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Plasma medicine is a field that utilizes reactive species generated from atmospheric low-temperature plasmas for applications such as sterilization, blood coagulation, and cancer therapy. Commercial plasma devices are available for wound healing, but research on the chemical modifications induced by these plasmas is scarce. This study explores the chemical modifications in hemoglobin when exposed to a helium plasma dielectric barrier discharge, with the aim of explaining the potential mechanisms through which it contributes to blood coagulation and enhances wound healing. Optical microscopy of cold atmospheric plasma (CAP) treated whole capillary blood showed an increase in red blood cell (RBC) size and the formation of rouleaux structures. The treatment of whole blood leads to hemolysis of RBCs and the release of intracellular protein content. We then treated purified hemoglobin protein at physiological concentrations, which led to the formation of aggregates that could be observed using ion mobility mass spectrometry (IM–MS), size exclusion chromatography, and optical microscopy. The aggregates formed fibril-like structures as observed using atomic force microscopy. The formation of hemoglobin aggregates is hypothesized to be the result of new intermolecular interactions formed following the CAP-mediated protein oxidation. We studied the changes to hemoglobin structure after treatment with a CAP using high-resolution MS and found that the hemoglobin subunits are oxidized with the addition of at least 4 oxygen atoms each. The intact tetrameric hemoglobin structure remains unchanged; however, the monomeric and dimeric proteins adopt a more compact structure, as observed by IM–MS. We propose that CAP treatment of fresh blood leads to hemolysis, and that the extracellular protein, primarily hemoglobin, is oxidized leading to the formation of aggregates. [ABSTRACT FROM AUTHOR]

Low-temperature plasma (LTP) jets are Food and Drug Administration (FDA)-approved medical devices to remove cancerous tissue and aid in wound healing. However, reports on their reaction with proteins are conflicting, ranging from fragmentation, oxidation, aggregation, or a combination thereof. In this study we bridge the gap between plasma-treatment of short peptides to proteins at physiologically relevant concentrations. The LTP in this study is based on a helium dielectric barrier discharge that forms a plasma-jet, which is directed at the solution without direct contact with the plasma, and results in the formation of reactive oxygen species (ROS) OH• and O2•− in solution. The longer the solution is treated, the more solution-phase ROS form. Treating peptide- and protein-containing solutions leads to extensive oxidation. The ROS led to the same oxidative modifications for peptide M with increasing chain length (9, 18, 37, 76 amino acids), which could be identified with high-resolution mass spectrometry. Oxidized species M + x O led to conformational changes such as compaction and elongation, while the unmodified peptide M remained unaltered, as found by ion mobility spectrometry and size exclusion chromatography. For proteins at high concentration, insoluble aggregates formed and could be identified by UV/V is light scattering and atomic force microscopy. The formation of aggregates is dependent on the amino acid chain length, the peptide concentration, and the time for aggregate formation. These findings highlight the importance of both peptide chain length and concentration in determining the fate of peptides following the exposure to LTP, while also offering valuable insights for the field of plasma medicine. [ABSTRACT FROM AUTHOR]

The concentration of harmful metal ions is growing globally, which raises the risk to both human and ecological health. The use of "adsorption" technique has been found to be very effective, for the removal of toxic metal ions. Among other things, hydrogels as an adsorbent work effectively for the removal of toxic metal ions and other aquatic pollutants. The newly designed potato starch‐xanthate (SX) based hydrogel (SX‐modified hydrogel) has been synthesized using a mixture of acrylamide (AAm) and acrylic acid (AA) monomers, with the help of free‐radical graft copolymerization technique. The synthesized SX‐modified hydrogel has been characterized by several analytical techniques, namely, UV–visible (UV–vis) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, gel permeation chromatography (GPC), thermogravimetric (TG) analysis, point of zero charge (ΔpHPZC) analysis, x‐ray diffraction (XRD) analysis, and scanning electron microscope (SEM) analysis. The main objective of the current work is to remove the Cu2+ and Co2+ ions from wastewater using SX‐modified hydrogel as well as to study the swelling and water retention properties of the SX‐modified hydrogel. The swelling ratio of SX‐modified hydrogel has been found to be 312.31, 374.01, and 410.20 g/g at optimum pH 10, temperature 35°C, time 675 min in gray wastewater, tap water, and distilled water, respectively. The maximum percentage removal of Cu2+ and Co2+ ions by SX‐modified hydrogel has been found as 97.7% and 94.2%, respectively, at optimum conditions. The Langmuir isotherm model fits best with the experimental data, with maximum adsorption capacity of 515.46 mg/g for Cu2+ and 483.09 mg/g for Co2+ ions, respectively. The kinetic studies suggest that the adsorption process is governed by the second order kinetic model with rate constant of 2.06 × 10−4 g/(mg min) for Cu2+ and 1.79 × 10−4 g/(mg min) for Co2+ ions, respectively. The negative ΔG values suggest the adsorption process is spontaneous in nature. In addition, the positive ΔH values support the adsorption process is endothermic in nature. The SX‐modified hydrogel showed a remarkable desorption efficiency with 96.7% for Cu2+ and 92.5% for Co2+ ions and reusability for four consecutive adsorption–desorption cycles. It can be concluded that the SX‐modified hydrogel has showed an effective, economical, easy, low energy consuming, and significant potential in the treatment of wastewater containing heavy metal ions. [ABSTRACT FROM AUTHOR]

Diblock copolymers consisting of higher hydrophobic styrene and hydrophilic N-hydroxyethylacrylamide (HEAAm) segments were successfully synthesized via direct two-step atom transfer radical polymerization. Polystyrene (PS) homopolymers were synthesized using methyl 4-(bromo-methyl) benzoate initiator in N,N′- dimethylformamide (DMF) in the presence of Cu(I)Br/pentamethyldiethylene tetramine (PMDETA) catalyst system at 90 and 110 °C in nitrogen atmosphere. PS was used as macro-initiator for the synthesis of PS-b-PHEAAm polymer. Block copolymerizations were carried out in pure DMF in the presence of the same catalyst system at 110 °C while argon was used for deoxygenation and inert environment. Block copolymer was purified through dialysis into deionized water using a dialysis tubing (MWCO 3500, cellulose membrane). The number average molecular weight of PS polymers (Mn = 2240, 3250 and 10,800 Da) was determined by size exclusion chromatography using tetrahydrofuran (THF) as eluent. The chemical structures, functional groups and actual compositions of copolymer were determined using instrumental data, c.f. elemental analysis, attenuated total reflectance infra-red and proton—nuclear magnetic resonance (1H-NMR) spectroscopy analysis. Thermo-gravimetric analysis confirms that diblock copolymer has higher thermal stability than PS homopolymer. Co-solvent effect on particle formation was investigated by regulating dielectric constant. It is revealed that THF/DMF (9:1) provides susceptible environment for the formation of smallest size (13 ± 4 nm) particle. This synthetic route would establish a direct synthesis of diblock copolymer with antagonistic segments for advanced technological applications. [ABSTRACT FROM AUTHOR]

A CO2‐based thermoplastic elastomer, poly(propylene carbonate)‐multiblock‐polyamide6 (PPC‐mb‐PA6) copolymer, was constructed through coupling amorphous/soft PPC blocks with crystalline/hard PA6 blocks. The PPC‐mb‐PA6 copolymers with different block length pairs were designed and synthesized, and then fully characterized by structural, thermal, and mechanical analysis. The proton nuclear magnetic resonance (1H‐NMR), diffusion ordered spectroscopy (DOSY), heteronuclear multiple‐bond correlation (HMBC) and gel permeation chromatography (GPC) results confirm that the multiblock sequence structure of PPC‐mb‐PA6. The differential scanning calorimetry (DSC) tests show that all PPC‐mb‐PA6s possess melting points around 179–206°C. The thermal and mechanical analysis indicates improved service temperatures (T5% up to 270°C) and tensile strength ranging from 15.8 to 39.6 MPa with higher PA6 content. The dynamic mechanical analysis demonstrates excellent shape memory effect of PPC‐mb‐PA6 with thermal stimuli responsiveness derived from the micro phase separation between soft PPC and hard PA6 domains. And the characteristic strain fixity parameter (Rf) and deformation recovery rate (Rr) are 98% and 100%, respectively, which are comparable with other excellent shape memory polymers. [ABSTRACT FROM AUTHOR]

