Bacillus sp. QDR3-1 and its yield was 4.8 g/L (pH 8.0, 12 % glucose, 30 °C and 6 % NaCl). The pure fraction (named EPS-M1) was purified by DEAE-cellulose and Sephadex G-100 column. EPS-M1 was a heteropolysaccharide composed of Man, Glc, Gal, and Fuc with a molecular weight of 33.8 kDa. Scanning electron microscopy (SEM) observed a rough surface and reticular structure of EPS-M1, and EPS-M1 formed spherical aggregates in aqueous solution observed in atomic force microscopy (AFM). Thermal analysis revealed that the degradation temperature of EPS-M1 was 306 °C. Moreover, methylation and NMR analysis determined that EPS-M1 was consisted of →3)-Manp-(1→, →2,6)-Manp-(1→, →4,6)-Glcp-(1→, →3)-Glcp-(1→, →4)-Galp-(1→, →4)-Fucp-(1→, and T-Manp-(1→. Furthermore, the cytotoxicity and the repair ability of UVR-mediated cell damage of EPS-M1 were studied with L929 cells. The results showed that EPS-M1 had good biocompatibility and it could mitigate UVR-mediated cell damage by regulating the levels of cellular reactive oxygen species (ROS), depolarization of mitochondrial membrane potential (MMP) and Caspase-3/7 activity. Overall, the structure analysis and the protective effects of EPS against L929 cells exposed to UVR provided an experimental basis for EPS in practical applications.

The aim of this study was to establish a method to improve the biological activity of polysaccharides. Three acid-treated polysaccharides (BGPS-2, BGPS-3 and BGPS-4) were obtained by treating black garlic polysaccharides (BGPS-1) with sulfuric acid at different intensities. The structure was characterized using the sulfuric acid-carbazole assay, IC, HPSEC-MALLS and FT-IR. The biological functions were evaluated using antioxidant and melanin biosynthesis inhibition assays. Compared with BGPS-1, the molecular weight of acid-treated polysaccharides significantly decreased, and the uronic acid content significantly increased. Antioxidant capacity negatively correlated with molecular weight, whereas melanin inhibition activity positively correlated with uronic acid content. BGPS-4 had the highest antioxidant capacity and the lowest molecular weight (1.25 × 10

Bioconjugation reactions, such as protein PEGylation, generally require excess reagents because of their inefficiency. Intriguingly, few reports have investigated the fundamental causes of this inefficiency. This study demonstrates that the excluded volume effect (EVE)─caused by the mutual repulsion of methoxy poly(ethylene glycol) (mPEG) and proteins under typical PEGylation conditions─causes proteins and protein-reactive mPEG (5 kDa) to self-associate into separate "protein-rich" and "mPEG-rich" nano-domains (i.e., soluble self-assemblies). To overcome this obstacle to reaction, "unreactive" low-molecular-weight mPEG was added as a co-solvent to promote the association between the larger protein and the reactive mPEG molecules by harnessing the same EVE. The near complete PEGylation of lysozyme could be achieved with close to stoichiometric amounts of reactive mPEG, and beneficial effects were observed for other proteins. Considering the general nature of the EVE (e.g., salting-out and PEGying-out), this study provides important perspectives on enhancing bioconjugation reactions, which are relevant to many nanoscale systems.

Dissolved organic matter (DOM) such as fulvic acid (FA) and humic acid (HA) in soil considerably affects the fate of per- and polyfluoroalkyl substances (PFASs). However, the effect of DOM on their behavior in plants remains unclear. Herein, hydroponic experiments indicate that FA and HA reduce the accumulation of an emerging PFAS of high concern, 6:2 chlorinated polyfluoroalkyl ether sulfonate (6:2 Cl-PFESA), in wheat roots by reducing its bioavailability in the solution. Nevertheless, FA with low molecular weight (MW) promotes its absorption and translocation from the roots to the shoots by stimulating the activity and the related genes of the plasma membrane H

Protein tyrosine phosphatases constitute an important class of drug targets, whose potential has been limited by the paucity of drug-like small molecule inhibitors. We recently described a class of active site-directed, moderately selective and potent inhibitors of the low molecular weight protein tyrosine phosphatase (LMW-PTP). Here we report our extensive structure-based design and optimization effort that afforded inhibitors with vastly improved potency and specificity. The leading compound inhibits LMW-PTP potently and selectively (K(i) = 1.2 nM, >8,000-fold selectivity). Many compounds exhibit favorable drug-like properties, such as low molecular weight, weak CYP inhibition, high metabolic stability, moderate to high cell permeability (Papp> 0.2 nm/s), and moderate to good oral bioavailability (%F from 23% to 50% in mice) and therefore can be used as in vivo chemical probes to further dissect the complex biological as well as pathophysiological roles of LMW-PTP and for the development of therapeutics targeting LMW-PTP.

