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Egg white proteins (EWPs) are valuable ingredients in the food industry due to their unique functional properties, including gelling, foaming, and emulsifying. The gelation properties of EWPs are particularly important, as they serve as the foundation for many food applications. The denaturation and aggregation of EWPs lead to the formation of a dense three-dimensional gel network structure, which is influenced by both intrinsic and extrinsic factors. EWPs' gels can be formed through physical and chemical methods, with heating being the most common approach. Researchers have recently explored novel processes to modify the gelation properties of EWPs, which have shown promising results. Importantly, EWPs are crucial components of various food formulations, including surimi and other meat- and plant-based food products. Previous studies have focused on the basic gelation properties of EWPs, but the effect of advanced technologies and intrinsic and extrinsic factors on their gelation potential have not been systematically reviewed. Hence, this review paper provides a systematic overview of the research around the gelation properties of EWPs and their application, serving as a guide for researchers and industry professionals to explore the versatility and value of EWPs as substantial food ingredients. [ABSTRACT FROM AUTHOR]

[Display omitted] Amino acids are among the most commercially promising additive solutions for achieving stable zinc anodes. However, greater attention should be given to the limitation arising from the protonation effects induced by high isoelectric point amino acids in the weakly acidic electrolytes of aqueous zinc-ion batteries (AZIBs). In this study, we introduce histidine (HIS) and ethylenediaminetetraacetic acid (EDTA) as hybrid additives into the aqueous electrolyte. Protonated HIS is adsorbed onto the anode interface, inducing uniform deposition and excluding H 2 O from the inner Helmholtz plane (IHP). Furthermore, the addition of EDTA compensates for the limitation of protonated HIS in excluding solvated H 2 O. EDTA reconstructs the solvation structure of Zn2+, resulting in a denser zinc deposition morphology. The results demonstrate that the Zn||Zn battery achieved a cycling lifespan exceeding 1480 h at 5 mA cm−2 and 5 mAh cm−2. It also reached over 900 h of cycling at a zinc utilization rate of 70 %. This study provides an innovative perspective for advancing the further development of AZIBs. [ABSTRACT FROM AUTHOR]

BACKGROUND: The increasing demand for sustainable alternatives to traditional protein sources, driven by population growth, underscores the importance of protein in a healthy diet. Pecan (Carya illinoinensis (Wangenh.) K. Koch) nuts are currently underutilized as plant‐based proteins but hold great potential in the food industry. However, there is insufficient information available on pecan protein, particularly its protein fractions. This study aimed to explore the physicochemical and functional properties of protein isolate and the main protein fraction glutelin extracted from pecan nuts. RESULTS: The results revealed that glutelin (820.67 ± 69.42 g kg−1) had a higher crude protein content compared to the protein isolate (618.43 ± 27.35 g kg−1), while both proteins exhibited amino acid profiles sufficient for adult requirements. The isoelectric points of protein isolate and glutelin were determined to be pH 4.0 and pH 5.0, respectively. The denaturation temperature of the protein isolate (90.23 °C) was higher than that of glutelin (87.43 °C), indicating a more organized and stable conformation. This is further supported by the fact that the protein isolate had a more stable main secondary structure than glutelin. Both proteins demonstrated improved solubility, emulsifying, and foaming properties at pH levels deviating from their isoelectric points in U‐shaped curves. Compared to the protein isolate, glutelin displayed superior water and oil absorption capacity along with enhanced gelling ability. CONCLUSION: The protein isolate and glutelin from pecan nuts exhibited improved stability and competitive functional properties, respectively. The appropriate utilization of these two proteins will support their potential as natural ingredients in various food systems. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The current study focused on optimizing the extraction parameters of terebinth seed proteins from cold‐pressed terebinth oil byproducts to maximize protein purity and protein yield. The isolated proteins were characterized to evaluate their properties; thus revealing the valorization potential of these byproducts. Response surface methodology was used to detect the effect of three extraction parameters (pH, temperature, and time). The protein isolates were studied for their physicochemical and functional characteristics. The results indicated that an extraction pH of 8, a temperature of 50°C, and an extraction period of 60 min are optimum conditions for obtaining protein isolates with the highest purity. On the other hand, it was demonstrated that an extraction pH of 12, a temperature of 46.4°C, and an extraction duration of 102.4 min were optimum conditions for the maximum protein yield. The proteins produced under these two sets of conditions, referred to as TRP (terebinth protein with maximum purity) and TRY (terebinth protein with maximum yield), respectively, exhibited comparable oil absorption capacity (OAC), foaming, emulsifying capabilities, and stability. Both proteins showed the highest solubility at pH 11, and their zeta potentials approached zero at pH 4, indicating proximity to their isoelectric points. However, FRAP and DPPH assays showed that TRP and TRY offered low antioxidative capacity. The high β‐sheet content in TRP and TRY suggests enhanced thermal stability but reduced digestibility of these proteins. Therefore, in addition to protein enrichment, TRP and TRY protein isolates can be utilized in muffins and other food applications thanks to their favorable oil absorption, foaming and emulsifying capacities, and thermal stabilities. [ABSTRACT FROM AUTHOR]

Eimeria maxima (APU1 and APU2) differ in virulence for chickens, due in part to the greater fecundity of the former. In a previous study, RNA-seq was used to identify a transcripts upregulated in E. maxima APU1 compared to E. maxima APU2. In this study, 2 of these upregulated genes (EMWEY 23530 and EMWEY 48910) were characterized by first confirming upregulation using quantitative RT-PCR. For both EMWEY 23530 and EMWEY 48910, RNA transcription was fairly consistent during sporulation. The extent of differential expression was about 2-fold log2 higher in APU-1 compared to APU-2 (peaking at 18 h for EMWEY 23530 and 0 h for EMWEY 48910). EMWEY 23530 and EMWEY 48910 cDNA were cloned and expressed as polyHis-fusion proteins in Escherichia coli. The observed size of recombinant EMWEY 23530 was 24 kDa; the observed size of recombinant EMWEY 48910 was 35 kDa, which are consistent with the predicted size based on the coding sequences. Immunostaining 2D gel blots of E. maxima APU1 and APU2 oocyst/sporocyst protein with antisera specific for EMWEY 23530 identified a 33.5 kDa protein with a pH 7.4 isoelectric point (Emax p33.5). Similar 2D gel blot analysis with EMWEY 48910 identified a 41 kDa protein with a pH 7.2 isoelectric point (Emax p41). The intensity of Emax p33.5 and Emax p41 was noticeably greater in oocyst/sporocyst proteins from E. maxima APU1 compared to E. maxima APU2. This was corroborated by ELISA wherein equal amounts of total E. maxima APU1 and APU2 protein were probed with serial dilutions of anti-rEmax p33.5 or anti-rEmax p41. Immunofluorescence (IFA) staining of permeabilized unsporulated E. maxima APU1 and APU2 oocysts revealed Emax p33.5 to be localized in one end of oocysts, while Emax p41 appeared on the surface of oocysts. After sporulation, the p33.5 and p41 antigens appeared loosely associated with sporocysts. Taken together, these data confirm excess expression of two proteins in the E. maxima strain characterized by greater fecundity and virulence, and may provide insight into basis for phenotypic differences among different E. maxima. [ABSTRACT FROM AUTHOR]