Size exclusion chromatography with UV detection (SEC‐UV) is a precise method for quantifying polyacrylamides in water. However, its accuracy can be compromised in complex matrices due to chemical degradation of the polymer. This study focuses on the impact of hydrolysis on quantifying sulfonated polyacrylamide copolymers (PAM‐AMPS) via SEC‐UV, analyzing key influencing factors. Using controlled hydrolysis, hydrolyzed copolymer (HPAM‐AMPS) standards with varying hydrolysis degree (HD) (HD = 7%–33%) were prepared without significant changes in polymer chain size. SEC‐UV analysis revealed that quantification accuracy is highly dependent on hydrolysis extent. Errors are minimal for HD up to 10%, but significantly higher for HDs of 22%–33%. Furthermore, this effect is modulated by polymer concentration. Our findings indicate that there are two compensating factors for the decrease in molar absorptivity due to amide‐to‐carboxylate substitution: the sulfonate group acting as an additional chromophore, and a redshift in absorption spectra due to hydrogen bonding at higher concentrations. This interplay between HD and concentration in this type of sulfonated polyacrylamide can help reduce quantification errors in complex aqueous samples prone to chemical degradation, like those found in environmental and subsurface applications. [ABSTRACT FROM AUTHOR]

Polygalacturonases (PG) are the enzymes that cause depolymerization of pectin. In current study, bacterial strain was isolated from rotten sample and then purified. It was screened for endo-polygalacturonase activity using PSAM media. It had shown positive response for endo-PG activity. Bacterial DNA was isolated and 16 S rRNA gene amplification was done using universal primer pair of 8 F and 1492 R. Bacterial strain was also amplified by 16 S rRNA gene for sequencing. BLAST of gene sequence on NCBI database had shown that bacterial isolate was identified as Bacillus pumilus. Endo-polygalacturonase enzyme was produced by submerged fermentation to optimize culture conditions by RSM in JMP-12 software. The optimized parameters had shown maximum endo-polygalacturonase activity of 153.2 U/mL/min after using 3 g of orange peels as substrate, 3 mL of inoculum size, 6.5 pH buffer, 40°C temperature and 3 days of incubation. After optimizing fermentation conditions, endo-polygalacturonase was precipitated using 70 % concentration of ammonium sulfate. This partially precipitated sample was dialyzed with dialysis tube and used to find activity of endo-polygalacturonase (1620.6 U/mL/min). This sample was applied to gel filtration chromatography for further purification. Endo-polygalacturonase activity increased many times 2231.44 U/mL/min after gel filtration. The Molecular weight of endo-polygalacturonase was 46 kDa after SDS PAGE characterization. High Vmax value and alkaliphilic nature of endo-polygalacturonase will make it good candidate for food and feed industry near future. [Display omitted] • A bacterial strain was isolated from rotten sample. • Bacillus pumilus was identified through 16 S rRNA gene sequencing. • Maximum enzyme activity of 153.2 U/mL/min was achieved under optimized conditions. • The characterized endo-polygalacturonase has great potential in industrial applications. • Polygalacturonase had shown maximum activity after column chromatography. [ABSTRACT FROM AUTHOR]

Plant biomass, in particular forestry wastes, is a promising renewable feedstock for deep chemical processing. Organosolv methods allow the use of underutilized lignin. The synthesis of modified polymers by azo coupling with the use of aspen (Populus tremula) ethanol lignin and its sulfated modification is studied. The success of the synthesis has been proven and the features of the structure and properties of the synthesized samples were studied by the physicochemical techniques, including Fourier transform infrared and nuclear magnetic resonance spectroscopy, gel permeation chromatography and thermogravimetric analysis. It was shown that the new azopolymers have the ability to photoisomerize, which opens up prospects for their high-tech applications. The modified lignins are proven to be bioactive antioxidants. [ABSTRACT FROM AUTHOR]

In this study, isothermal decomposition kinetic of a triple-base propellant (TBP) has been investigated by vacuum stability test (VST). The volumes of the evolved gas have been recorded step by step to monitor gas evolution in VST at 80°C, 90°C, 100°C, 110°C, 120°C, and 150°C. This semi-continuous data-recording mode makes it possible to use the Berthelot equation for shelf-life prediction of TBP. Activation energy has been calculated in both model-free and model-fitting methods, and F0/R1 is determined as the best model for thermal decomposition of TBP at VST. High-performance liquid chromatography has been used to determine stabilizer (methyl centralite) content of the TBP at various extents of reactions (α). The critical amount of stabilizer was specified at 2% α. Arrhenius average temperatures have been calculated at various climate categories and used as actual storage temperatures. Molecular weight (Mw) reductions of nitrocellulose were determined by gel permeation chromatography during aging. According to the results, the shelf-life of TBP has been estimated using the Berthelot equation at 2% α in various climate categories. Shelf-life of studied TBP differs from 14 to about 36 (years) in extremely hot dry (A1) to intermediate (A3) climate categories. [ABSTRACT FROM AUTHOR]

Background: Microecological equilibrium is essential for human health. Previous research has demonstrated that Streptococcus strain A, the main bacterial group in the respiratory tract, can suppress harmful microbes and protect the body. In this study, Streptococcus strain D19T was isolated from the oral and pharyngeal cavities of healthy children. Its antibacterial mechanism against Acinetobacter baumannii was examined, as well as its potential to prevent inflammatory damage to cells. We evaluated the effect of the fermentation conditions of D19T on inhibition of Acinetobacter baumannii growth; Isolation and purification of antibacterial active components of strain D19T and molecular mechanism of inhibition of Acinetobacter baumannii; Molecular mechanism of D19T antibacterial protein reversing cellular inflammatory injury induced by Acinetobacter baumannii. Results: The supernatant of fermentation broth of Streptococcus D19T was the active component against Acinetobacter baumannii, but the bacteria had no antibacterial activity. The supernatant of D19T fermentation broth was precipitated by (NH4)2SO4 solution, and the protein was the active antibacterial component. After gel filtration chromatography and anion gel filtration chromatography, the molecular weight of antibacterial protein was 53kD. D19T antibacterial protein can improve cell membrane permeability, limit extracellular soluble protein release, inhibit Acinetobacter baumannii biofilm formation, and prevent Acinetobacter baumannii adhesion. Acinetobacter baumannii induces inflammatory damage to respiratory cells via ferroptosis, and the D19T antibacterial protein can counteract this damage, protecting the respiratory tract. Conclusion: Streptococcus strain D19T, as a potential probiotic, inhibits the growth of Acinetobacter baumannii and the inflammatory damage of respiratory cells, playing a protective role in human respiratory health. [ABSTRACT FROM AUTHOR]

Objective Methods Results Interpretation Recombinant monoclonal therapeutic antibodies like lecanemab, which target amyloid beta in Alzheimer's disease, offer a promising approach for modifying the disease progression. Due to its relatively short half‐life, lecanemab administered as a bi‐monthly infusion (typically 10 mg/kg) has a relatively brief half‐life. Interaction with abundant plasma proteins binder in the bloodstream can affect pharmacokinetics of drugs, including their half‐life. In this study, we investigated potential plasma protein binding (PPB) interaction to lecanemab using lecanemab biosimilar.Lecanemab biosimilar used in this study was based on publicly available sequences. ELISA and western blotting were used to assess lecanemab biosimilar immunoreactivity in the fractions of human plasma obtained through size exclusion chromatography. The binding of lecanemab biosimilar to candidate plasma binders was confirmed by western blotting, ELISA, and surface plasmon resonance analysis.Using a combination of equilibrium dialysis, ELISA, and western blotting in human plasma, we first describe the presence of likely PPB partners to lecanemab biosimilar and then identify fibrinogen as one of them. Utilizing surface plasmon resonance, we confirmed that lecanemab biosimilar does bind to fibrinogen, although with lower affinity than to monomeric amyloid beta.In the context of lecanemab therapy, these results imply that fibrinogen levels could impact the levels of free antibodies in the bloodstream and that fibrinogen might serve as a reservoir for lecanemab. More broadly, these results indicate that PPB may be an important consideration when clinically utilizing therapeutic antibodies in neurodegenerative disease. [ABSTRACT FROM AUTHOR]

Background: Canine mammary tumours (CMT) are among the most common types of tumours in female dogs. Diagnosis currently requires invasive tissue biopsies and histological analysis. Tumour cells shed extracellular vesicles (EVs) containing RNAs and proteins with potential for liquid biopsy diagnostics. We aimed to identify CMT subtype-specific proteome profiles by comparing the proteomes of EVs isolated from epithelial cell lines derived from morphologically normal canine mammary tissue, adenomas, and carcinomas. Methods: Whole-cell protein lysates (WCLs) and EV-lysates were obtained from five canine mammary cell lines: MTH53A (non-neoplastic); ZMTH3 (adenoma); MTH52C (simple carcinoma); 1305, DT1406TB (complex carcinoma); and their proteins identified by LC-MS/MS analyses. Gene Ontology analysis was performed on differentially abundant proteins from each group to identify up- and down-regulated biological processes. To establish CMT subtype-specific proteomic profiles, weighted gene correlation network analysis (WGCNA) was carried out. Results: WCL and EVs displayed distinct protein abundance signatures while still showing the same increase in adhesion, migration, and motility-related proteins in carcinoma-derived cell lines, and of RNA processing and RNA splicing factors in the adenoma cell line. WGCNA identified CMT stage-specific co-abundant EV proteins, allowing the identification of adenoma and carcinoma EV signatures not seen in WCLs. Conclusions: EVs from CMT cell lines exhibit distinct protein profiles reflecting malignancy state, allowing us to identify potential biomarkers for canine mammary carcinomas, such as biglycan. Our dataset could therefore potentially serve as a basis for the development of a less invasive clinical diagnostic tool for the characterisation of CMT. [ABSTRACT FROM AUTHOR]

