Two-dimensional Nuclear Magnetic Resonance Research Articles

Isolation of a Marker Olean-12-en-28-butanol Derivative from Viscum continuum E. Mey. Ex Sprague and the Evaluation of Its Antioxidant and Antimicrobial Potentials.

Viscum continuum E. Mey. Ex Sprague (Mistletoe) is a semi-parasitic plant that grows on the branches of other trees with reported numerous biological activities. This study was aimed at isolating a compound/s that will be used as a standard reference for quality control of South African-based commercialized mistletoe products and to further perform antioxidant and antimicrobial tests on the isolated compound. A dried sample of mistletoe was ground and extracted successively with hexane, dichloromethane (DCM), acetone and methanol using a serial exhaustive cold maceration procedure. The compound was isolated using column chromatography, and its chemical structure was elucidated using two-dimensional nuclear magnetic resonance (2D NMR) and ultrahigh-performance liquid chromatography-mass spectrometry (UPLC-MS). The antioxidant activity of the compound was determined using DPPH, hydrogen radical scavenging activity and reducing power assays, whereas antimicrobial activity was assessed using the minimum inhibitory concentration (MIC) method. Subjection of the DCM extract to column chromatography resulted in the isolation of a compound elucidated as olean-12-en-28-butanol-1-one, 3-hydroxy-4,4,10, 14, 20-pentamethyl (D4). Both the DPPH, H2O2 radical scavenging activity and reducing power assays revealed a significant antioxidant potential of compound D4 with an IC50 of 0.701 mg/mL, lower than that of gallic acid (0.793 mg/mL) for the H2O2 radical scavenging assay. The results also indicated good antibacterial activity of D4 with an IC50 of 0.25 mg/mL, compared to ciprofloxacin with an IC50 of 0.0039 mg/mL, against two Gram-negative (Pseudomonas aeruginosa, Escherichia coli) and three Gram-positive (Streptococcus pyogenes, Bacillus cereus and Staphylococcus aureus) bacteria. This study is the first to report on the isolation of the olean-12-en-28-butanol derivative from mistletoe of the South African ecotype.

Relaxation Editing in NMR Using a New Two-Dimensional Long-Lived Coherence Method for Mixture Analysis.

The exploration of metabolomics and targeted segments of proteins stands as a pivotal facet of Nuclear Magnetic Resonance (NMR) analysis, furnishing valuable insights into molecular architectures and potential therapeutic applications. The issue of spectral congestion frequently presents challenges in ascribing distinct peaks within the confines of both one-dimensional (1D) and two-dimensional (2D) NMR spectra. Numerous strategies have been proposed to resolve specific resonances in NMR spectra differentially. Among these approaches, relaxation editing emerges as a viable solution. In the realm of relaxation phenomena within NMR, Long-Lived States (LLS) and Long-Lived Coherences (LLC) manifest as promising phenomena, offering enhanced relaxation lifetimes in comparison to the traditional longitudinal (T1) and transverse (T2) relaxation times for coupled nuclear spins. Notably, LLC presents a pathway to attenuate uncoupled high-intensity peaks, effectively diminishing their impact. The foundation of this technique rests upon the premise that the relaxation lifetime in the rotating frame (T1ρ) remains smaller than TLLC. In pursuit of refining spectral assignments within complex mixtures, we introduce a new pulse sequence tailored for LLC Total Correlation Spectroscopy (LLC-TOCSY). This demonstrates efficacy in extracting LLC signals within configurations involving multiple coupled spins, thereby decluttering the spectrum and enhancing the accuracy of peak assignments. To validate the effectiveness of this method, a collection of samples was subjected to scrutiny, yielding compelling results.

A Joint Inversion Method Algorithm for T2-Pc two Dimensional NMR based on Logistic Functions

Data processing is a key step in the analysis of nuclear magnetic resonance (NMR) experimental data, and efficient and accurate computer inversion algorithms are the core of the T2-Pc two-dimensional NMR experiment. T2-Pc two-dimensional NMR experiment is an advanced experimental method that characterizes reservoir connectivity through two dimensions: relaxation time and capillary pressure, and can obtain irreducible water saturation under different pressure differences. However, the algorithm currently used for T2-Pc two-dimensional spectrum inversion uses a mutation kernel function, resulting in low accuracy of calculation results. This paper uses the Logistic function as the two-dimensional inversion kernel function, rewrites the inversion algorithm, and obtains more accurate inversion results. Numerical simulations have proven that the T2-Pc two-dimensional map obtained by this method not only has higher resolution, but also has greater applicability in the case of low signal-to-noise ratio and a small number of centrifugal echo groups. Practice has found that the proposed method can reduce the number of centrifugations during the experiment and significantly improve the efficiency of T2-Pc two-dimensional nuclear magnetic resonance experiments.

