Soluble Protein Research Articles

Stingless bee honey: Nutritional, physicochemical, phytochemical and antibacterial validation properties against wound bacterial isolates.

With the rise of AMR the management of wound infections are becoming a big challenge. This has been attributed to the fact that most wound bacterial isolates have been found to possess various virulence factors like enzymes, toxins & biofilms production. Therefore, need for discovery of new lead compounds is paramount as such factors make these microbes to be resistant to already existing arsenal of antibiotics or even the immune system. This study aimed at documenting the nutritional, physicochemical, phytochemical and antibacterial properties of stingless bee honey. Isolation and characterization of bacterial isolates from 34 samples obtained from wounds of outpatients and surgical wards of Nakuru County Referral Hospital, Kenya was done. Various bacterial isolates (43) were isolated Staphylococcus aureus (34.8%) being predominant, followed by Pseudomonas aeruginosa (27.9%), Klebsiella pneumoniae (23.3%) and Escherichia coli (14.0%). A total of 36 out of the total isolates were genotypically characterized using molecular techniques detecting the prevalence of the following virulence genes; 16 srRNA (756 bp), hla (229 bp), cnf1 (426 bp), cnf2 (543 bp), hlyA (1011 bp), rmpA (461 bp), lasL (600 bp), gyrB (411 bp), khe (77 bp) and magA (128 bp). An assessment of the in vitro antibacterial activity of 26 stingless bee honey samples collected from their cerumen egg-shaped pots in Marigat sub-County, Baringo County, Kenya was done. Antibacterial properties of the stingless bee honey was done with varying susceptibility patterns being observed at different concentrations of honey impregnated discs (10x104, 20x104, 50x104 and 75x104 ml μg/ ml) giving mean inhibition diameters of 18.23 ± 0.4 mm (Staphylococcus aureus), 17.49 ± 0.3 mm (Pseudomonas aeruginosa), 16.05 ± 0.6 mm (Klebsiella pneumoniae) and 10.19 ± 0.5 mm (Escherichia coli) with a mean range of 14.54 ± 2.0 mm to 17.58 ± 3 mm. Higher susceptibility to honey was recorded across all the bacterial isolates compared to conventional antibiotics while the mean MIC and MBC of the honey were recorded at 62.5 ml μg/ ml and 250 ml μg/ ml respectively. Control bacterial isolates Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 27736 and Pseudomonas aeruginosa ATCC 27858 were used in the analysis. The stingless bee honey was found to be rich in various nutritive components like sugar (89.85 ± 5.07 g/100 g) and moisture (81.75 ± 10.35 mg/g) with a significant difference of P <0.05 as the main antibacterial components. Additionally, the stingless honey did possess water soluble vitamins, proteins and minerals of which potassium was the most dominant one. In regard to phytochemicals, on our preliminary analysis phenolic, flavonoid and carotenoid compounds were found to be present with phenolic compounds being the most dominant one. Stingless bee honey from Marigat, has antimicrobial properties which could be attributed to the rich phytochemicals it possesses and its physicochemical properties in addition to its high nutritive value.

Analysis of AlphaMissense data in different protein groups and structural context

Single amino acid substitutions can profoundly affect protein folding, dynamics, and function. The ability to discern between benign and pathogenic substitutions is pivotal for therapeutic interventions and research directions. Given the limitations in experimental examination of these variants, AlphaMissense has emerged as a promising predictor of the pathogenicity of missense variants. Since heterogenous performance on different types of proteins can be expected, we assessed the efficacy of AlphaMissense across several protein groups (e.g. soluble, transmembrane, and mitochondrial proteins) and regions (e.g. intramembrane, membrane interacting, and high confidence AlphaFold segments) using ClinVar data for validation. Our comprehensive evaluation showed that AlphaMissense delivers outstanding performance, with MCC scores predominantly between 0.6 and 0.74. We observed low performance on disordered datasets and ClinVar data related to the CFTR ABC protein. However, a superior performance was shown when benchmarked against the high quality CFTR2 database. Our results with CFTR emphasizes AlphaMissense’s potential in pinpointing functional hot spots, with its performance likely surpassing benchmarks calculated from ClinVar and ProteinGym datasets.