The novel bioactive polysaccharide, Sacha inchi shell polysaccharide‐3 (SISP‐3), was isolated from the Sacha inchi (Plukenetia volubilis L.) shell using anion exchange and gel filtration chromatography. This neutral polysaccharide fraction, characterized by a triple‐helix structure and a molecular weight of 17,684 Da, was successfully isolated. The composition analysis of SISP‐3 revealed the presence of Rha, Ara, Gal, Glc, and Fru in molar ratios of 7.2%:41.7%:40.3%:4.1%:6.7%. The dominant glycosidic linkage in SISP‐3 was identified as ⟶4)‐Galp‐(1⟶, accompanied by a complex branching pattern. The results from RAW264.7 cell experiments showed that SISP‐3 significantly enhanced cell viability, phagocytosis, and NO production. Additionally, it significantly stimulated the production of cytokines, including TNF‐α, IL‐1β, and IL‐6, in RAW264.7 cells. These findings suggest that SISP‐3 serves as an immune active agent with immunomodulatory properties, indicating its potential as a natural immunomodulatory agent for various applications. [ABSTRACT FROM AUTHOR]

Polyethylene (PE) is the most abundantly sourced plastic and significant efforts are needed for its end-of-life management. The deconstruction of PE is an uphill task and requires the breaking of highly stable C–C bonds. Here we demonstrate that PE can be deconstructed to value-added dodecene, along with other long-chain alkenes. The PCP–iridium complex catalyzes the dehydrogenation of commercial and post-consumer polyethylene waste to produce dehydrogenated polyethylene (DHP) with 0.5–1.0% unsaturation. The DHP was subjected to an ethylene cross-metathesis reaction in the presence of suitable catalysts. Through meticulous optimization of reaction parameters, 63% selectivity toward dodecene, with 26% overall yield, was achieved. The practical significance of our method has been demonstrated by subjecting post-consumer plastic waste to dehydrogenation followed by ethylene metathesis to produce dodecene as a major product, together with long-chain alkenes. The PE deconstruction has been confirmed by recording molar mass before and after depolymerization using high-temperature gel permeation chromatography. The existence of dodecene has been unambiguously ascertained using GC, GC-MS, NMR, and IR spectroscopy. Thus, these results demonstrate the conversion of waste PE to value-added dodecene and long-chain alkenes under mild reaction conditions. [ABSTRACT FROM AUTHOR]

During the last years, there has been an increasing research interest in the analysis of biological fluids requiring non-invasive sampling for biomedical and clinical applications. In this work, we have focused on the nasal exudate with the aim of investigating the potential use of this fluid to know the role of iron in stroke and also for diagnosis. Potential differences in the nasal exudate, collected in swabs, from diagnosed hemorrhagic stroke, ischemic stroke, and control groups were investigated with regard to total iron by inductively coupled plasma-mass spectrometry, iron fractionation studies by size exclusion chromatography together with post-column isotope dilution analysis, and four proteins containing iron (ferritin, transferrin, lactoferrin, and ferroportin) with ELISA kits. All these analyses represent an analytical challenge, considering the rather limited amount of sample (10–40 mg) available, being the nasal exudate extracted from the swab with 300 µL 10 mM Tris/HCl, pH = 7.4. Studies to obtain reliable analytical information, such as the blank contribution of the sampling step, evaluation of the extraction efficiency of the nasal exudate from the swab, and normalization strategies for data treatment, have been carried out. Results showed that despite the limited number of investigated samples, fractionation studies as well as the concentrations of ferritin and ferroportin obtained with ELISA kits showed a differential behavior between the different cohorts. [ABSTRACT FROM AUTHOR]

The self‐plasticization, i.e., the increase in the polymer chains' mobility by including its monomer, has a major impact on a polymer's structural, thermal, and mechanical properties. In this study, differential scanning calorimetry (DSC), optical and Raman microscopies, thermo‐mechanical analysis (TMA), size exclusion chromatography equipped with a multi‐angle light scattering detector (SEC‐MALS), and X‐ray diffraction analysis (XRD) are used to investigate the effect of thermally induced self‐plasticization of poly‐(p‐dioxanone), PDX, on the crystal growths from the amorphous and molten states. Significant changes in the crystallization behavior and mechanical properties of PDX are found only for samples self‐plasticized at the depolymerization temperature (Td) above 150 °C. The intense self‐plasticization leads to the decrease of the crystallization temperature, increase of the crystal growth rapidity, disappearance of the distinct α→α' polymorphic transition, reduction of the overall melting temperature, and segregation of the redundant monomer. Although the morphology of the crystalline phase has a major impact on the mechanical properties of PDX, the self‐plasticization itself does not seem to result in any major changes in the magnitude, localization, or morphology of formed crystallites (these are primarily driven by the temperature of crystal growth). The manifestation of the variable activation energy concept is discussed for the present crystallization data. [ABSTRACT FROM AUTHOR]

African swine fever (ASF), caused by the African swine fever virus (ASFV), is a deadly, highly contagious disease in both domestic pigs and wild boar. With mortality up to 100%, the disease has been making a serious impact on the swine industry worldwide. Because no effective antiviral treatment has been observed, proactive prevention such as vaccination remains the key to controlling the outbreak. In the pursuit of expediting vaccine development, our current work has made the first report for heterologous production of the viral outer envelope glycoprotein CD2v extracellular domain (CD2v ED), a proposed promising vaccine antigen candidate in the "green" synthetic host Nicotiana benthamiana. Protein oligomerization strategies were implemented to increase the immunogenicity of the target antigen. Herein, the protein was expressed in oligomeric forms based on the C-terminally fused GCN4pII trimerization motif and GCN4pII_TP oligomerization motif. Quantitative western blot analysis showed significantly higher expression of trimeric CD2v ED_GCN4pII with a yield of about 12 mg/100 g of fresh weight, in comparison to oligomeric CD2v ED_GCN4pII_TP, revealing the former is the better choice for further studies. The results of purification and size determination by size exclusion chromatography (SEC) illustrated that CD2v ED_GCN4pII was successfully produced in stable oligomeric forms throughout the extraction, purification, and analysis process. Most importantly, purified CD2v ED_GCN4pII was demonstrated to induce both humoral and cellular immunity responses in mice to extents equivalent to those of the live attenuated vaccine ASFV-G-∆I177L, suggesting it as the potential subunit vaccine candidate for preventing ASFV. [ABSTRACT FROM AUTHOR]

Here, we describe the preparation of bridged ethylene polyethylene oxide (BE‐PEO) surface‐modified silica packing materials for size exclusion chromatography. BE‐PEO surface‐modified silica was hydrolyzed and subsequent 1H nuclear magnetic resonance analysis of hydrolysis products confirmed the successful formation of BE‐PEO bonded surface. Silica particles exhibiting 3 µm diameters and 1000 Å nominal pore diameters were selected as a base material for this work out of the critical need to improve analytical capabilities for the testing of cell and gene therapy drug products. Accelerated high pH aging study revealed significant enhancement in column stability. Multi‐angle light scattering noise measurements showed inordinately lower baseline noise. Moreover, we evaluated the chromatographic performance of BE‐PEO silica‐packed columns through separations of a protein test mixture, DNA ladder, monoclonal antibody‐based therapeutics, and adeno‐associated viruses. BE‐PEO silica columns demonstrated high resolution, high recovery separations that were confirmed to be reproducible and capable of extended column lifetimes and exhibited low ionic and hydrophobic secondary interactions. In summary, BE‐PEO silica particles have yielded a new level of performance, improved base stability, and inherently lower baseline noise. These novel widepore particles will facilitate more sensitive size‐based detection and characterization of large biologics in the form of advanced gene therapy products. [ABSTRACT FROM AUTHOR]