Rapid industrialization has led to significant environmental challenges, including the disposal of effluents with high zinc ion concentrations. This study investigates the use of hydroxyapatite nanoparticles as an adsorbent for Zn (II) ions from aqueous solutions at neutral pH. These nanoparticles are characterized by their high purity, mesoporous structure, and a specific surface area of 60.97 ± .40 m2 g−1. Their thermal stability was confirmed by thermogravimetric analysis, and zeta potential measurements indicated a surface charge close to the isoelectric point. Adsorption experiments showed that the kinetics fit well with the pseudo‐second‐order model, with a rate constant of 2.58 ± .49 min−1 mg−1 and a determination coefficient (R2) of 1.00. Isotherm analyses using Redlich–Peterson, Langmuir, and Freundlich models revealed the highest adsorption capacity and best fitting with the Redlich–Peterson model, showing maximum capacities of 30.36 mg g−1 (linear fitting) and 32.11 mg g−1 (nonlinear fitting). These fits achieved R2 values of .9949 and .9923, respectively, suggesting efficient and reliable adsorption profiles. This research highlights the potential of hydroxyapatite nanoparticles for effective zinc ion removal, supporting their application in sustainable industrial practices and environmental remediation. [ABSTRACT FROM AUTHOR]

Melon is an important cash crop grown worldwide, with China leading the world in melon acreage. Melon wilt is becoming more common as the area under melon cultivation gradually increases. Therefore, it is crucial to understand the resistance pattern of Fusarium wilt in melons. In this work, among the four test melon cultivars, YG was highly resistant to Fusarium wilt and 1455 was disease‐resistant. However, 5657 was susceptible, while 5455 was highly susceptible to the disease. We detected 71,852 differentially expressed genes in 16 combinations, including 37,382 with up‐regulation and 3470 with down‐regulation, and we investigated 11 genes associated with resistance to Fusarium wilt through bioinformatics and transcriptome analysis. We identified two differentially expressed cinnamyl alcohol dehydrogenases (CAD) that were down‐regulated in the susceptible melon material in contrast to the resistant variety. Nine chitinase (CHT) genes were differentially expressed, with four showing up‐regulation and four showing down‐regulation in the susceptible compared to the resistant melon material. The MeloCHT genes were located on chromosomes 2, 6, 8, 11 and 12, with four of them on chromosome 8. MeloCAD1 was located on chromosome 1 and MeloCAD2 on chromosome 8. The physicochemical properties of the encoded proteins were predicted by bioinformatics, including theoretical isoelectric point, instability coefficient, relative molecular mass, lipid solubility index and hydrophilicity index. The genes encode proteins with α‐helix, extended strand, random coils and β‐turns. This study provides fundamental knowledge on the transcriptome of melons and establishes a foundation for future research on the detection and application of Fusarium wilt‐resistant genes in melons. [ABSTRACT FROM AUTHOR]

• Developed a novel, reproducible cellular assay to directly quantify non-specific endocytosis of therapeutic proteins. • A previous clinical candidate monoclonal antibody with rapid target-independent clearance in mice and humans possessed extensive non-specific endocytosis that was due to exposed positive charge features. • Demonstration of distinct rates of endocytosis into mammalian cells for disparate monoclonal antibodies, even those with common specificity for targets or isoelectric points. • Cell-based assay to quantify the potential impact of non-specific endocytosis on target-independent clearance and/or subcutaneous bioavailability of monoclonal antibodies. Past studies have demonstrated higher clearance for monoclonal antibodies possessing increased rates of non-specific endocytosis. However, this metric is oftentimes evaluated indirectly using biophysical techniques or cell surface binding studies that may not provide insight into the specific rates of cellular turnover. Furthermore, few examples evaluating non-specific endocytosis have been reported for a therapeutic antibody that reached clinical assessment. In the current report, we evaluated a therapeutic human immunoglobulin G2 monoclonal antibody targeted against the interleukin-4 receptor alpha chain (IL-4Rα) that exhibited elevated target independent clearance in previous Phase 1 and 2 studies. We confirmed high non-specific clearance of the anti-IL-4Rα antibody as compared to a reference antibody during pharmacokinetic assessments in wild type mice where target-mediated disposition was absent. We then developed a cell-based method capable of measuring cellular protein endocytosis and demonstrated the anti-IL-4Rα antibody exhibited marked non-specific uptake relative to the reference compound. Antibody homology modeling identified the anti-IL-4Rα antibody possessed positive charge patches whose removal via targeted mutations substantially reduced its non-specific endocytosis. We then expanded the scope of the study by evaluating panels of both preclinical and clinically relevant monoclonal antibodies and demonstrate those with the highest rates of non-specific uptake in vitro exhibited elevated target independent clearance, low subcutaneous bioavailability, or both. Our results support the observation that high non-specific endocytosis is a negative attribute in monoclonal antibody development and demonstrate the utility of a generic cell-based screen as a quantitative tool to measure non-specific endocytosis of protein therapeutics at the single-cell level. [ABSTRACT FROM AUTHOR]

The β-sandwich domain 1 (SD1) of islandisin is a stable thermophilic protein with surface loops that can be redesigned for specific target binding, architecturally comparable to the variable domain of immunoglobulin (IgG). SD1's propensity to aggregate due to incorrect folding and subsequent accumulation in Escherichia coli inclusion bodies limits its use in biotechnological applications. We rationally designed SD1 for improved variants that were expressed in soluble forms in E. coli while maintaining the intrinsic thermal stability of the protein (melting temperature (Tm) = 73). We used FoldX's ΔΔG predictions to find beneficial mutations and aggregation-prone regions (APRs) using Tango. The S26K substitution within protein core residues did not affect protein stability. Among the soluble mutants studied, the S26K/Q91P combination significantly improved the expression and solubility of SD1. We also examined the effects of the surface residue, pH, and concentration on the solubility of SD1. We showed that the surface polarity of proteins had little or no effect on solubility, whereas surface charges played a substantial role. The storage stability of several SD1 variants was impaired at pH values near their isoelectric point, and pH levels resulting in highly charged groups. We observed that mutations that create an uneven distribution of charged groups on the SD1 surface could enhance protein solubility by eliminating favorable protein–protein surface charge interactions. Our findings suggest that SD1 is mutationally tolerant to new functionalities, thus providing a novel perspective for the application of rational design to improve the solubility of targeted proteins. [ABSTRACT FROM AUTHOR]

Cardiovascular diseases remain the leading cause of mortality worldwide, with percutaneous coronary interventions (PCI) using drug-eluting stents (DES) as the standard treatment for coronary artery disease. Despite advancements in DES technology, challenges such as restenosis and stent thrombosis still necessitate further interventions and carry considerable risks, highlighting an urgent need for a novel therapeutic approach with lower restenosis rates and reduced reintervention risks. Drug-eluting balloons (DEBs) coated with anti-proliferative agents offer a novel method of local drug delivery for reducing restenosis by directly administering drugs to the lesion site. However, efficient delivery of drugs within the brief procedural window while minimizing drug loss during maneuvering to the target site remains a challenge. Herein, we developed a pressure-responsive and pH-triggered, paclitaxel (PTX) coated DEB catheter for rapid and high-dose delivery of paclitaxel to the lesion sites, overcoming the limitations of drug dispersion and inefficiencies associated with conventional DEBs. Using the charge reversion of albumin around its isoelectric point, paclitaxel-loaded human serum albumin nanoparticles (PTX-HSA-NPs) and heparin were coated using a layer-by-layer deposition method at pH 4.0. Upon exposure to physiological pH (i.e., 7.4), the coating is rapidly released from the balloon surface. Combined with a protective shellac coating, the DEB achieved controlled release of high-dose paclitaxel to the arterial tissue in vivo (up to 400 µg per gram tissue), exceeding current DEBs in the clinic. These results suggest the pH-responsive, multi-layer coated DEB's potential for superior procedural outcomes, addressing a critical unmet need in PCI. [ABSTRACT FROM AUTHOR]

Copyright of Food & Fermentation Industries is the property of Food & Fermentation Industries and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Corrosion of carbon steel rebars in concrete structures significantly compromises their safety, reliability, and environmental performance. This work focuses on enhancing rebar corrosion resistance by repairing the defects of oxide scale. Here, sol-gel was employed as a carrier, by which Ce3+ and Ni2+ were transferred to the oxide scale defects based on the isoelectric point theory, and deposited at these defects with the pH variation as temperature. The repaired oxide scale showed enhanced uniform elemental distribution and improved electrochemical properties, maintaining integrity even under prolonged exposure to Cl−-rich environments. Notably, the impedance modulus at 0.01 Hz (|Z | 0.01Hz) of treated samples was six times higher than untreated ones, indicating superior performance against high electric fields. This strategy shows great potential for enhancing the durability of concrete structures. [ABSTRACT FROM AUTHOR]