Fast protein translocations often lead to bandwidth-limited amplitude-attenuated event signatures. In this study, we developed a protein- and electrolyte chemistry-centric pathway to construct a readily executable decision tree for the detection of non-attenuated protein translocations using conventional electronics. Each optimization encompasses increasing capture rate (

There is an increasing demand for rapid, affordable, in-field screening methods for low molecular weight (LMW) compound detection. Anti-idiotypes (Ab2s) are biologically derived surrogates that can replace LMW compounds and their protein conjugates in immunoassays. Substitution with anti-idiotypes can improve assay standardisation, reduce cost, and contribute to environmental safety. Their application has been limited by difficult generation processes and varied effects on assay performance. This review examines a recent resurgence in the use of Ab2s within LMW compound detection, driven by the application of phage display and nanobodies. The methods used for Ab2 production are critically discussed and their potential role in improving LMW compound immunoassays is highlighted. Finally, forward-looking ideas for the production of anti-idiotypes are provided, along with barriers to their generation.

The bacterial strain HDE-9 was isolated from sauerkraut and identified as Levilactobacillus brevis. An exopolysaccharide (EPS) was isolated and purified from L. brevis HDE-9, and a preliminary investigation of its structural characteristics and biological activity was conducted. The molecular weight of the EPS was1.0 × 10

Diabetes leading to brain glucose metabolism disorders and cerebrovascular complications. Fucoidan is a kind of sulfated polysaccharides which found in brown algae, has multiply bioactivities and considered to be a promising therapeutic agent. Despite the increasing amount of evidence suggesting the diabetes protective role of fucoidans, the effect of fucoidan on brain abnormalities in type 2 diabetes mellitus patients remains unclear. In this study a low molecular weight fucoidan (LMWF) was obtained from Saccharina japonica and its effect on the cerebrovascular damage in db/db mice was investigated. Results were shown after LMWF treatment, the degree of cerebrovascular damage, the number of apoptotic neuronal cells and the inflammation were all decreased in db/db mice. Moreover, LMWF could up-regulates CD34 and VEGFA expression in db/db mice brain, and the subintestinal vessel angiogenesis in zebrafish was also promoted by LMWF. Moreover, the lumen formation of HUVEC endothelial cells was rescued by LMWF which was destroyed in high glucose treated endothelial cells. Further study found, LMWF alleviates vascular injury by up-regulating the expression level of phosphorylated PI3K and phosphorylated AKT. Our study indicates that LMWF has the potential to develop a cerebrovascular protection agent for type 2 diabetes patients.

A glycoprotein (MGP2) from mountain-cultivated ginseng (MCG) was purified by Tris-HCl extraction followed by DEAE-52 ion exchange chromatography and Sephadex G-100 gel filtration chromatography. The approximate molecular weight (27.0 kDa) and monomeric nature were determined by reduced and non-reduced SDS-PAGE. The structure of MGP2 was characterized by a practical and reliable "protein-polysaccharide analyzed by spectroscopy combined with chemical analysis" strategy. The results showed that MGP2 belonged to Arabinogalactan proteins (AGPs) which contained high amount of Glc (35.1 %). The hemagglutination test concluded that MGP2 was not a lectin. In addition, the MGP2 exhibited antioxidant activity by scavenging radical capacity tests and the ability to protect human erythrocytes and RAW264.7 cells from oxidative damage induced by AAPH. Therefore, these results suggested that glycoprotein MGP2 could be used as a natural antioxidant in drug and food industry.

For the last few decades, many efforts have been made in developing cell culture methods in order to overcome the biological limitations of the conventional two-dimensional culture. This paradigm shift is driven by a large amount of new hydrogel-based systems for three-dimensional culture, among other systems, since they are known to mimic some living tissue properties. One class of hydrogel precursors has received interest in the field of biomaterials, low-molecular-weight gelators (LMWGs). In comparison to polymer gels, LMWG gels are formed by weak interactions upon an external trigger between the molecular subunits, giving them the ability to reverse the gelation, thus showing potential for many applications of practical interest. This study presents the use of the nucleoside derivative subclass of LMWGs, which are glyco-nucleo-bola-amphiphiles, as a proof of concept of a 3D cell culture scaffold. Physicochemical characterization was performed in order to reach the optimal features to fulfill the requirements of the cell culture microenvironment, in terms of the mechanical properties, architecture, molecular diffusion, porosity, and experimental practicality. The retained conditions were tested by culturing glioblastoma cells for over a month. The cell viability, proliferation, and spatial organization showed during the experiments demonstrate the proof of concept of nucleoside-derived LMWGs as a soft 3D cell culture scaffold. One of the hydrogels tested permits cell proliferation and spheroidal organization over the entire culture time. These systems offer many advantages as they consume very few matters within the optimal range of viscoelasticity for cell culture, and the thermoreversibility of these hydrogels permits their use with few instruments. The LMWG-based scaffold for the 3D cell culture presented in this study unlocked the ability to grow spheroids from patient cells to reach personalized therapies by dramatically reducing the variability of the lattice used.