Iodinated contrast media (ICMs) used in X-ray imaging for medical diagnostics are released into wastewater and then encountered in river water at concentrations ranging from several dozen to hundreds of µg/L, and even thousands of µg/L in hospital effluents. ICMs are considered as emerging pollutants as their occurrence and impact on ecosystems and the environment are poorly documented. Even if they are considered inert for humans, aquatic organisms are continuously exposed to ICMs, and their potential deleterious effects are therefore questioned as we have recently demonstrated that they enter into organisms such as the zebra mussels. To answer this question, Dreissena polymorpha were exposed to two ICMs of different osmolality, diatrizoic acid and iohexol, at an environmental concentration (100 µg/L) for 21 days before a depuration phase of 4 days. The occurrence, fate, and impact of both ICMs in these organisms were studied using a metallomic approach. Thus, iodine as well as endogenous copper and zinc were quantified and analyzed in cytosolic extracts of digestive glands, gills, and gonads of mussels by size exclusion chromatography coupled to ICP MS. This work shows that a subcellular fractionation is necessary to distinguish variations in total element content. The cytosolic iodoprotein chromatographic pattern was consistent for the three organs and confirmed the presence of ICMs in cytosols. Additionally, this exploratory work tends to show a weak biological effect of ICMs with a substantial variation of the profile of Cu-binding proteins mostly in the gill cytosol and to a lesser extent, in the digestive gland cytosol. [ABSTRACT FROM AUTHOR]

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Sahiwal cattle, known for their high milk yield, are propagated through artificial insemination (AI) using male germplasm, largely contingent on semen quality. Spermatozoa, produced in the testes, carry genetic information and molecular signals essential for successful fertilization. Seminal plasma, in addition to sperm, contains nano-sized lipid-bound extracellular vesicles (SP-EVs) that carry key biomolecules, including fertility-related miRNAs, which are essential for bull fertility. The current study focused on miRNA profiling of SP-EVs from highfertile (HF) and low-fertile (LF) Sahiwal bulls. SP-EVs were isolated using size exclusion chromatography (SEC) and characterized by dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). Western blotting detected the EVspecific protein markers TSG101 and CD63. The DLS analysis showed SP-EV sizes of 170-180 nm in HF and 130-140 nm in LF samples. The NTA revealed particle concentrations of 5.76 × 1010 to 5.86 × 1011 particles/mL in HF and 5.31 × 1010 to 2.70 × 1011 particles/mL in LF groups, with no significant differences in size and concentration between HF and LF. High-throughput miRNA sequencing identified 310 miRNAs in SP-EVs from both groups, with 61 upregulated and 119 downregulated in HF bull. Further analysis identified 41 miRNAs with significant fold changes and p-values, including bta-miR-1246, bta-miR-195, bta-miR-339b, and bta-miR-199b, which were analyzed for target gene prediction. Gene Ontology (GO) and KEGG pathway analyses indicated that these miRNAs target genes involved in transcription regulation, ubiquitindependent endoplasmic reticulum-associated degradation (ERAD) pathways, and signalling pathways. Functional exploration revealed that these genes play roles in spermatogenesis, motility, acrosome reactions, and inflammatory responses. qPCR analysis showed that bta-miR-195 had 80% higher expression in HF spermatozoa compared to LF, suggesting its association with fertility status (p < 0.05). In conclusion, this study elucidates the miRNA cargoes in SP-EVs as indicators of Sahiwal bull fertility, highlighting bta-miR-195 as a potential fertility factor among the various miRNAs identified. [ABSTRACT FROM AUTHOR]

We have recently made the strikingly discovery that upon a muscle injury, Wnt7a is upregulated and secreted from new regenerating myofibers on the surface of exosomes to elicit its myogenerative response distally. Despite recent advances in extracellular vesicle (EVs) isolation from diverse tissues, there is still a lack of specific methodology to purify EVs from muscle tissue. To eliminate contamination with non-EV secreted proteins and cytoplasmic fragments, which are typically found when using classical methodology, such as ultracentrifugation, we adapted a protocol combining Tangential Flow Filtration (TFF) and Size Exclusion Chromatography (SEC). We found that this approach allows simultaneous purification of Wnt7a, bound to EVs (retentate fraction) and free non-EV Wnt7a (permeate fraction). Here we described this optimized protocol designed to specifically isolate EVs from hind limb muscle explants, without cross-contamination with other sources of non-EV bounded proteins. The first step of the protocol is to remove large EVs with sequential centrifugation. Extracellular vesicles are then concentrated and washed in exchange buffer by TFF. Lastly, SEC is performed to remove any soluble protein traces remaining after TFF. Overall, this procedure can be used to isolate EVs from conditioned media or biofluid that contains EVs derived from any cell type or tissue, improving reproducibility, efficiency, and purity of EVs preparations. Our purification protocol results in high purity EVs that maintain structural integrity and thus fully compatible with in vitro and in vivo bioactivity and analytic assays. [ABSTRACT FROM AUTHOR]

This document presents a study on the ultrasensitive detection of pathological protein aggregates associated with neurodegenerative diseases. The study focuses on a reagent called Brij-58, which enhances the seed amplification assay (SAA) without inhibiting it. The researchers found that Brij-58 significantly increased the sensitivity of detecting TAR DNA-binding protein 43 (TDP-43) seed fibrils in patient-derived brain tissues. The study suggests that Brij-58 has the potential to be used in the early diagnosis and monitoring of neurodegenerative diseases. However, further research is needed to apply these findings in clinical practice. [Extracted from the article]

Extracellular vesicles (EVs) are valuable targets for liquid biopsy. However, attempts to introduce EV‐based biomarkers into clinical practice have not been successful to the extent expected. One of the reasons for this failure is the lack of reliable methods for EV baseline purification from complex biofluids, such as cell‐free plasma or serum. Because available one‐step approaches for EV isolation are insufficient to purify EVs, the majority of studies on clinical samples were performed either on a mixture of EVs and lipoproteins, whilst the real number of EVs and their individual specific biomarker content remained elusive, or on a low number of samples of sufficient volume to allow elaborate 2‐step EV separation by size and density, resulting in a high purity but utmost low recovery. Here we introduce Fast Protein Liquid Chromatography (FPLC) using Superose 6 as a matrix to obtain small EVs from biofluids that are almost free of soluble proteins and lipoproteins. Along with the estimation of a realistic number of small EVs in human samples, we show temporal resolution of the effect of the duration of postprandial phase on the proportion of lipoproteins in purified EVs, suggesting acceptable time frames additionally to the recommendation to use fasting samples for human studies. Furthermore, we assessed a potential value of pure EVs for liquid biopsy, exemplarily examining EV‐ and tumour‐biomarkers in pure FPLC‐derived fractions isolated from the serum of patients with pancreatic cancer. Consistent among different techniques, showed the presence of diseases‐associated biomarkers in pure EVs, supporting the feasibility of using single‐vesicle analysis for liquid biopsy. [ABSTRACT FROM AUTHOR]

The morphogenesis of higher plants requires communication among distant organs throughout vascular tissues (xylem and phloem). Numerous investigations have demonstrated that phloem also act as a distribution route for signalling molecules being observed that different macromolecules translocated by the sap, including nucleic acids and proteins, change under stress situations. The participation of extracellular vesicles (EVs) in this communication has been suggested, although little is known about their role. In fact, in the last decade, the presence of EVs in plants has originated a great controversy, where major concerns arose from their origin, isolation methods, and even the appropriate nomenclature for plant nanovesicles. Phloem sap exudates from melon plants, either aphid‐free or infested with Aphis gossypii, were collected by stem incision. After sap concentration (Amicon), phloem EVs (PhlEVs) were isolated by size exclusion chromatography. PhlEVs were characterised using Nanoparticle Tracking Analysis, Transmission electron microscopy and proteomic analysis. Here we confirm the presence of EVs in phloem sap in vivo and the detection of changes in the particles/protein ratio and composition of PhlEVs in response to insect feeding, revealing the presence of typical defence proteins in their cargo as well as components of the proteasome complex. PhlEVs from infested plants showed lower particles/protein ratio and almost two times more proteolytic activity than PhlEVs from aphid‐free plants. In both cases, such activity was inhibited in a dose‐dependent manner by the proteasome inhibitor MG132. Our results suggest that plants may use this mechanism to prepare themselves to receive infectious agents and open up the possibility of an evolutionary conserved mechanism of defence against pathogens/stresses in eukaryotic organisms. [ABSTRACT FROM AUTHOR]