Insights into the effect of nitrate photolysis on short-chain fatty acids production from waste activated sludge in anaerobic fermentation system: Performance and mechanisms

Nitrate photolysis has become an efficient, low-cost and promising technology for emerging contaminants removal, while its performance and mechanism for waste activated sludge (WAS) treatment is still unknown. This study innovatively introduced nitrate photolysis for WAS disintegration, and investigated the effect of nitrate addition (150–375 mg N/L) for short-chain fatty acids (SCFAs) production during anaerobic fermentation (AF). The results showed that nitrate photolysis significantly promoted the SCFAs production from WAS, and peaked at 280.7 mg/g VSS with 7-d fermentation with 150 mg N/L addition (150N-UV), which increased by 8.8–35.0 % and 10.7–23.3 % compared with other photolysis groups and sole nitrate groups. Effective release of the soluble organics was observed in the nitrate photolysis groups during AF, especially soluble proteins, reaching 1505.4 mg COD/L at 9 d in 150N-UV group, promoted by 7.0∼15.7 % than nitrate/nitrate photolysis groups. The model compounds simulation experiment further demonstrated the positive effect of nitrate photolysis on organics hydrolysis and SCFAs accumulation. The result of the radical capture and quenching verified the reactive oxygen species contributed more compared with reactive nitrogen species. Functional group analysis confirmed the effective bioconversion of the macromolecular organics during the fermentation. Moreover, the nitrate photolysis enhanced the enrichment of the functional consortia, including anaerobic fermentation bacteria (AFB), e.g., Fnoticella, Romboutsia, Gracilibacter and Sedimentibacter, and nitrate reducing bacteria (NRB), e.g., Acinerobacter and Ahniella. The macrogenetic analysis further revealed that glycolysis, amino acid metabolism, acetate metabolism and nitrogen metabolism were the dominating metabolic pathways during fermentation, and the abundance of the relevant genes were enhanced in 150N-UV group.

Solid-state fermentation using wheat bran to produce glucose syrup and functional cereal bars.

Wheat bran is one of the most abundant by-products from grain milling, which can be used as substrate for solid-state fermentation (SSF) to obtain enzymes able to convert this agro-industrial waste into glucose syrup, which in turn can be applied for the production of different food products. The present study aimed to determine centesimal composition of wheat bran, obtain enzymatic extract that converts wheat bran into wheat glucose syrup (WGS), produce rice flakes cereal bars (RFCB), and evaluate their nutritional composition and the presence of functional compounds, as well as their antioxidant potential. Determination of centesimal composition of wheat bran demonstrated its nutritional potential. Enzymatic extract was obtained and it converted wheat bran into WGS, which were applied to rice flakes producing RFCB. These cereal bars proved to be a source of dietary fiber (1.8g) and soluble protein (7.2g) while RCFB produced with corn glucose syrup did not present these nutritional components. In addition, RFCB produced with WGS showed polyphenolic compounds, among them flavonoids, which exhibited antioxidant activity by DPPH and ABTS radical scavenging (47.46% and 711.89μM Trolox Equivalent/g, respectively), and iron ion reduction (71.70μM Trolox equivalent/g). Final product showed a decrease in caloric value and sodium content. Therefore, the present study showed that the bioprocess of SSF yields a nutritional, ecological, and functional food product, which might be of great interest for food industry, adding nutritional and functional value to a well-stablished product.

Aggrescan4D: structure-informed analysis of pH-dependent protein aggregation.

