Rational design of multiple structural optimization is a favorable approach to boost the activity and application of photoelectrochemical devices. Developing more extensive and multi-functional applications is also urgently necessary to fully exploit the potential of the photoelectrochemical catalytic system. Herein, we combined interface effects and defect engineering to obtain a molybdenum sulfide/polymeric carbon nitride composite system that simultaneously possesses photoelectrocatalytic water splitting and antibiotic sensing performances. This system first in-situ deposits molybdenum sulfide on polymeric carbon nitride. The partitioning effect of the polymer matrix contributes to the high dispersion and amorphous structure of molybdenum sulfide. Subsequent defect engineering provides abundant nitrogen defects and generates cyano groups. The synergistic effect of multiple advantages, including the more active sites of amorphous molybdenum sulfide, the interface effect between molybdenum sulfide and carbon nitride, the band modulation of nitrogen defects, and the electron-withdrawing effect of the cyano groups, in promoting photoelectrochemical performance of the system should be of significant responsibility. This composite system exhibits markedly boosted photoelectrochemical activities in both hydrogen and oxygen evolution, thereby highlighting the effectiveness of the bifunctional catalytic design. In order to expand the application of photoelectrochemical functional materials in sensors, this system, in combination with aptamers, constructs aptasensors with photoelectrochemical response, which can selectively detect the antibiotic enrofloxacin. This work demonstrates the potential of the synergy between interface effects and defect engineering in photoelectrochemical systems, providing new possibilities of a two-in-one multi-functional application for constructing efficient photocatalysts and photoelectrochemical sensors.

Quercetin exhibits a variety of biological activities, but its application is restricted due to its low solubility and instability. Nanoparticle technology can enhance the solubility and stability of quercetin, thereby enhancing its biological activities and potential applications. However, the structure and properties of nanoparticles are closely related to the materials used and the preparation methods employed. This study investigated the effect of preparation method (pH-driven method or antisolvent precipitation method) on the structure and properties of quercetin-loaded soy protein isolate-rhamnolipid nanoparticles (R-S-Q). Hydrophobic interactions, hydrogen bonds, and electrostatic interactions were the main driving forces for the formation and stabilization of nanoparticles. Nanoparticles prepared using the pH-driven method (PD/R-S-Q (2:1)) exhibited smaller particle size (195.60 +/- 0.55 nm), larger absolute value of zeta potential (38.86 +/- 0.22 mV), higher encapsulation efficiency (89.87 +/- 0.30 %), and loading capacity (2.90 +/- 0.04 %) than the ones prepared using the antisolvent precipitation method (ASP/R-S-Q (2:1)). PD/R-S-Q (2:1) demonstrated better stability (pH, ionic, light, thermal, and storage stability), water solubility of quercetin, re-dispersibility, and antioxidant activity than ASP/R-S-Q (2:1). Compared with nanoparticles encapsulating quercetin using soy protein isolate (SPI), ASP/R-S-Q (2:1) and PD/R-S-Q (2:1) showed better sustained release behavior in simulated digestion fluids. In conclusion, the pH-driven method was found to be more suitable for preparing SPI-surfactant nanoparticles for the delivery of quercetin and other hydrophobic bioactive substances. These results provided insights into the selection of nanoparticle preparation methods and the design of novel nanoparticle delivery systems.

