Soluble Protein Research Articles

Biochar Increases Rice Yield in Soda Saline-Alkali Paddy Fields by Improving Saline-Alkali Stress and Phosphorus Use Efficiency

Soda saline-alkali significantly hinders rice growth, phosphorus utilization efficiency, and yield formation. The application of biochar can alleviate the adverse effects of saline-alkali stress on crops. However, there is limited research on the interaction between biochar and phosphorus fertilizer concerning ionic accumulation, phosphorus utilization efficiency, and rice yield in soda saline-alkali soils. A two-year field experiment was conducted to study the combined effects of biochar and phosphate fertilizer on ionic accumulation, physiological status, phosphorus utilization efficiency, and rice yield in soda saline-alkali soil. Four treatments were established for the study: NK (225 kg N, 75 kg K ha−1 year−1), NPK (225 kg N, 70 kg P, 75 kg K ha−1 year−1), NK + B [225 kg N, 75 kg K ha−1 year−1, 1.5% biochar (w/w)], and NPK + B [225 kg N, 70 kg P, 75 kg K ha−1 year−1, 1.5% biochar (w/w)]. The findings indicated that the combined application of biochar and phosphorus fertilizer (NPK + B) significantly reduced the Na+ concentration, Na+/K+ ratio, malondialdehyde (MDA), superoxide anion (O2−), and hydrogen peroxide (H2O2) levels in rice plants. Furthermore, it resulted in a significant increase in K+ concentration and elevated the levels of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), proline (Pro), soluble protein (SP), soluble sugar (SS), and acid phosphatase (ACP). The NPK + B treatment exhibited a significant difference compared to the other treatments (p < 0.05). Compared with NK, phosphorus accumulation and phosphorus utilization efficiency under NPK + B were significantly increased (p < 0.05). The average of biomass yield, grain yield, and harvest index of NPK + B, NK + B, and NPK significantly surpassed those of NK by 6.28–12.25%, 19.80–42.13%, and 11.59–24.64%, respectively. Moreover, a significant difference was observed between NPK + B and the other treatments (p < 0.05). Principal component analysis of the two-year mean data revealed a strong positive correlation of 89.5% for PC1 and a minor negative correlation of 4.4% for PC2. Our research findings demonstrate that the combination of biochar and phosphorus fertilizer effectively enhances salt and alkali tolerance in rice plants, resulting in increased yield through improved ionic balance and physiological status.

2-Keto-L-Gulonic Acid Enhances Cold Stress Tolerance in Arabidopsis thaliana via Augmentation of Ascorbic Acid Biosynthesis and Antioxidant Defense Systems

Cold stress severely impairs plant growth, development, and yields. L-ascorbic acid (ASA), a crucial antioxidant, is pivotal in mitigating stress-induced damage. Previous research found that 2-keto-L-gulonic acid (2KGA), a precursor of ASA in its industrial production, effectively enhances the endogenous ASA content in plants. We hypothesized that 2KGA might alleviate chilling stress and tried to verify it through a cultivation experiment of Arabidopsis thaliana. The results demonstrate that the application of 2KGA significantly increased ASA content (24.58%) and up-regulated ASA biosynthetic genes in Arabidopsis at 4 °C for 24 h. Furthermore, 2KGA alleviated the decrease in fresh weight (17.05%) and total chlorophyll content (15.85%) caused by low temperatures. The contents of proline, soluble sugar (SS), soluble protein (SP), and the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were significantly increased under the 2KGA treatment at low temperatures, while the malondialdehyde (MDA) content was reduced. Moreover, 2KGA up-regulated the ICE-CBF-COR signaling pathway in response to cold stress. These collective findings strongly support the involvement of 2KGA in enhancing cold tolerance in Arabidopsis, presenting an innovative approach for agricultural practices aimed at enhancing crop resilience to environmental stresses.

An efficient protocol for the extraction of pigment-free active polyphenol oxidase and soluble proteins from plant cells.