Implementation of microalgae has been considered for enhancing effluent wastewater quality. However, it can cause environmental issues due to the release of extracellular and algal organic matter in the biological process. This study aimed to investigate the characteristics of dissolved effluent as algae- and bacteria-derived organic matter during the oxidation ditch process. Furthermore, experiments were conducted under three combinations filled by Spirulina platensis, Chlorella vulgaris, and without microalgae. The results showed that dissolved effluent organic matter was more aromatic and hydrophobic than before treatment. Fluorescence spectroscopy identified two components-aromatic protein-like and soluble microbial product-like components-at excitation/emission of 230/345 nm and 320/345 nm after treatment, instead of fulvic acid-like at 230/420 nm and humic acid-like at 320/420 nm in raw wastewater. These components were fractionated based on the average of molecular weight cut-offs (MWCOs), and high (MWCOs 50,000 Da), medium (MWCOs 50,000-1650 Da), and low molecular weights (MWCOs 1650 Da) were reported. Biological oxidation ditch under symbiosis algal bacteria generated humic and fulvic acid with a higher MWCOs than the process without algal. The quality and quantity of dissolved effluent organic matter in an oxidation ditch reactor were significantly affected by algal-bacteria symbiotic.

Multi-angle light scattering (MALS) was used to determine the absolute molar mass of fluorescent macromolecules. It is standard protocol to install bandwidth filters before MALS detectors to suppress detection of fluorescent emissions. Fluorescence can introduce tremendous error in light scattering measurements and is a formidable challenge in accurately characterizing fluorescent macromolecules and particles. However, we show that for some systems, bandwidth filters alone are insufficient for blocking fluorescence in molar mass determinations. For these systems, we have devised a post-collection correction procedure to calculate the amount of fluorescence interference in the filtered signal. By determining the intensity of fluorescent emission not blocked by the bandwidth filters, we can correct the filtered signal accordingly and accurately determine the true molar mass. The transmission rates are calculated before MALS experimentation using emission data from standard fluorimetry techniques, allowing for the characterization of unknown samples. To validate the correction procedure, we synthesized fluorescent dye-conjugated proteins using an IR800CW (LI-COR) fluorophore and Bovine Serum Albumin protein. We successfully eliminated fluorescence interference in MALS measurements using this approach. This correction procedure has potential application toward more accurate molar mass characterizations of macromolecules with intrinsic fluorescence, such as lignins, fluorescent proteins, fluorescence-tagged proteins, and optically active nanoparticles.

This work aims to evaluate the structural characteristics and study the oxidative depolymerization of lignins obtained from hardwood and softwood sulfite liquors. Lignins were obtained after ultrafiltration and freeze- drying of the sulfite liquors and characterized based on inorganic content, nitrobenzene oxidation, 13C NMR, and molecular weight determination. The structural characteristics achieved allow evaluating the potential of each lignin through oxidative depolymerization to produce added-value phenolic monomers. Hardwood and softwood lignins were submitted to alkaline oxidation with oxygen and the reaction conditions optimized to obtain a final oxidation mixture with the maximum yield of phenolic monomers. Through oxidation with O2, hardwood lignin generates mostly syringaldehyde while lignin from softwood biomass mainly produces vanillin; moreover, a lower reaction time and the interruption of O2 admission avoid the degradation of the oxidation products in the final mixture for both lignins, more evidenced to hardwood lignin due to its higher reactivity. From the results, it is possible to conclude that a phenolic aldehyde-rich oxidation mixture could be obtained, confirming the viability of lignin as raw material to produce added-value products as vanillin and syringaldehyde. This work was financially supported by LA/P/0045/2020 (ALiCE), UIDB/50020/2020 and UIDP/50020/2020 (LSRE-LCM), funded by national funds through FCT/MCTES (PIDDAC); Base Funding - UIDB/00690/2020 of CIMO - Centro de Investigação de Montanha - funded by national funds through FCT/MCTES (PIDDAC); COST Action LignoCOST (CA17128). Carlos Vega-Aguilar thanks the Costa Rican Science, Technology and Telecommunications Ministry for the PhD. Scholarship MICITT-PINN-CON-2-1-4-17-1-002. Authors are grateful to Company A and Caima - Indústria de Celulose, S.A. (Portugal) for the supply of the samples of sulfite liquor info:eu-repo/semantics/publishedVersion

Three signal peptides from α-mating factor (α-MF), inulinase (INU) and native levansucrase (LS) were compared for secretion efficiency of Bacillus subtilis levansucrase SacB-T305A in Pichia pastoris GS115. The first complete secretion of bacterial levansucrase in yeasts under methanol induction was achieved while using α-MF signal. The secreted recombinant Lev(α-MF) proved to be glycosylated by combination of NanoLC-MS/MS and Endo H digestion. Interestingly, glycosylation not only improved significantly the polymerase thermostability, but also reversed the products profiles to favor synthesis of high molecular weight (HMW) levan which accounted for approximately 73 % to total levan-type polysaccharides. It indicated for the first time that the glycosylation of recombinant B. subtilis levansucrase affected significantly the products molecular weight distribution. It also provided a promising enzymatic way to effectively product HMW levan from sucrose resources.