The objective of this study was to explore the effect of the assembly sequences of wall materials on the structure and properties of Antarctic krill peptide (AKP)-loaded ovalbumin (OVA)–chitosan (CS) nanoparticles (NPs). Two AKP-loaded NPs (CS/OVA-AKP and OVA/CS-AKP) were prepared by changing the sequences of OVA and CS. The results confirmed that CS/OVA-AKP had a smaller particle size (291 nm vs. 320 nm), lower polydispersity index (0.233 vs. 0.282), higher absolute zeta potential (34.4 mV vs. 32.1 mV), and higher encapsulation efficiency (81.6% vs. 75.4%) than OVA/CS-AKP. X-ray diffraction analysis confirmed that AKP was encapsulated in an amorphous state within the NPs. Fourier transform infrared spectroscopy and three-dimensional (3D) fluorescence spectroscopy revealed that electrostatic interactions, hydrogen bonding, and hydrophobic interactions were the primary driving forces for nanoparticle formation, with CS/OVA-AKP demonstrating a stronger OVA fluorescence quenching effect. Compared with OVA/CS-AKP, CS/OVA-AKP exhibited better redispersibility, and CS/OVA-AKP showed greater stability under various environmental factors (thermal treatment, salt concentration, pH, and storage time). During simulated gastrointestinal digestion, CS/OVA-AKP effectively protected AKP from gastric degradation and showed a higher AKP release rate in simulated intestinal fluid (61.1%) than OVA/CS-AKP (53.0%). The release followed the Korsmeyer–Peppas model, with OVA/CS-AKP exhibiting non-Fickian diffusion (n = 0.7500), and CS/OVA-AKP approached Case II transport (n = 0.9889), indicating erosion-controlled release behavior. CS/OVA-AKP also demonstrated higher hypoglycemic activity, with inhibition rates of 41.1%, 37.5%, and 36.1% for α-glucosidase, α-amylase, and DPP-IV, respectively. These findings underscore the important influence of wall-material assembly sequences on the structure and properties of AKP-loaded NPs, offering valuable insights for the development of bioactive peptide delivery systems.

Developing highly efficient light-responsive antibacterial hydrogels is an effective way to achieve low health risks and environmentally friendly antibacterial effects. Herein, a bismuth sulfide/polymeric carbon nitride/ chitosan/polyacrylamide (Bi2S3/PCN/CS/PAM) composite hydrogel with near-infrared response (NIR) photocatalytic antibacterial activity was prepared through photoinitiated cross-linking polymerization. The highly dispersed Bi2S3 loading onto PCN not only acts as a photoinitiator but also plays a role in absorbing visible and near-infrared light. The maximum antibacterial rate of this composite hydrogel against Escherichia coli under near-infrared light irradiation can reach 95.55 %. Under non-contact and illumination conditions, this composite hydrogel can extend the fish shelf life and grapes to the 8th and 14th days respectively. This work offers new opportunities for the application of photoresponsive antibacterial hydrogels in food preservation, especially non-contact preservation.

BACKGROUND:Kaempferol (KAE), a bioactive flavonoid, has limited solubility and stability in water. Zein-gum arabic (GA) nanoparticles (NPs) are promising carriers for KAE, but the influence of preparation methods on their structure and properties remains unclear. This study investigated the effect of preparation method on the structure and properties of KAE-loaded zein-GA NPs. RESULTS:The results showed that the NPs prepared by the antisolvent co-precipitation method (GA-zein-KAE) had a smaller particle size, a lower protein dispersibility index, a higher absolute zeta potential value, and greater encapsulation efficiency and loading capacity than NPs prepared by antisolvent precipitation (GA-zein/KAE). The superior performance of GA-zein-KAE likely resulted from the simultaneous solvent displacement of zein, GA, and KAE during co-precipitation, which promoted ternary synergistic co-assembly, homogeneous component distribution, and a stabilized composite matrix. Scanning electron microscopy revealed a smoother and more uniform surface for GA-zein-KAE, and X-ray diffraction demonstrated that KAE was encapsulated in an amorphous state within the NPs. Fluorescence spectroscopy and Fourier transform infrared spectroscopy confirmed hydrogen bonding, hydrophobic interactions, and electrostatic attractions among KAE, zein, and GA. The GA-zein-KAE also demonstrated superior re-dispersibility and stability across varying pH, ionic strength, thermal treatment, and storage conditions as well as higher antioxidant activity and faster KAE release in simulated intestinal fluid compared with GA-zein/KAE. CONCLUSION:The preparation method had a significant effect on the structure and properties of KAE-loaded zein-GA NPs, in which antisolvent co-precipitation promoted the ternary co-assembly of zein, GA, and KAE, yielding NPs with improved performance. © 2025 Society of Chemical Industry.