The elimination of brownish pigments from plant protein extracts has been a challenge in plant biochemistry studies. Although numerous approaches have been developed to reduce pigments for enzyme assays, none has been able to completely remove pigments from plant protein extracts for biochemical studies. A simple and effective protocol was developed to completely remove pigments from plant protein extracts. Proteins were extracted from red anthocyanin-rich transgenic and greenish wild-type tobacco cells cultured on agar-solidified Murashige and Skoog medium. Protein extracts from these cells were brownish or dark due to the pigments. Four approaches were comparatively tested to show that the diethylaminoethyl (DEAE)-Sephadex anion exchange gel column was effective in completely removing pigments to obtain transparent pigment-free protein extracts. A Millipore Amicon® Ultra 10K cut-off filter unit was used to effectively desalt proteins. Moreover, the removal of pigments significantly improved the measurement accuracy of total soluble proteins. Furthermore, enzymatic assays using catechol as a substrate coupled with high-performance liquid chromatography analysis demonstrated that the pigment-free proteins not only showed polyphenol oxidase (PPO) activity but also enhanced the catalytic activity of PPO. Taken together, this protocol is effective for extracting pigment-free plant proteins for plant biochemistry studies. A simple and effective protocol was successfully developed to not only completely and effectively remove anthocyanin and polyphenolics-derived quinone pigments from plant protein extracts but also to decrease the effects of pigments on the measurement accuracy of total soluble proteins. This robust protocol will enhance plant biochemical studies using pigment-free native proteins, which in turn increase their reliability and sensitivity.

All‐Atom Protein Sequence Design Based on Geometric Deep Learning

Designing sequences for specific protein backbones is a key step in creating new functional proteins. Here, we introduce GeoSeqBuilder, a deep learning framework that integrates protein sequence generation with side chain conformation prediction to produce the complete all‐atom structures for designed sequences. GeoSeqBuilder uses spatial geometric features from protein backbones and explicitly includes three‐body interactions of neighboring residues. GeoSeqBuilder achieves native residue type recovery rate of 51.6%, comparable to ProteinMPNN and other leading methods, while accurately predicting side chain conformations. We first used GeoSeqBuilder to design sequences for thioredoxin and a hallucinated three‐helical bundle protein. All the 15 tested sequences expressed as soluble monomeric proteins with high thermal stability, and the 2 high‐resolution crystal structures solved closely match the designed models. The generated protein sequences exhibit low similarity (minimum 23%) to the original sequences, with significantly altered hydrophobic cores. We further redesigned the hydrophobic core of glutathione peroxidase 4, and 3 of the 5 designs showed improved enzyme activity. Although further testing is needed, the high experimental success rate in our testing demonstrates that GeoSeqBuilder is a powerful tool for designing novel sequences for predefined protein structures with atomic details. GeoSeqBuilder is available at https://github.com/PKUliujl/GeoSeqBuilder

Two-way Dispatched function in Sonic hedgehog shedding and transfer to high-density lipoproteins.

The Sonic hedgehog (Shh) signaling pathway controls embryonic development and tissue homeostasis after birth. This requires regulated solubilization of dual-lipidated, firmly plasma membrane-associated Shh precursors from producing cells. Although it is firmly established that the resistance-nodulation-division transporter Dispatched (Disp) drives this process, it is less clear how lipidated Shh solubilization from the plasma membrane is achieved. We have previously shown that Disp promotes proteolytic solubilization of Shh from its lipidated terminal peptide anchors. This process, termed shedding, converts tightly membrane-associated hydrophobic Shh precursors into delipidated soluble proteins. We show here that Disp-mediated Shh shedding is modulated by a serum factor that we identify as high-density lipoprotein (HDL). In addition to serving as a soluble sink for free membrane cholesterol, HDLs also accept the cholesterol-modified Shh peptide from Disp. The cholesteroylated Shh peptide is necessary and sufficient for Disp-mediated transfer because artificially cholesteroylated mCherry associates with HDL in a Disp-dependent manner, whereas an N-palmitoylated Shh variant lacking C-cholesterol does not. Disp-mediated Shh transfer to HDL is completed by proteolytic processing of the palmitoylated N-terminal membrane anchor. In contrast to dual-processed soluble Shh with moderate bioactivity, HDL-associated N-processed Shh is highly bioactive. We propose that the purpose of generating different soluble forms of Shh from the dual-lipidated precursor is to tune cellular responses in a tissue-type and time-specific manner.