Lipid-modified low molecular weight branched polyethyleneimines (PEIs) are promising non-viral gene delivery systems that have been successfully explored for treatment of various diseases. The present study aims to determine in vitro safety of these delivery systems based on assessment of cytotoxicity with peripheral blood mononuclear cells (PBMCs), hemolysis with human red blood cells (RBC) and cytokine secretion from several sources of PBMCs. The viability of cells treated with lipopolymer/pDNA complexes was dependent on the polymer:pDNA ratio used but remained low at therapeutically relevant concentrations for most lipopolymers, except for the propionic acid substituted PEIs. The extent of hemolysis was minimal and below the accepted safety levels with most of the lipopolymers; however, some linoleic acid substituted PEIs yielded significant hemolysis activity. Unlike strong cytokine secretion from PMA/IO stimulated cells, most lipopolymer/pDNA complexes remained non-responsive, showing minimal changes in cytokine secretion (TNF-α, IL-6 and IFN-γ) irrespective of the lipopolymer/pDNA formulations. The 0.6 kDa PEI with lauric acid substituent displayed slight cytokine upregulation, however it remained low relative to the positive controls. This study demonstrated that the lipid modified LMW PEIs are expected to be safe in contact with blood components. However, close attention to lipopolymer concentration and ratio of polymer to pDNA in formulations might be required for individual lipopolymers for optimal safety response in nucleic acid therapies. STATEMENT OF SIGNIFICANCE: This manuscript investigated the safety aspects of various lipid modified low molecular weight polyethylenimine (LMW-PEI) polymers employed for pDNA delivery through in vitro studies. Using peripheral blood mononuclear cells (PBMCs) from multiple sources, we show that the hemolysis ability was minimal for most polymers, although a particular lipid substituent (linoleic acid) at specific ratios exhibited hemolysis. The levels of pro-inflammatory cytokines (TNF-α, IL-6 and IFN-γ) were slightly upregulated only with a lauric acid substituted 0.6PEI, but remained low relative to positive control treatments. We further report the beneficial effect of polyacrylic acid additives on hemolysis and cytokine secretion to a reasonable extent. This study confirms the feasibility of using LMW-PEI as safe delivery agents for various therapeutic purposes.

Aims The discovery of antiviral substances to respond to COVID-19 is a global issue, including the field of drug development based on natural materials. Here, we showed that chitosan-based substances have natural antiviral properties against SARS-CoV-2 in vitro. Methods and Results The molecular weight of chitosan-based substances was measured by the gel permeation chromatography analysis. In MTT assay, the chitosan-based substances have low cytotoxicity to Vero cells. The antiviral effect of these substances was confirmed by quantitative viral RNA targeting the RdRp and E genes and plaque assay. Among the substances tested, low molecular weight chitooligosaccharide decreased the fluorescence intensity of SARS-CoV-2 nucleocapsid protein of the virus-infected cells in a dose-dependent manner. Conclusions In conclusion, the chitooligosaccharide, a candidate for natural treatment, has antiviral effects against the SARS-CoV-2 virus in vitro. Significance and Impact of Study In this study, it was suggested for the first time that chitosan-based substances such as chitooligosaccharide can have an antiviral effect on SARS-CoV-2 in vitro.

Mtb8.4, a secretory T-cell antigen of Mycobacterium tuberculosis, is important for providing an antigen-specific immune response. In this study, we showed Mtb8.4 to have both heme-binding and fibril-forming properties, using experimental and in silico methods. High absorbance at 410 nm and interaction with hemin-agarose demonstrated its heme-binding nature. Titration of Mtb8.4 with heme resulted in 1 : 1 stoichiometry. The heme-binding pocket in Mtb8.4 was identified by molecular modeling, and binding residues were predicted using molecular docking. The molecular dynamics simulations of apo- and heme-bound Mtb8.4 confirmed that the heme group forms a stable complex. Transmission electron microscopy analyses and dye-binding assays showed that Mtb8.4 forms fibers. Computational studies predicted that the C-terminal sequence (

