Protein Research Articles

Selenium Biofortification of Soybean Sprouts: Effects of Selenium Enrichment on Proteins, Protein Structure, and Functional Properties.

Selenium (Se) biofortification during germination is an efficient method for producing Se-enriched soybean sprouts; however, few studies have investigated Se distribution in different germinated soybean proteins and its effects on protein fractions. Herein, we examined Se distribution and speciation in the dominant proteins 7S and 11S of raw soybean (RS), germinated soybean (GS), and germinated soybean with Se biofortification (GS-Se). The effects of germination and Se treatment on protein structure, functional properties, and antioxidant capacity were also determined. The Se concentration in GS-Se was 79.8-fold higher than that in GS. Selenomethionine and methylselenocysteine were the dominant Se species in GS-Se, accounting for 41.5–80.5 and 19.5–21.2% of the total Se with different concentrations of Se treatment, respectively. Se treatment had no significant effects on amino acids but decreased methionine in 11S. In addition, the α-helix contents decreased as the Se concentration increased; the other structures showed no significant changes. The Se treatment also had no significant effects on the water and oil-holding capacities in protein but increased the foaming capacity and emulsion activity index (EAI) of 7S, but only the EAI of 11S. The Se treatment also significantly increased the antioxidant capacity in 7S but not in 11S. This study indicates that the dominant proteins 7S and 11S have different Se enrichment abilities, and the protein structures, functional properties, and antioxidant capacity of GS can be altered by Se biofortification.

Cerebral venous sinus thrombosis and dural arteriovenous fistula associated with protein S deficiency: a case series study

ObjectiveTo describe the characteristics of patients with cerebral venous sinus thrombosis (CVST) and dural arteriovenous fistula (AVF) associated with protein S (PS) deficiency.MethodsWe conducted a search of medical records in Hainan General Hospital from January 2000 to December 2020 for coexistence of CVST and dural AVF associated with PS deficiency and searched PubMed、Embase and Chinese biomedical databases (CBM) for all literature describing CVST and dural AVF with PS. We analyzed clinical characteristics, location, sequence of CVST and dural AVF, level of PS, therapeutic methods and prognosis.ResultsWe presented 1 patient in our hospital’s database combined CVST and dural AVF associated with PS, plus 5 cases reported in literature. The most common symptoms were headache, generalized seizure, disturbance of consciousness. The most frequent location of CVST was at internal cerebral vein, while transverse sinus, sigmoid sinus, parietal region in dural AVF. Two patients developed dural AVF several months or years after CVST. Clinical characteristics and level of PS were summarized.ConclusionThese findings alert physicians to consider PS deficiency in patients who suffer from CVST, especially those combined with dural AVF.

Term Infant with Bilateral Parenchymal Brain Hemorrhages.

Term Infant with Bilateral Parenchymal Brain Hemorrhages.

Analyzing the roles of cyclic dimeric guanosine monophosphate (c-di-GMP) on the formation of autotrophic granules and autotrophic biofilm in integrated fixed-film activated sludge (IFAS) reactor

This study is the first to reveal the important roles of c-di-GMP on the formation of autotrophic granular sludge (AUGS) and autotrophic biofilm (AUB) in integrated fixed-film activated sludge (IFAS) system. The results revealed that the partial nitrification/anammox (PN/A) performance was successfully developed in the IFAS system and high nitrogen removal efficiency was obtained. Notably, it was clearly indicated that the intracellular c-di-GMP content was highly linearly correlated with the concentration of protein (PN) in tightly bound extracellular polymeric substances (TB-EPS) during AUGS formation. It was also found that the c-di-GMP content was highly linearly correlated with the PN concentration during AUB formation, and also had a good linear correlation with the polysaccharide (PS) content in TB-EPS. This proves that intracellular c-di-GMP played a vital role in regulating the TB-EPS secretion, thereby further affecting the aggregation ability of AUGS and AUB. In addition, when the c-di-GMP regulates the content of PN in TB-EPS, the secondary structure of the PN also changes significantly correspondingly, affecting the hydrophobicity of AUGS and AUB. In long-term operation, Candidatus_Kuenenia, Nitrosomonas and Flavobacterium gradually became dominant species, which were considered to favor the formation of AUGS and AUB by promoting the production of c-di-GMP and TB-EPS.