Two-way Dispatched function in Sonic hedgehog shedding and transfer to high-density lipoproteins

The Sonic hedgehog (Shh) signaling pathway controls embryonic development and tissue homeostasis after birth. This requires regulated solubilization of dual-lipidated, firmly plasma membrane-associated Shh precursors from producing cells. Although it is firmly established that the resistance-nodulation-division transporter Dispatched (Disp) drives this process, it is less clear how lipidated Shh solubilization from the plasma membrane is achieved. We have previously shown that Disp promotes proteolytic solubilization of Shh from its lipidated terminal peptide anchors. This process, termed shedding, converts tightly membrane-associated hydrophobic Shh precursors into delipidated soluble proteins. We show here that Disp-mediated Shh shedding is modulated by a serum factor that we identify as high-density lipoprotein (HDL). In addition to serving as a soluble sink for free membrane cholesterol, HDLs also accept the cholesterol-modified Shh peptide from Disp. The cholesteroylated Shh peptide is necessary and sufficient for Disp-mediated transfer because artificially cholesteroylated mCherry associates with HDL in a Disp-dependent manner, whereas an N-palmitoylated Shh variant lacking C-cholesterol does not. Disp-mediated Shh transfer to HDL is completed by proteolytic processing of the palmitoylated N-terminal membrane anchor. In contrast to dual-processed soluble Shh with moderate bioactivity, HDL-associated N-processed Shh is highly bioactive. We propose that the purpose of generating different soluble forms of Shh from the dual-lipidated precursor is to tune cellular responses in a tissue-type and time-specific manner.

Effect of Drought and Rehydration on Physiological Characteristics of Agriophyllum squarrosum (L.) Moq. in Different Habitats.

Agriophyllum squarrosum (L.) Moq. is a highly prevalent xerophytic species found throughout northern China. It is suitable for cultivation in semi-arid sandy environments and may establish roots in arid desert locations. This species plays a pioneering and exploratory role in the colonization of desert plants. In this study, we selected A. squarrosum from the Urat desert steppe (UD) and Horqin sandy land (HS) to explore their adaptation mechanisms to drought and rehydration environments by using the pot weighing control method to simulate an arid environment. The findings showed that the control (watering to 60-65% of field capacity) exceeded its required amount and the leaves turned yellow. The chlorophyll content was lower than those under moderate and severe drought, and rehydration caused a decrease. However, the contents of malondialdehyde, soluble sugar, and proline in the drought treatment were higher than those in the control. Under moderate and severe drought, the chlorophyll content and the quantum efficiency of photosystem II (Fv/Fm) of A. squarrosum from UD were higher than those from HS. During drought and rehydration processes, the proline content was relatively lower, while the activities of peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) and the content of soluble sugar and soluble protein were higher. However, antioxidant enzymes and osmoregulators from UD were higher than those from HS. The results suggest that the stronger ability of A. squarrosum to endure drought environments in UD is due to the high level of antioxidant enzymes and osmoregulators, which are conducive to relieving cell membrane damage when subjected to drought and rehydration.