A new exopolysaccharide (EPS) producing Gram-positive bacterium was isolated from the rhizosphere of Bouteloua dactyloides (buffalo grass) and its EPS product was structurally characterized. The isolate, designated as LB1-1A, was identified as Bacillus paralicheniformis based on 16S rRNA gene sequence and phylogenetic tree analysis. The EPS produced by LB1-1A was identified as a levan, having β(2 → 6) linked backbone with β(2 → 1) linkages at the branch points (4.66%). The isolate LB1-1A yielded large amount (~ 42 g/l) of levan having high weight average molecular weight (Mw) of 5.517 × 10

The non-equilibrium dynamics of linear and star-shaped

Signs of metabolic syndrome and prediabetes preceding type 2 diabetes are modelled in an experiment using a high-fat diet (HFD). The aim of this work was to study the effect of a low molecular weight systemically active nerve growth factor mimetic, compound GK-2 (hexamethylenediamide bis[N-monosuccinyl-L-glutamyl-L-lysine]), on indicators of abdominal obesity, basal blood glucose level, glucose tolerance, cholesterol and triglyceride blood levels, as well as the morphological structure of the liver in male Wistar rats fed a HFD. Rats were divided into three groups: one of them received standard food (control) and two others were fed a HFD containing 45% fat, 35% carbohydrates and 20% protein, with a total caloric value of 516 kcal/100 g, over 12 weeks. Starting from the ninth week, for the next 4 weeks, one of the HFD groups was treated orally with saline whilst the other group was treated orally with GK-2 at a dose of 5 mg/kg. GK-2 was found to reduce the basal glycaemia level and improve glucose tolerance, as well as to reduce the blood level of cholesterol by 30% and that of triglycerides by 28% in comparison with the saline-treated HFD animals. GK-2 reduced the degree of abdominal obesity to the level of the healthy animals and eliminated morphological abnormalities in the liver caused by the HFD. The results of the study determine the feasibility of further GK-2 research as a potential agent for prediabetes treatment.

Polyolefins represent the largest class of commodity materials due to their excellent material properties; however, they have limited pathways to chemical recycling and are often difficult to mechanically recycle. Here we demonstrate a new catalyst for the isoselective copolymerization of propylene and butadiene capable of favoring 1,4-insertion over 1,2-insertion while maintaining good molecular weights and turnover frequencies. This isotactic propylene copolymer with main-chain unsaturation was depolymerized to a telechelic macromonomer using an olefin metathesis catalyst and 2-hydroxyethyl acrylate. After hydrogenation, the telechelic macromonomer was repolymerized to form an ester-linked polypropylene material. This polymer shows thermal and mechanical properties comparable to linear low-density polyethylene. Finally, the telechelic macromonomer could be regenerated through the depolymerization of the ester-linked polypropylene material, which allows for the chemical recycling to macromonomer. This process provides a route to transform partially unsaturated polyolefins to chemically recyclable materials with similar properties to their parent polymers.

A novel polysaccharide (MSP-1) was isolated from the fruiting body of Morchella sextelata and purified using DEAE-52 and Sephadex G-75. The molecular weight of MSP-1 was 1.17 × 10

Many common polymers, especially vinyl polymers, are inherently difficult to chemically recycle and are environmentally persistent. The introduction of low levels of cleavable comonomer additives into existing vinyl polymerization processes could facilitate the production of chemically deconstructable and recyclable variants with otherwise equivalent properties. Here, we report thionolactones that serve as cleavable comonomer additives for the chemical deconstruction and recycling of vinyl polymers prepared through free radical polymerization, using polystyrene (PS) as a model example. Deconstructable PS of different molar masses (∼20-300 kDa) bearing varied amounts of statistically incorporated thioester backbone linkages (2.5-55 mol %) can be selectively depolymerized to yield well-defined thiol-terminated fragments (10 kDa) that are suitable for oxidative repolymerization to generate recycled PS of nearly identical molar mass to the parent material, in good yields (80-95%). A theoretical model is provided to generalize this molar mass memory effect. Notably, the thermomechanical properties of deconstructable PS bearing 2.5 mol % of cleavable linkages and its recycled product are similar to those of virgin PS. The additives were also shown to be effective for deconstruction of a cross-linked styrenic copolymer and deconstruction and repolymerization of a polyacrylate, suggesting that cleavable comonomers may offer a general approach toward circularity of many vinyl (co)polymers.