AbstractThe venom of the Odontobuthus doriae scorpion, prevalent in East Asia and Iran, has not been fully characterized. This study provides the first proteomic profile of O. doriae venom to explore its potential as a medical. 2D-PAGE analysis revealed 96 protein spots with isoelectric points from 3 to 9 and molecular weights between 6.6 to 205 kDa. Fourteen toxin fractions were isolated via HPLC, and SDS-PAGE showed seven protein bands ranging from 3.8 to 182 kDa. MALDI-TOF MS identified Peptide 1 and Peptide 2, resembling Hemoglobin beta-2 chain and Chaperonin HSP60 and suggest potential therapeutic applications for P1 and P2. [ABSTRACT FROM AUTHOR]

Cat calcium oxalate monohydrate kidney stone matrix proteome showed great similarity to human calcium oxalate monohydrate stone matrix proteome, but inference of mechanistic similarity was limited by the absence of cat urine proteomic data. In this study, urine proteome distributions were measured by the same methods in 7 healthy cats for comparison to both the published human urine and cat calcium oxalate stone matrix proteomes to assess for similar enrichment patterns in both species. Furthermore, proteomic distributions were determined in cat struvite stone matrix to test for similarity to calcium oxalate monohydrate stone matrix and urine proteomes. Cat urine proteins demonstrated a similar distribution of abundance as a function of isoelectric points or net charge to human urine samples, and consequently the similarly altered patterns of protein distributions seen in calcium oxalate monohydrate stone matrix seen from both cat and human stones likely derives from the same preferential adsorption mechanism. Furthermore, the fact that protein abundance patterns seen in cat struvite stone matrix samples differ from both urine and calcium oxalate monohydrate stone matrix proteomes in systematic ways suggests that a combination of protein–protein and protein crystal interactions underly the formation of the crystal aggregates that comprise stones. [ABSTRACT FROM AUTHOR]

Lepidolite, crucial for lithium extraction, is primarily processed through flotation. However, conventional flotation inhibitors pose environmental challenges. This study introduces Fe(III) as a selective and eco-friendly inhibitor in lepidolite flotation. We investigated its impact on flotation performance and interaction mechanisms with feldspar, quartz, and lepidolite. Adsorption studies revealed that dodecylamine (DDA) selectively adsorbs onto lepidolite when Fe(III) is present. Consistent contact angle and flotation results showed reduced hydrophobicity and recovery rates for feldspar and quartz, with minimal impact on lepidolite. Zeta potential measurements indicated lower potentials for feldspar and quartz compared to lepidolite. Notably, the addition of Fe(III) altered the isoelectric points of quartz and feldspar, suggesting stronger Fe(III) adsorption on these minerals. Infrared spectroscopy and X-ray photoelectron spectroscopy confirmed reduced DDA adsorption on feldspar and quartz surfaces due to Fe(III) adsorption, while DDA adsorption on lepidolite remained largely unaffected. The mechanism underlying Fe(III)'s selective inhibition on feldspar and quartz involves their more negative surfaces compared to lepidolite, which facilitates Fe(III) adsorption and inhibits DDA adsorption. This study offers insights into mechanisms relevant to systems using metal ions as depressants, providing valuable references for similar research. [ABSTRACT FROM AUTHOR]

The maize Snf2 gene family plays a crucial role in chromatin remodeling and response to environmental stresses. In this study, we identified and analyzed 35 members of the maize Snf2 gene family (ZmCHR1 to ZmCHR35) using the Ensembl Plants database. Each protein contained conserved SNF2-N and Helicase-C domains. Phylogenetic analysis revealed six groups among the Snf2 proteins, with an uneven distribution across subfamilies. Physicochemical analysis indicated that the Snf2 proteins are hydrophilic, with varied amino acid lengths, isoelectric points, and molecular weights, and are predominantly localized in the nucleus. Chromosomal mapping showed that these genes are distributed across all ten maize chromosomes. Gene structure analysis revealed diverse exon–intron arrangements, while motif analysis identified 20 conserved motifs. Collinearity analysis highlighted gene duplication events, suggesting purifying selection. Cis-regulatory element analysis suggested involvement in abiotic and biotic stress responses. Expression analysis indicated tissue-specific expression patterns and differential expression under various stress conditions. Specifically, qRT-PCR validation under drought stress showed that certain Snf2 genes were upregulated at 12 h and downregulated at 24 h, revealing potential roles in drought tolerance. These findings provide a foundation for further exploration of the functional roles of the maize Snf2 gene family in development and stress responses. [ABSTRACT FROM AUTHOR]

The Bax inhibitor-1 (BI-1) gene family, which is important for plant growth, development, and stress tolerance, remains largely unexplored in cauliflower. In this study, we identified and characterized cauliflower BI-1 family genes. Based on aligned homologous sequences and collinearity with Arabidopsis genes, we identified nine cauliflower BI-1 genes, which encode proteins that varied in length, molecular weight, isoelectric point, and predicted subcellular localization, including the Golgi apparatus, plasma membrane, and various compartments within the chloroplast. Phylogenetic analyses detected evolutionary conservation and divergence among these genes. Ten structural motifs were identified, with Motif 5 found to be crucial for inhibiting apoptosis. According to the cis-regulatory elements in their promoters, these genes likely influence hormone signaling and stress responses. Expression profiles among tissues highlighted the functional diversity of these genes, with particularly high expression levels observed in the silique and root. Focusing on BobBIL4, we investigated its role in brassinosteroid (BR)-mediated root development and salt stress tolerance. BobBIL4 expression levels increased in response to BR and salt treatments. The functional characterization of this gene in Arabidopsis revealed that it enhances root growth and salinity tolerance. These findings provide insights into BI-1 gene functions in cauliflower while also highlighting the potential utility of BobBIL4 for improving crop stress resistance. [ABSTRACT FROM AUTHOR]

This study provides a comprehensive analysis of pathogenesis-related (PR) proteins, focusing on PR1, PR5, and PR10, in three plant species: Arabidopsis thaliana (At), Solanum lycopersicum (Sl), and Solanum tuberosum (St). We investigated various physico-chemical properties, including protein length, molecular weight, isoelectric point (pI), hydrophobicity, and structural characteristics, such as RMSD, using state-of-the-art tools like AlphaFold and PyMOL. Our analysis found that the SlPR10-StPR10 protein pair had the highest sequence identity (80.00%), lowest RMSD value (0.307 Å), and a high number of overlapping residues (160) among all other protein pairs, indicating their remarkable similarity. Additionally, we used bioinformatics tools such as Cello, Euk-mPLoc 2.0, and Wolfpsort to predict subcellular localization, with AtPR1, AtPR5, and SlPR5 proteins predicted to be located in the extracellular space in both Arabidopsis and S. lycopersicum, while AtPR10 was predicted to be located in the cytoplasm. This comprehensive analysis, including the use of cutting-edge structural prediction and subcellular localization tools, enhances our understanding of the structural, functional, and localization aspects of PR proteins, shedding light on their roles in plant defense mechanisms across different plant species. [ABSTRACT FROM AUTHOR]