Investigation of biomolecular dynamics by sensitivity-enhanced 1H–2H CPMAS NMR using matrix-free dynamic nuclear polarization

Molecular dynamics of functional groups contain valuable information about structural properties and functional activities in biomolecules. NMR spectroscopy is a sensitive tool for the investigation of molecular dynamics over a wide range of timescales and thus may deepen the understanding of the biomolecules of interest. Here, we present an approach to use DNP-enhanced 2H NMR to study dynamics of selectively deuterated methyl groups in insoluble proteins such as amyloid beta (Aβ) fibrils. We adopted and optimized the matrix-free DNP approach by varying the amount of added polarizing agent as well as the rehydration level of model proteins. We show that the DNP enhancement obtained in 1H–2H cross-polarization (CP) MAS spectra may increase the sensitivity for selectively deuterated Aβ fibril samples by more than one order of magnitude, accelerating the collection of spin-lattice relaxation data in the DNP-accessible temperature range between 100 and 150 K by up to 400-fold. However, below the coalescence temperature, which describes the transition from the fast to the slow exchange regime, the experimentally obtained relaxation time constants suffer from a paramagnetic relaxation enhancement effect due to the presence of the polarizing agent. This seems to be a general effect for biomolecules as it is also confirmed for two other protein model systems. Our demonstration opens the possibility to extend the scope of 2H NMR for dynamics measurements to effective concentrations and/or temperatures below what is currently accessible; however, the observed interplay between paramagnetic relaxation and molecular dynamics also emphasizes the necessity for a better understanding of relaxation effects in DNP-enhanced NMR.

Effects of High-Pressure Homogenization on the Structure and Functional Properties of Solenaia oleivora Proteins.

Solenaia oleivora, a rare freshwater shellfish with high protein quality, is unique to China. However, the poor hydrosolubility and functional properties of Solenaia oleivora proteins hinder their utilization in food products. Herein, the alkaline dissolution-isoelectric precipitation method was used for the extraction of Solenaia oleivora proteins. Furthermore, the impact of high-pressure homogenization (HPH) treatment varying from 0 to 100 MPa on the structure and functional properties of Solenaia oleivora proteins was investigated. The obtained results indicated that HPH treatment decreased the α-helix content and enhanced the β-sheet and random coil content. Furthermore, the HPH caused the unfolding of protein structure, exposing aromatic amino acids, increasing the free thiol group content, and enhancing surface hydrophobicity. As the homogenization pressure increased from 0 to 100 MPa, the particle size of Solenaia oleivora proteins decreased from 899 to 197 nm with the polymer dispersity index (PDI) value decreased from 0.418 to 0.151, the ζ-potential increased from -22.82 to -43.26 mV, and the solubility increased from 9.54% to 89.96%. Owing to the significant changes in protein structure and solubility, the emulsifying, foaming, and digestive properties of Solenaia oleivora proteins have been significantly improved after treatment with HPH.

Investigation of physico-chemical and microbiological parameters of traditional mongolian cheese

The traditional technology of Mongolian cheese is almost the same in different parts of the country, it differs from other types of cheeses, there is no microbiological process in it. Within the framework of this study, physico-chemical and microbiological parameters were determined in order to assess the quality and safety of Mongolian cheese sold on the market in Ulanbaatar. The article is devoted to the traditional Mongolian cheese, which belongs to the group of fresh fermented milk cheeses produced by thermal acid coagulation of milk. The main stages of its production include pasteurization of milk, coagulation, molding and pressing of cheese mass. Unlike other types of cheese, the production process of Mongolian cheese does not include clot processing, salting and maturation, which limits its shelf life. However, this cheese is an important source of casein and whey proteins for the Mongolian population. The method of cheese production is based on thermic acid coagulation, which uses high temperature. Mongolian cheese, like other cheeses of this group, is absorbed by the body by 96-98%, being an important source of calcium, phosphorus and amino acids. Milk proteins in such cheeses are not decomposed by microorganisms, which reduces the content of soluble proteins. Studies of the microflora of Mongolian cheese have shown the presence of various microorganisms, mainly lactic acid bacteria, and the absence of pathogenic bacteria when properly stored. The addition of preservatives such as nisin and natamycin can extend the shelf life of cheese up to 15 days.

