Soluble Protein Research Articles

Evaluation of Alpha-Synuclein and Tau Antiaggregation Activity of Urea and Thiourea-Based Small Molecules for Neurodegenerative Disease Therapeutics.

Alzheimer's disease (AD) and Parkinson's disease (PD) are multifactorial, chronic diseases involving neurodegeneration. According to recent studies, it is hypothesized that the intraneuronal and postsynaptic accumulation of misfolded proteins such as α-synuclein (α-syn) and tau, responsible for Lewy bodies (LB) and tangles, respectively, disrupts neuron functions. Considering the co-occurrence of α-syn and tau inclusions in the brains of patients afflicted with subtypes of dementia and LB disorders, the discovery and development of small molecules for the inhibition of α-syn and tau aggregation can be a potentially effective strategy to delay neurodegeneration. Urea is a chaotropic agent that alters protein solubilization and hydrophobic interactions and inhibits protein aggregation and precipitation. The presence of three hetero atoms (O/S and N) in proximity can coordinate with neutral, mono, or dianionic groups to form stable complexes in the biological system. Therefore, in this study, we evaluated urea and thiourea linkers with various substitutions on either side of the carbamide or thiocarbamide functionality to compare the aggregation inhibition of α-syn and tau. A thioflavin-T (ThT) fluorescence assay was used to evaluate the level of fibril formation and monitor the anti-aggregation effect of the different compounds. We opted for transmission electron microscopy (TEM) as a direct means to confirm the anti-fibrillar effect. The oligomer formation was monitored via the photoinduced cross-linking of unmodified proteins (PICUP). The anti-inclusion and anti-seeding activities of the best compounds were evaluated using M17D intracellular inclusion and biosensor cell-based assays, respectively. Disaggregation experiments were performed with amyloid plaques extracted from AD brains. The analogues with indole, benzothiazole, or N,N-dimethylphenyl on one side with halo-substituted aromatic moieties had shown less than 15% cutoff fluorescence obtained with the ThT assay. Our lead molecules 6T and 14T reduced α-syn oligomerization dose-dependently based on the PICUP assays but failed at inhibiting tau oligomer formation. The anti-inclusion effect of our lead compounds was confirmed using the M17D neuroblastoma cell model. Compounds 6T and 14T exhibited an anti-seeding effect on tau using biosensor cells. In contrast to the control, disaggregation experiments showed fewer Aβ plaques with our lead molecules (compounds 6T and 14T). Pharmacokinetics (PK) mice studies demonstrated that these two thiourea-based small molecules have the potential to cross the blood-brain barrier in rodents. Urea and thiourea linkers could be further improved for their PK parameters and studied for the anti-inclusion, anti-seeding, and disaggregation effects using transgenic mice models of neurodegenerative diseases.

Pancreatic β-cells package double C2-like domain beta protein into extracellular vesicles via tandem C2 domains.

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). 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. 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. 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. 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.

Analysis of the Physiological Parameters of Cold Resistance in Core Winter and Spring Wheat Cultivars

We selected 46 core winter–spring wheat cultivars from China’s main wheat-producing areas as experimental materials to clarify the differences in the physiological parameters of their cold resistance and provide a theoretical basis and high-quality germplasm for cold resistance breeding. We investigated differences in their cold resistance under field conditions for two consecutive years, and determined the physiological parameters of the cold resistance, yield, and quality indicators of different winter–spring wheat cultivars. Our results showed that the cold resistance of winter wheat cultivars was higher than that of spring wheat cultivars. The chlorophyll (Chl), soluble sugar (SS), soluble protein (SP), and free proline (Pro) contents of different winter–spring wheat cultivars were positively correlated with cold resistance, and malondialdehyde (MDA) content was negatively correlated with cold resistance. The five physiological parameters can be used as physiological indicators for the breeding of cold-resistant cultivars. The cold resistance, yield, and quality indicators of different spring and winter wheat cultivars were comprehensively evaluated by using the average membership value and comprehensive score. It was found that the average membership value and comprehensive score of winter wheat cultivars were higher than those of spring wheat cultivars. Through classification using the K-means method, the cold-resistant, high-yield, and high-quality cultivars were screened out, namely, Jimai23 (JM23), Jimai44 (JM44), Shannong57 (SN57), and Jinmai 919 (JM919).

