Molecular insight into binding behavior of caffeine with lactoferrin: Spectroscopic, molecular docking, and simulation study.

The majority of bioactive substances in the human diet come from polyphenols. Here, we use spectroscopy, molecular docking, molecular dynamics simulations, and in vitro digestion to look at the relationship between caffeine (CAF) and bovine lactoferrin (BLF). The correlation analysis of the CAF-BLF fluorescence quenching process revealed that the reaction was spontaneous and that the CAF-BLF fluorescence quenching process may have been static. The predominant intrinsic binding forces were hydrogen bonds and van der Waals forces, which were also supported by molecular docking and molecular dynamics simulations. Through Fourier infrared and circular dichroism spectroscopy experiments, it was found that CAF changed the secondary structure of BLF and might bind to the hydrophobic amino acids of BLF. Compared with BLF, CAF-BLF showed inhibitory effects on digestion in simulated in vitro digestion. It will be helpful to better understand the interaction between CAF and BLF and provide the basis for the development of innovative dairy products.

Comparison of Non-Covalent and Covalent Interactions between Lactoferrin and Chlorogenic Acid.

Adding polyphenols to improve the absorption of functional proteins has become a hot topic. Chlorogenic acid is a natural plant polyphenol with anti-inflammatory, antioxidant, and anticancer properties. Bovine lactoferrin is known for its immunomodulatory, anticancer, antibacterial, and iron-chelating properties. Therefore, the non-covalent binding of chlorogenic acid (CA) and bovine lactoferrin (BLF) with different concentrations under neutral conditions was studied. CA was grafted onto lactoferrin molecules by laccase catalysis, free radical grafting, and alkali treatment. The formation mechanism of non-covalent and covalent complexes of CA-BLF was analyzed by experimental test and theoretical prediction. Compared with the control BLF, the secondary structure of BLF in the non-covalent complex was rearranged and unfolded to provide more active sites, the tertiary structure of the covalent conjugate was changed, and the amino group of the protein participated in the covalent reaction. After adding CA, the covalent conjugates have better functional activity. These lactoferrin-polyphenol couplings can carry various bioactive compounds to create milk-based delivery systems for encapsulation.

Lipid reassembly in asymmetric Langmuir-Blodgett/Langmuir-Schaeffer bilayers.

Molecular-reorganization-induced morphology alteration in asymmetric substrate-supported lipid bilayers (SLBs) was directly visualized by means of atomic force microscopy (AFM) and total internal reflection fluorescence (TIRF) microscopy. SLB samples were fabricated on mica-on-glass and glass substrates by Langmuir-Blodgett (LB)/Langmuir-Schaeffer (LS) using binary lipid mixtures, namely, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)/1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and ternary mixtures DOPC/DPPC/1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), labeled with 0.2 mol % Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (TR-DHPE) dye. Phase segregations were characterized by TIRF imaging, and DPPC-enriched domain structures were also observed. Interestingly for ∼40% (n = 6) of the samples with binary mixtures in the LB leaflet and a single component in the LS leaflet, that is, (DOPC/DPPC)(LB)+DOPC(LS), the contrast of the DPPC domains changed from the original dark (without dye) to bright (more TR dye partitioning) on TIRF images, returning to dark again. This contrast reverse was also correlated to AFM height images, where a DPPC-DPPC gel phase was spotted after the TIRF image contrast returned to dark. The rupture force mapping results measured on these binary mixture samples also confirmed unambiguously the formation of DPPC-DPPC gel domain components during the contrast change. The samples were tracked over 48 h to investigate the lipid molecule movements in both the DPPC domains and the DOPC fluid phase. The fluorescence contrast changes from bright to dark in SLBs indicate that the movement of dye molecules was independent of the movement of lipid molecules. In addition, correlated multimodal imaging using AFM, force mapping, and fluorescence provides a novel route to uncover the reorganization of lipid molecules at the solid-liquid interface, suggesting that the dynamics of dye molecules is highly structure dependent.

Molecular dynamics simulation and in vitro digestion to examine the impact of theaflavin on the digestibility and structural properties of myosin.