Comparison of rice bran characteristics fermented by mono- or di-strain probiotics based on omics technology

Rice bran is a premium agricultural byproduct rich in nutrients and bioactive substances. The aim of this study was to compare the effects of fermentation with Bacillus subtilis, Lactobacillus plantarum, and a mixture of both on the bacterial communities and metabolic functions of rice bran based on omics techniques. The results showed that probiotic fermentation decreased crude fiber (CF) content, phytic acid, and trypsin inhibitor activity (TIA), and increased trichloroacetic acid soluble protein (TCA-SP), reducing sugar (RS), and total phenol (TP) in rice bran. Ten dominant genera with relative abundances greater than 1% were successfully identified, and probiotic fermentation significantly reduced the relative abundances of Enterobacter, Kasakonia, and Pantoea. A total of 63 differentially expressed metabolites (DEMs) were identified and further analysis revealed that probiotic fermentation increased essential amino acids, phenolic compounds, and organic acids in rice bran. Additionally, the di-strain probiotic fermentation of Bacillus subtilis and Lactobacillus plantarum facilitated the growth of beneficial microbiota (Limosilactobacillus, Prevotella and Dialister) and the production of bioactive substances (lactic acid, catechol, beta-tyrosine, etc.). In conclusion, co-fermentation of di-strain probiotics is a valuable method to produce functional compounds and enhance the health-promoting properties of rice bran.

Hydrogen sulfide alleviates cadmium stress in germinating carrot seeds by promoting the accumulation of proline

Carrot (Daucus carota L.), a widely cultivated economically vegetable from the Apiaceae family, is grown globally. However, carrots can be adversely impacted by cadmium (Cd) pollution in the soil due to its propensity to accumulate in the fleshy root, thus impeding carrot growth and posing health hazards to consumers. Given the potential of hydrogen sulfide (H2S) to improve plant resistance against Cd stress, we treated germinating carrot seeds with varying concentrations of sodium hydrosulfide (NaHS), aiming to alleviate the toxic impacts of Cd stress on carrot seed germination. The results revealed that carrot seeds treated with a concentration of 0.25 mM NaHS displayed better seed germination-associated characteristics compared to seeds treated with NaHS concentrations of 0.1 mM and 0.5 mM. Further investigation revealed a rise in the expression levels of L-cysteine desulfhydrase and D-cysteine desulfhydrase, along with enhanced activity of L-cysteine desulfhydrase and D-cysteine desulfhydrase among the NaHS treatment group, thereby leading to H2S accumulation. Moreover, NaHS treatment triggered the expression of pyrroline-5-carboxylate synthase and pyrroline-5-carboxylate reductase and promoted the accumulation of endogenous proline, while the contents of soluble sugar and soluble protein increased correspondingly. Interestingly, since the application of exogenous proline did not influence the accumulation of endogenous H2S, suggesting that H2S served as the upstream regulator of proline. Histochemical staining and biochemical indices revealed that NaHS treatment led to elevated antioxidant enzyme activity, alongside a suppression of superoxide anion and hydrogen peroxide generation. Furthermore, high performance liquid chromatography analysis revealed that NaHS treatment reduced Cd2+ uptake, thereby promoting germination rate, seed vitality, and hypocotyl length of carrot seeds under Cd stress. Overall, our findings shed light on the application of NaHS to enhance carrot resistance against Cd stress and lay a foundation for exploring the regulatory role of H2S in plants responding to Cd stress.

Substituting partial chemical nitrogen fertilizers with organic fertilizers maintains grain yield and increases nitrogen use efficiency in maize

