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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]

Introduction: Double C2-like domain beta (DOC2B) is a vesicle priming protein critical for glucose-stimulated insulin secretion in β-cells. Individuals with type 1 diabetes (T1D) have lower levels of DOC2B in their residual functional β-cell mass and platelets, a phenotype also observed in a mouse model of T1D. Thus, DOC2B levels could provide important information on β-cell dys(function). Objective: Our objective was to evaluate the DOC2B secretome of β-cells. In addition to soluble extracellular protein, we assessed DOC2B localized within membrane-delimited nanoparticles – extracellular vesicles (EVs). Moreover, in rat clonal β-cells, we probed domains required for DOC2B sorting into EVs. Method: Using Single Extracellular VEsicle Nanoscopy, we quantified EVs derived from clonal β-cells (human EndoC-βH1, rat INS-1 832/13, and mouse MIN6); two other cell types known to regulate glucose homeostasis and functionally utilize DOC2B (skeletal muscle rat myotube L6-GLUT4myc and human neuronal-like SH-SY5Y cells); and human islets sourced from individuals with no diabetes (ND). EVs derived from ND human plasma, ND human islets, and cell lines were isolated with either size exclusion chromatography or differential centrifugation. Isolated EVs were comprehensively characterized using dotblots, transmission electron microscopy, nanoparticle tracking analysis, and immunoblotting. Results: DOC2B was present within EVs derived from ND human plasma, ND human islets, and INS-1 832/13 β-cells. Compared to neuronal-like SH-SY5Y cells and L6-GLUT4myc myotubes, clonal β-cells (EndoC-βH1, INS-1 832/13, and MIN6) produced significantly more EVs. DOC2B levels in EVs (over whole cell lysates) were higher in INS-1 832/13 β-cells compared to L6-GLUT4myc myotubes; SH-SY5Y neuronal-like cells did not release appreciable DOC2B. Mechanistically, we show that DOC2B was localized to the EV lumen; the tandem C2 domains were sufficient to confer sorting to INS-1 832/13 β-cell EVs. Discussion: Clonal β-cells and ND human islets produce abundant EVs. In cell culture, appreciable DOC2B can be packaged into EVs, and a small fraction is excreted as a soluble protein. While DOC2B-laden EVs and soluble protein are present in ND plasma, further studies will be necessary to determine if DOC2B originating from β-cells significantly contributes to the plasma secretome. [ABSTRACT FROM AUTHOR]

Giredestrant (GDC-9545) is a selective estrogen receptor degrader (SERD) that was developed for treatment of ER+/HER2− metastatic breast cancer. An anhydrous crystalline tartrate salt was identified as the solid form suitable for clinical development. An early clinical batch of the active pharmaceutical ingredient (API)/drug substance failed to pass the GMP purity specifications owing to the presence of a substantial amount of high molecular weight impurities (oligomers), as determined by size exclusion chromatography. Several trial rework batches were manufactured using various re-slurry and recrystallization conditions to purge impurities in the drug substance to adhere to purity specifications. Based on the melting point depression of the API in presence of oligomers in these rework batches, a differential scanning calorimetry method was developed to quantify impurity content as a function of melting point onset of the API. This thermal analysis method was used as a surrogate for chromatography as a rapid, effective in-process check method for impurity quantitation to enable the timely release of the final reworked clinical batch. Post release, the % w/w oligomer value determined by calorimetry was in excellent agreement to that obtained by size exclusion chromatography. [ABSTRACT FROM AUTHOR]