The effect of the degree of substitution (DS, 0.7, 0.9, 1.2) of carboxymethylcellulose (CMC) on the structure and properties of curcumin-loaded gliadin-carboxymethylcellulose nanoparticles (Cur/G-CMC NPs) was explored in our study. The primary interactions in Cur/G-CMC NPs were hydrogen bonding, electrostatic forces, and hydrophobic interactions. As the DS of CMC increased from 0.7 to 1.2, the particle size of Cur/G-CMC NPs decreased from 284.9 nm to 273.8 nm and 260.5 nm, while their zeta-potential values changed from -46.41 mV to -47.34 mV and -48.62 mV, respectively. The phenomenon indicated enhanced steric hindrance and electrostatic repulsion among NPs, resulting in optimal pH, ionic, photostability, thermal, and storage stability for Cur/G-CMC 1.2 NPs. Interestingly, during in vitro simulated intestinal digestion, Cur/G-CMC 1.2 NPs showed the highest cumulative release rate of Cur (72.24 %). Moreover, the bioaccessibility of Cur/G-CMC 1.2 NPs (51.33 %) was superior to that of Cur/G-CMC 0.7 and Cur/G-CMC 0.9 NPs. Furthermore, with the DS of CMC increasing, the antioxidant activity of Cur/G-CMC NPs was also enhanced. In summary, the structure and properties of Cur/ G-CMC NPs could be affected by the DS of CMC, providing a reference for constructing nanoparticle delivery systems for active substances with better performance.

In this study, the impact of three types of surfactants (anionic surfactant rhamnolipid, amphoteric surfactant lecithin, and nonionic surfactant tea saponin) on the structure and properties of quercetin-loaded soy protein isolate (SPI) nanoparticles (Q-S NPs) was evaluated. Hydrophobic interactions, electrostatic interactions, and hydrogen bonds were the dominant forces to form quercetin-loaded SPI/surfactant composite nanoparticles. Compared to quercetin-loaded SPI/lecithin composite nanoparticles (Q-S/L1:4 NPs) and quercetin-loaded SPI/tea saponin composite nanoparticles (Q-S/T1:3 NPs), quercetin-loaded SPI/rhamnolipid composite nanoparticles (QS/R1:2 NPs) exhibited the smallest particle size (195.6 nm) and the largest absolute value of zeta potential (33.21 mV), which may be attributed to the amphiphilic plate-like structure and the higher negative charge of rhamnolipid. Additionally, Q-S/R1:2 NPs exhibited the best water solubility of quercetin, re-dispersibility, and stability (ionic, photo, thermal, and storage). Compared to Q-S/L1:4 and Q-S/T1:3 NPs, Q-S/R1:2 NPs showed the best stability and sustained release during simulated digestion. In addition, Q-S/R1:2 NPs also exhibited the best antioxidant activity. In summary, this study provided some references for the preparation of nanoparticles with different surfactants to deliver bioactive substances.

Photocatalytic water splitting has emerged as a vital technology for producing clean hydrogen fuel, addressing global energy needs. However, most research has focused on half-water splitting with sacrificial agents, which limits its practical application in fuel production. In this study, we present an advanced co-catalyst system for polymeric carbon nitride (CN) designed for efficient overall water splitting, a crucial step toward scalable hydrogen fuel generation. The co-catalyst system includes full loading of CoOx, Zn-Pt, and Cr2O3, with Zn strategically inserted between Pt and CN. This configuration enhances the spatial separation of photoexcited carriers, promoting efficient segregation of oxidation and reduction sites while minimizing electron-hole recombination. The optimized system achieves a stoichiometric H2 and O2 ratio of 2:1, with a hydrogen evolution rate that is nine times higher than when Pt alone is used as the co-catalyst. This advancement provides a pathway for efficient solar-driven hydrogen production, addressing the critical challenge of moving beyond half-water splitting and contributing to the development of clean fuel technologies.

The use of chitosan-based sponge materials for Hg(II) removal has gained attention recently due to their effectiveness. However, the complex preparation, limited performance, and poor acid resistance remained major drawbacks. Herein, a nitrogen-sulfur functionalized macroporous chitosan sponge was successfully synthesized via two mild amidation reactions and exhibited abundant interconnected mesopores. These features endowed the functionalized chitosan-based sponge with high adsorption capacity (1227.15 mg g- 1 ), fast reaction rate (8.27 x 10-3 g mg- 1 & sdot;min- 1 ), broad pH adaptability (1-7), and high selectivity, even in the artificial chlor-alkali wastewater. Furthermore, the impressive saturation capacity of 1329.24 mg g- 1 was achieved in various heights and injection rates in the fixed-bed column test, and the good removal efficiency (>85 %) was maintained after six dynamic regeneration cycles. The excellent performance was primarily attributed to the chemisorption of C-S groups. Among the three machine learning models, the ANFIS algorithm owned the best results of the smallest RMSE (0.00315) and highest R2 (0.9752) for predicting dynamic adsorptive behaviors. Overall, this research provided a reference for preparing a promising mesoporous sponge as an alternative recyclable and efficient candidate for industrial wastewater treatment and offered a machine learning model to predict the dynamic adsorptive performance.