IntroductionLong-term application of excessive nitrogen (N) not only leads to low N use efficiency (NUE) but also exacerbates the risk of environmental pollution due to N losses. Substituting partial chemical N with organic fertilizer (SP) is an environmentally friendly and sustainable fertilization practice. However, the appropriate rate of SP in rainfed maize cropping systems in semi-arid regions of China is unknown.MethodsTherefore, we conducted a field experiment between 2021 and 2022 in a semi-arid region of Northern China to investigate the effects of SP on maize growth, carbon and N metabolism (C/NM), and NUE. The following treatments were used in the experiment: no N application (CK), 100% chemical N (SP0, 210 kg N ha–1), and SP substituting 15% (SP1), 30% (SP2), 45% (SP3), and 60% (SP4) of the chemical N. The relationship between these indicators and grain yield (GY) was explored using the Mantel test and structural equation modeling (SEM).Results and discussionThe results found that the SP1 and SP2 treatments improved the assimilates production capacity of the canopy by increasing the leaf area index, total chlorophyll content, and net photosynthetic rate, improving dry matter accumulation (DMA) by 6.2%–10.6%, compared to the SP0 treatment. SP1 and SP2 treatments increased total soluble sugars, starch, free amino acids, and soluble protein contents in ear leaves via increasing the enzymatic reactions related to C/NM in ear leaves during the reproductive growth stage compared with SP0 treatment. The highest plant nitrogen uptake (PNU) and nitrogen recovery efficiency were obtained under the SP2 treatment, and the GY and nitrogen agronomic efficiency were higher than the SP0 treatment by 9.2% and 27.8%. However, SP3 and SP4 treatments reduced DMA and GY by inhibiting C/NM in ear leaves compared to SP0 treatment. Mantel test and SEM results revealed that SP treatments indirectly increased GY and PNU by directly positively regulating C/NM in maize ear leaves. Therefore, in the semi-arid regions, substituting 30% of the chemical N with SP could be considered. This fertilizer regime may avoid GY reduction and improve NUE. This study provides new insights into sustainable cultivation pathways for maize in semi-arid regions.

Nectin1 is a pivotal host factor involved in attachment and entry of red-spotted grouper nervous necrosis virus in the early stages of the viral life cycle.

Nervous necrosis virus (NNV) is a highly neurotropic virus that poses a persistent threat to the survival of multiple fish species. However, its inimitable neuropathogenesis remains largely elusive. To rummage potential partners germane to the nervous system, we investigated the interaction between red-spotted grouper NNV (RGNNV) and grouper brain by immunoprecipitation coupled with mass spectrometry and discerned Nectin1 as a novel host factor subtly involved in viral early invasion events. Nectin1 was abundant in neural tissues and implicated in the inception of tunnel nanotubes triggered by RGNNV. Its overexpression not only dramatically potentiated the replication dynamics of RGNNV in susceptible cells, but also empowered non-sensitive cells to expeditiously capture free virions within 2 min. This potency was impervious to low temperatures but was dose-dependently suppressed by soluble protein or specific antibody of Nectin1 ectodomain, indicating Nectin1 as an attachment receptor for RGNNV. Mechanistically, efficient hijacking of virions by Nectin1 strictly depended on intricate linkages to different modules of viral capsid protein, especially the direct binding between the IgC1 loop and P-domain. More strikingly, despite abortive proliferation in Nectin1-reconstructed CHSE-214 cells, a non-sensitive cell, RGNNV could gain access to the intracellular compartment by capitalizing on Nectin1, thereby inducing canonical cytoplasmic vacuolation. Altogether, our findings delineate a candidate entrance for RGNNV infiltration into the nervous system, which may shed unprecedented insights into the exploration and elucidation of RGNNV pathogenesis.IMPORTANCENervous necrosis virus (NNV) is one of the most virulent pathogens in the aquaculture industry, which inflicts catastrophic damage to ecology, environment, and economy annually around the world. Nevertheless, its idiosyncratic invasion and latency mechanisms pose enormous hardships to epidemic prevention and control. In this study, deploying grouper brain as a natural screening library, a single-transmembrane glycoprotein, Nectin1, was first identified as an emergent functional receptor for red-spotted grouper NNV (RGNNV) that widely allocated in nervous tissues and directly interacted with viral capsid protein through distinct Ig-like loops to bridge virus-host crosstalk, apprehend free virions, and concomitantly propel viral entry. Our findings illuminate the critical role of Nectin1 in RGNNV attachment and entry and provide a potential target for future clinical intervention strategies in the therapeutic race against RGNNV.