While bioactivity and a favorable safety profile for biotherapeutics is of utmost importance, manufacturability is also worth of consideration to ease the manufacturing process. Manufacturability in the scientific literature is mostly related to stability of formulated drug substances, with limited focus on downstream process‐related manufacturability, that is, how easily can a protein be purified. Process‐related impurities or biological impurities like viruses and host cell proteins (HCP) are present in the harvest which have mostly acid isoelectric points and need to be removed to ensure patient safety. Therefore, during molecule design, the surface charge of the target molecule should preferably differ sufficiently from the surface charge of the impurities to enable an efficient purification strategy. In this feasibility study, we evaluated the possibility of improving manufacturability by adapting the surface charge of the target protein. We generated several variants of a GLP1‐receptor‐agonist‐Fc‐domain‐FGF21‐fusion protein and demonstrated proof of concept exemplarily for an anion exchange chromatography step which then can be operated at high pH values with maximal product recovery allowing removal of HCP and viruses. Altering the surface charge distribution of biotherapeutic proteins can thus be useful allowing for an efficient manufacturing process for removing HCP and viruses, thereby reducing manufacturing costs. [ABSTRACT FROM AUTHOR]

Increasing the use of plant proteins in foods requires improving their physical and chemical properties, such as emulsification, gelation capacity, and thermal stability. These properties determine the acceptability and functionality of food products. Higher protein solubility significantly impacts these properties by affecting denaturation and the stability of emulsifiers or gels. Therefore, developing plant-based protein ingredients requires accurately and conveniently measuring their solubility. Colorimetric solubility methods overcome many issues of more robust combustion and titration methods, but complicated chemical mechanisms limit their applicability for certain proteins. This study aims to compare the effectiveness of four common colorimetric solubility measurement methods for pulse and non-pulse legume proteins and hydrolysates. Pea, chickpea, lentil, and soy protein isolates were made from defatted flour and their solubility at a range of pHs was measured using the Bradford, Lowry, bicinchoninic acid (BCA), and biuret methods. Solubility was also measured for chickpea and soy protein hydrolysates made using Alcalase and Flavourzyme. A comparison of the methods for solubility quantification revealed that the Bradford and Lowry methods most closely match the expected results for the unhydrolyzed protein, with the BCA and biuret methods underestimating solubility by 30%. The Lowry method was the preferred method for hydrolysate solubility measurement, with the Bradford method measuring 0% solubility at the isoelectric point due to an inability to interact with peptides that are soluble at this pH. This study identifies reliable methods for measuring plant protein solubility that establish uniform outcomes and enable a better comparison across studies, giving a consensus for key functional properties in food applications. [ABSTRACT FROM AUTHOR]

[Display omitted] The structural details of foams made with pea albumins are affected by the pH of the initial solution and followed heat treatment. An in situ, time-resolved investigation of foams prepared with pea albumins was conducted using small-angle neutron scattering (SANS) in combination with imaging and conductance measurements. Solutions were tested at pH three pH values (3, 4.5, and 8) before and after heating (90 °C for 1 and 5 min). The characteristic structures present in the foam from the nano to the meso-scale differed during drainage depending on solution pH. Foams obtained at pH 3, had the largest bubble radius and thinnest plateau border, as well as the highest extent of liquid drainage. At pH 4.5, close to the isoelectric point of the proteins, foams displayed similar bubbles' behavior to those at pH 8, but with the largest film thickness. In this case, the proteins were extensively aggregated. Heating of the solutions prior to foaming did not significantly affect the foam aging regardless of pH. The quantification of specific surface areas and film thickness over time without sample disruption shows to be a powerful approach to designing foam structures. [ABSTRACT FROM AUTHOR]

An efficient synthesis of spiro[2.3]hexane‐ and spiro[3.3]heptane‐derived α‐amino acids is reported. The used methods are divided on two different approaches. First of them involves modification of commercially available spirocyclic diesters via monohydrolysis, Curtius rearrangement and functional groups deprotection sequence. Alternative approach was based on Bucherer ‐ Bergs hydantoine synthesis with its subsequent cleavage to the target amino acid fragment. The proposed synthetic schemes offer the desired product in 31–52 % overall yield on up to 22.5 g scale. Acid‐base titration of the obtained spirocyclic amino acids revealed lowered pI values compared to the parent 1‐amino‐cyclohexane‐ and ‐cycloheptanecarboxylic acids. [ABSTRACT FROM AUTHOR]

Immunoglobulin G (IgG) molecules that bind antigens on the membrane of target cells spontaneously form hexameric rings, thus recruiting C1 to initiate the complement pathway. However, our previous report indicated that a mouse IgG mutant lacking the Cγ1 domain activates the pathway independently of antigen presence through its monomeric interaction with C1q via the CL domain, as well as Fc. In this study, we investigated the potential interaction between C1q and human CL isoforms. Quantitative single-molecule observations using high-speed atomic force microscopy revealed that human Cκ exhibited comparable C1q binding capabilities with its mouse counterpart, surpassing the Cλ types, which have a higher isoelectric point than the Cκ domains. Nuclear magnetic resonance and mutation experiments indicated that the human and mouse Cκ domains share a common primary binding site for C1q, centred on Glu194, a residue conserved in the Cκ domains but absent in the Cλ domains. Additionally, the Cγ1 domain, with its high isoelectric point, can cause electrostatic repulsion to the C1q head and impede the C1q-interaction adjustability of the Cκ domain in Fab. The removal of the Cγ1 domain is considered to eliminate these factors and thus promote Cκ interaction with C1q with the potential risk of uncontrolled activation of the complement pathway in vivo in the absence of antigen. However, this research underscores the presence of potential subsites in Fab for C1q binding, offering promising targets for antibody engineering to refine therapeutic antibody design. [ABSTRACT FROM AUTHOR]

The objective of this study was to investigate the effect of verjuice on beef M. biceps femoris (BF). BF blocks were marinated with 30%, 70%, and 100% verjuice solutions containing 2% NaCl, for different marination times (12, 24, 48, and 72 h). Verjuice marination reduced the pH values of BF samples from 6.77 in control sample to 3.66 in 100% of verjuice for 72 h. The decreased values of water holding capacity (from 54.06% to 47.46%) with increasing verjuice concentration (from 30% to 100% for 72 h) confirmed the drop of proteins isoelectric point of the muscle due to salt presence preventing fibers swelling. Less cookout was observed with increasing acid concentration. Marination time had no significant effect on L* and a* coordinates of uncooked samples while acidification made the samples lighter and less red. Enzymatic proteolysis of myosin and troponin‐T concomitant with increase in myofibrillar fragmentation index contributed to the decrease of shear force in a way dependent on verjuice concentration and marination time. Sensory panelists gave the highest score to cooked samples marinated with 70% verjuice solution. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Journal of Chinese Institute of Food Science & Technology is the property of Journal of Chinese Institute of Food Science & Technology Periodical Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this study, proteins were prepared from Schizochytrium pombe residue after oil extraction using isoelectric point precipitation, and their physicochemical and emulsifying properties were investigated. Our objective was to assess the suitability of these proteins for functional ingredient applications. Through a one-way experiment and optimization using response surface design, the effects of time, temperature, pH, and the material–liquid ratio of NaOH alkaline extraction were explored. The isoelectric point is verified by isoelectric point precipitation; the results revealed that crude protein from Schizochytrium (SCP) is minimally soluble at pH 4.2. Compared with whey protein (WP), it promotes better emulsion stability through the emulsification test. This study suggests that Schizochytrium oil-processing byproducts represent a promising source of protein, with potential applications as functional ingredients, offering implications for the usage of these byproducts in various industries. [ABSTRACT FROM AUTHOR]