Molecular level insights into the dynamic evolution of forward osmosis fouling via thermodynamic modeling and quantum chemistry calculation: Effect of protein/polysaccharide ratios

While forward osmosis (FO) shows broad application prospects in wastewater reuse and desalination, it is still hindered by membrane fouling issues. From the perspective of molecular structure and energy, this work investigated the effects of protein (PN)/polysaccharide (PS) ratios of organic foulants on the dynamic evolution of membrane fouling in FO. Fouling tests showed two interesting membrane fouling phenomena: Ⅰ) when PN content increased from 0.2 to 0.8 g/L with fixed PS content of 0.2 g/L, membrane fouling decreased for filtration of the same volume of foulants; Ⅱ) with the increase of filtration volume, the membrane fouling caused by 1:1 PN/PS foulants increased almost linearly, while the membrane fouling caused by 4:1 PN/PS foulants increased significantly at first, and then rapidly reached a stable fouling state. Thermodynamic analyses and quantum chemical calculations revealed that the attractive interaction energy between 1:1 PN/PS foulants and membrane was 1.90 times greater than that for 4:1 PN/PS foulants, and the protein and polysaccharide molecules with a ratio of 1:1 were more likely to form a tight cross-linked structure. Furthermore, the reverse solute diffusion significantly reduced the electrostatic repulsion energy among the polymer chains for 4:1 PN/PS foulants, which induced the transformation of foulant morphology from gel to floc and reduced the adhesion energy to membrane surface by 61%. As a result, in the later fouling stage, few 4:1 PN/PS foulants were newly attached to the membrane, leading to a pseudo-stable flux. Based on the aforementioned findings, this study reveals a unified thermodynamic framework for the effects of the PN/PS ratios of organic foulants on the dynamic evolution of FO membrane fouling. The proposed mechanism is beneficial in formulating “tailored” fouling mitigation strategies and elucidating the appropriate application fields for FO technology.

Analysis of Effects of PTEN-Mediated TGF-β/Smad2 Pathway on Osteogenic Differentiation in Osteoporotic Tibial Fracture Rats and Bone Marrow Mesenchymal Stem Cell under Tension

Purpose. To discuss effects of phosphatase and tensin homolog protein (PTEN)-mediated transforming growth factor-β (TGF-β)/Smad homologue 2 (Smad2) pathway on osteogenic differentiation in osteoporotic (OP) tibial fracture rats and bone marrow mesenchymal stem cell (BMSC) under tension. Methods. A tibial fracture model was established. The rats were divided into sham-operated group and model group, and tibia tissue was collected. Purchase well-grown cultured rat BMSC, and use the Flexercell in vitro cell mechanics loading device to apply tension. The expression of PTEN was detected by qRT-PCR. After the BMSCs were transfected with si-PTEN and oe-PTEN, the force was applied to detect cell differentiation. The expression of TGF-β/Smad2 protein was detected by Western blot. The formation of calcium nodules in BMSC was detected by alkaline phosphatase (ALP) staining and alizarin red (AR) staining. Results. The expression of PTEN was higher in the model group and tension MSC group, and the expression of TGF-β and Smad2 protein was lower. The expression of TGF-β and Smad2 protein in oe-PTEN group was lower than the oe-NC group and control group. The expression of TGF-β and Smad2 protein in si-PTEN group was higher than the si-NC group and control group. The results of ALP staining and AR staining also confirmed the above results. Conclusion. PTEN-mediated TGF-β/Smad2 pathway may play a key role in the osteogenic differentiation of OP tibial fracture rats. Downregulation of PTEN and upregulation of TGF-β/Smad2 signal can promote the osteogenic differentiation of BMSC under tension.