Osmolytes and CsAQP expression jointly influence water physiology in the peel and pulp of orange (Citrus sinensis (L.) Osbeck) fruit during postharvest water loss.

Water loss is a serious issue affecting the quality of postharvest horticultural products. Aquaporins (AQPs) regulate the transport of water across biological membranes, along the gradient of water potential, and may play a role in water loss. In this study, matured orange fruits (Citrus sinensis) stored at ambinent temperature (RH 85-95%) for 105 d showed that the weight loss persistently increased, and its rate peaked at 45-60 d and 90-105 d. Both water content and potential were higher in the pulp than in the peel. Water content rose before 60 d, and peel water potential fell with an increased gradient after 60 d. Comparing with peel, osmolytes such as soluble sugar, sucrose, glucose, fructose, and organic acids showed higher accumulation, and their levels were the lowest around 60 d. In contrast, soluble protein and inorganic minerals showed low levels of accumulation in the pulp. In total, 31 CsAQP genes were expressed in the fruit, and most of them were down-regulated in the peel but up-regulated in the pulp during storage. These genes were subsequently classified into four clusters based on their expression patterns. Genes in Cluster I - including CsNIP1;1/2;1/2;2/2;3/3;1/4;1/6;1, CsTIP1;3/2;2/2;3/5;1/6;1, CsXIP1;1/1;2, CsSIP1;2, and CsPIP1;2 - were persistently up-regulated in the pulp for the 105 d of storage, especially at day 60, when some genes showed 103-fold higher expression. Pearson's correlation and principal component analysis further revealed a significant positive correlation among weight loss rate, water content, and water potential gradient (R2 = 0.85). Indexes positively correlated with osmolyte content and Cluster I gene expression in pulp samples suggest that increased CsAQP gene expression in pulp is linked to faster water loss in oranges, particularly at 60 days postharvest.

Mechanistic effects of lipid binding pockets within soluble signaling proteins: Lessons from acyl-CoA-binding and START-domain-containing proteins.

While lipids serve as important energy reserves, metabolites, and cellular constituents in all forms of life, these macromolecules also function as unique carriers of information in plant communication given their diverse chemical structures. The signal transduction process involves a sophisticated interplay between messengers, receptors, signal transducers, and downstream effectors. Over the years, an array of plant signaling proteins have been identified for their crucial roles in perceiving lipid signals. However, the mechanistic effects of lipid binding on protein functions remain largely elusive. Recent literature has presented numerous fascinating models that illustrate the significance of protein-lipid interactions in mediating signaling responses. This review focuses on the category of lipophilic signaling proteins that encompass a hydrophobic binding pocket located outside of cellular membranes and provides an update on the lessons learned from two of these structures, namely the acyl-CoA-binding and START domains. It begins with a brief overview of the latest advances in understanding the functions of the two protein families in plant communication. The second part highlights five functional mechanisms of lipid ligands in concert with their target signaling proteins.

HIP is involved in NaCl and endoplasmic reticulum stress resistance in Arabidopsis

Heat shock protein 70 (HSP70)-interacting proteins (HIPs) have been studied in animals. HIPs perform diverse cellular functions, ranging from alleviating stress to suppressing the formation of insoluble protein, but how their orthologs function in plants is less known. This study shows that Arabidopsis HIP is a cytosolic and nuclear protein associated with HSP70. The hip mutants showed compromised tolerance to NaCl and endoplasmic reticulum (ER) stress, although they grew normally under standard growth conditions. Furthermore, hip mutants presented a decreased HSP70 level compared with the wild type under NaCl and ER stress conditions. These findings suggest the involvement of HIP in NaCl and ER stress tolerance.