In this study, the interaction mechanism between theaflavin and myosin was explored to confirm the potential application of theaflavin in the meat protein system. A series of theaflavin and myosin solutions were prepared for spectroscopic studies. Spectroscopy results showed that theaflavins formed complexes with myosin and affected the microenvironment of myosin. And that addition of theaflavin cause static quenching of the myosin solution. Theaflavin and bovine myosin combined through hydrophobic interaction to form a complex, and gradually increasing the temperature was conducive to the binding of theaflavin and bovine myosin. This interaction results in a decrease in the α -helix content of myosin. Molecular dynamics simulation results confirmed that hydrophobic interactions and hydrogen bonds made the protein structure more compact and stable. And the in vitro digestion process was simulated. The results showed that the addition of theaflavin could significantly reduce the digestibility of myosin.

High-efficiency degradation catalytic performance of a novel Angelica sinensis polysaccharide-silver nanomaterial for dyes by ultrasonic cavitation.

Currently, the polluted wastewater discharged by industry accounts for the major part of polluted bodies of water. As one of the industrial wastewaters, dye wastewater is characterized by high toxicity, wide pollution, and difficulty in decolorization degradation. In this paper, a novel composite nanomaterial catalyst of silver was prepared by using Angelica sinensis polysaccharide (ASP) as a reducing and stabilizing agent. And the optimum reaction conditions explored are VAgNO3 = 5 mL (300 mM) and vASP = 7% (w/v) for 6 h at 90 °C. In addition, the ASP-Ag nanocatalyst was characterized by several techniques. The results demonstrated that ASP-Ag nanoparticles were successfully synthesized. Degradation rate, which provides a numerical visualization of the percentage reduction in pollutant concentration. With the wrapping of ASP, the ultrasonic catalytic degradation rates of different organic dyes including rhodamine B (RB), methylene blue (MB), and methyl orange (MO) were from 88.2%, 88.7%, and 85.2% to 96.1%, 95.2% and 93.5% at room temperature, respectively. After the experiments, when cdyes = 10 mg/L, the highest degradation rate can be observed under cAPS-AgNPs = 10 mg/L with the most powerful cavitation frequency f = 59 kHz. The effect of ultrasonic frequency on the acoustic pressure distribution in the reactor was investigated by using COMSOL Multiphysis@ software to propose the mechanism of ultrasonic degradation and the mechanism was confirmed by OH radical trapping experiments. It indicates that OH produced by the ultrasonic cavitation effect plays a determinant role in the degradation. And then, the intermediate products of the dye degradation process were analyzed by gas chromatography and mass spectrometry (GC-MS), and the possible degradation processes of dyes were proposed. The resulting products of degradation are SO42-, NH4+, NO3-, N2, CO2 and H2O. Finally, the recycling degradation experiments showed that catalyst maintains a high degradation rate within reusing 5 cycles. Thus, this catalyst is highly efficient and recyclable.

Horizontal comparison of "red or blue shift" and binding energy of six fluoroquinolones: Fluorescence quenching mechanism, theoretical calculation and molecular modeling method.

In this article, the interaction between six fluoroquinolones (FQs) and bovine serum albumin (BSA) was initially studied at 298 K, 303 K and 310 K respectively under simulated physiological conditions by fluorescence spectroscopy. At the same time, the sub-structural domains on BSA that may bind to FQs were investigated by molecular docking simulation technique. A combination of quantitative and qualitative approaches was used in the analysis of the binding constants, binding sites and corresponding thermodynamic parameters in the interaction system, it was found that FQs forms a complex with BSA and undergoes static quenching, which is the main cause of fluorescence quenching. The results indicated that hydrogen bonds, Van der Waals force and electrostatic interaction were the main binding forces between the complexes, it also showed that these six fluoroquinolones mainly bound to the IIA and IIIA structural domains of BSA, while DANO and SARA may be more toxic than other antibiotics. Based on Foster's non-radiative energy transfer theory, the binding distance between FQs and BSA was calculated to be less than 7 nm, indicating the existence of energy transfer between small molecule drugs and proteins. Synchronous fluorescence and UV-Vis absorption spectroscopy further confirmed that FQs can alter the secondary conformational change of BSA. Lomefloxacin has a different effect from the other five fluoroquinolone antibiotics because it causes a decrease in polarity and an increase in hydrophobicity around tryptophan residues, while the other five FQs have the opposite effect. Together, the study of FQs-BSA is of great significance to elucidate the pharmacokinetics and pharmacodynamics of FQs.