Atherosclerotic lesions mainly form in arterial areas exposed to low shear stress (LSS), where endothelial cells express a senescent and inflammatory phenotype. Conversely, areas exposed to high shear stress (HSS) are protected from plaque development. Endothelial extracellular vesicles (EVs) have been shown to regulate inflammation and senescence, and therefore play a crucial role in vascular homeostasis. Whilst previous studies have shown links between hemodynamic forces and EV release, the effects of shear stress on the release and uptake of endothelial EVs remains elusive. We aim to decipher the interplay between these processes in endothelial cells exposed to atheroprone or atheroprotective shear stress. Confluent HUVECs were exposed to LSS or HSS for 24 h. Large and small EVs were isolated from conditioned medium by centrifugation and size exclusion chromatography. They were characterised by TEM, Western blot, tunable resistive pulse sensing, flow cytometry and proteomics. Uptake experiments were performed using fluorescently‐labelled EVs and differences between groups were assessed by flow cytometry and confocal microscopy. We found that levels of large and small EVs in conditioned media were fifty and five times higher in HSS than in LSS conditions, respectively. In vivo and in vitro uptake experiments revealed greater EV incorporation by cells exposed to LSS conditions. Additionally, endothelial LSS‐EVs have a greater affinity for HUVECs than HSS‐EVs or EVs derived from platelets, erythrocytes and leukocytes. Proteomic analysis revealed that LSS‐EVs were enriched in adhesion proteins (PECAM1, MCAM), participating in EV uptake by endothelial cells. LSS‐EVs also carried mitochondrial material, which may be implicated in elevating ROS levels in recipient cells. These findings suggest that shear stress influences EV biogenesis and uptake. Given the major role of EVs and shear stress in vascular health, deciphering the relation between these processes may yield innovative strategies for the early detection and treatment of endothelial dysfunction. [ABSTRACT FROM AUTHOR]

Multiple mitochondrial dysfunctions syndrome type 3 (MMDS3) is a rare autosomal recessive mitochondrial leukoencephalopathy caused by biallelic pathogenic variants in the IBA57 gene. The gene protein product, IBA57, has an unknown role in iron–sulfur (Fe-S) cluster biogenesis but is required for the maturation of mitochondrial [4Fe-4S] proteins. To better understand the role of IBA57 in MMDS3, we have investigated the impact of the pathogenic p.Gly104Cys (c.310G > T) variant on the structural and functional properties of IBA57. The Gly104Cys variant has been associated with a severe MMDS3 phenotype in both compound heterozygous and homozygous states, and defects in the activity of mitochondrial respiratory complexes and lipoic acid-dependent enzymes have been demonstrated in the affected patients. Size exclusion chromatography, also coupled to multiple angle light scattering, NMR, circular dichroism, and fluorescence spectroscopy characterization has shown that the Gly104Cys variant does not impair the conversion of the homo-dimeric [2Fe-2S]–ISCA22 complex into the hetero-dimeric IBA57–[2Fe-2S]–ISCA2 but significantly affects the stability of IBA57, in both its isolated form and in complex with ISCA2, thus providing a rationale for the severe MMDS3 phenotype associated with this variant. [ABSTRACT FROM AUTHOR]

Vinyl acetate (VAc)-based emulsions represent one of the main media used by modern and contemporary artists. Their long-term behaviour is still not completely understood, especially due to the scarce knowledge on the influence of other compounds in the formulation, which may impact ageing over time. Besides the polymer backbone based on vinyl acetate, other co-monomers and additives can be added to the emulsion to alter the final film's physical, chemical, and optical properties. By extension, the formulation will also impact the long-term stability of artworks and objects on which it has been applied, as well as possible current and future conservation interventions such as cleaning. For those reasons, studies shedding light on the correlation between composition and long-term stability are largely necessary. In this study, different emulsions, including homopolymers, copolymers, plasticised, and un-plasticised compositions, were gathered and artificially aged. A multivariate analyses approach based on the application of principal component analyses (PCA) and hierarchical cluster analyses (HCA) was employed for the first time on the combination of data obtained by pH, contact angle (CA), colour measurements, Fourier transform infrared spectroscopy in attenuated total reflection (FTIR-ATR), and size exclusion chromatography (SEC). This approach helped to highlight the changes that occurred during ageing and find correlations with the formulation compositions. The results further sustain the thesis that not all vinyl acetate-based emulsions are chemically the same and that their formulation deeply impacts their long-term behaviour. [ABSTRACT FROM AUTHOR]

Waste cooking oil (WCO) recycled asphalt is facing issues regarding insufficient thermal oxidation stability and aging resistance. In this research, glycerol esterification was adopted to pretreat WCO, and the consequences of this treatment on the aging resistance and thermal stability of WCO were analyzed. The impacts of varying levels of esterification of WCO on the high-temperature, low-temperature performances, fatigue properties, and aging resistance of recycled asphalt were investigated. Furthermore, the mechanisms of regeneration and the anti-aging of deeply esterified WCO recycled asphalt were revealed by Fourier transform infrared spectroscopy (FTIR) and gel permeation chromatography (GPC) tests. The results indicated that variations in the physical properties of WCO during the aging process were reduced, and its aging resistance was improved following glycerol esterification therapy. The initial thermal decomposition temperature was increased by approximately 115 °C, which resulted in the enhancement of thermal stability significantly. Recycled asphalt obtained from deeply esterified WCO exhibited superior high-temperature, low-temperature performances, and fatigue properties. Moreover, the thermal oxidation stability and aging resistance of recycled asphalt with deep-esterified WCO could be promoted by reducing the oxidation and volatilization of light components during the aging process, with the complex modulus ageing resistance index decreasing by 13.27% and the phase angle ageing resistance index increasing by 14.71%. [ABSTRACT FROM AUTHOR]

L‐lactide (LA) is polymerized with ethyl L‐lactate or trifluoroethanol as initiators (LA/In = 30/1), and the resulting poly(L‐lactide) esters are acetylated with acetic anhydride. The acetylated PLA esters are mixed with SnOct2, dibutyltin bis(4‐chlorophenoxide), or dibutyltin bis(pentafluorophenoxide) in solution and crystallized at 120 °C. The doped crystals are annealed at 140 °C or at 160 °C for 7, 14, and 28 days, and the chemical modifications that occurred in the solid state are monitored by matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI TOF) mass spectrometry, gel permeation chromatography (GPC), and differential scanning calorimetry (DSC) measurements. All catalysts promoted polycondensation, while no significant formation of cycles is observed. However, in the presence of SnOct2, disproportionation of chains occurred upon annealing at 120 or 140 °C, and crystallites consisting of extended chains with Mn values ≈3500 – 3600 and low dispersities (Ð < 1.5) are formed. [ABSTRACT FROM AUTHOR]

[Display omitted] • Fasciola hepatica extracellular vesicles (Hp EVs) induce extracellular matrix secretion. • Hp EVs reduce proliferation of hepatic stellate cells (HSCs) • Dicrocoelium dendriticum EVs (Dd EVs) trigger a global inflammatory response in HSCs and hepatocytes. • Dd EVs have a more potent anti-proliferative effect than Fh EVs. • Fh EVs are taken up faster and in higher amounts than Dd EVs by HSCs and hepatocytes. Fasciola hepatica and Dicrocoelium dendriticum are parasitic trematodes residing in the bile ducts of mammalian hosts, causing, in some cases, impairment of liver function and hepatic fibrosis. Previous studies have shown that extracellular vesicles released by F. hepatica (Fh EVs) and D. dendriticum (Dd EVs) induce a distinct phenotype in human macrophages, but there is limited information on the effect of parasitic EVs on liver cells, which interact directly with the worms in natural infections. In this study, we isolated Fh EVs and Dd EVs by size exclusion chromatography and labeled them with a lipophilic fluorescent dye to analyze their uptake by human hepatic stellate cells (HSC) and hepatocytes, important cell types in liver pathology, using synthetic liposomes as internal labeling and uptake control. We analyzed EV uptake and the proteome profiles after the treatment with EVs for both cell types. Our results reveal that EVs establish unique and specific interactions with stellate cells and hepatocytes, suggesting a different role of EVs derived from each parasite, depending on the migration route to reach their final niche. Fh EVs have a cytostatic effect on HSCs, but induce the extracellular matrix secretion and elicit anti-inflammatory responses in hepatocytes. Dd EVs have a more potent anti-proliferative effect than Fh EVs and trigger a global inflammatory response, increasing the levels of NF-κB and other inflammatory mediators in both cell types. These interactions may have a major influence on the progression of the disease, serving to generate conditions that may favor the establishment of the helminths in the host. [ABSTRACT FROM AUTHOR]