Combination of maleic hydrazide and coumarin inhibits rice seed germination involving reactive oxygen species accumulation, ABA metabolism and starch degradation

Pre-harvest sprouting (PHS) in cereal crops is a prevalent phenomenon that impacts grain yield and quality. Several PHS inhibitory compounds were screened and identified in previous studies, such as eugenol (EUG), maleic hydrazide (MH), coumarin (COU), etc. However, few studies have focused on the combination of PHS inhibitors, and the inhibitory mechanism remains unclear. Here, through combination tests of EUG, MH, and COU, the optimal combination of PHS inhibitors was selected as MH 20 mg L−1 + COU 100 mg L−1, which presented the lowest germination percentages. The optimal combination treatment significantly decreased the germination rate, α-amylase activity, content of soluble sugar and soluble protein, enhanced ABA content and the activity of superoxide dismutase (SOD) and peroxidase (POD), inhibited the production of superoxide anion (O2−) and hydrogen peroxide, and reduced the content of malondialdehyde (MDA); conversely, this trend is precisely the opposite in normal germination. Furthermore, gene expression analysis revealed that the optimal combination of MH and COU significantly decreased the expression level of OsAmy1A and OsAmy3D at 12 and 48 h after imbibition (HAI); and promoted the expression of OsRbohs (OsRbohA, OsRbohC, OsRbohD, OsRbohE, OsRbohH) and ABA biosynthetic genes OsNCED1, OsNCED2, and OsNCED5, especially OsNCED2 at 12 HAI, but down-regulated expression of OsRbohs and ABA catabolic genes OsABA8ox1-3 at 48 HAI. These results demonstrated that the delay in seed germination induced by MH and COU involved in ROS, ABA, and sugars; the optimal combination of MH and COU inhibited the germination process by promoting ABA biosynthesis and reducing ABA catabolism, and restraining the α-amylase activity to lower soluble sugar content. Intriguingly, although the expression of OsRbohs, which play a crucial role in generating ROS, increased in early imbibition (12h), the activity of the antioxidant enzymes SOD and POD also increased with the optimal combination treatment of MH and COU, which lead to the delay in ROS accumulation and inhibition of germination. These results have deepened our understanding of the regulatory mechanism of PHS inhibitors and provided theoretical support for the application of MH and COU in preventing sprouting before crop harvesting.

Metabolic pathways regulated by strigolactones foliar spraying enhance osmoregulation and antioxidant defense in drought-prone soybean

BackgroundDrought stress is a significant abiotic stressor that hinders growth, development, and crop yield in soybeans. Strigolactones (SLs) positively regulate plant resistance to drought stress. However, the impact of foliar application of SLs having different concentrations on soybean growth and metabolic pathways related to osmoregulation remains unknown. Therefore, to clarify the impact of SLs on soybean root growth and cellular osmoregulation under drought stress, we initially identified optimal concentrations and assessed key leaf and root indices. Furthermore, we conducted transcriptomic and metabolic analyses to identify differential metabolites and up-regulated genes.ResultsThe results demonstrated that drought stress had a significant impact on soybean biomass, root length, root surface area, water content and photosynthetic parameters. However, when SLs were applied through foliar application at appropriate concentrations, the accumulation of ABA and soluble protein increased, which enhanced drought tolerance of soybean seedlings by regulating osmotic balance, protecting membrane integrity, photosynthesis and activating ROS scavenging system. This also led to an increase in soybean root length, lateral root number and root surface area. Furthermore, the effects of different concentrations of SLs on soybean leaves and roots were found to be time-sensitive. However, the application of 0.5 µM SLs had the greatest beneficial impact on soybean growth and root morphogenesis under drought stress. A total of 368 differential metabolites were screened in drought and drought plus SLs treatments. The up-regulated genes were mainly involved in nitrogen compound utilization, and the down-regulated metabolic pathways were mainly involved in maintaining cellular osmoregulation and antioxidant defenses.ConclusionsSLs enhance osmoregulation in soybean plants under drought stress by regulating key metabolic pathways including Arachidonic acid metabolism, Glycerophospholipid metabolism, Linoleic acid metabolism, and Flavone and flavonol biosynthesis. This study contributes to the theoretical understanding of improving soybean adaptability and survival in response to drought stress.