The impact of zwitterionic graphene oxide (GO‐Arg) nanostructures into polyethersulfone (PES) membranes was studied, aiming to perform mixed matrix membranes (MMMs) with combined antibacterial and antifouling properties. For this purpose, 0.25, 0.50, and 1.0 wt% of GO‐Arg were incorporated through the phase inversion method. The chemical composition reveals that the electrostatic interactions of PES, polyvinylpyrrolidone, and GO‐Arg occur by enhancing the CH/NH vibrations associated with the intrinsic MMMs and their combined antibacterial and antifouling performance. Well‐defined finger‐like morphology was observed, performing the pore distribution and the finger‐cavity inclination induced by the electrostatic charge of GO‐Arg. As a result, GO‐Arg MMM's hydrophobic properties decreased from 77.8° to 52.3°, inducing a partially hydrophilic surface and improved water flux. The isoelectric point of nanostructured membranes slightly changes by the charge modulation produced by zwitterionic GO‐Arg. The antibacterial activity against Gram (−) Escherichia coli was improved, closing the complete bacterial inhibition after 24 h. As a result, antifouling performance was improved, avoiding bacterial adhesion into the membrane surface and the possibility of promoting biofilm formation. Our results demonstrate that zwitterionic nanomaterials in MMMs perform the multifunctional capacity of materials, emerging as an effective alternative with potential application in wastewater treatment. Highlights: The incorporation of zwitterionic GO‐Arg improves the mechanical properties of MMMs.GO‐Arg‐enhanced well‐defined porous finger‐like morphology in MMMs.GO‐Arg improves the antibacterial/antifouling performance of PES‐based MMMs. [ABSTRACT FROM AUTHOR]

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By means of molecular dynamics computer simulation, the conformational space of polyampholyte macromolecules with various distributions of the charged groups along the chain is studied. A coarse‐grained model where each monomer unit of the chain is presented as a non‐charged group in the backbone of the macromolecule connected with a charged side pendant is considered. A limiting case of fully charged chains in the isoelectric point is investigated. The oppositely charged monomer units are distributed in various patterns: regular alternating, multiblock, or random sequences. It is found that the chains with random unit distribution adopt much more compacted conformations than the chains with regular distributions with comparable block lengths. Calculating the chain size and its fluctuation along with the spatial density distribution, coil, and globular conformations are distinguished and arranged on the diagrams in terms of chain length, block length, and Bjerrum length. [ABSTRACT FROM AUTHOR]

Many food proteins can be assembled into nanofibrils under pH conditions far from the isoelectric point and with a low ionic strength by heating them for a long period. These food protein nanofibrils (FPN) have outstanding functional and biological properties and are considered sustainable biomaterials in many fields. In this study, we review the recent developments in FPN gels and introduce the key factors in promoting food protein self-assembly in order to create functional gels. The major variables discussed are the morphology of nanofibrils, protein concentration, heating time, and the type and concentration of salts. We also highlight current advances in the formation and properties of different types of FPN gels. In addition, the various applications of FPN gels in bioactive and nutrient delivery, adsorbents for CO2 and toxic pollutants, cell scaffolding biomaterials, biosensors, and others are introduced and discussed. [ABSTRACT FROM AUTHOR]

Protein‐based hydrogels have great potential to be used as bioinks for biofabrication‐driven tissue regeneration strategies due to their innate bioactivity. Nevertheless, their use as bioinks in conventional 3D bioprinting is impaired due to their intrinsic low viscosity. Using embedding bioprinting, a liquid bioink is printed within a support that physically holds the patterned filament. Inspired by the recognized microencapsulation technique complex coacervation, crystal self‐healing embedding bioprinting (CLADDING) is introduced based on a highly transparent crystal supporting bath. The suitability of distinct classes of gelatins is evaluated (i.e., molecular weight distribution, isoelectric point, and ionic content), as well as the formation of gelatin‐gum arabic microparticles as a function of pH, temperature, solvent, and mass ratios. Characterizing and controlling this parametric window resulted in high yields of support bath with ideal self‐healing properties for interaction with protein‐based bioinks. This support bath achieved transparency, which boosted light permeation within the bath. Bioprinted constructs fully composed of platelet lysates encapsulating a co‐culture of human mesenchymal stromal cells and endothelial cells are obtained, demonstrating a high‐dense cellular network with excellent cell viability and stability over a month. CLADDING broadens the spectrum of photocrosslinkable materials with extremely low viscosity that can now be bioprinted with sensitive cells without any additional support. [ABSTRACT FROM AUTHOR]

Ophibolin A, a fungal sesterterpene, exerts a pivotal influence in a diverse array of biological processes, encompassing herbicidal, bactericidal, fungicidal, and cytotoxic activities. Sixty genes associated with sesterterpene compound biosynthesis were obtained from Bipolaris eleusines via transcriptome sequencing, and those closely linked to ophiobolin A biosynthesis were subsequently filtered. A gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) that catalyzes the first committed step of ophiobolin biosynthesis in the mevalonic acid (MVA) pathway was isolated and characterized using RACE (Rapid Amplification of cDNA Ends) technology from ophiobolin A-producing fungus, B. eleusines. The full-length cDNA of the B. eleusines HMGR gene (BeHMGR) was 3906 bp and contained a 3474 bp open reading frame (ORF) encoding 1157 amino acids. Sequence analysis revealed that deduced BeHMGR had high homology to the known HMGRs from Pyrenophora tritici-repentis and Leptosphaeria maculans. It had a calculated molecular mass of about 124.65 kDa and an isoelectric point (pI) of 6.90. It contained two putative HMG-CoA-binding motifs and two NADP(H)-binding motifs. Induced expression analysis of the BeHMGR gene by methyl jasmonate treatment using quantitative fluorescence PCR showed that it significantly elevated after 3 h of methyl jasmonate treatment, peaked at 6 h, and then gradually decreased. This demonstrates that BeHMGR gene expression is induced by methyl jasmonate. [ABSTRACT FROM AUTHOR]

The present study examines the impacts of saponin, extracted from the fruits of Acacia Concinna, in its effectiveness as a dispersant and stabilizer during the pipeline conveyance of iron ore particles. The basic fundamental characteristics of iron ore, which comprises a majority of hematite and has a size of (≤75 µm), have been examined and characterized. Iron ore water slurry (IWS) rheological behavior has been investigated as a function of saponin doses, pH, particle concentration, and the shear rate-shear stress correlation. The change in zeta potential with pH is used to compute the isoelectric point of IWS. An appropriate mechanism has been presented to explicate the interaction between saponin and particles of Iron ore. The surface-modified particles exhibit improved dispersion characteristics in the aqueous solution due to the modification of electrostatic force that the surface charge on each particle causes. Investigation indicates that the optimal proportion of saponin had a significant favorable effect on the fluidity and stability of the suspension. The analysis of the economic impact of the dispersant has been conducted through the evaluation of slurry head loss and specific energy consumption (SEC) pertaining to the implementation of the IWS pipeline in its intended commercial application. [ABSTRACT FROM AUTHOR]

Expansin(EXP)plays an important role in plant response to environmental stress by regulating cell wall relaxation. To explore the role of EXP genes in soybean response to abiotic stress, two soybean EXP genes (GmEXPB5 and GmEXPB7) and their protein sequences were analyzed by bioinformatics, and their expression levels were detected by real-time fluorescent quantitative PCR(qRT-PCR). The results were as follows: (1)The GmEXPB5 and GmEXPB7 were located on chromosomes 10 and 12 of soybean, and encoded proteins containing 272 and 267 amino acids, respectively. The molecular weight of GmEXPB5 protein was 29.07 kD and the theoretical isoelectric point was 7.51. The molecular weight of GmEXPB7 protein was 29.09 kD and the theoretical isoelectric point was 8.66. Both GmEXPB5 and GmEXPB7 were stable hydrophilic proteins, and localized in the cell wall. Both GmEXPB5 and GmEXPB7 proteins contained a signal peptide sequence and a conserved DPBB_1 structural domain. (2)GmEXPB5 protein had the closest affinity with CaEXPB15 protein of Cicer arietinum, and GmEXPB7 protein was closely related to EXPB3 proteins of Spatholobus suberectus, Vigna angularis and V. unguiculata. (3)GmEXPB5 and GmEXPB7 were expressed in root, stem and leaf of soybean, and their expression levels in root and leaf were significantly higher than those in stem. (4)GmEXPB5 and GmEXPB7 could respond to salt, drought and cold stresses in soybean seedlings. (5)The promoter region of GmEXPB5 contained two types of stress-related cis-elements(ABRE and ARE). The promoter region of GmEXPB7 contained five types of stress-related cis-elements(ABRE, ARE, CGTCA-motif, TC-rich repeats and MBS). In conclusion, these results indicate that GmEXPB5 and GmEXPB7 can participate in the response of soybean to abiotic stress. [ABSTRACT FROM AUTHOR]