The Effects of Silveroxide on mRNA Level of Key Genes Involved In the Biosynthesis of Rebaudioside-A in Stevia Rebaudiana

Stevia rebaudiana bertoni is a plant with sweetening properties. This medicinal plant is perennial and from Asteraceae family. Its leaves contain glycoside compounds of a sugar part and non-sugar sectors. One of the glycosides compounds is Rebaudioside¬A which has a greater importance in market. Several key regulating genes including copalyl diphosphate synthase (CPPS) (AF034545.1), geranylgeranyl diphosphate synthase (GGDPS) (DQ432013.3), (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase (HDS) (FJ755689.1), UDP glucosyltranserase 85C2(UGT85C2)(AY345978.1), UDPglucosyltranserase–74G1 (UGT74G1) (AY345982.1) and UDP glucosyltranserase–76G1 (UGT76G1) (KC631816.1) are involved in the biosynthesis Rebaudioside A. This experiment was conducted to evaluate the effect of silver oxide Ag2O on the mRNA level of these genes in the stevia rebaudiana. The experiments repeated 3 times and with concentrations of 50, 100 and 200 µM. Increasing concentrations of 50 micromoles of silver oxide up to 100 micromoles leads to an increase in the expression levels of all the studied genes. Also according to the expression profile of these genes and the results of HPLC there is a significant increase on the expression level of the genes and production of Rebaudioside¬A under Ag2O treatment. In general, it was found that increasing the concentration of Ag2O can lead to an increase in the level of mRNA for the chosen genes. On the other hand, the low expression of the genes studied under control conditions (No Treatment), compared to the treatment with Ag2O, revealed that the treatment can lead to higher sweetener glycoside components in the Stevia leaves. The physiological assay showed that Ag2O treatment in concentrations of 100 and 200 µM have more positive effect on chlorophyll, protein, carbohydrates and carotenoids in Stevia.

Quantifying the thermochemical pathways of soluble organics in sewage sludge flocs during pyrolysis for precursor optimization and by-product control

The composition of sewage sludge is complex, and understanding the product contribution and pyrolysis pathways of typical components in sludge are important for product quality improvement and contaminant volatiles control. Here, three major sludge organics, i.e., protein (PR), polysaccharide (PS), and humic acid (HA), were extracted and pyrolyzed individually to quantify the correlation between sludge components and products. The temperature-response and sludge inorganics-response of pyrolysis processes were elucidated simultaneously. The main contributor to the carbonaceous components in char shifted from PR (from 51% to 7%) to HA (from 27% to 78%) as temperature increased from 250℃ to 850℃. Over 95% tar and 70% gas were originated from PR and PS at temperature above 550℃. Regarding the effect of inorganic matters, Ca/Si/Al-containing inorganics were the major inducement for the formation of nitrogenous tars and NOx, whereas polyferric chloride retained sludge organics in the carbon matrix in the form of C–C/C = C and heterocyclic-N/S, thus reducing the generation of volatiles. Based on pyrolysis product evolution and elemental flow analysis, sludge rich in HA and low in Ca/Si/Al-containing inorganics was found to be a promising source for carbonization at 550-850℃, while PR/PS-rich sludge was more suitable for pyrolysis or liquefaction at below 550℃. These findings provided reliable strategies for pyrolysis precursor selection, gas emission control, and potential application of pyrolysis product.

Detection of Unknown and Rare Pathogenic Variants in Antithrombin, Protein C and Protein S Deficiency Using High-Throughput Targeted Sequencing.

The deficiency of natural anticoagulants—antithrombin (AT), protein C (PC), and protein S (PS)—is a highly predisposing factor for thrombosis, which is still underdiagnosed at the genetic level. We aimed to establish and evaluate an optimal diagnostic approach based on a high-throughput sequencing platform suitable for testing a small number of genes. A fast, flexible, and efficient method involving automated amplicon library preparation and target sequencing on the Ion Torrent platform was optimized. The cohort consisted of a group of 31 unrelated patients selected for sequencing due to repeatedly low levels of one of the anticoagulant proteins (11 AT-deficient, 13 PC-deficient, and 7 PS-deficient patients). The overall mutation detection rate was 67.7%, highest in PC deficiency (76.9%), and six variants were newly detected—SERPINC1 c.398A > T (p.Gln133Leu), PROC c.450C > A (p.Tyr150Ter), c.715G > C (p.Gly239Arg) and c.866C > G (p.Pro289Arg), and PROS1 c.1468delA (p.Ile490fs) and c.1931T > A (p.Ile644Asn). Our data are consistent with those of previous studies, which mostly used time-consuming Sanger sequencing for genotyping, and the indication criteria for molecular genetic testing were adapted to this process in the past. Our promising results allow for a wider application of the described methodology in clinical practice, which will enable a suitable expansion of the group of indicated patients to include individuals with severe clinical findings of thrombosis at a young age. Moreover, this approach is flexible and applicable to other oligogenic panels.