Variation pattern in the macromolecular (cellulose, hemicelluloses, lignin) composition of cell walls in Pinus tabulaeformis tree trunks at different ages as revealed using multiple techniques

The plant cell wall is a complex, heterogeneous structure primarily composed of cellulose, hemicelluloses, and lignin. Exploring the variations in these three macromolecules over time is crucial for understanding wood formation to enhance chemical processing and utilization. Here, we comprehensively analyzed the chemical composition of cell walls in the trunks of Pinus tabulaeformis using multiple techniques. In situ analysis showed that macromolecules accumulated gradually in the cell wall as the plant aged, and the distribution pattern of lignin was opposite that of polysaccharides, and both showed heterogenous distribution patterns. In addition, gel permeation chromatography (GPC) results revealed that the molecular weights of hemicelluloses decreased while that of lignin increased with age. Two-dimensional heteronuclear single quantum coherence nuclear magnetic resonance (2D-HSQC NMR) analysis indicated that hemicelluloses mainly comprised galactoglucomannan and arabinoglucuronoxylan, and the lignin types were mainly comprised guaiacyl (G) and p-hydroxyphenyl (H) units with three main linkage types: β-O-4, β-β, and β-5. Furthermore, the C-O bond (β-O-4) signals of lignin decreased while the C-C bonds (β-β and β-5) signals increased over time. Taken together, these findings shed light on wood formation in P. tabulaeformis and lay the foundation for enhancing the processing and use of wood and timber products.

Undescribed secondary metabolites derived from Cinnamomum migao H. W. Li, showcasing anti-inflammatory, antioxidant, and in silico properties

Inflammatory processes are renowned as the fundamental basis for numerous pathological conditions. The objective of this study was to assess the antioxidant, anti-inflammatory, molecular docking, and molecular dynamics simulation properties of 3 previously undescribed and 1 known guaiane-type sesquiterpenoids isolated from Cinnamomum migao. The structures of these compounds were confirmed using numerous chromatographic and spectroscopic methods, such as high-resolution electrospray ionization mass spectrometry (HRESIMS), one-dimensional nuclear magnetic resonance (1D NMR), two-dimensional nuclear magnetic resonance (2D NMR), and experimental circular dichroism (ECD). Additionally, in oxidatively stressed mouse hepatocytes, all tested compounds at a concentration of 50 µM increased the levels of natural antioxidant enzymes such as CAT, SOD, POD, and Nrf-2 compared to those in untreated cells. Compounds 1 and 2 strongly suppressed NO generation and proinflammatory cytokine production, with IC50 values of 8.76 and 5.45 µM, respectively, against TNF-α mRNA. Furthermore, the molecular docking and simulations explore the high affinity of the selected compounds and best fit in the TNF-α active site. Compounds 1 and 2 show prominent binding energies and bring significant confirmational changes in the TNF-α protein. The results of this study provide further validation for the traditional use of C. migao in the treatment of inflammation and pain, providing novel evidence regarding the therapeutic potential of the identified guaiane-type sesquiterpenoids.

Enhancing the efficacy of monolaurin against SARS-CoV-2 and influenza A (H1N1) with a nanoemulsion formulation

Monolaurin was utilized to formulate temperature-driven phase inversion nanoemulsions containing lesser galangal essential oil, fixed oil, and Cremophor RH40, with aim for eradicating enveloped viruses. Results showed that the droplet size of the nanoemulsion depended on lesser galangal essential oil–fixed oil ratio, monolaurin concentration, and oil concentration. Nanoemulsions prepared from lesser galangal essential oil–perilla oil (60:40) exhibited approximately 50-nm nanosized droplets and high entrapment efficiency (98.68 % ± 2.45 %). After storage at 25 °C for 1 year, droplet size did not vary significantly from the initial size, and monolaurin content was >95 %, indicating good physical and chemical stability. The monolaurin was located at the oil–water interface as indicated by a two-dimensional nuclear Overhauser effect nuclear magnetic resonance spectroscope and computer simulation. The 0.2% w/v monolaurin nanoemulsion inhibited SARS-CoV-2 and influenza A (H1N1) viruses with efficacy more than 3 log reduction (99.90 %) and low cytotoxicity. Hence, the monolaurin nanoemulsion can successfully eradicate enveloped viruses, especially SARS-CoV-2 and influenza A (H1N1).