In the present work, nanowire (NW) structured TiO2 nanoparticles were prepared using the hydrothermal method and characterized by X-Ray Diffraction (XRD), Scanning Electron Microscope (SEM), and BET specific surface area techniques. They were obtained in the anatase phase and presented a high surface area (ca. 300 m2/g). A commercial TiO2 (anatase, Merck) was used for comparison. The TiO2 catalysts were tested for photocatalytic oxidation of 3-pyridinemethanol to 3-pyridinemethanal and vitamin B3 in water under UVA irradiation. The effects of acid treatment and subsequent calcination for TiO2 catalysts after the hydrothermal synthesis were also investigated. The sample, subjected to acid treatment and calcined at 300 °C (NW-HCl-300), showed the highest photocatalytic activity and selectivity towards the products. Consequently, this sample and Merck TiO2 were used to prepare polystyrene (PS)/TiO2 composites using the hydrothermal method. They were characterized by XRD, SEM–EDX, Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FT-IR), Thermogravimetric analysis (TGA), Differential Scanning Calorimetry (DSC), UV–Vis, Gel Permeation Chromatography (GPC), and contact angle measurements and tested for PS (present in the composite) photodegradation. The results indicated that NW-HCl-300 had a high surface area, and was highly hydroxylated, favouring a good distribution of PS in the composite. The composite presented high thermal stability, but under UVA irradiation the polymer underwent solid-state photocatalytic degradation due to the contact with TiO2. The composite photodegradation was investigated using gravimetric, GPC, FT-IR, UV–Vis, and SEM techniques. [ABSTRACT FROM AUTHOR]

Macroprolactinemia is one of the major causes of hyperprolactinemia. The aim of this study was to clarify the origin of macroprolactin (macro-PRL). We examined macro-PRL in the sera of 826 pregnant women and in those of their babies' umbilical cords at delivery. Macro-PRL was evaluated by precipitation with polyethylene glycol (PEG), gel filtration chromatography (GFC), and absorption with protein G (PG). We detected macro-PRL in 16 out of the 826 pregnant women (1.94 %) and in 14 of their babies, which may indicate the possibility of hereditary origin of macro-PRL. However, the macro-PRL ratios of the babies correlated positively with those of their mothers (r=0.72 for GFC, p<0.001 and r=0.77 for PG, p<0.001), suggesting that the immunoglobulin (Ig)G-type anti-PRL autoantibodies might be actively transferred to babies via the placenta and form macro-PRL by binding to their babies' PRL or PRL-IgG complexes may possibly pass through the placenta. There were two cases in which only mothers had macro-PRL, indicating that the mothers had autoantibodies that did not pass through the placenta, such as IgA, PRL bound to the other proteins or PRL aggregates. No cases were found in which only the babies had macro-PRL and their mothers did not, suggesting that macro-PRL might not arise by non-hereditary congenital causes. Macro-PRL in women of reproductive age might be mostly IgG-type anti-PRL autoantibody-bound PRL. The likely origin of macro-PRL in babies is the transplacental transfer of IgG-type anti-PRL autoantibodies or PRL-IgG complexes from the mothers to their babies. [ABSTRACT FROM AUTHOR]

Human sialic acid-binding immunoglobulin-like lectins (Siglecs) are expressed on subsets of immune cells. Siglec-8 is an immune inhibitory Siglec on eosinophils and mast cells, which are effectors in allergic disorders including eosinophilic esophagitis. Inhibition occurs when Siglec-8 is crosslinked by multivalent Siglec ligands in target tissues. Previously we discovered a high-affinity Siglec-8 sialoglycan ligand on human airways composed of terminally sialylated keratan sulfate chains carried on a single protein, DMBT1. Here we extend that approach to another allergic inflammatory target tissue, human esophagus. Lectin overlay histochemistry revealed that Siglec-8 ligands are expressed predominantly by esophageal submucosal glands, and are densely packed in submucosal ducts leading to the lumen. Expression is tissue-specific; esophageal glands express Siglec-8 ligand whereas nearby gastric glands do not. Extraction and resolution by gel electrophoresis revealed a single predominant human esophageal Siglec-8 ligand migrating at >2 MDa. Purification by size exclusion and affinity chromatography, followed by proteomic mass spectrometry, revealed the protein carrier to be MUC5B. Whereas all human esophageal submucosal cells express MUC5B, only a portion convert it to Siglec-8 ligand by adding terminally sialylated keratan sulfate chains. We refer to this as MUC5B S8L. Material from the esophageal lumen of live subjects revealed MUC5B S8L species ranging from ~1–4 MDa. We conclude that MUC5B in the human esophagus is a protein canvas on which Siglec-8 binding sialylated keratan sulfate chains are post-translationally added. These data expand understanding of Siglec-8 ligands and may help us understand their roles in allergic immune regulation. [ABSTRACT FROM AUTHOR]

Production of phytases from Pichia kudriavzevii FSMP-Y17 yeast was enhanced by optimization of the fermentation variables under submerged fermentation conditions. Maximum 3.67 U/mL of phytases was produced using 2.5% orange peel flour as a substrate, at pH 5.0 and temperature 50°C, by supplementing fermentation medium with 0.2% (wt/vol) galactose as carbon source and 0.2% (wt/vol) ammonium nitrate as a nitrogen source. Addition of sodium phytate to the production medium, however, did not cause any enhancing effect on phytase production by P. kudriavzevii FSMP-Y17 under submerged fermentation. The yeast enzyme purified to near homogeneity in three steps (ammonium sulphate precipitation, anion exchange chromatography and gel filtration) was found showing thermostability (from 40 to 70°C, with highest activity at 55°C) and stable at acidic pH (from 4.0 to 7.0, with highest activity at pH from 5.0 to 6.0). The purified enzyme was added to the diets of the broilers. The diets enriched with yeast phytase showed increased feed intake in the birds, while food conversion rate was lowered. The phytase supplementation caused better phosphorus retention in the birds consequently resulting in enhanced growth of the broilers. The augmented diets also showed improved egg production and egg quality in the hens. [ABSTRACT FROM AUTHOR]

A novel endo -polygalacturonase (PGC-AN64) from Aspergillus niger HO32 was produced (88.22 U/mL) on orange peel by-product (OPP), purified, biochemically characterized, and its utility in clarification of orange juice was elucidated. The enzyme was purified by ammonium sulfate fractionation (80 %) and gel filtration chromatography on Sephacryl® S-200 HR column. Its pH and temperature optima were 6.5 and 70 °C, respectively, and was stable over a pH range of 5–7 and temperature 60–80 °C. The 26-residue NH 2 -terminal sequence of PGC-AN64 showed high homology with those of Aspergillus pectinases. Metal ions (Na+, Ca2+, Mg2+, Mn2+, and Zn2+) positively affect PGC-AN64 activity, while Ba2+, Cd2+, and Cr3+ negatively affect its original activity. Interestingly, PGC-AN64 exhibited a considerable substrate specificity and catalytic efficiency compared to the commercial enzymes PECLYVE V (P1) and PECLYVE CP (P2). More interestingly and compared to P1 and P2, PGC-AN64 showed that the clarification process had a significant effect on the transmittance percentages with 84.68 ± 2.08, 88.05 ± 1.79, and 93.13 ± 3.58 % for PGC-AN64, P1, and P2, respectively. Furthermore, color parameters after the clarification process increase in the L* and b* values, and led to a decrease in the a* value. The obtained results suggest that PGC-AN64 revealed its potential in orange juice clarification. [Display omitted] • Orange peel waste is used as substrate for pectinase production from A. niger HO32. • A novel thermostable endo -polygalacturonase PGC AN64 was purified and characterized. • PGC AN64 is a monomer of 64 kDa and its optimal activity was at pH 6.5 and 70 °C. • It exhibited a good substrate specificity and k cat /K m than PECLYVE V and PECLYVE CP. • This pectinase is considered as possible candidate for clarification of orange juice. [ABSTRACT FROM AUTHOR]

Liquid rubber (LR) has been proven to be a valuable asphalt modifier capable of addressing the existing deficiencies associated with crumb rubber–modified asphalt (CRMA), including poor workability and storage stability. Therefore, an accessible preparation method based on olefin metathesis was explored to produce optimal LR from crumb rubber (CR) in the laboratory. This research established the optimal laboratory preparation conditions for LR and investigated the performance of liquid rubber–modified asphalt (LRMA) mixture when used in pavement. A total of 32 LR samples were generated by olefin metathesis and physical swelling under various catalytic and swelling conditions. The physical properties, viscosity, and storage ability of LRMA were evaluated to select the optimal preparation parameters for LR. A comprehensive analysis was performed to compare the physicochemical characteristics of LR and the original CR through microscopic examinations with scanning electron microscopy (SEM), Fourier-transform infrared spectrometry (FTIR), and gel permeation chromatography (GPC). An in-depth evaluation assessed the stability at high temperatures, the properties at low temperatures, and the moisture stability of LRMA and CRMA mixtures. Experimental results showed that the optimal laboratory preparation conditions for LR were a catalyst dosage between 3‰ and 5‰, a catalytic duration of 48 h, a swelling temperature of 160°C, and a swelling time of 8 h. Moreover, LR possessed a smooth surface without rubber particles. The molecular weight and distribution of LR were considerably reduced compared with those of CR after olefin metathesis and physical swelling. The introduction of LR significantly enhanced the cracking resistance of the base asphalt mixture. However, the enhancement effect of LR on the base asphalt mixture to resist high-temperature rutting and moisture damage was inferior to that of CR. [ABSTRACT FROM AUTHOR]