Protein aggregation is behind the genesis of incurable diseases and imposes constraints on drug discovery and the industrial production and formulation of proteins. Over the years, we have been advancing the Aggresscan3D (A3D) method, aiming to deepen our comprehension of protein aggregation and assist the engineering of protein solubility. Since its inception, A3D has become one of the most popular structure-based aggregation predictors because of its performance, modular functionalities, RESTful service for extensive screenings, and intuitive user interface. Building on this foundation, we introduce Aggrescan4D (A4D), significantly extending A3D's functionality. A4D is aimed at predicting the pH-dependent aggregation of protein structures, and features an evolutionary-informed automatic mutation protocol to engineer protein solubility without compromising structure and stability. It also integrates precalculated results for the nearly 500,000 jobs in the A3D Model Organisms Database and structure retrieval from the AlphaFold database. Globally, A4D constitutes a comprehensive tool for understanding, predicting, and designing solutions for specific protein aggregation challenges. The A4D web server and extensive documentation are available at https://biocomp.chem.uw.edu.pl/a4d/. This website is free and open to all users without a login requirement.

Dysregulated Coagulation and Fibrinolysis Are Present in Patients Admitted to the Emergency Department with Acute Hypoxemic Respiratory Failure: A Prospective Study.

Acute hypoxemic respiratory failure (AHRF) is defined as acute and progressive, and patients are at a greater risk of developing acute respiratory distress syndrome (ARDS). Until now, most studies have focused on prognostic and diagnostic biomarkers in ARDS. Since there is evidence supporting a connection between dysregulated coagulant and fibrinolytic pathways in ARDS progression, it is plausible that this dysregulation also exists in AHRF. The aim of this study was to explore whether levels of soluble endothelial protein C receptor (sEPCR) and plasminogen differentiate patients admitted to the emergency department (ED) with AHRF. sEPCR and plasminogen levels were measured in 130 AHRF patients upon ED presentation by ELISA. Our results demonstrated that patients presenting to the ED with AHRF had elevated levels of sEPCR and plasminogen. It seems that dysregulation of coagulation and fibrinolysis occur in the early stages of respiratory failure requiring hospitalisation. Further research is needed to fully comprehend the contribution of sEPCR and plasminogen in AHRF.

Investigating the growth promotion potential of biochar on pea (Pisum sativum) plants under saline conditions

Pea, member of the plant family Leguminosae, play a pivotal role in global food security as essential legumes. However, their production faces challenges stemming from the detrimental impacts of abiotic stressors, leading to a concerning decline in output. Salinity stress is one of the major factors that limiting the growth and productivity of pea. However, biochar amendment in soil has a potential role in alleviating the oxidative damage caused by salinity stress. The purpose of the study was to evaluate the potential role of biochar amendment in soil that may mitigate the adverse effect of salinity stress on pea. The treatments of this study were, (a) Pea varieties; (i) V1 = Meteor and V2 = Green Grass, Salinity Stress, (b) Control (0 mM) and (ii) Salinity (80 mM) (c) Biochar applications; (i) Control, (ii) 8 g/kg soil (56 g) and (iii) 16 g/kg soil (112 g). Salinity stress demonstrated a considerable reduction in morphological parameters as Shoot and root length decreased by (29% and 47%), fresh weight and dry weight of shoot and root by (85, 63%) and (49, 68%), as well as area of leaf reduced by (71%) among both varieties. Photosynthetic pigments (chlorophyll a, b, and carotenoid contents decreased under 80 mM salinity up to (41, 63, 55 and 76%) in both varieties as compared to control. Exposure of pea plants to salinity stress increased the oxidative damage by enhancing hydrogen peroxide and malondialdehyde content by (79 and 89%), while amendment of biochar reduced their activities as, (56% and 59%) in both varieties. The activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) were increased by biochar applications under salinity stress as, (49, 59, and 86%) as well as non-enzymatic antioxidants as, anthocyanin and flavonoids improved by (112 and 67%). Organic osmolytes such as total soluble proteins, sugars, and glycine betaine were increased up to (57, 83, and 140%) by biochar amendment. Among uptake of mineral ions, shoot and root Na+ uptake was greater (144 and 73%) in saline-stressed plants as compared to control, while shoot and root Ca2+ and K+ were greater up to (175, 119%) and (77, 146%) in biochar-treated plants. Overall findings revealed that 16 g/kg soil (112 g) biochar was found to be effective in reducing salinity toxicity by causing reduction in reactive oxygen species and root and shoot Na+ ions uptake and improving growth, physiological and anti-oxidative activities in pea plants (Fig. 1).Figure 1A schematic diagram represents two different mechanisms of pea under salinity stress (control and 80 mM NaCl) with Biochar (8 and 16 g/kg soil).