The influence of protease hydrolysis of lactic acid bacteria on the fermentation induced soybean protein gel: Protein molecule, peptides and amino acids

The purpose explored the influence of protease hydrolysis of lactic acid bacteria and its hydrolysates on the fermentation induced soybean protein gel. The changes of protein molecule, peptides and amino acids after hydrolysis were analyzed. With the extension of fermentation time, the molecules (≤30 kDa) of L. paracasei-gel and L. plantarum-gel increased to 68.8% and 63%, the surface hydrophobicity of protein was increased, internal sulfhydryl groups and soluble proteins were reduced. These results indicated physicochemical properties of protein molecules were changed, small molecular proteins produced by hydrolysis were aggregated through intermolecular forces to promote gel formation. Peptide profiles and amino acid profiles showed that the peptides (≤10AA) of L. casei-gel increased by 3.32%, the peptides derived from 7S protein increased by more than 5%, the hydrophobic peptides and hydrophobic amino acids increased, these enzymatic hydrolysates may form insoluble aggregate through intermolecular forces to promote gel formation.

Choline Chloride Induces Growth Performance of Indian Major Carps and Air-Breathing Fish Species with an Outcome of Quality Food-Fish under a Semi-Intensive Culture System: A Biochemical Investigation.

The present study was intended to determine the possible influences of direct field application of choline chloride into pond water in addition to farm-made-aqua-feed under a semi-intensive culture system on the growth and biochemical parameters of two Indian major carps (IMCs), Catla catla (Catla) and Labeo rohita (Rahu), and two air-breathing species, Clarias batrachus (Magur) and Anabas testudineus (Koi), cultured in a ratio of 2:5:1:1 (Catla/Rahu/Magur/Koi) in three experimental ponds for a period of 90 days during the breeding season (June to August). Results were compared with control (C: fed only with farm-made-aqua-feed) and treatment (T: P1 and P2: farm-made-aqua-feed plus choline chloride into pond water directly at the rate of 350 g bigha–1 fortnightly or 350 g per 1600 square meter fortnightly). A significant increasing trend was observed in the growth parameters including total length-final (TLF), standard length-final (SLF), mean weight-final (MWF), % gain of mean total length (MTL), % gain of mean standard length (MSL), % weight gain (WG), specific growth rate (SGR) % per day, and survivability %. However, a reverse pattern was noticed in the food conversion ratio (FCR) both in IMCs and air-breathing fish species under choline supplementation. Serum biochemical responses, e.g., total protein (PRO), lactate dehydrogenase (LDH), glucose (GLU), and calcium (Ca) showed significant enhancement, and alkaline phosphatase (ALP), alanine amino transaminase (ALT), aspartate amino transaminase (AST), cholesterol (CHOLES), and triglycerides (Trig) showed gradual significant reduction during the breeding season under choline exposure. Treated fishes showed prevention from liver dysfunction and fatty liver formation, and increased body crude protein content. Results indicated favorable growth and yield, which may benefit fish farmers during their culture practices, and the output fish species under choline supplementation resulted in quality food-fish for human consumption.

Pten heterozygosity restores neuronal morphology in fragile X syndrome mice

Genetic studies of hippocampal granule neuron development have been used to elucidate cellular functions of Pten and Fmr1. While mutations in each gene cause neurodevelopmental disorders such as autism and fragile X syndrome, how Pten and Fmr1 function alone or together during normal development is not known. Moreover, Pten mRNA is bound by the fragile X mental retardation protein (FMRP) RNA binding protein, but how this physical interaction impinges on phosphatase and tensin homolog protein (PTEN) expression is not known. To understand the interaction of PTEN and FMRP, we investigated the dentate gyrus granule neuron development in Pten and Fmr1 knockout (KO) mice. Interestingly, heterozygosity of Pten restored Fmr1 KO cellular phenotypes, including dendritic arborization, and spine density, while PTEN protein expression was significantly increased in Fmr1 KO animals. However, complete deletion of both Pten and Fmr1 resulted in a dramatic increase in dendritic length, spine density, and spine length. In addition, overexpression of PTEN in Fmr1 KO Pten heterozygous background reduced dendritic length, arborization, spine density, and spine length including pS6 levels. Our findings suggest that PTEN levels are negatively regulated by FMRP, and some Fmr1 KO phenotypes are caused by dysregulation of PTEN protein. These observations provide evidence for the genetic interaction of PTEN and FMRP and a possible mechanistic basis for the pathogenesis of Fmr1-related fragile X neurodevelopmental disorders.