Vortex-assisted dispersive low-density liquid–liquid microextraction of xanthydrol derivatized acrylamide in processed chips and water samples for gas chromatographic analysis

Acrylamide, a probable human carcinogen present in heat-processed foods and environmental contaminants, requires sample extraction and preconcentration before chromatographic analysis. The method developed in this study employed derivatization with xanthydrol and dispersive liquid–liquid microextraction utilizing low-density anisole. Durian or potato chips were combined with deionized water, defatted with hexane, and subjected to precipitation of soluble carbohydrates and proteins using clarification reagents. Water samples were filtered through a membrane filter. Acrylamide was derivatized by introducing an acidic methanolic solution of xanthydrol at 50 °C. The derivatized acrylamide was extracted with 70 µL of anisole and vortexed, with the methanol from the xanthydrol solution serving as the disperser solvent. The anisole layer was analyzed using gas chromatography with both flame ionization and mass spectrometric detection. Linear calibration plots exhibited coefficients of determination >0.9997. The precision was measured at <10% RSD, and recoveries ranged from 84% to 107%. The quantitation limit varied from 2 to 10 µg kg−1 for processed chips and from 0.05 to 0.10 µg L−1 for water samples. Acrylamide was detected in all processed chip samples, with some concentrations exceeding the benchmark value of 750 μg kg−1. However, no acrylamide was identified in any of the water samples.

Advances in Research on the Improvement of Low-Salt Meat Product Through Ultrasound Technology: Quality, Myofibrillar Proteins, and Gelation Properties.

The high sodium content in meat products poses health risks to consumers and does not align with modern green and healthy living standards. Current strategies for directly reducing the sodium content in meat products are limited by their negative impact on the sensory or quality attributes of the products. In recent years, there has been great interest in applying ultrasound technology to reduce sodium content. This paper discusses the advantages and disadvantages of current mainstream strategies for reducing the sodium content in meat products, as well as the potential mechanisms by which ultrasound-assisted marination improves the quality of low-salt meat products. The main findings indicate that ultrasound, through its cavitation and mechanical effects, facilitates the transition of proteins from stable insoluble aggregates to stable soluble complexes, exposing more hydrophilic groups and, thus, enhancing protein solubility. At the same time, ultrasound promotes a greater number of proteins to participate in the formation of interfacial layers, thereby increasing emulsifying activity. Furthermore, ultrasound treatment promotes the interaction between proteins and water, leading to partial unfolding of protein chains, which allows polar residues to more readily capture water in the gel, thereby improving the water-holding capacity of the gel. These effects will contribute to the formation of high-quality low-salt meat products. However, variations in the frequency, intensity, and duration of ultrasound treatment can lead to differing effects on the quality improvement of low-salt meat products.

Cardiac Amyloidosis: State-of-the-Art Review in Molecular Pathology.

Amyloidosis refers to a group of diseases caused by extracellular deposits of misfolded proteins, which alter tissue function and structure, potentially affecting any organ. The term "amyloid" was introduced in the 19th century and later associated with pathological protein deposits. Amyloid fibrils, which are insoluble and resistant to degradation, originate from soluble proteins that undergo misfolding. This process can be triggered by several factors, such as aging, elevated protein concentrations, or pathogenic variants. Amyloid deposits damage organs both by disrupting tissue architecture and through direct cytotoxic effects, leading to conditions such as heart failure. Amyloidosis can be classified into acquired or inherited forms and can be systemic or localized. Diagnosing cardiac amyloidosis is complex and often requires tissue biopsies, which are supported by Congo Red dye staining. In some cases, bisphosphonate bone scans may provide a less invasive diagnostic option. In this state-of-the-art review, we focus on the most common forms of cardiac amyloidosis, from epidemiology to therapy, emphasizing the differences in molecular mechanisms and the importance of pathological diagnosis for appropriate treatment using a multidisciplinary approach.