Metastable Liquid–Liquid Phase Separation and Aging Lead to Strong Processing Path Dependence in Mini‐Spidroin Solutions

AbstractRecombinant silk proteins provide a route toward sustainable and biocompatible materials. For making such materials, the assembly process from dilute protein into a functional material is central. The assembly mechanism in engineered materials is by necessity different from the natural ones—this poses challenges but also opens opportunities for scaling up and for developing novel properties. The phase behavior of a mini‐spidroin, NT‐2Rep‐CT is studied, which is a widely studied variant of recombinant silk. NT‐2Rep‐CT can be triggered to assemble by lowering the pH, but even at high pH—considered as storage conditions—it can be in various states, such as forming condensates, clusters, gels, and soluble protein. It is shown how its assembly phases evolve through both metastable and dynamically arrested states. The observed behavior of silk protein solutions is highly complex, and elements thereof from phase diagrams associated with polymers, colloidal systems, and globular proteins are found. Based on the characterization of cluster formation and structural intermediates, a minimalist phase diagram is proposed for NT‐2Rep‐CT and argues that the understanding and insight into silk assembly via its phase behavior, and especially the arrested states, is central for designing recombinant silk proteins and their processing for materials applications.

Alterations in circulating immunoregulatory proteins discriminate poor CD4 T lymphocyte trajectories in people with HIV on suppressive antiretroviral therapy.

It is essential to track immune perturbations related to insufficient CD4 T-cell recovery in PWH on suppressive ART as those with incomplete reconstitution are at a greater risk of non-AIDS-related morbidity and mortality. Several inflammatory soluble mediators have associated with poor immune reconstitution and adverse morbid outcomes in PWH, yet their implementation into routine clinical care to guide management remains inconsistent. Circulating immune checkpoint proteins have been linked to dysregulated immune pathways during suppressive ART and may serve as improved surrogate markers of clinical relevance. Here we investigate soluble lymphocyte-associated immunoregulatory proteins in virally suppressed PWH with no reported co-morbid outcomes and varying CD4 T-cell counts, to reveal underlying pathways that remain perturbed despite ART. This novel signature of immunoregulatory markers pertaining to poor CD4 T-cell trajectories uncover previously overlooked immune checkpoints as important targets for clinical monitoring of PWH in the setting of durable viral suppression by ART.

A High-Throughput Screening Platform for Engineering Poly(ethylene Terephthalate) Hydrolases.

The ability of enzymes to hydrolyze the ubiquitous polyester, poly(ethylene terephthalate) (PET), has enabled the potential for bioindustrial recycling of this waste plastic. To date, many of these PET hydrolases have been engineered for improved catalytic activity and stability, but current screening methods have limitations in screening large libraries, including under high-temperature conditions. Here, we developed a platform that can simultaneously interrogate PET hydrolase libraries of 104-105 variants (per round) for protein solubility, thermostability, and activity via paired, plate-based split green fluorescent protein and model substrate screens. We then applied this platform to improve the performance of a benchmark PET hydrolase, leaf-branch compost cutinase, by directed evolution. Our engineered enzyme exhibited higher catalytic activity relative to the benchmark, LCC-ICCG, on amorphous PET film coupon substrates (∼9.4% crystallinity) in pH-controlled bioreactors at both 65 °C (8.5% higher conversion at 48 h and 38% higher maximum rate, at 2.9% substrate loading) and 68 °C (11.2% higher conversion at 48 h and 43% higher maximum rate, at 16.5% substrate loading), up to 48 h, highlighting the potential of this screening platform to accelerate enzyme development for PET recycling.

Enhanced release of volatile halocarbons of microalgae in response to antibiotic-induced stress: Based on laboratory and ship-field experiments

This study investigated the impacts of sulfamethazine (SMZ) and oxytetracycline (OTC) antibiotics on the marine microalgae Nitzschia closterium and its release of volatile halocarbons (VHCs), which contribute to ozone depletion and climate change. High concentrations of SMZ and OTC suppressed cell density, reduced chlorophyll a content, and hindered Fv/Fm elevation in N. closterium, indicating its growth was inhibited. The exposure of N. closterium to antibiotics led to increased reactive oxygen species (ROS), reduced soluble protein content, and heightened catalase (CAT) activity, indicative of increased oxidative stress. This stress increased the release of three VHCs (CHBrCl2, CHBr2Cl, and CHBr3). Ship-borne experiments showed that high phytoplankton biomass was linked to high VHC release. Notably, the production and release of VHCs were significantly higher in the high-concentration antibiotic group (100 μg/L) than the low-concentration group (0.1 μg/L). These findings suggested that antibiotics induce excess ROS in algal cells, stimulating VHC production and release.