Efecto hepatoprotector de una mezcla de plantas medicinales y vitaminas en Rattus norvegicus con intoxicación hepática

In the present investigation, the hepatoprotective effect of a powder mixture of leaves of Gentianella alborosea(Gilg) Fabris, Cynara scolymus, Berberis vulgaris, Peumus boldus, Taraxacum officinaleroots, and vitamins B6 and B12 (PROTHEPA®) was studied in Rattus norvegicus. Sprague Dawley with paracetamol-induced liver intoxication. In the experiment, 25 female rats were randomly distributed into five groups, G1: physiological serum (SF) 4mL/kg; G2 (negative control): SF 4mL/kg + paracetamol 400mg/kg; G3 (positive control): Silymarin 100 mg/kg + paracetamol 400 mg/kg; G4: PROTHEPA® mixture 100 mg/kg + paracetamol 400 mg/kg; and G5: PROTHEPA® mixture 500 mg/Kg + paracetamol 400 mg/kg. Biochemical analyzes were performed to determine the liver profile (glutamic oxalacetic transaminase (GOT), glutamic pyruvic transaminase (GTP), total bilirubin (BT), and total protein (PT)), as well as pathological evaluations. The results showed that G2 presents a significant increase in TGP (80.20 ±1.3), BT (0.39 ±0.23), anda significant reduction in PT (4.82 ±0.46). As for G3, G4 and G5; all tests remained within normal values, similar to G1. These results demonstrate that the PROTHEPA® mixtures of 100mg/kg and 500mg/kg have a hepatoprotective effect, which is corroborated by the anatomo-pathological evaluations.

Application of an airlift internal circulation membrane bioreactor for the treatment of textile wastewater

A large amount of water is used in the textile industry during the finishing and dyeing processes leading to the production of what is known as textile wastewaters. Textile wastewater is highly rich in COD and color and is characterized by relatively low biodegradability. This study aimed to investigate the treatability of reactive red dye-rich textile wastewater with the application of an airlift internal circulation membrane bioreactor (AIC-MBR). Experimental results demonstrated that high removal efficiencies of COD, NH3-N, and reactive red up to 99.70%, 97.83%, and 97.23%, respectively, can be achieved using the AIC-MBR system. Besides, EPS and SMP analyses reflected an SMP polysaccharide (PS) and protein (PN) membrane rejection that reached 88% and 72.6%, respectively. Finally, the capillary suction time measurement highlighted a good dewatering capacity of the sludge with a low membrane fouling tendency at the end of the operating period.

Comparative structural and emulsifying properties of ultrasound-treated pea (Pisum sativum L.) protein isolate and the legumin and vicilin fractions

The structural properties, interfacial behavior, and emulsifying ability of ultrasound-treated pea protein isolate (PPI) and the legumin (11S) and vicilin (7S) globulin fractions prepared with a salt-solubilization procedure were investigated. Of the three protein groups, PPI was strongly responsive to ultrasound perturbation (20 kHz, 57–60 W·cm−2) showing the greatest solubility increase, particle size reduction, structure destabilization, and conformational change. Similar but less remarkable effects were observed on 11S globulins; 7S proteins, already highly soluble (>99%), were generally less sensitive to ultrasound. The ultrasound treatment significantly improved emulsifying activity, which resulted in greater emulsifying capacity and stronger interfacial adsorption for all protein samples. PPI exhibited the higher activity increase (70.8%) compared to approximately 30% for 11S and 7S. For both control and ultrasound treated proteins, the emulsifying capacity was in the order of 7S > 11S > PPI, inversely related to the trend of protein loading at the interface, indicating efficiency differences. The latter was attributed to emulsion clusters formed through protein–protein interaction in PPI and 11S emulsions which were visibly absent in 7S emulsions.