We have previously shown that semicrystalline polymers can be reinforced by adding nanoparticles (NPs) and then ordering them into specific motifs using the crystallization process. A key result we have found is that when the spherulite growth rate is slowed below a critical value, then, NPs can order into the amorphous interlamellar regions of the semicrystalline structure. The effects of spherulite growth rate in this context have previously been examined, and here we focus on the role of NP diffusivity. We achieve this goal by changing the poly(ethylene oxide) (PEO) molecular weight as a route to altering the matrix viscosity. In particular, four molecular weights of PEO were employed ranging from 5.4-46 kDa. Each sample was loaded with 10 vol % of bare 14 nm diameter silica NPs. After initially studying spherulite growth rates, experiments were designed to fix the spherulite growth rate across sample molecular weights to study particle ordering, induced by polymer crystallization. We find that, at the fastest growth rate studied (12 μm/s), the lowest molecular weight sample showed the highest order, presumably due to enhanced particle mobility. However, as the spherulite growth rate is slowed, the maximum ordering behavior is observed at intermediate molecular weights. The trend observed at slow growth rates is explained by the large-scale segregation of NPs (presumably into the grain boundaries, i.e., the interspherulitic regions); evidence for this is the observed transition of spherulite growth to diffusion-control at slow growth rates in the lowest molecular weight PEO sample studied.

Shear-thickening fluids that absorb the impact energy of high-velocity projectiles are of great interest for aerospace and body-armor applications. In such a frame, we investigate transient states of neat and aqueous polyelectrolytes (PE) having low molecular weights and containing poly([2-(methacryloyloxy)ethyl]trimethylammonium) as polycations and poly(acrylamide

Low molecular weight gels are formed by the self-assembly of small molecules into anisotropic structures that form a network capable of immobilizing the solvent. Such gels are common, with a huge number of different examples existing, and they have many applications. However, there are still significant gaps in our understanding of these systems and challenges that need to be addressed if we are to be able to fully design such systems. Here, a number of these challenges are discussed.

A novel acidic polysaccharide named SSPA50-1 was isolated from Scapharca subcrenata using a simulated gastric fluid extraction method. SSPA50-1 is a heteropolysaccharide with an average molecular weight of 44.7 kDa that is composed of galacturonic acid, glucose, galactose, mannose, ribose, rhamnose, fucose, xylose and arabinose at a molar ratio of 1.00:5.40:9.04:3.10:1.59:4.01:2.10:2.21:2.28. The structural characterization based on the methylation and 1D/2D NMR analyses indicated that SSPA50-1 is composed of →3)-β-L-Rhap-(1→, →3)-β-L-2-O-Me-Fucp-(1→, →2)-α-D-Xylp-(1→, →5)-α-L-Araf-(1→, →3)-β-D-Galp-(1→, →6)-α-D-Glcp-(1→, →3,4)-β-D-Manp-(1→, →3,4)-β-D-Galp-(1→, β-D-Ribf-(1→, α-D-Glcp-(1→, and α-D-GalAp6Me-(1→. Furthermore, SSPA50-1 possessed potent immunoregulatory activity by enhancing the phagocytosis and NO, iNOS, TNF-α and IL-6 secretion capacity of RAW264.7 cells. Otherwise, SSPA50-1 significantly promoted the proliferation of splenic lymphocytes and RAW264.7 macrophages. These results indicated that SSPA50-1 could be developed as a potential ingredient for immunostimulatory agents.

This study aimed to investigate the chemical structure, physicochemical properties, antioxidant capacity, antibacterial ability and anti-biofilm formation activity of an exopolysaccharide (EPS) produced by Lactobacillus curvatus SJTUF 62116 from the fish Gymnocypris przewalskii. The purified EPS, denoted as EPS-1, was mainly composed of glucose and mannose at a relative molar ratio of 1:1.05 with molecular weight of 31.9 kDa. The chemical structure of EPS-1 was consisted of →2)-α-D-Manp-(1→, →4)-α-D-Manp-(1→, →3,6)-α-D-Manp-(1→, T-β-D-Glcp-(1→, →6)-β-D-Glcp-(1→, and →3)-β-D-Glcp-(1→ glycosidic bonds. A sheet-like structure of dried EPS-1 was determined by scanning electron microscope (SEM), whilst a peak-shaped structure of EPS-1 was observed by atomic force microscope (AFM). The degradation temperature of EPS-1 was determined as 300.21 °C using thermogravimetric analysis (TGA). Moreover, the antioxidant capacity of EPS-1 at a concentration of 5.0 mg/mL against DPPH and ABTS was 84.50% and 92.53%, respectively. Furthermore, EPS-1 exhibited acceptable bacteriostatic efficacy against S. Enteritidis, E. coli, and S.aureus with significant inhibition of S. Enteritidis biofilm formation.

The addition of arabinoxylans (AXs) is important for improving the structure of wheat starch-AX gels, which further influences the functionality of starch-based products. The properties of wheat starch-AX gels (including rheology, texture, water distribution, microstructure, and degree of crystallinity) were studied. AX with high molecular weight (Mw) significantly decreased the swelling and leached amylose, while increasing the solubility of amylose. The AX with high Mw also clearly reduced the apparent viscosity, elasticity, and viscosity of wheat starch-AX gels. The Mw of AX was positively correlated to the hardness of the gels and negatively correlated to adhesiveness to a certain extent. The spin-spin relaxation time of the gels increased with an increase in Mw, which resulted in more free water. Scanning electron microscopy showed that AX with high Mw clearly reduced the degree of starch gelatinization while forming a fragile gel structure. In summary, AX with high Mw from natural wheat grains can effectively affect wheat starch gelling properties. These results may be useful for the application of natural AXs in wheat starch-based functional foods.