Physiological and Transcriptomic Analyses Reveal the Role of the Antioxidant System and Jasmonic Acid (JA) Signal Transduction in Mulberry (Morus alba L.) Response to Flooding Stress

In recent decades, the frequency of flooding has increased as a result of global climate change. Flooding has become one of the major abiotic stresses that seriously affect the growth and development of plants. Mulberry (Morus alba L.) is an important economic tree in China. Flooding stress is among the most severe abiotic stresses that affect the production of mulberry. However, the physiological and molecular biological mechanisms of mulberry responses to flooding stress are still unclear. In the present study, reactive oxygen species (ROS) metabolism, antioxidant mechanism, and plant hormones in mulberry associated with the response to flooding stress were investigated using physiological and transcriptomic analysis methods. The results showed significant increases in the production rate of superoxide anion (O2•−) and the content of hydrogen peroxide (H2O2) in leaves on the 5th day of flooding stress. This led to membrane lipid peroxidation and elevated malondialdehyde (MDA) levels. Antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) exhibited enhanced activities initially, followed by fluctuations. The ascorbic acid–glutathione (AsA-GSH) cycle played a crucial role in scavenging ROS, promoting the reduction of oxidized glutathione (GSSG) to reduced glutathione (GSH). Transcriptomic analysis revealed the up-regulation of the gene-encoding antioxidant enzymes (APX, MDHAR, GPX, GR, GST) involved in ROS scavenging and stress tolerance mechanisms. Jasmonic acid (JA) levels and the expression of JA synthesis-related genes increased significantly in mulberry leaves under flooding stress. This activation of the JA signaling pathway contributed to the plant’s adaptability to flooding conditions. Proline (Pro) and soluble sugar (SS) contents increased notably in response to flooding stress. Proline helped maintain cell turgor and protected enzymes and membranes from damage, while soluble sugars supported anaerobic respiration and energy supply. However, soluble protein (SP) content decreased, suggesting inhibition of protein synthesis. The study provides insights into mulberry’s flooding tolerance mechanisms, guiding future molecular breeding efforts. This summary captures the key findings and implications of the study on mulberry’s response to flooding stress, focusing on physiological and molecular mechanisms identified in the research.

Biochemical Analysis of Different Wheat Varieties Grown under Various Zones of India

Background: The study aimed to evaluate the flour quality of different wheat varieties (Triticum durum L.) for various end-use products such as chapati, bread, biscuits and pasta. The focus was on identifying varieties with superior nutritional and processing qualities by analyzing various qualitative parameters. Methods: The study assessed multiple wheat varieties using a range of qualitative parameters, including moisture, ash, total soluble sugar, starch, protein, lysine, tryptophan, dry and wet gluten, gluten index, water binding capacity, β-carotene content, sedimentation value, thousand kernel weight and mineral content (Fe and Zn). The wheat varieties evaluated included A-206, GW-1, HI 8498 (Kota), PDW 291 (Hissar) and NIDW 295 (Pune). Result: The study identified A-206 and GW-1 as wheat varieties with superior protein content, gluten quality and water binding capacity, making them ideal for various end-use products. A-206 also had the highest lysine and tryptophan levels, while GW-1 and HI 8498 (Kota) had elevated ash content. PDW 291 (Hissar) was notable for its higher total soluble sugar and NIDW 295 (Pune) excelled in starch content, β-carotene and thousand kernel weight. Overall, NIDW 295 showed the most promise for pasta production due to its comprehensive quality traits.