An accurate characterization of the fine molecular structure of starch in terms of the chain length distribution (CLD) of amylopectin requires complete dispersion of the starch in a solvent, either buffer or water and the non-interference of this on the subsequent enzymatic debranching of starch with isoamylase (pretreatments before determining the CLD). In this study, the HYLON V and HYLON VII starches (commercial high-amylose corn starches) were selected, since high-amylose starches typically present difficulties in their dispersion and, therefore, the study of their fine structure can commonly present errors. The goal of this research was to study two water dispersion treatments (1. boiling water for 1 h and 2. autoclaving at 120 °C and 15 Psi for 15 min) to achieve a fast and reliable method of the fine structure of two high-amylose corn starches using high-performance size exclusion chromatography coupled with a refractive index detector (HPSEC-RID) and high-performance anion-exchange chromatography coupled a pulsed amperometric detector (HPAEC-PAD). The HPSEC-RID chromatograms of both starches showed better resolution using autoclaving than boiling water treatment. The two starches tested had higher amylose and degree of polymerization (DP) values utilizing the autoclaving than the boiling water treatment. However, no appreciable differences were observed in the CLD values employing HPAEC-PAD for either of the two water dispersion treatments. Lastly, the water dispersion treatment by autoclaving before enzymatic debranching is recommended to obtain a more accurate quantification of the amylose content of high-amylose starches. [ABSTRACT FROM AUTHOR]

Injectable, in situ-forming hydrogels, both biocompatible and biodegradable, have garnered significant attention in tissue engineering due to their potential for creating adaptable scaffolds. The adaptability of these hydrogels, made from natural proteins and polysaccharides, opens up a world of possibilities. In this study, sodium alginate was used to synthesize alginate di-aldehyde (ADA) through periodate oxidation, resulting in a lower molecular weight and reduced viscosity, with different degrees of oxidation (54% and 70%). The dual-crosslinking mechanism produced an injectable in situ hydrogel. Initially, physical crosslinking occurred between ADA and borax via borax complexation, followed by chemical crosslinking with gelatin through a Schiff's base reaction, which takes place between the amino groups of gelatin and the aldehyde groups of ADA, without requiring an external crosslinking agent. The formation of Schiff's base was confirmed by Fourier-transform infrared (FT-IR) spectroscopy. At the same time, the aldehyde groups in ADA were characterized using FT-IR, proton nuclear magnetic resonance (¹H NMR), and gel permeation chromatography (GPC), which determined its molecular weight. Furthermore, borax complexation was validated through boron-11 nuclear magnetic resonance (¹¹B NMR). The hydrogel formulation containing 70% ADA, polyethylene glycol (PEG), and 9% gelatin exhibited a decreased gelation time at physiological temperature, attributed to the increased gelatin content and higher degree of oxidation. Rheological analysis mirrored these findings, showing a correlation with gelation time. The swelling capacity was also enhanced due to the increased oxidation degree of PEG and the system's elevated gelatin content and hydrophilicity. The hydrogel demonstrated an average pore size of 40–60 µm and a compressive strength of 376.80 kPa. The lower molecular weight and varied pH conditions influenced its degradation behavior. Notably, the hydrogel's syringeability was deemed sufficient for practical applications, further enhancing its potential in tissue engineering. Given these properties, the 70% ADA/gelatin/PEG hydrogel is a promising candidate and a potential game-changer for injectable, self-crosslinking applications in tissue engineering. Its potential to revolutionize the field is inspiring and should motivate further exploration. [ABSTRACT FROM AUTHOR]

Polymer modification of titania nanomaterials can provide media dispersibility and various functionalities onto the titania surface. Herein, we report the direct in situ polymer modification of the surface of titania nanotubes (TNTs) and titania nanoparticles (TNPs) via ultraviolet (UV)‐irradiated radical polymerization without any pretreatment of titania. The resulting polymer‐modified TNTs and TNPs dispersed well in solvents. The characterization of the products using various techniques including Fourier transform infrared spectroscopy, thermogravimetric analysis, and scanning electron microscopy‐energy dispersive X‐ray spectroscopy confirmed the successful modification of the TNT and TNP surfaces by the polymers produced via UV‐irradiated radical polymerization. The polymers on the titania surface were isolated by dissolving titania using hydrofluoric acid and analyzed by means of size exclusion chromatography and matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry. The polymer‐modified TNTs and TNPs maintained their photocatalytic activity in dye degradation under UV irradiation. Moreover, glycopolymer‐modified TNTs were successfully prepared using the UV‐irradiated polymerization system. The glycopolymer retained its lectin biding affinity on the TNT surface. [ABSTRACT FROM AUTHOR]

To improve the utilization of by-products of the Hyriopsis cumingii, this study investigated the effect of Hyriopsis cumingii polysaccharide (HCP) on the proliferation and migration of normal cells, as well as its reparative effects on ultraviolet radiation-B (UVB)-damaged cells. First, HCP was isolated and purified using DEAE Cellulose-52 column chromatography, Sephadex G-100 column chromatography and nanofiltration. Then, structural analysis was conducted using high-performance liquid chromatography, high-performance gel permeation chromatography, and infrared spectroscopy. Following this, cell proliferation experiments, cell scratch tests, UVB damage experiments, and protein expression analyses were employed to explore the anti-UVB damage effects and corresponding mechanisms of polysaccharide components. The results indicated the three substances obtained by isolation and purification were all homogeneous polysaccharide components. Among them, HCP-2 played a major role in promoting cell proliferation in HCP and facilitated the migration of HaCaT cells. The recovery of UVB-damaged cell vitality was raised by the increasing concentration of HCP-2. Furthermore, the reparative effect was more substantial than the protective effect at the same sample concentration. It was also observed that HCP-2 up-regulated the expression of the anti-apoptotic protein bcl-2 while inhibited the expression of apoptosis-related proteins casp9, p-Akt, and p-p38, indicating that HCP-2 could repair HaCaT cells by regulating apoptosis of cells damaged by UVB. In conclusion, HCP had the ability to inhibit cell apoptosis which provide a theoretical basis for its future application in the development of food and health products. [ABSTRACT FROM AUTHOR]

A series of biobased aliphatic‐aromatic copolyesters, poly(pentylene dodecanoate‐co‐furandicarboxylates) (PPeDFs) were synthesized via an esterification and polycondensation melt process. The copolyesters were characterized using gel permeation chromatography, Fourier transform infrared spectroscopy, 1H NMR spectroscopy, differential scanning calorimetry, thermogravimetric analysis, wide angle x‐ray scattering, and tensile testing. The thermal transition behavior was strongly dependent on composition, with the melting and glass transition temperatures reaching a minimum at approximately equimolar ratio of D to F. All copolyesters were stable below 300°C with their R600 (the weight of material remaining at 600°C) values increasing with F fraction. PPeD to PPeDF30 (e.g., mole ratio D/F = 7:3) show sharp PPeD crystalline reflections only while broad PPeF reflections are shown in PPeF and PPeDF90. PPeDF40 to PPeDF80 showed both crystal structures. The fractional crystallinity for the PPeD was much higher than PPeF and the fractional crystallinity of the copolymers showed a minimum at D/F ratios closer to the latter. The stress at break and modulus both exhibited maxima at D/F ratios that were either high or low, but somewhat surprisingly a strong maximum in percent elongation at break of over 600% occurs at PPeDF40. For this composition, a typical plastic behavior curve was found including a yield point, high elongation at break, and strain hardening. Highlights: Poly(pentylene dodecanoate‐co‐furandicarboxylates) (PPeDFs) are synthesizedMechanical properties, a function of D:F ratio. High elongation at 40 mol% F.Two melting temperatures/crystal structures, one PPeD, the other PPeFGlass transition temperature minimum at intermediate D:F ratios [ABSTRACT FROM AUTHOR]

A novel diester monomer synthesized from 4-hydroxybenzoic acid, a compound which can be derived from lignin, was used to obtain aliphatic-aromatic copolyesters (P1–P4) by melt polymerization with 1,4-cyclohexanedimethanol, and either 1,4-butanedioic acid, 1,6-hexanedioic acid, 1,8-octanedioic acid, or 1,12-dodecanedioic acid as aliphatic diacid. Structure-property relations for the copolyesters were established using FTIR and 1H NMR spectroscopy, gel permeation chromatography, differential scanning calorimetry, thermogravimetric analysis, dynamic mechanical analysis and tensile testing. The weight-average molecular weight (Mw) of the samples ranged from 41,400 to 48,300 g/mol. As the spacer length in the aliphatic diacid was increased, the glass transition temperature (Tg) decreased from 90 to 51 °C, the melting point (Tm) from 175 to 147 °C, the tensile modulus from 1800 to 980 MPa, and the yield strength from 76 to 54 MPa, while the elongation at break increased from 270 to 320%. The thermal stability of the copolyesters also decreased for longer aliphatic diacid spacers. The copolyester derived from 1,12-dodecanedioic acid displayed the highest degradation rate in artificial soil, with 4.4% degradation after 30 weeks, and low ecotoxicity in an earthworm viability test. These findings contribute to understanding structure-property relationships and the environmental impact of aliphatic-aromatic copolyesters as potential sustainable materials. [ABSTRACT FROM AUTHOR]