Brønsted Acid-Promoted Cyclodimerization of α,β-Unsaturated γ-Ketoesters: Construction of Fused Pyrano-ketal-lactones and γ-Ylidene-butenolides.

Unprecedented MsOH-promoted diastereoselective cascade dimerization and intramolecular lactonization of readily accessible α,β-unsaturated γ-ketoesters are presented. The results obtained in this work, control experiments, and density functional theory (DFT) calculations suggested that the initial enolization and E to Z isomerization/equilibration of olefin (C=C) of substrate α,β-unsaturated γ-ketoesters give a Z-isomer preferentially over an E-isomer. Subsequently, the Z-isomer undergoes intermolecular annulation with α,β-unsaturated γ-ketoesters via domino Michael addition/ketalization/lactonization steps to furnish fused tetracyclic pyrano-ketal-lactone. However, the Z-isomer prefers intramolecular trans-esterification in a competing pathway and gives bicyclic γ-ylidene-butenolide. The key features of this work include simple Brønsted acid catalysis, the formation of three bonds, two rings, and three contiguous stereogenic centers in a single step, DFT calculations, and the assignment of relative stereochemistry through X-ray diffraction (XRD) and two-dimensional (2D) nuclear magnetic resonance (NMR) analyses.

Isolation, characterization, identification and quantification of 6-F oxyphenisatin dipropionate, a novel illegal additive, from a fruit-flavored jelly

ObjectiveThis study is aimed to screen, identify and detect illegal additives from healthcare products which claim or imply to have weight-loss effects. MethodUltra-high performance liquid chromatography-quadruple-time-of-flight mass spectroscopy (UPLC-Q-TOF/MS) was employed to perform non-targeted screening of illegal additives from a total of 26 batches of healthcare products with weight-loss effects. A novel oxyphenisatin dipropionate analog was discovered in a fruit-flavored jelly that was not clearly labeled as containing added drugs. After being separated and purified by silica gel column chromatography, the analog was unambiguously characterized by one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) spectroscopies. The molecular structure of the analog was finally identified by comparing the spectra of the analog with those of suspected candidates prepared by de novo synthesis strategy. Thereafter, a sensitive and precise reversed phase ultra performance liquid chromatography coupled with photodiode array (UPLC-PDA) detection method was developed and verified for the determination of the analog in 15 batches of real samples. ResultsIn the MS/MS spectra, the signal intensity of mass/charge ratios (m/z, 242 and 214) of the novel analog fragments was highly similar to that of mass/charge ratios (m/z, 224 and 196) of oxyphenisatin dipropionate fragments. Additionally, the 1D NMR spectrum of the analog was completely consistent with that of one of the suspected candidates prepared by the de novo synthesis strategy. Based on the above analysis, the structure of the analog was determined as 3,3-bis[4'-(propionyloxy)phenyl]-6-fluoro-2-oxoindoline, which was briefly named 6-F oxyphenisatin dipropionate. A developed quantitative method showed good linearity (R2 > 0.999) in a concentration range of 1.0–100 μg/mL. The limits of detection (LOD) and quantification (LOQ) for the analog was 3 mg/kg and 10 mg/kg, respectively. The average recoveries of the analog from spiked three different matrix samples in low (1 time of LOQ), medium (2 times of LOQ), and high (10 times of LOQ) concentrations were varied from 93.9 % to 107.8 % with a precision of 0.03–1.56 %. Results of quantitative analysis in 15 batches of healthcare products revealed that the content of 6-F oxyphenisatin dipropionate in a fruit-flavored jelly and a solid beverage was 118 mg/kg and 330 mg/kg, respectively. ConclusionIn terms of its structure, 6-F oxyphenisatin dipropionate replaces hydrogen atom by the fluorine atom at position 6 on the indolinone fragment in oxyphenisatin dipropionate. To our best knowledge, 6-F oxyphenisatin dipropionate has never been detected as an illegal additive in foods. Such illegal addition of the analog to foods is more concealing, thus the supervision and testing departments should attach great importance to its application in food markets.