Bottlebrush polymers are characterized by an expansive parameter space, including graft length and spacing along the backbone, and these features impact various structural and physical properties such as molecular diffusion and bulk viscosity. In this work, we report a synthetic strategy for making grafted block polymers with poly(propylene oxide) and poly(ethylene oxide) side chains, bottlebrush analogues of poloxamers. Combined anionic and sequential ring-opening metathesis polymerization yielded low dispersity polymers, at full conversion of the macromonomers, with control over graft length, graft end-groups, and overall molecular weight. A set of bottlebrush poloxamers (BBPs), with identical graft lengths and composition, was synthesized over a range of molecular weights. Dynamic light scattering and transmission electron microscopy were used to characterize micelle formation in aqueous buffer. The critical micelle concentration scales exponentially with overall molecular weight for both linear and bottlebrush poloxamers; however, the bottlebrush architecture shifts micelle formation to a much higher concentration at a comparable molecular weight. Consequently, BBPs can exist in solution as unimers at significantly higher molecular weights and concentrations than the linear analogues.

The carbohydrate hyaluronan (or hyaluronic acid, HA) is found in all human tissues and biofluids where it has wide-ranging functions in health and disease that are dictated by both its abundance and size. Consequently, hyaluronan evaluation in physiological samples has significant translational potential. Although the analytical tools and techniques for probing other biomolecules such as proteins and nucleic acids have become standard approaches in biochemistry, those available for investigating hyaluronan are less well established. In this review, we survey methods related to the assessment of native hyaluronan in biological specimens, including protocols for separating it from biological matrices and technologies for determining its concentration and molecular weight.

The filterability of barley malt is a critical quality parameter in beer brewing. The effects of two mashing processes (processes A and B) on the filterability of the three barley malts and their macromolecule contents were investigated. Filtration volume increased by 4%, 9%, and 13% for the Baudin, Ganpi, and Gangpi malts, respectively, and the final filtration volume of Gangpi was still poorer than that of Baudin. A downward mashing process (process C) was applied to measure the β-glucan, arabinoxylans (AX), the polymeric arabinoxylan (PAX), and high molecular weight nitrogen (HMWN) content. The β-glucan degradation rate of well-modified malt during malting was higher than that of poorly modified malt, whereas the PAX and HMWN solubilization rates during malting were lower in well-modified malt than in poorly modified malt. The filterability of poorly modified malt did not effectively improve with an initial mashing phase at 37℃. β-Glucan degradation and PAX and HMWN solubilization during malting were critical for ensuring malt quality. Thus, the goal of predicting the filterability of malt was achieved by applying a downward mashing process. PRACTICAL APPLICATION: Quality of wort and beer as well as production efficiency is affected by the malt quality. The filterability of barley malt can affect the production efficiency and quality of wort. The change in the macromolecule contents during malting is important to ensure the production of high-quality malt. The results of this study can provide a good method for the detection of malt filtration performance, and it also may contribute to the purpose of prediction the changes in barley malt and the resulting barley malt filterability.

The time-domain transients in the Fourier transform mass spectrometry (FTMS) analysis of monoclonal antibodies (mAbs) are known to exhibit characteristic isotopic beat patterns. These patterns are defined by the isotopic distributions of all gaseous mAb ions present in the FTMS mass analyzer, originating from single or multiple charge states, and from single or multiple proteoforms. For an isolated charge state of a single proteoform, the mAb isotopic beat pattern resembles narrow splashes of signal amplitude (beats), spaced periodically in the time-domain transient, with broad (often exceeding 1 s) "valleys" between them. Here, we reinforce the importance of isotopic beat patterns for the accurate interpretation and presentation of FTMS data in the analysis of mAbs and other large biopolymers. An updated, mAb-grade version of the transient-mediated FTMS data simulation and visualization tool, FTMS Simulator is introduced and benchmarked. We then apply this tool to evaluate the charge-state dependent characteristics of isotopic beats in mAbs analyses with modern models of Orbitrap and ion cyclotron resonance (ICR) FTMS instruments, including detection of higher-order harmonics. We demonstrate the impact of the isotopic beat patterns on the analytical characteristics of the resulting mass spectra of individual and overlapping mAb proteoforms. The results reported here detail highly nonlinear dependences of resolution and signal-to-noise ratio on the time-domain transient period, absorption or magnitude mode spectra representation, and apodization functions. The provided description and the demonstrated ability to routinely conduct accurate simulations of FTMS data for large biopolymers should aid the end-users of Orbitrap and ICR FTMS instruments in the analysis of mAbs and other biopolymers, including viruses.