GPSFun: geometry-aware protein sequence function predictions with language models.

Knowledge of protein function is essential for elucidating disease mechanisms and discovering new drug targets. However, there is a widening gap between the exponential growth of protein sequences and their limited function annotations. In our prior studies, we have developed a series of methods including GraphPPIS, GraphSite, LMetalSiteand SPROF-GO for protein function annotations at residue or protein level. To further enhance their applicability and performance, we now present GPSFun, a versatile web server for Geometry-aware Protein Sequence Function annotations, which equips our previous tools with language models and geometric deep learning. Specifically, GPSFun employs large language models to efficiently predict 3D conformations of the input protein sequences and extract informative sequence embeddings. Subsequently, geometric graph neural networks are utilized to capture the sequence and structure patterns in the protein graphs, facilitating various downstream predictions including protein-ligand binding sites, gene ontologies, subcellular locationsand protein solubility. Notably, GPSFun achieves superior performance to state-of-the-art methods across diverse tasks without requiring multiple sequence alignments or experimental protein structures. GPSFun is freely available to all users at https://bio-web1.nscc-gz.cn/app/GPSFun with user-friendly interfaces and rich visualizations.

SAPPα Peptide Promotes Damaged Microglia to Clear Alzheimer's Amyloid-β via Restoring Mitochondrial Function.

Alzheimer's disease (AD) is an age-related neurodegenerative disease with amyloid-β (Aβ) deposition as the main pathological feature. It's an important challenge to find new ways to clear Aβ from Brain. The soluble amyloid precursor protein α (sAPPα) is a neuroprotective protein and can attenuate neuronal damage, including toxic Aβ. However, the regulatory role of sAPPα in non-neuronal cells, such as microglia, is less reported and controversial. Here, we showed that sAPPα promoted the phagocytosis and degradation of Aβ in both normal and damaged microglia. Moreover, the function of damaged microglia was improved by the sAPPα through normalizing mitochondrial function. Furthermore, the results of molecular docking simulation showed that sAPPα had a good affinity with Aβ. We preliminarily reveal that sAPPα is similar to antibodies and can participate in the regulation of microglia phagocytosis and degradation of Aβ after binding to Aβ. sAPPα is expected to be a mild and safe peptide drug or drug carrier for AD.

The impacts of pullulan soaking on radish seed germination and seedling growth under salt stress.

Pullulan can not only provide a source of organic carbon but also has excellent properties. However, current research is mostly limited to the physical properties of the high molecular-weight components of pullulan, and little is known of the application of its low molecular-weight components. This study was designed to explore the impact of the pre-soaking of radish seeds in a pullulan solution on seed germination and subsequent seedling growth under salt stress conditions. Pullulan soaking was found to enhance the germination rates of radish seeds subjected to salt stress, while also enhancing the aboveground growth of radish seedlings. Pullulan soaking resulted in increases in chlorophyll, soluble protein, and soluble sugar concentrations in the leaves of these seedlings, together with greater peroxidase activity and root activity as well as decreases in Na+ and malondialdehyde concentrations. This provides an important reference for the application of pullulan in plant protection.

Helical reorganization in the context of membrane protein folding: Insights from simulations with bacteriorhodopsin (BR) fragments