Selective response is the key index to evaluate the performance of polymeric carbon nitride (PCN)-based heavy metal ion fluorescence sensors. Herein, to explore the role of cyano groups on selectivity, four kinds of PCN, including PCN-Cl, PCN-Ac, PCN-B and PCN-K were prepared by the molten salt method of sodium chloride and sodium acetate, the reduction method of sodium borohydride and the etching method of potassium hydroxide, respectively. These PCNs exhibited different surface cyano characteristics, but all of them had significant blue emission under ultraviolet excitation. It is proved that the assistant of sodium chloride or potassium hydroxide is an effective method to prepare PCNs with abundant surface cyano group. A series of fluorescence quenching experiments of metal ions showed that the cyano-rich degree of PCN is closely related to its selective response to mercury (II) ions. PCN-Cl and PCN-K emerged good selective quenching of mercury (II) ions, which may be related to the soft acid-soft base strong interaction between mercury (II) ions and cyano groups. Both PCN-Cl and PCN-K fluorescent probes for mercury (II) ions had a linear range of 5 similar to 50 mu mol L-1 , and PCN-Cl exhibited a lower detection limit of 0.38 mu mol L-1. This work confirmed the selective fluorescence response of cyano-rich PCN to mercury (II) ions, proposed the mechanism of selective fluorescence quenching response of mercury (II) ions, and provided a new idea for the design of efficient and accurate PCN-based fluorescence probes.

Rational design of antibacterial hydrogels is an important link to promote the multi-scenario application of this promising soft material. Herein, acrylamide and sodium alginate were selected as mixed monomers, silver deposited polymeric carbon nitride nanosheets were used as a photoinitiator, pre-gelation was achieved under UV irradiation, and then a composite hydrogel was obtained through calcium ion crosslinking. The composite hydrogels exhibit good mechanical stability, structural stability in water and degradability under simulated physiological conditions. The composite hydrogels have the activity of killing Escherichia coli with or without light. The visible light response and surface plasmon resonance effect of the silver-deposited polymeric carbon nitride boosted the photocatalytic antibacterial activity of the composite hydrogels, and the bactericidal rate of the optimal sample reached 95.5% after 120 min of visible light irradiation. This work provides a constructive and green roadmap for the design of composite dual-network photoresponsive antibacterial hydrogels.

The facile fabrication of doped carbon materials with multiple functions has been a challenging issue in materials science. Herein, nitrogen-doped carbon nanosheets with high doping level (32.05 wt%) were obtained by one-step calcination with urea and polyaniline as precursors. The presence of abundant edges and crimp, and the increase of the proportion of pyrrolic and graphitic nitrogen promote the exposure of the active site of the doped nanosheets. These structural advantages result in a favourable electrochemical response to organic pollutant malachite green. The linear range of detection of malachite green is 1 similar to 10 mu mol/L and the limit of detection is 13.2 nmol/L by differential pulse voltammetry electrochemical sensor constructed with high nitrogen-doped carbon nanosheets. The electrochemical sensor showed a satisfactory recovery rate for malachite green detection in actual tap water sample. This work offers a constructive solution for the facile fabrication and chemical sensor application of doped two-dimensional carbon-based materials.

As a protein extracted from soybeans, soy protein isolate (SPI) may undergo the Maillard reaction (MR) with co-existing saccharides during the processing of soy-containing foods, potentially altering its structural and functional properties. This work aimed to investigate the effect of mono- and polysaccharides on the structure and functional properties of SPI during MR. The study found that compared to oat β-glucan, the reaction rate between SPI and D-galactose was faster, leading to a higher degree of glycosylation in the SPI–galactose conjugate. D-galactose and oat β-glucan showed different influences on the secondary structure of SPI and the microenvironment of its hydrophobic amino acids. These structural variations subsequently impact a variety of the properties of the SPI conjugates. The SPI–galactose conjugate exhibited superior solubility, surface hydrophobicity, and viscosity. Meanwhile, the SPI–galactose conjugate possessed better emulsifying stability, capability to produce foam, and stability of foam than the SPI–β-glucan conjugate. Interestingly, the SPI–β-glucan conjugate, despite its lower viscosity, showed stronger hypoglycemic activity, potentially due to the inherent activity of oat β-glucan. The SPI–galactose conjugate exhibited superior antioxidant properties due to its higher content of hydroxyl groups on its molecules. These results showed that the type of saccharides had significant influences on the SPI during MR.