Introduction: Peptides from lactic acid bacteria provide health benefits and can inhibit the growth of pathogenic organisms. The present work aimed to isolate and characterize peptides with antibacterial activity from Lactobacillus plantarum 1407. Methods: Peptides were isolated and purified from L. plantarum 1407. The effect of various physiological parameters on the antibacterial activity of the isolated peptides was analyzed. The mode of action of the peptides on indicator organisms was observed using transmission microscopy analysis and flow cytometry analysis. Results: Antibacterial activity and mode of action of peptides isolated from L. plantarum 1407 against gram-positive and gram-negative bacteria have been studied. L. plantarum culture exhibited maximum antibacterial activity at 40 °C, pH 8, and 0.7% salt concentration. The cell-free supernatant (CFS) was concentrated using a 3 kDa ultrafiltration membrane and the peptide fractions (<3 kDa) were further fractionated using Sephadex G-25 gel filtration chromatography. The antibacterial activity of the eluted fractions (F1 to F4) was evaluated using flow cytometry and transmission electron microscopy. F3 fraction exhibited increased antibacterial activity than F1, F2, and F4 fractions against the indicator organisms. Cell membrane damage and leakage of cytoplasmic content of the bacterial cells treated with the antibacterial F3 fraction peptides were observed. Conclusion: The active peptides from L. plantarum 1407 can be potentially used for the treatment of bacterial infections. [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]

Adhesive formulations derived from sustainable feedstocks, like waste-activated sludge and biosolids from wastewater treatment plants, are developed due to protein-based adhesives receiving attention for their low-cost, resourcefulness, and minimal ecological footprint. The protein composition and associated dynamic changes of the adhesive formulations were studied via gel permeation chromatography, which detailed a molecular size distribution of 8.72 × 105 g/mol for the adhesive formulation and 6.89 × 103 g/mol for the dewatered biosolid base fraction, which confirms the formation of multiple protein functional groups combining to form the larger adhesive molecules. Further analysis determined the types of proteins present in the dewatered biosolids as glutelin, prolamin, globulin, and albumin proteins, with the glutelin proteins as the most prevalent, as thus likely responsible for adhesive formation. The rheological properties of the novel protein adhesive were also studied to interpret the structure of the adhesives, which detailed the findings of viscoelastic properties and flow behaviors of each adhesive in relation to the wastewater treatment plant sample location, which yielded higher flow points, storage moduli, and loss moduli for the dewatered biosolids in comparison to the waste-activated sludge and biosolid adhesives, which correlates with the higher solids content of the dewatered biosolids and potentially cell rupturing when exposed to filtration stress. [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]

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In recent years, fruit pulp products that fully retain the nutritional value and original flavor of fresh fruit have been favored by consumers. This paper investigates the changes in cell wall structure during the enzymatic preparation of Goji berry pulp and the impact of enzymatic treatment on pulp quality. The results showed that, compared to the control group, the yield of pulp after 95 min of enzymatic treatment increased from 74.90 ± 0.64% to 91.30 ± 0.21%, with a reduction in particle size and a 164.7% increase in conductivity. Additionally, the cell wall components of Lycium barbarum were also structurally characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), atomic force microscopy (AFM), high performance gel permeation chromatography and ion chromatography. The study revealed that enzymatic hydrolysis disrupted the cell wall structure, resulting in a 66.1% reduction in pectin molecular weight and degradation of the RG-I structure in monosaccharides. These structural changes led to an increase in the nutritional content and antioxidant capacity of Goji berry pulp as the enzyme treatment time was extended. Specifically, after 95 min of enzymatic hydrolysis, the protein and vitamin C content in the pulp increased by 13.79 and 52.17%, respectively, compared to the non-enzymatic pulp. This study underscores the potential of compound enzyme treatment as an effective approach for enhancing the processing and production of Goji berry pulp. It can improve the quality and nutritional value of the pulp to meet consumer demand for high-quality products and offers valuable insights for optimizing pulp processing techniques. [ABSTRACT FROM AUTHOR]

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 high-fertile (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 EV-specific 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 x 1010 to 5.86 x 1011 particles/mL in HF and 5.31 x 1010 to 2.70 x 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, ubiquitin-dependent 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]

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]

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]

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]

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]