Accurate diagnostic and serology assays are required for the continued management of the COVID-19 pandemic yet spike protein mutations and intellectual property concerns with antigens and antibodies used in various test kits render comparability assessments difficult. As the use of common, well-characterized reagents can help address this lack of standardization, the National Research Council Canada has produced two protein reference materials (RMs) for use in SARS-CoV-2 serology assays: biotinylated human angiotensin-converting enzyme 2 RM, ACE2-1, and SARS-CoV-2 Omicron BA.4/5 spike protein RM, OMIC-1. Reference values were assigned through a combination of amino acid analysis via isotope dilution liquid chromatography tandem mass spectrometry following acid hydrolysis, and ultraviolet–visible (UV–Vis) spectrophotometry at 280 nm. Vial-to-vial homogeneity was established using UV–Vis measurements, and protein oligomeric status, monitored by size exclusion liquid chromatography (LC-SEC), was used to evaluate transportation, storage, and freeze–thaw stabilities. The molar protein concentration in ACE2-1 was 25.3 ± 1.7 µmol L−1 (k = 2, 95% CI) and consisted almost exclusively (98%) of monomeric ACE2, while OMIC-1 contained 5.4 ± 0.5 µmol L−1 (k = 2) spike protein in a mostly (82%) trimeric form. Glycoprotein molar mass determination by LC-SEC with multi-angle light scattering detection facilitated calculation of corresponding mass concentrations. To confirm protein functionality, the binding of OMIC-1 to immobilized ACE2-1 was investigated with surface plasmon resonance and the resulting dissociation constant, KD ~ 4.4 nM, was consistent with literature values. [ABSTRACT FROM AUTHOR]

In this study, peptide‐calcium chelate was screened from antler bone hydrolysate, and its bioactivity on MC3T3‐E1 cells and its chelating mechanism were investigated. In vitro experiments showed that peptide‐calcium chelate promoted the differentiation and mineralization of MC3T3‐E1 cells. Subsequently, three novel calcium‐chelating peptides were obtained from antler bone hydrolysate using hydroxyapatite chromatography (HAC), Sephadex G‐25 gel filtration chromatography, and reversed‐phase high‐performance liquid chromatography (RP‐HPLC). Meanwhile, this work determined peptides' amino acid sequences as TKLGTQLQL, LETVILGLLKT, and KMVFLMDLLK based on LC–MS/MS. Then the present work prepared the three peptides, with the corresponding calcium‐chelating rates being verified as 87.68 ± 2.86%, 80.72 ± 0.93%, and 67.96 ± 0.98%, respectively. Fourier transform infrared (FTIR) spectroscopy, ultraviolet–visible absorption (UV–vis) spectroscopy, X‐ray diffraction (XRD), circular dichroism (CD), zeta potential, and molecular dynamics (MD) simulations were adopted to investigate the chelating mode of peptides with calcium ions. As a result, oxygen in the carboxyl group and the nitrogen in the amino group were related to calcium binding. In addition, the chelation site preferred the negatively charged carboxylate groups of Leu or Thr. The present work revealed that antler bone might be the new calcium‐chelating peptide source and elucidated their positive role in osteogenesis. [ABSTRACT FROM AUTHOR]

Previous studies demonstrated that EV71-infected cells secrete extracellular vesicles (EVs), facilitating the transfer of viral components to recipient cells and thereby promoting virus spread. Considering lipid signaling plays a crucial role in EVs-mediated cell-to-cell communication, we compared the lipid profile of EVs secreted from uninfected and EV71-infected cells (EVs-Mock and EVs-EV71) using the human rhabdomyosarcoma (RD) cell model. These two groups of EVs were purified by using size exclusion chromatography (SEC), respectively, and evaluated by transmission electron microscopy (TEM), nanoparticle tracking technology (NTA), and Western blotting (WB). In-depth lipidomic analysis of EVs identified 1705 lipid molecules belonging to 43 lipid classes. The data showed a significant increase in the lipid content of EVs after EV71 infection. Meanwhile, we deeply analyzed the changes in lipids and screened for lipid molecules with significant differences compared EVs-EV71 with EVs-Mock EVs. Altogether, we report the alterations in the lipid profile of EVs derived from RD-cells after EV71 infection, which may affect the function of the EVs in the recipient cells. [ABSTRACT FROM AUTHOR]

Hemp (Cannabis sativa L.) seeds are an interesting raw material for malting regarding its relatively high bioactive compounds concentration and proven advantageous properties in different food products and dietary supplements. In the first stage of the study, important seeds properties relevant to the malting process including moisture content, seed viability, and water absorption capacity were determined. However, a few parameters determining the seeds' usability for malt preparation, such as germination ability and water sensitivity, are different in comparison to typical malting raw materials such as barley or wheat. However, they make it possible to obtain high-quality hemp malt. In the next stage of research, spectroscopic and chromatographic analyses, including measurements of antioxidant activity and protein separation by SEC-HPLC, were conducted. The results showed that the malting process improved the total antioxidant potential of hemp seeds by 15%, leading to an increase in the concentration of lower molecular weight proteins and oligopeptides—below molecular mass of 10 kDa—responsible for this high antioxidant activity. The processing of hemp seeds reduced the phytate content while increasing phosphate fractions with fewer phosphate groups, which may have a beneficial effect on nutritional value. These results suggest that malting hemp seeds needs optimalization of the process but can increase its nutritional value as a promising raw material in the food industry. [ABSTRACT FROM AUTHOR]

Nanocomposites prepared with a terpolymer of poly(L–lactide) (PLLA)–poly(ε–caprolactone) (PCL)–poly(ethylene glycol) (PEG) and partially oxidized carbon nanotubes (CNTspo) were synthesized and characterized to evaluate their ability to act as an effective nanocarrier of the anticancer drug methotrexate. The homopolymers of PLLA and PCL were synthesized through ring-opening polymerization (ROP) and characterized through gel permeation chromatography (GPC). The PLLA–PCL–PEG terpolymers were synthesized through a four-step chemical route using oxalyl chloride as a linker agent and analyzed with 1H–NMR, 13C–NMR, and FTIR spectroscopies. Additionally, the nanocomposites were characterized through FTIR, and X-ray photoelectron spectroscopy (XPS), as well as the differential scanning calorimetry (DSC) technique. XPS analysis revealed that PLLA–PCL–PEG terpolymer chains are grafted onto CNTspo. Moreover, evaluations through FTIR and DSC strongly suggest that the PCL-rich domains are preferentially oriented toward CNTspo. The release tests exhibited a "burst effect" profile, which was more evident in the terpolymers than in the nanocomposites. Five models were used to assess methotrexate's in vitro release. For the nanocomposites, the best fit to the experimental data was obtained using the first-order model, whereas the results obtained from the Korsmeyer–Peppas model indicated that Fickian diffusion drives methotrexate's release. [ABSTRACT FROM AUTHOR]

Extracellular vesicle (EV) cargo is known to change in response to stimuli such as muscle damage. This study aimed to assess particle size, concentration and microRNA (miR) content within small EV‐enriched separations prepared from human blood taken before and after unaccustomed eccentric‐biased exercise‐induced muscle damage. Nine male volunteers underwent plyometric jumping and downhill running, with blood samples taken at baseline, 2, and 24 h post‐exercise. EVs were separated using size exclusion chromatography (SEC) and their characteristics evaluated by nanoparticle tracking. No changes in EV size or concentration were seen following the muscle‐damaging exercise. Small RNA sequencing identified 240 miRs to be consistently present within the EVs. RT‐qPCR analysis was performed: specifically, for known muscle‐enriched/important miRs, including miR‐1, −206, −133a, −133b, −31, −208b, −451a, −486 and − 499 and the immune‐important miR‐21, −146a and − 155. Notably, none of the immune‐important miRs within the EVs tested changed in response to the muscle damage. Of the muscle‐associated miRs tested, only the levels of miR‐31‐5p were seen to change with decreased levels at 24 h compared to baseline and 2 h, indicating involvement in the damage response. These findings shed light on the dynamic role of EV miRs in response to exercise‐induced muscle damage. [ABSTRACT FROM AUTHOR]