Growth, physiological and N, P, K accumulation responses of Erythropalum scandens Bl. Seedlings under different substrates

Erythropalum scandens Bl. is a medicinal woody vegetable found in southern China and parts of Southeast Asia. Studies have shown improper substrate hindered E. scandens seedling growth, causing water accumulation and nutrient deficiency. In pursuit of an ideal growth medium for E. scandens seedlings during the early stages, this study conducted a pot experiment to identify a mixed substrate with optimal water permeability and fertility. In this study, pure Alfisols soil treatment as the control (CK), and two soilless substrates (peat soil and perlite) were combined with Alfisols soil into different volume ratios, in order to better use soil resources from understory space and balance the texture of mixed substrates. The growth, physiological characteristics and nutrient status of 24-month-old E. scandens seedlings were determined after planting in different mixed ratios. The results showed that as the proportion of peat soil increased in the mix, most indexes exhibited an initial increase followed by a decline, while soluble protein content decreased consistently. Conversely, an increasing perlite ratio resulted in a general decline in most growth and physiological indexes. Root growth, biomass accumulation and chlorophyll content, peaked in the 66.67% Alfisols soil + 33.33% perlite (T4) treatment. Notably, T3 (66.67% Alfisols soil + 33.33% peat soil) showcased the best above-ground growth, while T1 (50.00% Alfisols soil + 50.00% peat soil) excelled in element content accumulation. In conclusion, the cultivation substrate should primarily consist of Alfisols soil, constituting at least 50%. The addition of peat soil enhances above-ground growth and nutrients accumulation, while perlite contributes to robust root development. One third of peat soil and a small amount of perlite can be added to the substrate during E. scandens seedling cultivation, and proper fertilization should also be used in order to increase nutrient accumulation in aboveground and underground parts. This research provides valuable insights into maximizing the potential of E. scandens seedlings through precise cultivation methods.

Combined effect of molybdenum and nitrogen fertilization on nitrogen metabolism and amino acid content in tobacco leaves

IntroductionThis study investigated the combined effects of molybdenum (Mo) and nitrogen (N) fertilization on N metabolism and amino acid content in the leaves of flue-cured tobacco (Nicotiana tabacum L.) during its mature stage through a pot experiment.MethodsDifferent application levels of Mo (0, 0.15, 0.30 mg/kg soil) and N (0.06, 0.24, 0.42 g/kg soil) were set to observe and analyze changes in leaf quality, N, and amino acid content in the tobacco plants.ResultsThe results revealed that the N fertilizer application level exhibited a primary effect on regulating the total nitrogen, nitrate nitrogen, soluble protein, and amino acid nitrogen concentrations within tobacco leaves, while the effectiveness of Mo fertilization was influenced by the level of N applied. Specifically, under the conditions of 0.24 g/kg soil N and 0.30 mg/kg soil Mo application, the N content, N accumulation, and dry matter mass of the tobacco plants increased significantly by 110%, 204%, and 48%, respectively. Concurrently, nitrate reductase activity increased by 107-fold, and the nitrate nitrogen content was relatively low, contributing to enhanced tobacco yield and safety. Moreover, this treatment led to a notable (170%) increase in free amino acid nitrogen content, with minimal impact on total amino acids and soluble proteins. Notably, it effectively increased the content of free amino acids beneficial to the sensory quality of tobacco (such as histidine, arginine, aspartic acid, isoleucine, and glutamic acid) without reducing the total amino acid content, while simultaneously reducing other amino acids that might affect quality. Therefore, the combined application of 0.30 mg/kg soil Mo and 0.24 g/kg soil N specifically optimized the amino acid composition in tobacco leaves, positively impacting overall quality and market competitiveness.DiscussionThis study provides a theoretical basis for the rational application of Mo fertilizer in Mo-deficient areas to improve the yield and quality of flue-cured tobacco.

Recombinant Plasminogen Activator of the Sandworm (Perinereis aibuhitensis) Expression in Escherichia coli.