Interaction of graphene oxide with lysozyme:Insights from conformational structure and surface charge investigations.

Lysozyme (Lyz) is an important antibacterial protein that exists widely in nature. In recent years, the application of graphene oxide (GO) in the field of biotechnology electronics, optics, chemistry and energy storage has been extensively studied. However, due to the unique properties of GO, the mechanism of its interaction with biomacromolecule proteins is very complex. To further explore the interaction between GO and proteins we explore the influence of different pH and heat treatment conditions on the interaction between GO and Lyz, the GO (0-20 µg/mL) was added at a fixed Lyz concentration (0.143 mg/mL) under different pHs. The structure and surface charge changes of Lyz were measured by spectroscopic analysis and zeta potential. The results showed that the interaction between GO and Lyz depends on temperature and pH, significant changes have taken place in its tertiary and secondary structures. By analyzing the UV absorption spectrum, it was found that lysozyme and GO formed a stable complex, and the conformation of the enzyme was changed. In acidic pH conditions (i.e., pH < pI), a high density of Lyz were found to adsorb on the GO surface, whereas an increase in pH resulted in a progressive decrease in the density of the adsorbed Lyz. This pH-dependent adsorption is ascribed to the electrostatic interactions between the negatively charged GO surface and the tunable ionization of the Lyz molecules. The secondary structure of Lyz adsorbed on GO was also found to be highly dependent on the pH. In this paper, we investigated the exact mechanism of pH-influenced GO binding to lysozyme, which has important guidance significance for the potential toxicity of GO biology and its applications in biomedical fields such as structure-based drug design.

The impact of fluoxetine and pH values on relaxation of the ternary lipid monolayers.

Fluoxetine (FLX), used in the clinic to treat depression, is a well-known cationic amphiphilic antidepressant. To get a deeper insight into the effect of FLX on Langmuir monolayers, in this study the stability and relaxation of 1,2-dioctadecanoyl-sn-glycero-3-phophocholine/1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine/cholesterol (DSPC/POPC/CHOL) monolayers without and with FLX at different pH values were studied. The experiments involved surface pressure-area (π-A) measurements, mean molecular area-time (A-t) measurements, and atomic force microscope (AFM) analysis. It was found that intermolecular interactions decreased after the addition of FLX in the subphase but increased with increasing pH values. The relaxation of the ternary lipid monolayers with FLX was dominated by dissolution steps, and the dissolution rates decreased with increasing pH values. These findings can be easily confirmed by the analysis of thermodynamic parameters calculated for the investigated films. The data obtained in this study help to understand the effect of drugs on the ternary lipid monolayers from the molecular point of view.

Physical field simulation of the ultrasonic radiation method: An investigation of the vessel, probe position and power.

In this paper, the effects of ultrasonic probe position, vessel shape, and ultrasonic input power on the sound pressure distribution in the reactor were investigated by solving the Helmholtz equation using COMSOL Multiphysis@ software. Three different types of glass containers were used in the study, which are beaker, Erlenmeyer flask, and round bottom flask. The maximum value of sound pressure in the three containers will gradually increase when the distance between the probe and the bottom of the container decreases. When the distance decreases, the area of the high acoustic pressure region in the round bottom flask does not change significantly, while the area of the high acoustic pressure region in the beaker and Erlenmeyer flask increases sharply, which means that the use of the round bottom flask can reduce the influence of the dead zone on the preparation of nanomaterials. In addition, the change in power increases the value of the peak negative acoustic pressure in the vessel, enhancing the response efficiency of ultrasonic cavitation.

Effect of ruptured cavitated bubble cluster on the extent of the cell deformation by ultrasound.