Membrane protein folding is distinct from folding of soluble proteins. Conformational acquisition in major membrane protein subclasses can be delineated into insertion and folding processes. An exception to the “two stage” folding, later developed to “three stage” folding, is observed within the last two helices in bacteriorhodopsin (BR), a system that serves as a model membrane protein. We employ a reductionist approach to understand interplay of molecular factors underlying the apparent defiance. Leveraging available solution NMR structures, we construct, sample in silico, and analyze partially (PIn) and fully inserted (FIn) BR membrane states. The membrane lateral C-terminal helix (CH) in PIn is markedly prone to transient structural distortions over microsecond timescales; a disorder prone region (DPR) is thereby identified. While clear transmembrane propensities are not acquired, the distortions induce alterations in local membrane curvature and area per lipid. Importantly, energetic decompositions reveal that overall, the N-terminal helix (NH) is thermodynamically more stable in the PIn. Higher overall stability of the FIn arises from favorable interactions between the NH and the CH. Our results establish lack of spontaneous transition of the PIn to the FIn, and attributes their partitioning to barriers that exceed those accessible with thermal fluctuations. This work paves the way for further detailed studies aimed at determining the thermo-kinetic roles of the initial five helices, or complementary external factors, in complete helical folding and insertion in BR. We comment that complementing such efforts with the growing field of machine learning assisted energy landscape searches may offer unprecedented insights.

Structural elucidation and functional characteristics of novel potential prebiotics produced from Limosilactobacillus reuteri N1 GtfB-treated maize starches

The recently characterized Limosilactobacillus reuteri N1 GtfB (LrN1 GtfB) from glycoside hydrolase family 70 is a novel 4,6-α-glucanotransferase acting on starch/maltooligosaccharides with high enzyme activity and soluble protein yield (in heterogenous system). In this study, the influence of the treatment by LrN1 GtfB on the fine structure and functional characteristics of three maize starches were furtherly investigated and elucidated. Due to the treatment of LrN1 GtfB, the starch molecules were transformed into reuterans containing linear and branched (α1 → 6) linkages with notably smaller molecular weight and shorter chain length. Moreover, the (α1 → 6) linkage ratios in the GtfB-modified high-amylose maize starch (GHMS)/normal maize starch (GNMS)/waxy maize starch (GWMS) increased by 18.3 %/12.6 %/9.0 % as compared to their corresponding controls. In vitro digestibility experiment revealed that the resistant starch content of GHMS, GNMS and GWMS increased by 16 %, 18 % and 25 % as compared to the starch substrates. Furthermore, the butyric acid yielded from GHMS, GNMS and GWMS in the in vitro fermentation experiments were 1.4, 1.5 and 1.4 times higher than those of commercial galactose oligosaccharides. These results indicated that the highly-branched short-clustered reuteran synthesized by LrN1 GtfB might serve as novel potential prebiotics, and provide insights for the synthesis of promising prebiotic dietary fiber from starch.

The tetraploid wheat (Triticum dicoccum (Schrank) Schuebl.) improves nitrogen uptake and assimilation adaptation to nitrogen-deficit stress.

The genetic diversity in tetraploid wheat provides a genetic pool for improving wheat productivity and environmental resilience. The tetraploid wheat had strong N uptake, translocation, and assimilation capacity under N deficit stress, thus alleviating growth inhibition and plant N loss to maintain healthy development and adapt to environments with low N inputs. Tetraploid wheat with a rich genetic variability provides an indispensable genetic pool for improving wheat yield. Mining the physiological mechanisms of tetraploid wheat in response to nitrogen (N) deficit stress is important for low-N-tolerant wheat breeding. In this study, we selected emmer wheat (Kronos, tetraploid), Yangmai 25 (YM25, hexaploid), and Chinese spring (CS, hexaploid) as materials. We investigated the differences in the response of root morphology, leaf and root N accumulation, N uptake, translocation, and assimilation-related enzymes and gene expression in wheat seedlings of different ploidy under N deficit stress through hydroponic experiments. The tetraploid wheat (Kronos) had stronger adaptability to N deficit stress than the hexaploid wheats (YM25, CS). Kronos had better root growth under low N stress, expanding the N uptake area and enhancing N uptake to maintain higher NO3- and soluble protein contents. Kronos exhibited high TaNRT1.1, TaNRT2.1, and TaNRT2.2 expression in roots, which promoted NO3- uptake, and high TaNRT1.5 and TaNRT1.8 expression in roots and leaves enhanced NO3- translocation to the aboveground. NR and GS activity in roots and leaves of Kronos was higher by increasing the expression of TANIA2, TAGS1, and TAGS2, which enhanced the reduction and assimilation of NO3- as well as the re-assimilation of photorespiratory-released NH4+. Overall, Kronos had strong N uptake, translocation, and assimilation capacity under N deficit stress, alleviating growth inhibition and plant N loss and thus maintaining a healthy development. This study reveals the physiological mechanisms of tetraploid wheat that improve nitrogen uptake and assimilation adaptation under low N stress, which will provide indispensable germplasm resources for elite low-N-tolerant wheat improvement and breeding.