The aim of this study was to construct a chitosan quaternary ammonium salt (HTCC)/sodium alginate (SA) nanoparticle delivery system for the encapsulation of Antarctic krill peptide (AKP), with the goal of enhancing its stability and post-digestive hypoglycemic activity. Additionally, the effect of the content and encapsulation sequence of HTCC/SA on the structure and properties of AKP-loaded nanoparticles (AKP-loaded NPs) was investigated. As the content of the outermost wall material (HTCC) increased, the particle size of nanoparticles initially increased and then decreased. Concurrently, the zeta potential showed a trend of increase, ranging from -38.77 to 46.07 mV. When SA was used as the outermost wall material, the nanoparticles exhibited a smaller particle size (240.1 nm) and better dispersibility. Electrostatic interaction and hydrogen bonding were the major forces involved in the formation of AKP-loaded NPs. X-ray diffraction revealed that AKP was successfully encapsulated in an amorphous state. Scanning electron microscopy showed that AKP-loaded NPs had an elliptical or blocky appearance. Moreover, AKP-loaded NPs demonstrated good storage and temperature stability, while nanoparticles with HTCC in the outermost layer (HTCC:SA = 2:1) showed good pH and ionic stability. During simulated digestion, AKP-loaded NPs inhibited the release of AKP in gastric fluid. Consequently, the hypoglycemic activity of digested AKP-loaded NPs was remarkably higher than unencapsulated AKP. Notably, the nanoparticles with HTCC in the outermost layer provided superior hypoglycemic activity. In summary, the polysaccharide-based delivery system exhibited great potential to improve the stability and post-digestive hypoglycemic activity of AKP, broadening its applications in the food and pharmaceutical industries.

The wettability of polymeric carbon nitride (PCN) is an important surface chemical factor affecting its catalytic and separation performance. However, due to the amphipathic characteristics of hydrophilic/lipophilic and porous powder properties, the contact angle (CA) of PCN measured by the common sessile drop method will be interfered by the substrate, resulting in randomness of the results. Herein, the pristine PCNs obtained by thermal polycondensation of different precursors in air or nitrogen atmosphere were used as the wettability research object. The CA of the PCNs was measured by a modified sessile drop method after the traditional powders were replaced by pressed-tablet samples. The influence of sample mass and applied pressure on specific surface area, pore volume, surface morphology and CA of the pressed-tablet samples was investigated carefully, and the optimal experimental conditions were determined. When the sample mass was 0.15 g and the applied pressure was 10 MPa, the measured water CA of PCNs was between 27 similar to 34 degrees, and the diiodomethane CA is between 24 similar to 29 degrees. The measurement results showed considerable stability and reproducibility. This improved CA measurement method provides a simple and standardized way to accurately explore the surface chemistry of PCN, and has the potential to be applied to other hydrophilic and/or oleophilic porous powder materials.

Pathogenic bacterial infections, even at extremely low concentrations, pose significant threats to human health. However, the challenge persists in achieving high-sensitivity bacterial detection, particularly in complex samples. Herein, we present a novel sandwich-type electrochemical sensor utilizing bacteria-imprinted polymer (BIP) coupled with vancomycin-conjugated MnO2 nanozyme (Van@BSA-MnO2) for the ultrasensitive detection of pathogenic bacteria, exemplified by Staphylococcus aureus (S. aureus). The BIP, in situ prepared on the electrode surface, acts as a highly specific capture probe by replicating the surface features of S. aureus. Vancomycin (Van), known for its affinity to bacterial cell walls, is conjugated with a Bovine serum albumin (BSA)-templated MnO2 nanozyme through EDC/NHS chemistry. The resulting Van@BSA-MnO2 complex, serving as a detection probe, provides an efficient catalytic platform for signal amplification. Upon binding with the captured S. aureus, the Van@BSA-MnO2 complex catalyzes a substrate reaction, generating a current signal proportional to the target bacterial concentration. The sensor displays remarkable sensitivity, capable of detecting a single bacterial cell in a phosphate buffer solution. Even in complex milk matrices, it maintains outstanding performance, identifying S. aureus at concentrations as low as 10 CFU mL-1 without requiring intricate sample pretreatment. Moreover, the sensor demonstrates excellent selectivity, particularly in distinguishing target S. aureus from interfering bacteria of the same genus at concentrations 100-fold higher. This innovative method, employing entirely synthetic materials, provides a versatile and low-cost detection platform for Gram-positive bacteria. In comparison to existing nanozyme-based bacterial sensors with biological recognition materials, our assay offers distinct advantages, including enhanced sensitivity, ease of preparation, and cost-effectiveness, thereby holding significant promise for applications in food safety and environmental monitoring.