In this paper, the bubble-cell model is presented. The effects of the spacing between the bubble population and the cell, the radius of the bubble and the bubble medium on the degree of cell deformation were investigated by solving the Helmholtz equation and the equilibrium of motion equation using COMSOL Multiphysis@ software. The ultrasonic transducer is applied in a round bottom flask with the bubble-cell model on the side of the ultrasonic transducer. When the distance between the bubble cluster and the cell gradually increases, the extent of deformation of the cell is reflected as first increasing and then decreasing, reaching the maximum deformation at D = 2. When the radius of the bubble is changed, there is a "constant frequency" at low frequency ultrasound in any distance case, at which the cell deformation will be violent. However, when the bubble medium is changed, there is no significant change in the degree of deformation of the cells. In other words, changes in the structure of the bubble-cell model affect the degree of cell deformation, but without structural changes, the degree of cell deformation changes very little.

The interaction of graphene oxide-silver nanoparticles with trypsin: Insights from adsorption behaviors, conformational structure and enzymatic activity investigations.

In this work, we synthesized graphene oxide-silver nanoparticles (GO-AgNPs) hybrids by one-pot method. Since there are relatively few reports on whether GO-AgNPs bind and change the structure and function of trypsin, A variety of methods were employed to systematically characterize the molecular interaction between GO-AgNPs and trypsin. Results exhibited that GO-AgNPs bound with trypsin to form a ground state complex. GO-AgNPs had higher adsorption capacity for trypsin compared with single GO. Langmuir-Blodgett assembly method was used to confirm that AgNPs did not interfere with the adsorption of trypsin by GO. The secondary structure and the microenvironment of amino acid residues of trypsin were altered after interacting with GO-AgNPs. In addition, GO-AgNPs can enhance the activity of trypsin and promote the hydrolysis of bovine serum protein (BSA) by trypsin. These findings provide important support for the application of GO-based nanocomposites in the efficient immobilization of enzymes.

Cholesterol-dependent nanomechanical stability of phase-segregated multicomponent lipid bilayers.

Cholesterol is involved in endocytosis, exocytosis, and the assembly of sphingolipid/cholesterol-enriched domains, as has been demonstrated in both model membranes and living cells. In this work, we explored the influence of different cholesterol levels (5-40 mol%) on the morphology and nanomechanical stability of phase-segregated lipid bilayers consisting of dioleoylphosphatidylcholine/sphingomyelin/cholesterol (DOPC/SM/Chol) by means of atomic force microscopy (AFM) imaging and force mapping. Breakthrough forces were consistently higher in the SM/Chol-enriched liquid-ordered domains (Lo) than in the DOPC-enriched fluid-disordered phase (Ld) at a series of loading rates. We also report the activation energies (DeltaEa) for the formation of an AFM-tip-induced fracture, calculated by a model for the rupture of molecular thin films. The obtained DeltaEa values agree remarkably well with reported values for fusion-related processes using other techniques. Furthermore, we observed that within the Chol range studied, the lateral organization of bilayers can be categorized into three distinct groups. The results are rationalized by fracture nanomechanics of a ternary phospholipid/sphingolipid/cholesterol mixture using correlated AFM-based imaging and force mapping, which demonstrates the influence of a wide range of cholesterol content on the morphology and nanomechanical stability of model bilayers. This provides fundamental insights into the role of cholesterol in the formation and stability of sphingolipid/cholesterol-enriched domains, as well as in membrane fusion.

Bovine hemoglobin adsorption onto modified silica nanoparticles: Multi-spectroscopic measurements based on kinetics and protein conformation.

Herein, the effects of bovine hemoglobin (BHb) binding to hydrophilic silica nanoparticles (SN1) and hydrophobic silica nanoparticles (SN2) were explored under physiological conditions. SEM and XRD were used to characterize silica nanoparticles (SNs). Zeta potential and DLS confirmed the formation of protein corona (PC), and SN2 showed more increase in their size after PC formation comparing with SN1. The adsorption isotherms were fitted well by the Freundlich model, and the kinetics tended to follow pseudo-second-order kinetics. Then, the second structure of BHb has been analyzed by UV-vis and FT-IR spectroscopy, which implied the impact of SN2 on the secondary structure of BHb was greater than that of SN1 on BHb. Moreover, fluorescence spectroscopy and Raman spectroscopy showed that SNs may induce heme degradation to form fluorescent heme product, resulting in increased fluorescence intensity. This investigation will be significant in exploring the toxicity profile of SNs for their in vivo.