Theory of 2D electronic spectroscopy of water soluble chlorophyll-binding protein (WSCP): Signatures of Chl b derivate.

We investigate how electronic excitations and subsequent dissipative dynamics in the water soluble chlorophyll-binding protein (WSCP) are connected to features in two-dimensional (2D) electronic spectra, thereby comparing results from our theoretical approach with experimental data from the literature. Our calculations rely on third-order response functions, which we derived from a second-order cumulant expansion of the dissipative dynamics involving the partial ordering prescription, assuming a fast vibrational relaxation in the potential energy surfaces of excitons. Depending on whether the WSCP complex containing a tetrameric arrangement of pigments composed of two dimers with weak excitonic coupling between them binds the chlorophyll variant Chl a or Chl b, the resulting linear absorption and circular dichroism spectra and particularly the 2D spectra exhibit substantial differences in line shapes. These differences between Chl a WSCP and Chl b WSCP cannot be explained by the slightly modified excitonic couplings within the two variants. In the case of Chl a WSCP, the assumption of equivalent dimer subunits facilitates a reproduction of substantial features from the experiment by the calculations. In contrast, for Chl b WSCP, we have to assume that the sample, in addition to Chl b dimers, contains a small but distinct fraction of chemically modified Chl b pigments. The existence of such Chl b derivates has been proposed by Pieper et al. [J. Phys. Chem. B 115, 4042 (2011)] based on low-temperature absorption and hole-burning spectroscopy. Here, we provide independent evidence.

Stress Responses and Ammonia Nitrogen Removal Efficiency of Oocystis lacustris in Saline Ammonium-Contaminated Wastewater Treatment.

The increasing concern over climate change has spurred significant interest in exploring the potential of microalgae for wastewater treatment. Among the various types of industrial wastewaters, high-salinity NH4+-N wastewater stands out as a common challenge. Investigating microalgae's resilience to NH4+-N under high-salinity conditions and their efficacy in NH4+-N utilization is crucial for advancing industrial wastewater microalgae treatment technologies. This study evaluated the effectiveness of employing nitrogen-efficient microalgae, specifically Oocystis lacustris, for NH4+-N removal from saline wastewater. The results revealed Oocystis lacustris's tolerance to a Na2SO4 concentration of 5 g/L. When the Na2SO4 concentration reached 10 g/L, the growth inhibition experienced by Oocystis lacustris began to decrease on the 6th day of cultivation, with significant alleviation observed by the 7th day. Additionally, the toxic mechanism of saline NH4+-N wastewater on Oocystis lacustris was analyzed through various parameters, including chlorophyll-a, soluble protein, oxidative stress indicators, key nitrogen metabolism enzymes, and microscopic observations of algal cells. The results demonstrated that when the Oocystis lacustris was in the stationary growth phase with an initial density of 2 × 107 cells/L, NH4+-N concentrations of 1, 5, and 10 mg/L achieved almost 100% removal of the microalgae on the 1st, 2nd, and 4th days of treatment, respectively. On the other hand, saline NH4+-N wastewater minimally impacted photosynthesis, protein synthesis, and antioxidant systems within algal cells. Additionally, NH4+-N within the cells was assimilated into glutamic acid through glutamate dehydrogenase-mediated pathways besides the conventional pathway involving NH4+-N conversion into glutamine and assimilation amino acids.