The illustration highlights thermally resilient, phase-invertible, ultra-stable water-in-water (W/W) droplets by pH-modulated casein colloidal particles. It includes radar plots contrasting the robustness of W/W droplets stabilized by casein colloidal particles with those stabilized by other protein/polymer particles reported in literature (see also Table S1). [Display omitted] The essence of compartmentalization in cells is the inspiration behind the engineering of synthetic counterparts, which has emerged as a significant engineering theme. Here, we report the formation of ultra-stable water-in-water (W/W) emulsion droplets. These W/W droplets demonstrate previously unattained stability across a broad pH spectrum and exhibit resilience at temperatures up to 80℃, overcoming the challenge of insufficient robustness in dispersed droplets of aqueous two-phase systems (ATPS). The exceptional robustness is attributed to the strong anchoring of micelle-like casein colloidal particles at the PEO/DEX interface, which maintains stability under varying environmental conditions. The increased surface hydrophobicity of these particles at high temperatures contributes to the formation of thermally-stable droplets, enduring temperatures as high as 80℃. Furthermore, our study illustrates the adaptable affinity of micelle-like casein colloidal particles towards the PEO/DEX-rich phase, enabling the formation of stable DEX-in-PEO emulsions at lower pH levels, and PEO-in-DEX emulsions as the pH rises above the isoelectric point. The robust nature of these W/W emulsions unlocks new possibilities for exploring various biochemical reactions within synthetic subcellular modules and lays a solid foundation for the development of novel biomimetic materials. [ABSTRACT FROM AUTHOR]

【Objective】The present paper aimed to investigate the important role of NAC gene in the growth and development of Cinnamomum camphora and its role in salt stress, and to clarify the response mechanism of C. camphora under salt stress. 【Method】The genome-wide NAC genes of C. camphora were identified and analyzed by bioinformatics to investigate the mode of response to abiotic stress in C. camphora. By identifying the members of NAC family of C. camphora,the paper aonducteda systematic analysis in terms of physicochemical properties, phylogenetic analysis, conserved motifs and gene structures, chromosomal localization and inter-species covariance, cis-acting element prediction and expression patterns.【Result】A total of 103 CcNAC genes were identified, the amino acid counts ranged from 137 to 1136, the isoelectric points from 4.54 to 10.00, the relative molecular masses from 15 644.40 to 129 859.29 u, and the mean hydrophobicity of all proteins was negative, indicating hydrophilicity. Subcellular localization was predicted to be in the nucleus and cytoplasm. A phylogenetic evolutionary tree was constructed with Arabidopsis thaliana NAC gene sequences, which classified CcNAC genes into 16 subfamilies. The NAM subfamily had the most members, with 17 CcNAC genes, while 25 CcNAC genes were not included. Conservative motif and gene structure analysis revealed that CcNAC genes were found to contain 2-8 introns and 10 motifs. Among these, motif3 appeared to be particularly stable in the genetic evolution of C. camphora genes, while motif9 and motif10 were distributed across the NAM subfamily. Chromosome localization and replication event analysis revealed that 103 genes were distributed across 12 chromosomes, with the highest number of genes（13） found in CcNAC genes distributed across chromosomes Chr02, with the lowest number of genes found on Chr10 and Chr11, which contained two genes each; Fragment duplication was the main mode of amplification for the CcNAC family. Multi-species co-linkage analysis revealed that CcNAC genes formed 162 pairs of co-linkage genes with CcNAC genes, indicating a closer relationship than with other species. The CcNAC genes were more closely related to the CcNAC genes and to those of other species than to any other. The CcNAC gene promoters contained several abiotic stress, growth and development, and hormone response elements. The results of the qRT-PCR showed that in leaves, the expression levels of the CcNAC058 and CcNAC092 genes were maximised under low salt stress. In roots, the expression of CcNAC007, CcNAC033, CcNAC058 and CcNAC092 genes increased with increasing salt concentration.【Conclusion】The NAC gene family may be involved in the process of salt stress response during the growth and development of C. camphora. The CcNAC092 and CcNAC058 genes exhibite significant responses in both leaves and roots under salt stress. It is hypothesised that CcNAC058 and CcNAC092 are the key genes contributing to the ability of the CcNAC to produce self-defensive behaviours under salt stress. [ABSTRACT FROM AUTHOR]

The physicochemical properties of high-molecular weight compounds based on N,N-diallyl-N′-acyllhydrazines were studied. Based on the results of thermal analysis it was established that sorbents are stable up to 160–210 °C; the destruction of the initial N,N-diallyl-N′-acyllhydrazines starts at lower temperatures. The stability of sorbents in various media, the values of static exchange capacities for hydrogen ions and hydroxide ions ( SEC H + and SEC OH − ), and isoelectric points of the surface were determined. Samples for further study of the rare earth metals sorption were selected. [ABSTRACT FROM AUTHOR]

Due to the growing demand for human-edible protein sources, microalgae are recognized as an economically viable alternative source of proteins. The investigation into the structural characteristics and functional properties of microalgin is highly significant for its potential application in the food industry as an alternative source of protein. In this research, we extracted protein from Euglena gracilis by using alkaline extraction and acid precipitation and investigated its structural characteristics and functional properties in different acidic and alkaline environments. The molecular weight distribution of Euglena gracilis protein (EGP), as revealed by the size exclusion chromatography results, ranges from 152 to 5.7 kDa. EGP was found to be rich in hydrophobic amino acids and essential amino acids. Fourier infrared analysis revealed that EGP exhibited higher α-helix structure content and lower β-sheet structure content in alkaline environments compared with acidic ones. EGP exhibited higher foaming properties, emulsifying activity index, solubility, free sulfhydryl, and total sulfhydryl in pH environments far from its isoelectric point, and lower fluorescence intensity (2325 A.U.), lower surface hydrophobicity, larger average particle size (25.13 µm), higher emulsifying stability index, and water-holding capacity in pH environments near its isoelectric point. In addition, X-ray diffraction (XRD) patterns indicated that different acidic and alkaline environments lead to reductions in the crystal size and crystallinity of EGP. EGP exhibited high denaturation temperature (Td; 99.32 °C) and high enthalpy (ΔH; 146.33 J/g) at pH 11.0, as shown by the differential scanning calorimetry (DSC) results. The findings from our studies on EGP in different acidic and alkaline environments provide a data basis for its potential commercial utilization as a food ingredient in products such as emulsions, gels, and foams. [ABSTRACT FROM AUTHOR]

Two‐dimensional electrophoresis (2DE) is a gel‐based protein separation method based on size and charge which is commonly used for the characterization of host cell proteins (HCPs) during drug development in biotech and pharmaceutical companies. HCPs are a heterogenous mixture of proteins produced by host cells during a biologics drug manufacturing process. Different gel electrophoresis methods including traditional 2D SDS‐PAGE with silver and SYPRO Ruby fluorescent dye staining as well as two‐dimensional difference gel electrophoresis (2D‐DIGE) were compared for their relative abilities to characterize HCPs. SYPRO Ruby was shown to be more sensitive than silver stain in the traditional 2D gels both with and without product protein present. Silver stain also displayed a significant preference for staining acidic proteins over basic ones while SYPRO Ruby was more consistent in imaging proteins across different isoelectric points. The non‐traditional method of 2D‐DIGE provides high resolution and reproducibility when comparing samples with similar protein profiles but was limited in imaging HCP spots due to its narrow dynamic range. Overall, 2DE is a powerful tool to separate and characterize HCPs and is optimized by choosing the best stain or method for each specific application. Using a combination of two or more different 2DE staining methods, when possible, provides the most comprehensive coverage to support the characterization of a complex mixture like HCPs. However, in instances where only one staining method can be used, SYPRO Ruby is shown to be the more reliable, more sensitive, and easier to use traditional staining method for most HCP‐based applications. [ABSTRACT FROM AUTHOR]