Cell wall water induced dimensional changes of beech and pine wood

Hygroexpansion in timber construction is a phenomenon that occurs as a result of changes in the cell wall structure of wood caused by water. In this study, beech (Fagus sylvatica) and pine (Pinus taeda L.) wood were conditioned at different relative humidities with saturated salt solutions and saturated through vacuum pressure impregnation. The moisture content and dimensions of the samples were evaluated under various adsorption equilibrium states, as well as during drying from water saturation to an air-dried state. The results obtained using the two-dimensional time-domain nuclear magnetic resonance technique revealed two distinct types of cell wall water in both wood species, designated as B-water and C-water, as they displayed different mobilities. With increasing moisture content, the mobility of cell wall water increased; however, B-water maintained a constant level of mobility and formed water clusters when it accumulated in the high humidity region and at slightly above the fiber saturation point (FSP), which is evidence of the occurrence of free-bound water. The B-water, located in the amorphous region of microfibrils and their surrounding hemicellulose, exerted a predominant influence on the dimensional changes of the wood both under equilibrium and nonequilibrium conditions, in comparison to the C-water which located in the matrix of hemicellulose and lignin.

The structure-activity relationship for the ABA triblock siloxane surfactant in aqueous-ethanol system

It is challenging to improve the foam stability in high concentration ethanol solution. In this paper, well-defined polyether-modified siloxane surfactants were synthesized, and their structures were characterized through infrared spectroscopy and one- and two-dimensional Nuclear magnetic resonance (NMR) spectroscopy. Furthermore, the relationships of between their chemical structure, surface activity, and foam property in high concentration ethanol solution were investigated in detail. The results showed that the target surfactant exhibited an ABA-type structure. They can effectively reduce the surface tension and produce the excellent foam in 75 % ethanol. In particular, these surfactants with loner siloxane chain confer strong foaming power in 75 % solution. This ability with good surface activity in water-ethanol system suggests a wide range of potential applications in low surface tension solvent system.

Influence of Reservoir Microstructure on the State of Residual Oil According to Nuclear Magnetic Resonance (NMR) Spectroscopy

The influence of core properties on the state of residual oil in the process of oil displacement by water at the micro level is investigated. The pore size distribution, core permeability, dynamics and morphology of residual oil were studied. The analysis of the available experimental approaches to the study of the properties of the core and residual oil in the core samples showed that the existing methods do not provide complete information about the studied parameters. To solve these problems, it is proposed to use a combination of innovative relaxation-diffusion spectroscopy technology of nuclear magnetic resonance with traditional technology. A combination of mercury injection and nuclear magnetic resonance is used to measure the pore size distribution. The core permeability was determined using the nuclear magnetic resonance method. Two-dimensional nuclear magnetic resonance spectroscopy makes it possible to study the microscopic state of residual oil in an undisturbed core during the displacement process. With the help of the proposed methodology, a core study of the Shengli deposit in China was carried out. Pore size distributions were obtained, permeability and residual oil saturation at different stages of displacement were studied. Four types of residual oil are distinguished: strip-shaped (island), film, mesh, continuous. The influence of permeability on the fraction content of different types of residual oil in the process of displacement is shown. The research results demonstrate the influence of the pore space structure and wettability on the state of residual oil.

Initial Occurrence State and Movability Evaluation of the Gulong Shale Oil Reservoir, Songliao Basin