Steel pickling rinse wastewater treatment by two-stage MABR system: Reactor performance, extracellular polymeric substances (EPS) and microbial community

A bench-scale two-stage membrane-aerated biofilm reactor (MABR) system was applied to treat steel pickling rinse wastewater with high salinity and refractory organic. The effects of salinity and aeration pressure on the treatment efficiency, extracellular polymeric substances (EPS) characteristics and microbial community structure were studied. The optimal removal efficiencies of COD, NH+ 4-N and TN reached to 62.84%, 99.57% and 51.65%, respectively. Shortcut nitrification was achieved at low aeration, and the salinity less than 4% did not remarkable affect system performance. Colorimetric determination, three-dimensional exaction-emission matrix (3D-EEM) and Fourier transform infrared spectrum (FTIR) were employed to characterize the content and composition of proteins (PN) and polysaccharides (PS) in EPS of the biofilm. The results indicated that PN, not PS, response to changes of environmental conditions played a key role. Moreover, EPS might alleviate intracellular and extracellular osmotic pressure imbalance induced by high salinity, which imparted the biofilm in MABR with prominent salt-tolerant. High-throughput sequencing displayed that nitrifiers (Nitrosomonas, Nitrospira), denitrifiers (Dechloromonas, Hyphomicrobium, Denitromonas, Denitratisoma, Candidatus_Competibacter) and aerobic denitrifiers (Pseudomonas, Thauera) were predominant salt-tolerant bacteria.

Enhanced aerobic granular sludge formation by applying Phanerochaete chrysosporium pellets as induced nucleus.

The long start-up period is a major challenging issue for the widespread application of aerobic granular sludge (AGS). In this study, a novel rapid start-up strategy was developed by inoculating Phanerochaete chrysosporium (P. chrysosporium) pellets as the induced nucleus in a sequencing batch airlift reactor (SBAR) to enhance activated sludge granulation. The results demonstrated that P. chrysosporium pellets could effectively shorten the aerobic granulation time from 32 to 20days. The AGS promoted by P. chrysosporium pellets had a larger average diameter (2.60-2.74mm) than that without P. chrysosporium pellets (1.78-1.88mm) and had better biomass retention capacity and sedimentation properties; its mixed liquor suspended solids (MLSS) and sludge volume index (SVI30) reached approximately 5.2g/L and 45mL/g, respectively. The addition of P. chrysosporium pellets promoted the secretion of extracellular polymeric substances (EPS), especially protein (PN). The removal efficiencies of chemical oxygen demand (COD), ammonia nitrogen (NH4+-N), total nitrogen (TN), and total phosphorus (TP) in P. chrysosporium pellets reactor were 98.91%, 89.17%, 64.73%, and 94.42%, respectively, which were higher than those in the reactor without P. chrysosporium pellets (88.73%, 82.09%, 55.75%, and 88.92%). High throughput sequencing analysis indicated that several functional genera that were responsible for the formation of aerobic granules and the removal of pollutants, such as Acinetobacter, Pseudomonas, Janthinobacterium, and Enterobacter, were found to be predominant in the mature sludge granules promoted by P. chrysosporium pellets.

Validation in the ESPOIR cohort of vitamin K-dependent protein S (PROS) as a potential biomarker capable of predicting response to the methotrexate/etanercept combination

BackgroundTo validate the ability of PROS (vitamin K-dependent protein S) and CO7 (complement component C7) to predict response to the methotrexate (MTX)/etanercept (ETA) combination in rheumatoid arthritis (RA) patients who received this therapeutic combination in a well-documented cohort.MethodFrom the ESPOIR cohort, RA patients having received the MTX/ETA or MTX/adalimumab (ADA) combination as a first-line biologic treatment were included. Serum concentrations of PROS and CO7 were measured by ELISA prior to the initiation of ETA or ADA, at a time where the disease was active (DAS28 ESR > 3.2). The clinical efficacy (response/non-response) of both combinations has been evaluated after at least 6 months of treatment, according to the EULAR response criteria with some modifications.ResultsThirty-two were treated by MTX/ETA; the numbers of responders and non-responders were 24 and 8, respectively. Thirty-three patients received the MTX/ADA combination; 27 and 5 patients were respectively responders and non-responders. While there were no differences for demographic, clinical, biological, and X-rays data, as well as for CO7, serum levels of PROS tended to be significantly higher in responders to the MTX/ETA combination (p = 0.08) while no difference was observed in the group receiving MTX/ADA. For PROS, the best concentration threshold to differentiate both groups was calculated at 40 μg/ml using ROC curve. The theranostic performances of PROS appeared better for the ETA/MTX combination. When considering the response to this combination, analysis of pooled data from ESPOIR and SATRAPE (initially used to validate PROS and CO7 as potential theranostic biomarkers) cohorts led to a higher theranostic value of PROS that became significant (p = 0.009).ConclusionPROS might be one candidate of a combination of biomarkers capable of predicting the response to MTX/ETA combination in RA patients refractory to MTX.Trial registrationClinicalTrials.gov identifiers: NCT03666091 and NCT00234234.