As an essential thrombolytic agent, the tissue plasminogen activator receives increasing attention due to its longer half-life, lower immunogenicity, and easier administration, which are superior to other thrombolytic agents. In this study, the isolated and purified plasminogen activator from the sandworm (Perinereis aibuhitensis) was expressed in E. coli (Escherichia coli) to investigate its potential for simplifying the development process. The sandworm plasminogen activator was previously successfully cloned and expressed in E. coli with low yield and activity in the culture supernatant. This low yield and activity prompted us to optimize its DNA sequence. Furthermore, to raise the efficiency in the separation of the target protein, the protein's solubility was enhanced by fusing it with maltose-binding protein (MBP) tags. Eventually, the fibrinolytic activity was successfully restored after digestion with tobacco etch virus (TEV) protease. This study provides an innovative method of efficiently expressing and purifying plasminogen activators from sandworm in E. coli and broadens its applications in therapeutic treatment of cardiovascular diseases, including thrombosis, stroke, and coronary atherosclerotic heart disease.

Mitigation of microplastic toxicity in soybean by synthetic bacterial community and arbuscular mycorrhizal fungi interaction: Altering carbohydrate metabolism, hormonal transduction, and genes associated with lipid and protein metabolism

Microplastics (MPs) have become ubiquitous environmental pollutants, eliciting concerns about their negative impacts on terrestrial and agroecosystems. However, employing sustainable remediation approaches holds the potential to mitigate these negative impacts and bolster MPs tolerance in both the seeds and vegetative structures of plants. The current investigation explores the impact of arbuscular mycorrhizal fungi (AMF) and synthetic bacterial community (SynCom) as potential alleviators of MP-induced stress. Results exhibited that co-applied SynCom and AMF, resulted in the highest plant biomass, leaf area, pod per plant, and 100-seed weight under control (no MPs) and MPs stress. The combined application of SynCom and AMF enhanced colonization, association, and arbuscule abundance in the non-stressed plants (no MPs) as compared to stressed plants. The sole or co-application of SynCom and AMF enhanced the primary metabolites content and mitigated the MPs-induced reduction in soluble sugars, lipids, protein, and oil contents. Soybean inoculation stimulated the genes expression involved in lipid and protein biosynthesis, while a contradictory drift was noted for genes associated with lipid and protein degradation, underpinning the observed increment in protein and lipid content. Soybean inoculated with AMF exhibited the utmost α-amylase and β-amylase activities, demonstrating enhanced osmolyte (soluble sugar) production, mainly under MPs stress. Remarkably, sole or co-application further reinforces the positive effect of MPs stress on the osmoprotectants and antioxidant levels, for instance, phenol, flavonoid, glycine betaine contents, and glutathione-S-transferase (GST) activities. Subsequent to the stress release, a stress hormones known as abscisic acid (ABA) reduced in the seeds of inoculated soybean while gibberellin (GA), trans-zeatin riboside (ZR), and indole acetic acid (IAA) were increased. Therefore, it is concluded that the combined application of SynCom and AMF protected the soybean plants from MPs-induced oxidative damage by upregulating osmoprotectants and antioxidant levels. This study provides insights into the potential of sustainable remediation approaches in alleviating the complex impact of MPs on soybean, contributing to future strategies for environmental sustainability. Future studies could explore the optimization of SynCom and AMF formulations for different crops and environmental conditions, potentially leading to practical applications in sustainable agriculture and pollution management.

Spray Drying of Inca Peanut Meal Protein Hydrolysate to Produce Protein Powder Drink Mix