BACKGROUND:Antarctic krill peptide (AKP) has gained considerable interest because of its multiple biological functions. However, its application may be limited by its poor stability and susceptibility to degradation. Encapsulation of AKP using a nanoparticle delivery system is an effective way to overcome these problems. In the present study, bovine serum albumin (BSA) and chitosan (CS) were used as delivery vehicles to encapsulate AKP. RESULTS:The results revealed that the particle size (83.3 ± 4.4-222.4 ± 32.7 nm) and zeta-potential (35.1 ± 0.7-45.0 ± 2.7 mV) of nanoparticles (NPs) increased with the increasing content of BSA, but the polydispersity index decreased (1.000 ± 0.002 to 0.306 ± 0.011). Hydrogen bonding, hydrophobic and electrostatic interactions were the main forces to form BSA/CS-AKP NPs. X-ray diffraction revealed that AKP was encapsulated by BSA/CS. Scanning electron microscopy images exhibited that the NPs were spherical in shape, uniform in size and tightly bound. BSA/CS-AKP NPs exhibited excellent stability in the pH range (2-5) and after 15 days of storage, and could hinder the release of AKP in simulated gastric environment and promote the release of AKP in simulated intestinal environment. After simulated digestion, the hypoglycemic activity of encapsulated AKP was better than that of unencapsulated AKP. CONCLUSION:Our results revealed that the BSA/CS showed great potential for protecting and delivering AKP. © 2024 Society of Chemical Industry.

BACKGROUND:The widespread use of quercetin is limited by its instability, low solubility and poor oral bioavailability. Encapsulation of quercetin using a nanoparticle delivery system is an effective way to overcome these drawbacks. RESULTS:The effect of the molecular weight (Mw) of chitosan (CS) (100, 200, 500 and 1000 kDa) on quercetin-loaded chitosan nanoparticles (QCNPs) was investigated. The structure, stability, release properties and antioxidant activities of the nanoparticles (QCNP-10, QCNP-20, QCNP-50 and QCNP-100) were assessed. Particle size of QCNPs decreased and polydispersity index increased with the increasing Mw of CS. The main forces involved in the formation of QCNPs were hydrogen bonding and hydrophobic interaction. X-ray diffraction verified that quercetin was loaded into CS nanoparticles. The photostability and thermal stability of QCNPs increased with increasing Mw of CS. QCNP-100 exhibited the lowest release rate in a mixture of water and anhydrous ethanol. The antioxidant activities of QCNPs were enhanced with increasing Mw of CS, and QCNP-100 possessed the highest antioxidant activities, which might be relevant to its smallest particle size. CONCLUSION:Overall, these results revealed that the Mw of CS affected the properties of QCNPs, and QCNP-100 possessed the smallest particle, best stability, lowest release rate and highest antioxidant activities. © 2024 Society of Chemical Industry.

Maillard reaction (MR) with oat β-glucan changed the structure of soybean protein isolate (SPI), further leading to the enhancement of its functional properties. SPI was unfolded by MR, and the SPI conjugates with high molecular weight were identified. The water solubility of SPI was improved by cross-linking with hydrophilic β-glucan, while the hydrophobicity also increased along with the unfolding of the SPI. Cross-linking with β-glucan elevated the viscosity of SPI, thus enhancing viscosity-related physiological activities, including bile acid binding ability, fat binding capacity, and hypoglycemic activity, and the functional properties increased as the βG content involved in MR increased.