We studied the influence of microstructure on the chromatographic behavior of gradient copolymers with different gradient strengths and block copolymer with completely segregated blocks by using gradient liquid adsorption chromatography (gLAC) and liquid chromatography at critical conditions (LCCC) for one of the copolymer constituents. The copolymers consist of repeating units of poly(propylene oxide) and poly(propylene phthalate) and have comparable average chemical composition and molar mass, and a narrow molar mass distribution to avoid as much as possible the influence of these parameters on the elution behavior of the copolymers. On both reversed stationary phases, the elution volume of gradient copolymers increases with the increasing strength of the gradient. The results indicate that for both modes of liquid interaction chromatography, it is important to consider the effect of microstructure on the elution behavior of the gradient copolymers in addition to the copolymer chemical composition and molar mass in the case of gLAC and the length of the chromatographically visible copolymer constituent in the case of LCCC.

Hyaluronic acid (HA) plays an important role in cellular and extracellular matrix (ECM) homeostasis. Recent studies demonstrate that low molecular weight (MW) HA has pro-inflammatory characteristics while high MW HA is considered anti-inflammatory and regenerative. In formulating a topical HA product, the possibility of creating a focused high MW HA technology was posed, combining external surface high MW HA constituents with active agents promoting fibroblast production of high MW in the depths of the dermis.Human dermal fibroblasts and keratinocytes were treated with various agents, and RNA sequencing (RNA-seq) was conducted to identify genes involved in HA synthesis. HA production by fibroblasts was assessed by collecting the culture supernatant, concentrating the protein, and conducting polyacrylamide gel electrophoresis (PAGE). The gel was stained with Stains-All to identify bands relative to known HA products of different MWs. Subsequently, the supernatants were treated with hyaluronidase to confirm the bands corresponded to HA.The RNA-seq results revealed a variety of agents upregulated HA-related genes. However, a potent upregulation of HA synthesis gene was observed by hexapeptide-11 in the keratinocytes and a newly identified proprietary octapeptide in the fibroblasts. PAGE demonstrated not only robust production of HA by octapeptide, but significantly, the HA produced was ~2 Mega Daltons in size. Octapeptide was the most potent stimulator among the tested agents.Comprehensive in vitro testing identified a group of active agents that stimulated high MW HA production. This novel approach to HA topical application with exclusively high MW HA production should maximize hydration capacity while encouraging regenerative activity within the ECM. Multi-center trials are underway.

The research focused on tuberculosis as it is one of the world's most serious health problems. The extracted chitin from the gladius of Sepioteuthis lessoniana converted into Chitosan (CH). The purified and freeze-dried CH was refined as Sulfated Chitosan (SCH). The SCH was converted into low molecular weight of SCH with various doses of Gamma Irradiation (GIR). Fluorescence characteristics of GIR-SCH and elemental analysis were confirmed. The structure and molecular weights of GIR-SCH were determined with FT-IR, NMR and MALDI-TOF/Mass Spectroscopy. 100 Gy of GIR-SCH significantly showed the minimum inhibitory concentration (MIC) against Mycobacterium smegmatis. The MIC against M. smegmatis was not affected by the varied sulfate levels in the identical molecular weight GIR-SCH. However, the lowest molecular weight GIR-SCH displayed a significantly MIC against M. smegmatis. In docking analysis, the 6ZT3 ligand had the lowest binding energy of -1.57 kcal/Mol indicating a superior binding interaction with GIR-SCH. The effect of molecular weight reduction by GIR on the anti-tuberculosis capacity of GIR SCH was investigated in this study which had antimicrobial implications.

In this study, we investigated the antioxidant properties and the changes of molecular weight (Mw), antioxidant activity, and microstructure of Chinese yam polysaccharides (CYP-A) during in vitro digestion. Results showed that the scavenging rate of 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical of CYP-A was approximately 79% at the concentration of 6 mg/mL. Furthermore, the antioxidant ability positively correlated with the concentration of CYP-A. In addition, the Mw of CYP-A decreased (p 0.05) after intestinal digestion. Fourier transform infrared showed that the degrees of esterification of CYP-A increased to 39.04% after 6 h of gastric digestion. Moreover, the structure of CYP-A changed after in vitro gastric digestion. The scanning electron microscope (SEM) images indicated that the surface morphology of CYP-A turned from smooth and irregular to a layered honeycomb after gastrointestinal digestion in vitro. Surprisingly, the activity of DPPH radical scavenging significantly increased (p 0.05) in the intestinal digested samples of CYP-A, which showed a positive correlation with the concentration of CYP-A. However, the reducing power significantly decreased (p 0.05) after in vitro intestinal digestion.