The escalating global emphasis on sustainability, coupled with stringent regulatory frameworks, has spurred the quest for environmentally viable alternatives to petroleum-derived materials. Within this context, the adhesives industry has been actively seeking renewable options and eco-friendly synthesis pathways. This study introduces geraniol, a monoterpenoid alcohol, in its unmodified form, as a key component in the production of waterborne pressure-sensitive adhesives (PSAs) based on acrylic latex through emulsion polymerization. Multiple formulations were developed at varying reaction times. The adhesives underwent comprehensive chemical characterization employing techniques such as Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), Nuclear Magnetic Resonance (NMR), Gel Permeation Chromatography (GPC), and dynamic light scattering (DLS). The viscosities of the formulations were measured between 4000 and 5000 cP. Adhesion tests showed peel strength values of 0.52 N/mm on cardboard and 0.32 N/mm on painted steel for the geraniol-based formulations. The results demonstrate the potential for geraniol-based PSAs to offer a sustainable alternative to petroleum-derived adhesives, with promising thermal and adhesive properties. [ABSTRACT FROM AUTHOR]

This study aims to investigate the comparative performance of ester- and ether-based polycarboxylate superplasticizers in maintaining the fluidity and controlling the setting time of alkali-activated slag (AAS) paste. The experiments employed rheological tests, mini-slump tests, ultrasonic pulse velocity (UPV) measurements, and gel permeation chromatography (GPC) analysis. The results indicate that ether-based superplasticizers maintain fluidity approximately 25% longer than their ester-based counterparts and extend the setting time by about 30%. The enhanced performance of ether-based superplasticizers is attributed to their superior molecular stability in highly alkaline environments, which mitigates early polymer degradation. Additionally, the Na2O/SiO2 ratio was maintained at 1:1 throughout the experiments to ensure consistency in the activation process. The relationship between fluidity loss and the onset of setting occurs more rapidly in AAS paste than in conventional cement-based systems. These findings provide valuable insights for the development of environmentally friendly construction materials by optimizing the use of superplasticizers in alkali-activated systems. [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]

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Featured Application: Microbial enzyme production falls into the so-called white biotechnology field, a part of modern biotechnology. Different applications of microbial enzymes have significant importance in regard to sustainable production, replacing hazardous chemicals. The newly isolated and purified haloalkalophilic pectinase enzyme revealed unique characteristics, making it suitable for application in the pretreatment of wastewaters from the textile, fabric, and paper industries, containing pectinecious wastes, in this way hindering environmental pollution. Its potential uses in the food-processing industry include improving the quality of fruit juices and wine in a cost-effective and sustainable manner. The interest in microbial pectinases is increasing due to their use in the biotechnological and food industries in response to global demand. In this study, among the 22 halophilic bacterial strains screened, 20 showed pectinase activity. The highest activity was measured for Virgibacillus salarius strain 434. As pectinase production meets many parameter optimization constraints, optimal production conditions for maximizing the yield of the pectinase synthesized using the investigated strain were followed. Enzyme purification was accomplished at up to 11.5-fold via ultrafiltration and gel filtration chromatography, until the enzyme's specific activity had increased 10-fold (104.3 U/mg). The enzyme's molecular weight was determined to be 68 kDa. Km and Vmax values of 0.38 mg/mL and 120 U/mg, respectively, were determined. The purified pectinase from Virgibacillus salarius strain 434 showed good thermal stability. A half-life of 20 min at 70 °C and high catalytic activity under haloalkaline conditions (pH 9 and 70 g L−1 NaCl) were observed. Its storage stability at −20 °C exceeded 2 months. This demonstrates its potential for application in industries where these extremes are present, as the harsh conditions in industrial processes require novel pectinases with unique properties instead of the use of hazardous chemicals. [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]

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]