Studies on the interfacial behavior of DPPC/DPPG mixed monolayers in the presence of fluoxetine.

In this study, the interfacial behavior of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine/1,2-dipalmitoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DPPC/DPPG) mixed monolayers with fluoxetine (FLX) in the subphase was investigated by a combination of the Langmuir-Blodgett technique and atomic force microscopy (AFM). It was found that DPPC/DPPG mixed monolayers showed different interfacial behaviors before and after addition of FLX in the subphase. The electrostatic interaction between FLX and lipids molecules destroys the homogeneity of the mixed monolayers and changes the arrangement of lipids molecules at the interface after addition of FLX in the subphase, thereby leading to an increase of compressibility and miscibility and a decrease in the stability of the mixed monolayers. The surface morphology of the mixed monolayers observed by AFM was different between without and with FLX in the subphase, indicating the penetration of FLX into the mixed monolayers. The present study has provided detailed information for further understanding the interactions of drugs with membrane lipids in other lipid monolayers.

Effect of fluoxetine at different concentrations on the adsorption behavior of Langmuir monolayers.

Fluoxetine (FLX), approved for the treatment of depression and anxiety by the FDA in 2002, is an amphiphilic antidepressant. In general, amphiphilic drugs have high membrane permeability. Therefore, the interactions between these drugs and monolayers have been widely concerned. In this study, the adsorption of FLX on dipalmitoylphosphatidylcholine (DPPC) monolayers at different concentrations and surface pressures have been investigated by pressure-area isotherms (π-A), adsorption curves, compression-expansion curves, and atomic force microscopy (AFM). Our data showed that the adsorption behavior was related to the surface pressures and FLX concentrations in the subphase. The FLX that was added in the subphase under lower surface pressure (π = 10 mN/m) was easily adsorbed on DPPC monolayers. The stability of the monolayers was strong. The adsorption of FLX on DPPC monolayers and the stability decreased when π = 20 mN/m. In addition, the adsorption behavior and stability increased with increasing FLX concentrations. The AFM images of the monolayers confirmed the results of fitted adsorption curves. This study will be critical to our understanding of the interactions between drugs and lipid monolayers.

Modulation of membrane properties by DNA in liposomes: A spectroscopic study.

Liposome mediated DNA transport possesses a number of preventing diseases in clinical trials, thus, the study of interaction between DNA and liposomes has become a hot research direction. In this paper, the adsorption behavior of DNA onto two representative lipids had been studied by the fluorescence spectrum measurement, Ultraviolet absorption spectrum and Langmuir-Blodgett technology. The results of fluorescence spectrum measurement indicated that the fluorescence liposomes were quenched statically by DNA at all three temperatures. Thermodynamic analysis displayed that the intermolecular forces between DNA and liposomes were van der Waals forces and Hydrogen bonding. The experimental results of Ultraviolet absorption spectrum and Langmuir-Blodgett technology further verified these mechanisms. This work provides useful theoretical basis for the development of novel DNA delivery materials.

Carbon quantum dots (CQDs) modified ZnO/CdS nanoparticles based fluorescence sensor for highly selective and sensitive detection of Fe(III).