Monoclonal antibodies (mAbs) are an essential class of biotherapeutics. A platform process is used for mAb development to ensure clinically safe and stable molecules. Regulatory authorities ensure that mAb production processes include sufficient viral clearance steps to achieve less than one virus particle per million doses of product. Virus filtration is used for size‐based removal of enveloped and nonenveloped viruses during downstream processing of mAbs. Process development in mAb purification relies on empirical approaches and often includes adsorptive prefiltration to mitigate virus filter fouling. Opportunities for molecular‐level prediction of mAb filterability are needed to plug the existing knowledge gap in downstream processing. A molecular‐level approach to understanding the factors influencing mAb filterability may reduce process development time, material loss, and processing costs due to oversized virus filters. In this work, pH step gradient fractionation was applied on polished bulk mAb feed to obtain concentrated pools of fractionated mAb variants. Biophysical properties and quality attributes of fractionated pools, including oligomeric state (size), isoelectric point profile, diffusion interaction parameters, and glycoform profile, were determined using bioanalytical methods. Filterability (loading and throughput) of fractionated pools were evaluated. Statistical methods were used to obtain correlations between quality attributes of mAb fractions and filterability on the Viresolve Pro virus filter. [ABSTRACT FROM AUTHOR]

Cytokines are small signaling proteins that regulate the immune responses to infection and tissue damage. Surface charges of cytokines determine their in vivo fate in immune regulation, e.g., half-life and distribution. The overall negative charges in the extracellular microenvironment and the acidosis during inflammation and infection may differentially impact cytokines with different surface charges for fine-tuned immune regulation via controlling tissue residential properties. However, the trend and role of cytokine surface charges has yet to be elucidated in the literature. Interestingly, we have observed that most pro-inflammatory cytokines have a negative charge, while most anti-inflammatory cytokines and chemokines have a positive charge. In this review, we extensively examined the surface charges of all cytokines and chemokines, summarized the pharmacokinetics and tissue adhesion of major cytokines, and analyzed the link of surface charge with cytokine biodistribution, activation, and function in immune regulation. Additionally, we identified that the general trend of charge disparity between pro- and anti-inflammatory cytokines represents a unique opportunity to develop precise immune modulation approaches, which can be applied to many inflammation-associated diseases including solid tumors, chronic wounds, infection, and sepsis. [Display omitted] • Majority of pro-inflammatory cytokines are negatively charged. • Majority of anti-inflammatory cytokines are positively charged. • Charge disparity may indirectly regulate cytokines in inflammation via altering their tissue residence and half-lives. • Targeting the charge disparity of cytokines may provide a novel mechanism for immune modulation. [ABSTRACT FROM AUTHOR]

Fructose-1,6-biphosphate aldolase (FBA) genes have important roles in plant stress responses. At the same time, these genes positively affect growth and development in plants. FBA is involved in gluconeogenesis, glycolysis, and the Calvin-Benson cycle, and it is an enzyme that plays an important role in signal transduction of these stages. This study aims to determine and characterize the FBA gene family in the bean genome. As a result of the study, 7 Pvul-FBA genes were determined in the bean (Phaseolus vulgaris L.) genome. The highest amino acid number of Pvul-FBA proteins was determined in the Pvul-FBA-1 gene (1374), and the highest molecular weight (43.03 kDa) was determined in the Pvul-FBA-7 gene. Again, the highest isoelectric point (8.03) was determined in the Pvul-FBA-3 gene. It has been determined that the Pvul-FBA-6/Pvul-FBA-7 genes are segmental duplicated genes. The main four groups were obtained according to the phylogenetic analysis consisting of FBA proteins of three plants (P. vulgaris, Glycine max, and Arabidopsis thaliana). As a result of interproscan analysis, Motif-1, 2, 3, 4 and 5 were found to contain the fructosebisphosphate aldolase domain. According to in silico gene expression analysis, it was determined that the expression rates of Pvul-FBA genes increased or decreased under salt and drought stress conditions. Synteny analyses of FBA genes in common bean and A. thaliana plants showed that these three plants have a relationship in terms of FBA genes. The results of this research will allow a better designation of the molecular structure of the FBA gene family in common bean. [ABSTRACT FROM AUTHOR]

Anthocyanidin reductase (ANR) is a key enzyme regulating anthocyanin synthesis and accumulation in plants. Here, lychee ANR genes were globally identified, their sequence and phylogenetic characteristics were analyzed, and their spatiotemporal expression patterns were characterized. A total of 51 ANR family members were identified in the lychee genome. The length of the encoded amino acid residues ranged from 87 aa to 289 aa, the molecular weight ranged from 9.49 KD to 32.40 KD, and the isoelectric point (pI) ranged from 4.83 to 9.33. Most of the members were acidic proteins. Most members of the LcANR family were located in the cytoplasm. The 51 LcANR family members were unevenly distributed in 11 chromosomes, and their exons and motif conserved structures were significantly different from each other. Promoters in over 90% of LcANR members contained anaerobically induced response elements, and 88% contained photoresponsive elements. Most LcANR family members had low expression in nine lychee tissues and organs (root, young leaf, bud, female flower, male flower, pericarp, pulp, seed, and calli), and some members showed tissue-specific expression patterns. The expression of one gene, LITCHI029356.m1, decreased with the increase of anthocyanin accumulation in 'Feizixiao' and 'Ziniangxi' pericarp, which was negatively correlated with pericarp coloring. The identified LcANR gene was heterologously expressed in tobacco K326, and the function of the LcANR gene was verified. This study provides a basis for the further study of LcANR function, particularly the role in lychee pericarp coloration. [ABSTRACT FROM AUTHOR]

[Objective] The expression pattern and protein characteristics of the B-galactosidase gene NIBGAL from Nicotiana tabacum were explored to provide a basis for the future development of tobacco as a versatile platform material. (Method) Through comparing the homologous between the gene sequences of N. tabacum K326 and NbBGALI gene of N. benthamiana, NIBGAL gene was cloned through PCR by taking cDNA of K326 as the template. Subsequently, bioinformatics tools were used to analyze its gene structure and protein physicochemical properties. The gene expression pattern in different tobacco tissues (roots, stems, leaves and flowers) was examined using qRT-PCR. Finally, the research on prokaryotic expression, protein purification and protein characterization of the gene was carried out. (Result) The cloned tobacco NIBGAL gene obtained in the study exhibited the highest homology with the NbBGALI gene in N. benthamiana, and both possessed identical structural domains (Motif). The gene was located on chromosome 9, and it contained 18 introns. The mRNA length of the gene was 6649 bp. The CDS was 2541 bp encoding 846 amino acids. The enzyme belonged to the GH35 family. The NtBGAL protein had a molecular weight of 92.44 kDa, a theoretical isoelectric point (pl) of 6.8, GRAVY score of -0.222, and was localized on the cell wall. There was a significant difference in gene expression levels of NIBGAL in different tissues of N. tabacum, followed by flower > leaf > stem > root. The prokaryotic expression was performed at 37 °C, NtBGAL protein had obvious expression, and the higher purified NIBGAL protein was obtained by purification method of inclusion body. The optimal temperature for the enzymatic reaction of NtBGAL protein was 30-40 °C and the optimal pH was 4.0-6.5. NIBGAL enzyme activity was enhanced at Mn2+ concentrations ranging from 0.05 to 0.15 mmol/L. [Conclusion] The study clarifies the tissue expression pattern and protein characterization of the NIBGAL gene, which lays the foundation for further modification of the NIBGAL gene to produce humanized glycoproteins in N. tabacum. [ABSTRACT FROM AUTHOR]