The optimal conditions (maltodextrin concentration and inlet air temperature) for spray drying Inca peanut meal protein hydrolysate (IMPH) were determined. The maltodextrin concentration was varied at 5, 10 and 15 % by weight, and inlet air temperature was varied at 150, 160 and 170 °C. The IMPH was subjected to spray drying, where the feed solution was atomized into fine droplets and exposed to a hot air stream, leading to rapid drying and the formation of powder. The moisture, protein, yield, water activity and water solubility index (WSI) of IMPH powder were analyzed. The optimal condition for spray drying IMPH powder utilized 10 % maltodextrin and an inlet air temperature of 160 °C. Under these conditions, the moisture, protein, yield and WSI of IMPH powder were 2.93, 36.93, 38.95 and 99.59 %, respectively, while water activity was 0.41. IMPH powder particles were spherical and non-agglomerate. Inca peanut meal protein powder drink mix (IMPD) with cocoa flavor was the most acceptable. The IMPD with cocoa flavor could be promoted as a high protein and high dietary fiber product. It could be kept at 35 ± 2 °C for 12 weeks, whilst retaining good microbiological properties. HIGHLIGHTS The optimal conditions for spray drying Inca peanut meal protein hydrolysate (IMPH) were determined to be 10% maltodextrin and an inlet air temperature of 160 °C. This resulted in high yield, protein content, and water solubility, while keeping moisture and water activity low. IMPH powder had spherical, non-agglomerated particles, and cocoa-flavored Inca peanut protein powder drink mix (IMPD) was the most preferred in sensory evaluations. The IMPD with cocoa flavor was identified as a high-protein, high-dietary-fiber product, suitable for storage at 35 °C for up to 12 weeks while maintaining good microbiological quality. The study highlights the nutritional benefits of the cocoa-flavored IMPD, which includes essential amino acids, omega-3 fatty acids, and antioxidant properties. GRAPHICAL ABSTRACT

Influence of Temperature and Host Plant on the Digestion of Frankliniella intonsa (Trybom) Revealed by Molecular Detection.

Frankliniella intonsa (Trybom) (Thysanoptera: Thripidae) is an important type of thrip and a polyphagous pest, which poses a serious threat to many crops, especially those in tropical regions of China. Its feeding behavior and the damage caused vary among different host plant species and are affected by ambient temperature and plant nutrients as well. The digestion rate is an important index for directly observing the digestion process, but there have been no studies directly measuring the digestion in thrips under the influence of different temperatures and host plants. Here, the digestion rate of F. intonsa was assessed by using a molecular diagnostic tool. We also determined the nutrient content in three host plant (mango, cowpea, and pepper), including soluble proteins, free fatty acids, soluble sugars, and water. The results showed that the high and low temperatures (16 °C and 32 °C) both seemed to accelerate the digestion of F. intonsa compared to the optimal temperature (26 °C) and the protein content of plants played an important role in the digestive response of F. intonsa to temperature changes. The findings can help reveal the feeding damage caused by F. intonsa to different plants and help to better understand its feeding ecology, according to its interaction with the host plant.

Effects of Salt Stress on Seed Germination and Embryo Growth of Oxytropis coerulea

Background: Oxytropis coerulea is considered to be an exceptional forage. The salinization of soil is a prevalent ecological issue worldwide. In order to study the effects of saline solution on seed germination and embryo growth of O. coerulea. Methods: An experiment was carried out using the culture dish filter paper method. Three salt stresses, namely, Na2CO3, NaCl and double salt were set, each of which was set to 0%, 0.3%, 0.6% and 0.9% concentration. Result: The results indicated that as the concentration of salt increased, there was a decrease in the germination rate, germination index and embryo growth. The activities of SOD, POD and CAT, as well as soluble protein content, initially increased and then decreased with increasing salt stress. The toxic effects of the three salts ranged from strong to weak as Na2CO3 greater than double salt greater than NaCl. The salt resistance indexes of 0.3% Na2CO3, NaCl and double salt were 53.87, 57.00 and 38.73, respectively and the salt resistance index of 0.6% NaCl was 50.21, indicating a high salt-tolerant degree. It showed certain resistance to 0.3% Na2CO3, 0.6% NaCl and 0.3% double salt. In short, the various soluble mechanisms of O. coerulea in different stages of salt stress and different salt stress levels have different reaction mechanisms, which reflect the salt stress of O. coerulea seeds. O. coerulea seeds can be planted in a suitable concentration of saline-alkali land, which can utilize the feed value and medicinal value of O. coerulea to improve economic benefits and utilize saline soil to improve the ecological environment.