Gamma Irradiation-Mediated Alterations in Chemical Composition and Antimicrobial Efficacy of Zubaidi, a Wild Edible Desert Truffle (Terfezia boudieri), with Implications for Pharmaceutical Applications

In exploring the impact of γ-radiation on the biochemical makeup and antimicrobial attributes of Zubaidi truffle (Terfezia boudieri), this study delved into the nuanced interplay between irradiation doses, chemical components, and antimicrobial efficacy. This research scrutinized the influence of γ-radiation on the chemical constitutes and antimicrobial characteristics of Zubaidi truffle (Terfezia boudieri). By applying four distinct doses of γ-radiation (2.5, 5.0, 7.5, and 10 kGy), the study assesses various parameters, including phenols, flavonoids, total and reduced soluble sugars, crude and soluble protein, total amino acids, antioxidant activity, and the susceptibility of examined strains to irradiated and non-irradiated truffle extracts in comparison to standard antibiotics. The findings elucidate that γ-irradiation induces moderate adjustments in the chemical composition, coupled with a dose-dependent escalation in antioxidant activity. Remarkably, irradiated truffle extracts highlight antibiotic efficacy comparable to standard antibiotics, unveiling a nuanced correlation influenced by both radiation dosage and bacterial strain.

Unveiling the Antioxidant Potential of Halophyte Plants and Seaweeds for Health Applications

Halophyte plants and seaweed are described in the literature as rich sources of antioxidant compounds that can be used in the pharmaceutical and food industries. In this work, we studied the antioxidant composition of five species of halophytic plants (Suaeda vera Forssk, Portulaca oleracea L., Inula crithmoides L., Salicornia ramosissima (Hook.f.) J. Woods and Sarcocornia perennis (Mill.) A.J.Scott) and three seaweeds (Gracilaria gracilis (Stackhouse) Steentoft, L.Irvine and Farnham, Fucus spiralis L. and Ulva rigida C. Agardh) collected in Sado Estuary, Portugal. In the case of the plants, different parts of the plant were also assessed. Various extraction procedures were also performed to understand which methods were most suitable for extracting the various antioxidant compounds. Therefore, the aim of this study was to characterize the antioxidant compounds in halophytes and seaweed using various methods (ABTS, DPPH and FRAP), as well as the phenolic (TPC) and flavonoid (TFC) contents in the different extracts obtained. The amount of soluble protein in each extract was also determined. The results show that methanolic extracts generally have a higher antioxidant capacity, while the highest soluble protein content was observed in aqueous extracts. The seaweed Fucus Spiralis showed the highest antioxidant content, while in halophytic plants the highest antioxidant content was detected in the leaves. In general, this work confirms the potential of halophytes and seaweed as sources of antioxidant compounds for use in the food and pharmaceutical industries.

Tumor-associated CD8+T cell tolerance induced by erythroid progenitor cells.

CD8+T cell tolerance plays an important role in tumor escape. Recent studies have shown that CD45+ erythroid progenitor cells (CD45+EPCs) generated through splenic extramedullary erythropoiesis suppress tumor immunity. However, the mechanism underlying how CD45+EPCs mediate CD8+T cell tolerance remains incompletely understood and requires further research. In this study, the antigen-processing abilities of CD45+EPCs was verified through both in vitro and in vivo experiments. We have used the method of co-culture in vitro and adoptive transfer experiments in vivo to explore the effects of CD45+EPCs on CD8+T cell tolerance. RNA-sequencing analysis and blocking experiments were used to evaluate the role of ROS in the CD45+EPC mediated tolerance of CD8+T cells. Finally, we incorporated uric acid into the adoptive transfer experiments to rescue the CD45+EPC mediated tumor-promoting effect. We found that CD45+EPCs take up soluble proteins, present antigenic epitopes on their surface, and induce antigen-specific CD8+T cell anergy. In addition, we found that CD45+EPC directly nitrates tyrosine within the TCR/CD8 complex via the production of reactive oxygen species and peroxynitrite, preventing CD8+ T cells from responding to their specific peptide antigens. Furthermore, uric acid treatment effectively abolished the immunosuppressive effects of CD45+EPCs during CD8+T cell adoptive transfer, thereby enhancing the anti-tumor efficacy. These results demonstrated that CD8+T cell tolerance in tumor-bearing mice is induced by CD45+EPCs. The results of this study have direct implications for tumor immunotherapy.