[Objective] The ZF-HD (Zinc finger-homeodomain) transcriptional factors of hemp were identified and their expression patterns were analyzed to provide theoretical reference for ZF-HD function research and improvement of hemp genetic traits. (Method) The gene structures, protein physicochemical properties, protein phylogenetic relationships and promoter cis-elements of five ZF-HD transcription factors in hemp were analyzed by bioinformatics, and the gene expression levels were analyzed by real-time fluorescent quantitative PCR (qRT-PCR). [Result] Five CsZF-HD genes were distributed on two chromosomes of hemp genome. Their coding sequence lengths were 783-1185 bp, encoding 260-394 amino acids residues. The molecular weights of five CsZF-HD proteins were 27.95-43.33 ku, and the theoretical isoelectric points were 7.46-8.87. Five CsZF-HD proteins all contained a conserved ZF-HD_dimer domain and were unstable hydrophilic proteins localized in the nucleus. The ZF-HD protein family was divided into three subfamilies (A, B and C). CsZF-HD1 and CsZF-HD2 be- long to the A subgroup, CsZF-HD3 belongs to the C subgroup, and CsZF-HD4 and CsZF-HD5 belong to the B subgroup. The expression levels of CsZF-HDI and CsZF-HD2 in stems and leaves were not significantly different, but the expression levels in roots were the lowest. The expression level of CsZF-HD3 was the highest in stems and the lowest in leaves. The expression of CsZF-HD4 was the highest in leaves and the lowest in roots. The expression level of CsZF-HD5 was the highest in roots and the lowest in stems. All the five CsZF-HD genes were responsive to salt and drought stresses, and the expression levels of CsZF-HD1 and CsZF-HD3 were most significantly increased under salt and drought stresses. The promoters of five CsZF-HD genes contained different types and quantities of stress-related cis-elements. [Conclusion] ZF-HD transcription factors may play a role in the transcriptional regulation of hemp in response to salt and drought stresses. [ABSTRACT FROM AUTHOR]

Graphitic carbon nitride (g-C3N4) is an intriguing nanomaterial that exhibits photoconductive fluorescence properties under UV–visible light. Dopamine (DA) coating of g-C3N4 prepared from melamine was accomplished via self-polymerization of DA as polydopamine (PDA). The g-C3N4 was coated with PDA 1, 3, and 5 times repeatedly as (PDA@g-C3N4) in tris buffer at pH 8.5. As the number of PDA coatings was increased on g-C3N4, the peak intensity at 1512 cm−1 for N–H bending increased. In addition, the increased weight loss values of PDA@g-C3N4 structures at 600 °C from TGA thermograms confirmed that the coating was accomplished. The band gap of g-C3N4, 2.72 eV, was reduced to 0.87 eV after five coatings with PDA. A pristine g-C3N4 was found to have an isoelectric point (IEP) of 4.0, whereas the isoelectric points of 1PDA@g-C3N4 and 3PDA@g-C3N4 are close to each other at 3.94 and 3.91, respectively. On the other hand, the IEP of 5PDA@g-C3N4 was determined at pH 5.75 assuming complete coating with g-C3N4. The biocompatibility of g-C3N4 and PDA@g-C3N4 against L929 fibroblast cell lines revealed that all PDA@g-C3N4 coatings were found to be biocompatible up to a 1000 mg/mL concentration, establishing that PDA coatings did not adversely affect the biocompatibility of the composite materials. In addition, PDA@g-C3N4 was screened for antioxidant potential via total phenol content (TPC) and total flavonoid content assays and it was found that PDA@g-C3N4 has recognizable TPC values and increased linearly with an increased number of PDA coatings. Furthermore, blood compatibility of pristine g-C3N4 is enhanced considerably upon PDA coating, affirmed by hemolysis and the blood clotting index%. Additionally, α-glucosidase inhibitory properties of PDA@g-C3N4 structures revealed that 67.6 + 9.8% of this enzyme was evenly inhibited by 3PDA@g-C3N4 structure. [ABSTRACT FROM AUTHOR]

The viscoelasticity of monoclonal antibodies (mAbs) is important during their production, formulation, and drug delivery. High concentration mAbs can provide higher efficacy therapeutics (e.g., during immunotherapy) and improved efficiency during their production (economy of scale during processing). Two humanized mAbs were studied (mAb-1 and mAb-2) with differing isoelectric points. Using high speed particle tracking microrheology, we demonstrated that the mAb solutions have significant viscoelasticities above concentrations of 40 mg/ml. Power law viscoelasticity was observed over the range of time scales ( 10 − 4 –1 s) probed for the high concentration mAb suspensions. The terminal viscosity demonstrated an exponential dependence on mAb concentration (a modified Mooney relationship) as expected for charged stabilized Brownian colloids. Gelation of the mAbs was explored by lowering the pH of the buffer and a power law scaling of the gelation transition was observed, i.e., the exponent of the anomalous diffusion of the probe particles scaled inversely with the gelation time. [ABSTRACT FROM AUTHOR]

Baby alpaca fur industry generates considerable wastewater during the soaking process, which contains high levels of total suspended solids (TSSs), proteins, and salts, among other components. The valorization of wastewater after precipitation, coagulation–flocculation, and aeration treatments was evaluated for use in irrigation water, fertigation, groundwater recharge, concrete construction, and disposal. The precipitation treatment sludge and the coagulation–flocculation treatment were evaluated as a protein source, soil quality improvement, and disposal. The treatment system included evaluations of nine pH levels, seven coagulant doses, and seven aeration times. The contents of TSSs, chemical oxygen demand (COD), total Kjeldalh nitrogen (TKN), ammonia nitrogen (N–NH3), and oils and fats (O&G), among other parameters, were determined in the treated and untreated wastewater. Before entering the treatment system, the physicochemical characterization of the wastewater showed a high concentration of parameters related to organic matter and dust, such as O&G, five-day biological oxygen demand (BOD5), COD, TSSs, TKN, and N–NH3. The optimal removal parameters were pH 12 for the chemical precipitation of proteins, a dose of 480 mg/L FeCl3 as a coagulant for TSSs removal, and 150 min of aeration; removal efficiencies of 99.02 %, 77.49 %, 79.93 %, and 64.62 % for TSSs, Cod, TKN, and N–NH3, respectively, were obtained. The wastewater after treatment can be used for groundwater recharge and concrete construction, and the wastewater with 2 % dilution can be used for irrigation water and fertigation. The sludge after precipitation is rich in protein and can be used as a protein source or soil quality improver.

ABSTRACTTherapeutic mAbs show a specific “charge fingerprint” that may affect safety and efficacy, and, as such, it is often identified as a critical quality attribute (CQA). Capillary iso-electric focusing (cIEF), commonly used for the evaluation of such CQA, provides an analytical tool to investigate mAb purity and identity across the product lifecycle. Here, we discuss the results of an analysis of a panel of antibody products by conventional and whole-column imaging cIEF systems performed as part of European Pharmacopoeia activities related to development of “horizontal standards” for the quality control of monoclonal antibodies (mAbs). The study aimed at designing and verifying an independent and transversal cIEF procedure for the reliable analysis of mAbs charge variants. Despite the use of comparable experimental conditions, discrepancies in the charge profile and measured isoelectric points emerged between the two cIEF systems. These data suggest that the results are method-dependent rather than absolute, an aspect known to experts in the field and pharmaceutical industry, but not suitably documented in the literature. Critical implications from analytical and regulatory perspectives, are herein thoughtfully discussed, with a special focus on the context of market surveillance and identification of falsified medicines.