The Qing-1 layer of the Gulong Depression in the northern Songliao Basin is a liquid-rich shale oil reservoir that has the characteristics of nanopores, high maturity, high gas/oil ratio (GOR), etc. The production performance of wells in the Gulong shale oil reservoir shows the characteristics of “single gas production followed by oil-gas production”. It is difficult to analyze the initial occurrence state and movability of fluid in the shale nanopores using conventional methods. In this study, a comprehensive method, including phase behavior analysis, physical experiments, and molecular simulation, was established to analyze the initial occurrence state and movability of fluid in the Gulong shale oil reservoir. The phase state of the fluid was calculated by the equation of state (EOS), considering nano-confinement effects, and the initial occurrence state was quantitatively evaluated by combining two-dimensional nuclear magnetic resonance (NMR) and molecular dynamics simulation. The movable fluid saturation was quantitatively determined by centrifugal experiments. The results show that the condensate gas state was in small pores, while the volatile oil state was in large pores. The occurrence states of oil were mainly adsorbed oil and free oil. The proportion of adsorbed oil in inorganic pores was about 24.4%, while the proportion of absorbed oil in organic pores was about 57.8%. Based on the cutoff value of T2 before and after the centrifuged laboratory experiments, the movable limit of oil was determined to be 4.5 nm, and the movable fluid saturation was about 11%. The research method proposed in this study has important guiding significance for the initial occurrence state and movability evaluation of similar liquid-rich shale reservoirs.

Characterization of the Pb Coordination Environment and Its Connectivity in Lead Silicate Glasses: Results from 2D 207Pb NMR Spectroscopy.

The Pb-O coordination environment in binary (PbO)x(SiO2)100-x glasses with 30 ≤ x ≤ 70 is probed by using two-dimensional 207Pb nuclear magnetic resonance (NMR) isotropic-anisotropic correlation spectroscopy. The isotropic 207Pb NMR spectra show little composition-dependent evolution of the Pb-O nearest-neighbor coordination environment. The systematic variation of the chemical shift tensor parameters offers a unique insight into their local site symmetry and suggests the presence of pyramidal PbO3 and PbO4 sites with sterically active electron lone pairs and with Pb-O bond lengths ranging between 0.23 and 0.25 nm. The PbO3/PbO4 ratio shows a small but monotonic increase from ∼70:30 to 80:20 as the PbO content increases from 30 to 70 mol %. When taken together, the isotropic and anisotropic 207Pb NMR spectra suggest that the majority of the PbOn (3 ≤ n ≤ 4) pyramids in these glasses are connected to the SiO4 tetrahedra via Pb-O-Si linkages. A significant fraction of Pb-O-Pb linkages, where the oxygen is linked only to Pb atoms, appears only in glasses with PbO ≥ 60 mol %. These oxygen atoms appear to be corner-shared between the PbOn pyramids in the structure, and no evidence for edge-sharing between these pyramids is observed in this composition range. We hypothesize that a substantial fraction of the constituent PbOn pyramids start to participate in edge-sharing only at higher PbO contents (>70 mol %), which diminishes the glass-forming ability of the network. This work illustrates the potential of isotropic-anisotropic correlation NMR spectroscopy in structural studies involving nuclides with large chemical shift ranges and anisotropy.

Cyclization-enhanced poly(β-amino ester)s vectors for efficient CRISPR gene editing therapy

Among non-viral gene delivery vectors, poly(β-amino ester)s (PAEs) are one of the most versatile candidates because of their wide monomer availability, high polymer flexibility, and superior gene transfection performance both in vitro and in vivo. Over two decades, PAEs have evolved from linear to highly branched structures, significantly enhancing gene delivery efficacy. Building on the proven efficient sets of monomers in highly branched PAEs (HPAEs), this work introduced a new class of cyclic PAEs (CPAEs) constructed via an A2 + B4 + C2 cyclization synthesis strategy and identified their markedly improved gene transfection capabilities in gene delivery applications. Two sets of cyclic PAEs (CPAEs) with rings of different sizes and topologies were obtained. Their chemical structures were confirmed via two-dimensional nuclear magnetic resonance and the photoluminescence phenomena, and their DNA delivery behaviours were investigated and compared with the HPAE counterparts. In vitro assessments demonstrated that the CPAEs with a macrocyclic architecture (MCPAEs), significantly enhanced DNA intracellular uptake and facilitated efficient gene expression while maintaining perfect biocompatibility. The top-performance MCPAEs have been further employed to deliver a plasmid coding dual single guide RNA-guided CRISPR-Cas9 machinery to delete COL7A1 exon 80 containing the c.6527dupC mutation. In recessive dystrophic epidermolysis bullosa (RDEB) patient-derived epidermal keratinocytes, MCPAEs facilitated the CRISPR plasmid delivery and achieved efficient targeted gene editing in multiple colonies.

High-Safety Electrolytes with an Anion-Rich Solvation Structure Tuned by Difluorinated Cations for High-Voltage Lithium Metal Batteries.