Utilizing a hydrophobic primary container surface to reduce the formation of subvisible particles in monoclonal antibody solution caused by fluid shear

The exposure of protein molecules to interfaces may cause protein aggregation and particle formation in protein formulations, especially hydrophobic interfaces, which may promote protein aggregation in solution. In this study, we found that modification of the surface properties by application of a hydrophobic Octadecyltrichlorosilane (OTS) could reduce the generation of protein aggregates and particles in protein solution induced by fluid shear. A stable protein adsorption layer was formed at the hydrophobic interface through the strong hydrophobic interaction between the protein and hydrophobic surface, which could prevent the aggregated protein from falling off into the bulk solution to form subvisible particles and insoluble protein aggregates. In addition, human complement enzyme linked immunosorbent assay results showed that the particles that were generated in the OTS-coated container did not activate human complement which indicated the OTS-coated container could be used as primary containers for certain types of monoclonal antibody formulation.

The Influence of Microalgae Fertilizer on Soil Water Conservation and Soil Improvement: Yield and Quality of Potted Tomatoes

We aim to study the impact of microalgae fertilizer on soil nutrients, water conservation and crop yield and quality while also determining the optimal ratio of microalgae fertilizer to chemical fertilizer. Using “Xinoufen No.9” tomatoes as the test subject, we conducted pot experiments with four different treatments: control with 100% chemical fertilizer (CK), T1 (25% microalgae fertilizer + 75% regular chemical fertilizer), T2 (75% microalgae fertilizer + 25% regular chemical fertilizer) and T3 (100% microalgae fertilizer). The results show that an increased application of microalgae fertilizer enhanced the soil organic matter, ammonium nitrogen, available phosphorus and potassium content. T3 showed the most improvement followed by T2. The co-application of microalgae fertilizer with chemical fertilizer can significantly increase the stem girth, plant height and yield of tomatoes. At the same time, microalgae fertilizer effectively regulates leaf stomatal conductance, promoting tomato leaf respiration. As the stomatal conductance increases, the transpiration rate and net photosynthesis rate of all treatments improve, followed by a decline in intercellular CO2 concentration, with T2 exhibiting the best performance. Among all treatments, T2 treatment yielded the highest per-plant production (0.630 kg), followed by T3 (0.521 kg). This is because the microalgae fertilizer promotes the distribution of photosynthetic products to the fruit, enhancing the yield and quality of tomatoes. Additionally, the microalgae fertilizer also increases the content of soluble sugars, soluble protein, vitamin C and lycopene in the fruit while reducing the nitrate content. Compared to the control group CK, T2 increases the content of soluble sugars, vitamins and lycopene by 26.74%, 39.29% and 158.31%, respectively. Microalgae fertilizer also helps to improve soil water and thermal conditions, enhancing the water-use efficiency of tomatoes. Compared to CK, the water-use efficiency of T2 treatment increased by 54.05%. Correlation analysis indicates that water and fertilizer factors significantly affect tomato yield, with a correlation exceeding 70%. The net photosynthesis and transpiration rates significantly influence fruit quality, with correlations above 80%. By applying microalgae fertilizer, the efficiency of water and fertilizer use can be effectively improved, thus achieving the goal of water conservation and quality enhancement. Therefore, through comprehensive analysis, using the membership function method of indicators such as soil environment, crop yield, fruit quality and water-use efficiency, it is concluded that T2 is the optimal fertilization treatment. This study provides theoretical support for the application of microalgae biofertilizer technology in the cultivation of tomatoes and other vegetables in the northern, cold and arid regions.