A simple and fast spectrofluorimetric method coupled with carbon quantum dots (CQDs) modified ZnO/CdS nanoparticles was developed for the detection of Ferric iron (Fe(III)). The fluorescence of CQDs/ZnO/CdS NPs was effectively quenched by Fe(III) due to the strong interaction between the CQDs/ZnO/CdS NPs and Fe(III). In addition, the detection limit of Fe(III) was about 1.72×10-7M. The effect of foreign ions on the fluorescence intensity of CQDs/ZnO/CdS NPs showed that the interference response in detecting of Fe(III) ions was low. Moreover, the quenching of Fe(III) and CQDs/ZnO/CdS NPs was discussed to be a static quenching procedure, which was proved by quenching constant KSV and fluorescence lifetime τ. The study of thermodynamics showed that the values of entropy change (ΔS) and enthalpy change (ΔH) were both positive, and the value of free energy (ΔG) was negative, which implied that the weak interaction of the molecular between CQDs/ZnO/CdS NPs and Fe(III) was hydrophobic force, and the quenching process was endothermic and spontaneous.

Fabrication of graphene oxide and sliver nanoparticle hybrids for fluorescence quenching of DNA labeled by methylene blue.

Since graphene oxide­silver nanoparticles (GO-AgNPs) have special affinities to DNA, it become increasingly important in fields of biological analysis in which GO-AgNPs nanocomposites universally functioned as a quencher. In this paper, GO-AgNPs nanocomposites with different GO to AgNPs ratios were synthesized as a fluorescence quencher to interact with DNA labeled by methylene blue (MB). The results showed that the fluorescence intensity of DNA-MB system decreased with the increasing of GO-AgNPs nanocomposites concentration. The quenching phenomenon of DNA-MB by AgNPs and GO was not a simple additive effect but a synergistic effect. The quenching efficiency of synthesized GO-AgNPs nanocomposites with different ratios (1:1, 1:3, 1:5, 1:10) increased with the decrease of GO/Ag ratio. Thermodynamic analysis was employed to investigate the interaction of GO-AgNPs and DNA-MB, it can be concluded that the intermolecular force between GO-AgNPs and DNA-MB was hydrogen bonding. Our works will provide important theoretical and experimental bases for fluorescence sensing of DNA.

[IR/UV spectroscopic analysis of gangliosides and their microstructures of polymeric aggregates observed by AFM technique].

Gangliosides, a kind of acid glycosphingolipid containing sialic acid, plays a very important physiological role in biomembrane as one of the important components of neurocyte membrane. They were extracted from bovine brain by the Folch method and purified by silica gel and DEAE-Sephadex A-25 column chromatograph. Their molecular functional groups and microstructures of polymeric aggregates were studied by infrared spectrum (IR), ultraviolet spectrum (UV) and atomic force microscope (AFM). The experimental results indicate that: 55.2 mg of Gls from 100 g of wet bovine brain had a certain purity, 62.84%. And their UV absorption spectra appeared at 195 nm, near to the results reported by other peoples. Compared with the IR spectra of sialic acid, the experimental results showed that the structures of the products had the units of sialic acid. In order to investigate the aggregate structures of ganglioside. AFM technique was applied in water, and the results showed that gangliosides can form spherical or ellipsoidal structures in water. It was determined that the size of polymeric aggregates of gangliosides varies between 55 and 380 nm, the average size is (148.9+/-66.7) nm; the height is between 1.0 and 5.0 nm, and the average height is (3.25+/-1.01) nm. The experimental results provide a theoretical and experimental basis for investigating biological activity and the exploitation and utilization of neural drugs.

Studied on the dynamic adsorption process of Lycium barbarum polysaccharide in the POPC/DPPC monolayers.

In this study, the interaction between Lycium barbarum polysaccharide (LBP) and unsaturated 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) or saturated 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was explored using the Langmuir films technique and atomic force microscopy (AFM). Comparing the pure lipid monolayer with the mixed monolayers, the π-A isotherms of the mixed monolayers shifted to larger molecular areas when LBP was added to the subphase. The compression modulus showed that the compressibility of the monolayer films decreased with the addition of LBP. Adsorption curves revealed that the variation in the surface pressure of LBP with POPC was larger than that with DPPC. This phenomenon was verified by the AFM images and the number of each lipid molecule combining with polysaccharide molecules in the mixed monolayer (Ap value), indicating that hydrophobic interactions between LBP and POPC are stronger than those of DPPC. These findings lay the foundation for exploring the pharmacological mechanism of LBP as an in vivo therapeutic.