As next-generation energy storage devices, lithium metal batteries (LMBs) must offer high safety, high-voltage resistance, and a long life span. Electrolyte engineering is a facile strategy to tailor the interfacial chemistry of LMBs. In particular, the solvation structure and derived solid electrolyte interphase (SEI) are crucial for a satisfactory battery performance. Herein, a novel middle-concentrated ionic liquid electrolyte (MCILE) with an anion-rich solvation structure tuned by difluorinated cations is demonstrated to achieve ultrahigh safety, high-voltage stability, and excellent ternary-cathode compatibility. Novel gem-difluorinated cations first synthesized for prestoring fluorine on positively charged species, not only preferentially adsorb in the inner-Helmholtz layers, but also participate in regulating the Li+ solvation structure, resulting in a robust interphase. Moreover, these weak interactions in the Li+ solvation structure including anion-solvent and ionic liquid (IL) cation-solvent pairs are first revealed, which are beneficial for promoting an anion-dominated solvation structure and the desolvation process. Benefiting from the unique anion-rich solvation structure, a stable hetero-SEI structure is obtained. The designed MCILE exhibits compatibility with Li metal anode and the high-voltage ternary cathode at high temperatures (60°C). This work provides a new approach for regulating the solvation structure and electrode interphase chemistry of LMBs via difluorinated IL cations.

Engineering Dual CO2- and Photothermal-Responsive Membranes for Switchable Double Emulsion Separation.

Stimulus-responsive membranes demonstrate promising applications in switchable oil/water emulsion separations. However, they are unsuitable for the treatment of double emulsions like oil-in-water-in-oil (O/W/O) and water-in-oil-in-water (W/O/W) emulsions. For efficient separation of these complicated emulsions, fine control over the wettability, response time, and aperture structure of the membrane is required. Herein, dual-coated fibers consisting of primary photothermal-responsive and secondary CO2-responsive coatings are prepared by two steps. Automated weaving of these fibers produces membranes with photothermal- and CO2-responsive characteristics and narrow pore size distributions. These membranes exhibit fast switching wettability between superhydrophilicity (under CO2 stimulation) and high hydrophobicity (under near-infrared stimulation), achieving on-demand separation of various O/W/O and W/O/W emulsions with separation efficiencies exceeding 99.6%. Two-dimensional low-field nuclear magnetic resonance and correlated spectra technique are used to clarify the underlying mechanism of switchable double emulsion separation. The approach can effectively address the challenges associated with the use of stimulus-responsive membranes for double emulsion separation and facilitate the industrial application of these membranes.

Nuclear magnetic resonance study on wettability of shale oil reservoir

In order to identify and evaluate reservoir wettability, so as to select suitable mining methods to improve oil recovery, wettability evaluation through one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) technology combined with physical simulation experiments was carried out in this study, and the feasibility of the experimental method was verified by contact Angle method. According to the principle of hydrophilic inorganic content and hydrophobic organic content of shale, the wettability can be evaluated through vacuum self-imbibition oil and self-imbibition water physical simulation experiments combined with 1D NMR technology, that is, total water absorption can be considered as inorganic content, organic content can be obtained by subtracting total water absorption from total oil absorption, and adsorbed oil content can be obtained by subtracting pore volume from total oil absorption. In addition to the wettability measurement by contact Angle method to verify the experimental method, the occurrence ratio of adsorbed oil can also be verified twice by 2D NMR spectrum. The results show that the error between the proportion of adsorbed oil measured by 2D NMR spectrum and that measured by self-imbibition method is within 4%. In the saturated oil-bound water state, the bound water is within the relaxation interval of 1<T1<10 ms and 1<T2<10 ms, and the oil signal is within the relaxation interval of T1>10 ms and T2>10 ms. And the relaxation time of aqueous phase moved to the right compared with that in the saturated water state, the interaction force between water and the pore wall was weakened, showing the characteristics of free fluid, and the rock sample became oil-wet, which was consistent with the wettability results measured by contact Angle. Therefore, the wettability of rock samples can be evaluated by 1D NMR technique combined with vacuum imbibition method, or by analyzing the changes of NMR spectra of aqueous phase in completely saturated water and saturated oil-bound water.

Strategy for the Enzymatic Acylation of the Apple Flavonoid Phloretin Based on Prior α-Glucosylation.