Long-term effects of Cu(II) on denitrification in hydrogen-based membrane biofilm reactor: Performance, extracellular polymeric substances and microbial communities

Divalent copper (Cu(II)) frequently coexists with nitrate (NO3−) in industrial wastewater and the effect of Cu(II) on the autotrophic denitrification system using H2 as the electron donor remains unknown. In this study, the hydrogen-based membrane biofilm reactor (H2-MBfR) was operated continuously over 150 days to explore the effect of Cu(II) on the performance of autotrophic denitrification system and understand the key roles of EPS and microbial community. More than 95% of 20 mg-N/L NO3− was removed at 1–5 mg/L Cu(II), and the removal rate of NO3−-N was stabilized to 82% at 10 mg/L Cu(II) after a short period, while NH4+ and NO2− in effluent were hardly detected, indicated that high concentration of Cu(II) did not permanently inhibit the denitrification performance in H2-MBfR. Colorimetric determination showed that Cu(II) stimulated the secretion of EPS, in which the protein (PN) content was much higher than polysaccharide (PS). The PN/PS ratios increased from 0.93 to 1.99, and the PN was more sensitive to copper invasion. The results of three-dimensional excitation-emission matrix illustrated that tryptophan was the main component of EPS chelating Cu(II) to reduce toxicity. The results of Fourier-transform infrared demonstrated that hydroxyl, carboxyl, and protein amide groups bound and reduced Cu(II). Furthermore, Cu(II) was effectively removed (>80%), and the results of distribution and morphology analysis of Cu(II) show that the electron-dense deposits of monovalent copper (Cu(I)) were found in EPS and biofilms and the reduction of Cu(II) to Cu(I) was an obvious self-defense reaction of biofilm to copper stress. The microbial richness and diversity decreased with the long-term exposure to Cu(II), while the relative abundance of denitrifiers Azospira and Dechloromonas increased. This study provides a scientific basis for the optimal design of treatment system for removal of nitrate and recovery of heavy metals simultaneously.

Association Between Dietary Protein Intake and Sleep Quality in Middle-Aged and Older Adults in Singapore.

Poor sleep has been associated with the increased risk of developing detrimental health conditions. Diet and certain nutrients, such as dietary protein (PRO) may improve sleep. This cross-sectional study aimed to investigate the relationship between PRO intake, their amino acid components, and sources with sleep quality in middle-aged and older adults residing in Singapore. A dataset of 104 healthy subjects between the age of 50 and 75 years old were used. Collected data included 3-day food record and sleep quality [sleep duration, global sleep score (GSS), sleep latency (SL), and sleep efficiency (SE)]. The collected 3-day food records were extracted for PRO, tryptophan (Trp), and large neutral amino acid (LNAA) intake. PRO intake was further categorized into plant and animal PRO. A multivariate multiple linear regression (MLR) was performed to assess the association between PRO intake and sleep quality. Dietary Trp:LNAA ratio was positively associated with sleep duration (βtotal: 108.234 h; p: 0.005) after multiple covariates adjustment. Similarly, plant Trp (βplant: 2.653 h/g; p: 0.020) and plant Trp:LNAA (βplant: 54.006 h; p: 0.008) was positively associated with sleep duration. No significant associations were observed for both SL and SE. Sleep duration in middle-aged and older Singaporean adults was positively associated with dietary Trp and Trp:LNAA, especially when obtained from plant sources.