Transition metal (TM) distribution through the phloem is an essential part of plant metabolism and is required for systemic signaling and balancing source-to-sink relationships. Due to their reactivity, TMs are expected to occur in complexes within the phloem sap; however, metal speciation in the phloem sap remains largely unexplored. Here, we isolated phloem sap from Brassica napus and analyzed it via size exclusion chromatography coupled online to sector-field ICP-MS. Our data identified known TM-binding proteins and molecules including metallothioneins (MT), glutathione, and nicotianamine. While the main peak of all metals was low MW (-1.5 kD), additional peaks -10 to 15 kD containing Cu, Fe, S, and Zn were also found. Further physicochemical analyses of MTs with and without affinity tags corroborated that MTs can form complexes of diverse molecular weights. We also identified and characterized potential artifacts in the TM-biding ability of B. napus MTs between tagged and non-tagged MTs. That is, the native BnMT2 binds Zn, Cu, and Fe, while MT3a and MT3b only bind Cu and Zn. In contrast, his-tagged MTs bind less Cu and were found to bind Co and Mn and aggregated to oligomeric forms to a greater extent compared to the phloem sap. Our data indicates that TM chemistry in the phloem sap is more complex than previously anticipated and that more systematic analyses are needed to establish the precise speciation of TM and TM-ligand complexes within the phloem sap. [ABSTRACT FROM AUTHOR]

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]

Exosomes are cell-derived, membrane-surrounded particles that deliver bioactive molecules to various cells. Due to their small size, low immunogenicity, extended blood circulation, and involvement in cellular communication, they hold potential as effective drug carriers. Exosomes are present in various biological fluids, including mare's milk, a traditional drink in Central Asia. This study aims to compare exosome isolation methodologies and determine the stability of mare's milk-derived exosomes as potential therapeutic carriers. Three extraction methods—immunoprecipitation, size exclusion chromatography, and total exosome isolation—were compared in terms of exosome characteristics, purity, and content. The isolated exosomes were then loaded with quercetin, and their ability to increase its bioavailability was tested in vitro and in vivo. Total exosome isolation was identified as the most efficient method for producing high-quality exosomes. These exosomes were loaded with quercetin and compared to free quercetin and exosomes alone. Exosomes loaded with 80 µM quercetin significantly restored β-galactosidase activity and cellular viability in doxorubicin-treated cells, exhibiting similar potency to 160 µM free quercetin. In aged model animals, treatment with quercetin-loaded exosomes resulted in significantly less acute and subacute damage to the myocardium, kidneys, and liver compared to untreated control animals. This study provides a proof-of-concept that mare's milk-derived exosomes can be effectively absorbed by cells and animal tissues, supporting their potential use as drug carriers. [ABSTRACT FROM AUTHOR]

Seminal extracellular vesicles (SemEVs) are repositories of biomolecules, including metabolites involved in the regulation of sperm function. The correlation between the metabolite profile of SemEVs and semen parameters, along with their role in regulating sperm function, is an unexplored area. This preliminary study evaluated the metabolomic content of SemEVs. Semen samples were obtained from 18 healthy men, and SemEVs were extracted from seminal plasma using the size exclusion chromatography qEV Gen 2–35 nm column coupled with an automatic fraction collector. The physical characterization of SemEVs was carried out with the ZetaView PMX-430-Z QUATT laser system. EV protein markers were detected using Western blot. In addition, these SemEVs were used for metabolomic profiling and functional bioinformatic analysis. The mean concentration of isolated SemEVs was 1.7 ± 1.1 × 1011/mL of seminal plasma, whereas SemEVs size and zeta potential were 129.5 ± 5.5 nm and −40.03 ± 3.99 mV, respectively. Western blot analysis confirmed the presence of EV specific markers such as CD81, ALIX, and TSG101. A total of 107 metabolites were identified using this untargeted metabolomic approach in SemEVs. Bioinformatics analysis further revealed that metabolites associated with tyrosine metabolism were highly enriched in these SemEVs. Ingenuity Pathway Analysis (IPA) also indicated that these metabolites present in SemEVs were involved in the regulation of the free radical scavenging pathway. Furthermore, our metabolomic results suggest that these SemEV-associated metabolites may play a pivotal role in the maintenance of seminal plasma redox homeostasis. [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]