The acylation of flavonoids serves as a means to alter their physicochemical properties, enhance their stability, and improve their bioactivity. Compared with natural flavonoid glycosides, the acylation of nonglycosylated flavonoids presents greater challenges since they contain fewer reactive sites. In this work, we propose an efficient strategy to solve this problem based on a first α-glucosylation step catalyzed by a sucrose phosphorylase, followed by acylation using a lipase. The method was applied to phloretin, a bioactive dihydrochalcone mainly present in apples. Phloretin underwent initial glucosylation at the 4'-OH position, followed by subsequent (and quantitative) acylation with C8, C12, and C16 acyl chains employing an immobilized lipase from Thermomyces lanuginosus. Electrospray ionization-mass spectrometry (ESI-MS) and two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) confirmed that the acylation took place at 6-OH of glucose. The water solubility of C8 acyl glucoside closely resembled that of aglycone, but for C12 and C16 derivatives, it was approximately 3 times lower. Compared with phloretin, the radical scavenging capacity of the new derivatives slightly decreased with 2,2-diphenyl-1-picrylhydrazyl (DPPH) and was similar to 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+). Interestingly, C12 acyl-α-glucoside displayed an enhanced (3-fold) transdermal absorption (using pig skin biopsies) compared to phloretin and its α-glucoside.

Integrative Molecular Structure Elucidation and Construction of an Extended Metabolic Pathway Associated with an Ancient Innate Immune Response in COVID-19 Patients.

We present compelling evidence for the existence of an extended innate viperin-dependent pathway, which provides crucial evidence for an adaptive response to viral agents, such as SARS-CoV-2. We show the in vivo biosynthesis of a family of novel endogenous cytosine metabolites with potential antiviral activities. Two-dimensional nuclear magnetic resonance (NMR) spectroscopy revealed a characteristic spin-system motif, indicating the presence of an extended panel of urinary metabolites during the acute viral replication phase. Mass spectrometry additionally enabled the characterization and quantification of the most abundant serum metabolites, showing the potential diagnostic value of the compounds for viral infections. In total, we unveiled ten nucleoside (cytosine- and uracil-based) analogue structures, eight of which were previously unknown in humans allowing us to propose a new extended viperin pathway for the innate production of antiviral compounds. The molecular structures of the nucleoside analogues and their correlation with an array of serum cytokines, including IFN-α2, IFN-γ, and IL-10, suggest an association with the viperin enzyme contributing to an ancient endogenous innate immune defense mechanism against viral infection.

Helical sulfonyl-γ-AApeptides modulating Aβ oligomerization and cytotoxicity by recognizing Aβ helix

In contrast to prevalent strategies which make use of β-sheet mimetics to block Aβ fibrillar growth, in this study, we designed a series of sulfonyl-γ-AApeptide helices that targeted the crucial α-helix domain of Aβ13-26 and stabilized Aβ conformation to avoid forming the neurotoxic Aβ oligomeric β-sheets. Biophysical assays such as amyloid kinetics and TEM demonstrated that the Aβ oligomerization and fibrillation could be greatly prevented and even reversed in the presence of sulfonyl-γ-AApeptides in a sequence-specific and dose-dependent manner. The studies based on circular dichroism, Two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) spectra unambiguously suggested that the sulfonyl-γ-AApeptide Ab-6 could bind to the central region of Aβ42 and induce α-helix conformation in Aβ. Additionally, Electrospray ionisation-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS) was employed to rule out a colloidal mechanism of inhibitor and clearly supported the capability of Ab-6 for inhibiting the formation of Aβ aggregated forms. Furthermore, Ab-6 could rescue neuroblastoma cells by eradicating Aβ-mediated cytotoxicity even in the presence of pre-formed Aβ aggregates. The confocal microscopy demonstrated that Ab-6 could still specifically bind Aβ42 and colocalize into mitochondria in the cellular environment, suggesting the rescue of cell viability might be due to the protection of mitochondrial function otherwise impaired by Aβ42 aggregation. Taken together, our studies indicated that sulfonyl-γ-AApeptides as helical peptidomimetics could direct Aβ into the off-pathway helical secondary structure, thereby preventing the formation of Aβ oligomerization, fibrillation and rescuing Aβ induced cell cytotoxicity.