Seed osmopriming with polyethylene glycol (PEG) enhances seed germination and seedling physiological traits of Coronilla varia L. under water stress.

Water stress can adversely affect seed germination and plant growth. Seed osmopriming is a pre-sowing treatment in which seeds are soaked in osmotic solutions to undergo the first stage of germination prior to radicle protrusion. Seed osmopriming enhances germination performance under stressful environmental conditions, making it an effective method to improve plant resistance and yield. This study analyzed the effect of seed osmopriming with polyethylene glycol (PEG) on seed germination and physiological parameters of Coronilla varia L. Priming treatments using 10% to 30% PEG enhanced germination percentage, germination vigor, germination index, vitality index, and seedling mass and reduced the time to reach 50% germination (T50). The PEG concentration that led to better results was 10%. The content of soluble proteins (SP), proline (Pro), soluble sugars (SS), and malondialdehyde (MDA) in Coronilla varia L. seedlings increased with the severity of water stress. In addition, under water stress, electrolyte leakage rose, and peroxidase (POD) and superoxide dismutase (SOD) activities intensified, while catalase (CAT) activity increased at mild-to-moderate water stress but declined with more severe deficiency. The 10% PEG priming significantly improved germination percentage, germination vigor, germination index, vitality index, and time to 50% germination (T50) under water stress. Across the water stress gradient here tested (8 to 12% PEG), seed priming enhanced SP content, Pro content, and SOD activity in Coronilla varia L. seedlings compared to the unprimed treatments. Under 10% PEG-induced water stress, primed seedlings displayed a significantly lower MDA content and electrolyte leakage than their unprimed counterparts and exhibited significantly higher CAT and POD activities. However, under 12% PEG-induced water stress, differences in electrolyte leakage, CAT activity, and POD activity between primed and unprimed treatments were not significant. These findings suggest that PEG priming enhances the osmotic regulation and antioxidant capacity of Coronilla varia seedlings, facilitating seed germination and seedling growth and alleviating drought stress damage, albeit with reduced efficacy under severe water deficiency.

Effect of limited proteolysis and CaCl2 on the rheology, microstructure and in vitro digestibility of pea protein-carboxymethyl cellulose mixed gel

Limited proteolysis, CaCl2 and carboxymethyl cellulose (CMC) have individually demonstrated ability to increase the gel strength of laboratory-extracted plant proteins. However, the syneresis effects of their combination on the gelling capacity of commercial plant protein remains unclear. This was investigated by measuring the rheological property, microstructure and protein–protein interactions of gels formed from Alcalase hydrolyzed or intact pea proteins in the presence of 0.1 % CMC and 0–25 mM CaCl2. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed the molecular weight of pea protein in the mixture were < 15 kDa after hydrolysis. The hydrolysates showed higher intrinsic fluorescence intensity and lower surface hydrophobicity than the intact proteins. Rheology showed that the storage modulus (G') of hydrolyzed pea protein (PPH)-based gels sightly decreased compared to those of native proteins. 5–15 mM CaCl2 increased the G’ for both PP and PPH-based gels and decreased the strain in the creep-recovery test. Scanning electron microscopy (SEM) showed the presence of smaller protein aggregates in the PPH-based gels compared to PP gels and the gel network became denser, and more compact and heterogenous in the presence of 15 and 25 mM CaCl2. The gel dissociation assay revealed that hydrophobic interactions and hydrogen bonds were the dominant forces to maintain the gel structure. In vitro digestion showed that the soluble protein content in PPH-based gels was 10 ∼ 30 % higher compared to those of the PP counterpart. CaCl2 addition reduced protein digestibility with a concentration dependent behavior. The results obtained show contrasting effects of limited proteolysis and CaCl2 on the gelling capacity and digestibility of commercial pea proteins. These findings offer practical guidelines for developing pea protein-based food products with a balanced texture and protein nutrition through formulation and enzymatic pre-treatment.