Interactions of indole alkaloids with myoglobin: A mass spectrometry based spectrometric and computational method.

RATIONALE: Interactions of drug molecules and proteins play important roles in physiological and pathological processes in vivo. It is of significance to establish a reliable strategy for studying protein-drug ligand interactions and would be helpful for the design and screening of new drugs in pharmacological research. METHODS: The interactions between four indole alkaloids (IAs) extracted from Ophiorrhiza japonica (O. japonica) and myoglobin (Mb) protein were investigated using a multi-spectrometric and computational method of native electrospray ionization mass spectrometry (native ESI-MS), hydrogen/deuterium exchange mass spectrometry (HDX-MS), circular dichroism (CD) and molecular docking (MD). RESULTS: The IA-bound Mb complexes were analyzed using native ESI-MS, with the obtained protein-to-ligand stoichiometry at 1:1, 1:2 and 1:3. Binding constants were measured according to the interpretation of MS spectra. MD complemented MS measurements, probing the binding sites and modes of the four IAs to Mb. Analyses involving CD and HDX-MS demonstrated that exposure to IAs could affect the conformation of Mb by decreasing the α-helix content and made Mb more susceptible to HDX at the backbone. CONCLUSIONS: A new MS-based integrated analysis method has been developed to successfully study the interactions of Mb and IAs extracted from O. japonica. The experimental and calculation results have good consistency, revealing all of the four IA molecules could bind to Mb to form 1:1, 1:2 and 1:3 Mb-IA complexes. The order of binding ability of these IAs to Mb was ophiorrhine B > compound C > ophiorrhine A > compound D. CD and HDX-MS results indicated that binding with IAs destabilizes Mb. HDX-MS analysis suggests that Mb becomes more susceptible to HDX, indicating that binding with IAs destabilizes the structure of Mb. In addition, the interaction with IAs affected the overall structure of Mb, ascribed to the decrease of α-helix content and less folding of the backbone.

A monolithic copolymer prepared from N-(4-vinyl)-benzyl iminodiacetic acid, divinylbenzene and N,N'-methylene bisacrylamide for preconcentration of cadmium(II) and cobalt(II) from biological samples prior to their determination by ICP-MS.

A capillary monolith consisting of poly[N-(4-vinyl)-benzyl iminodiacetic acid-co-divinylbenzene-co-N,N'-methylene bisacrylamide), referred to as poly(VBIDA-DVB-Bis), has been prepared. It is shown to be an efficient sorbent for the enrichment of Co(II) and Cd(II). The two ions are completely retained by the monolith in the pH range from 4.0 to 9.0. The breakthrough curve tests were adopted to evaluate the adsorption performance of the monolith towards Co(II) and Cd(II). A dose-response model was used to describe the breakthrough curves of the two ions at different initial concentrations. The adsorption capacities for Co(II) and Cd(II) are 1.54 and 1.73 mg·m-1 at a concentration level of 2.5 mg·L-1, respectively. The enrichment factor is 100, and the required sample volume is 5 mL. Following elution of the two ions with 0.5 M HNO3, they were quantified by ICP-MS. The limits of detection in a 1 mL sample are 0.35 ng·L-1 for Co(II) and 0.44 ng·L-1 for Cd(II). The method was applied to the determination of Co(II) and Cd(II) in spiked rice, human urine and seawater samples. Graphical abstract Schematic representation of a monolithic copolymer prepared from N-(4-vinyl)-benzyl iminodiacetic acid (VBIDA), divinylbenzene (DVB) and N,N'-methylene bisacrylamide (Bis) and its application for selective capturing of cadmium(II) and cobalt(II) from complex sample matrices prior to their determination by ICP-MS.

Molecular genetic analysis of the Jk(a-b-) phenotype in Chinese: A novel silent recessive JK allele.

The Jk(a-b-) phenotype, referred to as Jknull, is rare in most populations. This blood type is characterized by the absence of Kidd glycoprotein on the surface of red blood cells (RBCs) and moderately reduced ability to concentrate urine. The molecular basis for Jknull phenotype includes splice-site mutations, missense mutations, and a partial gene deletion in the JK(SLC14A1) gene that encodes the human urea transporter protein. In this study, we have analyzed 10 Chinese Jknull samples to determine their molecular bases. In addition to the well known Polynesian Jknull allele, three Jknull alleles were detected including one novel Jknull allele: JKA (130A, 220G).

Mechanism of alcohol-enhanced lucigenin chemiluminescence in alkaline solution.

The chemiluminescence (CL) of lucigenin (Luc(2+)) can be enhanced by different alcohols in alkaline solution. The effect of different fatty alcohols on the CL of lucigenin was related to the carbon chain length and the number of hydroxyl groups. Glycerol provides the greatest enhancement. UV/Vis absorption spectra and fluorescence spectra showed that N-methylacridone (NMA) was produced in the CL reaction in the presence of different alcohols. The peak of the CL spectrum was located at 470 nm in all cases, indicating that the luminophore was always the excited-state NMA. The quenching of lucigenin CL by superoxide dismutase (SOD) and the electron spin resonance (ESR) results with the spin trap of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) demonstrated that superoxide anions (O2 (•-)) were generated from dissolved oxygen in the CL reaction and that glycerol and dihydroxyacetone (DHA) can promote O2 (•-) production by the reduction of dissolved oxygen in alkaline solution. It was assumed that the enhancement provided by different alcohols was related to the solvent effect and reducing capacity. Glycerol and DHA can also reduce Luc(2+) into lucigenin cation radicals (Luc(•+) ), which react with O2 (•-) to produce CL, and glycerol can slowly transform into DHA, which is oxidized quickly in alkaline solution.

18F-FDG PET/CT findings in fatal Balamuthia Mandrillaris encephalitis in brain stem: A case report.

We presented a case of a 66-year-old female whose initial symptom was headache without obvious inducement. The patient's condition progressed rapidly to a semi-coma state after symptomatic treatment. The 18F-FDG PET/CT scan revealed circular FDG hypermetabolism and central metabolic defect of the pons and left frontal lobe lesions. The combination of clinical findings, MRI, and Metagenomic next-generation sequencing (NGS) of cerebrospinal fluid led to the diagnosis of Balamuthia mandrillaris encephalitis. The patient died 5 days after discharge.

[Study on the optical properties of ultra-thin metal films in the THz band].

Thin metal films are good candidates of terahertz detectors, reflectors, waveguides and terahertz quantum-cascade lasers (THz-QCLs). The optical parameter is the basis not only for designing the THz components but also for developing novel optoelectronic materials. In the present paper, the complex refractive indices of the ultra-thin metal (Cr, Ni and Ti) films in the THz band were obtained by the THz differential time-domain spectroscopy. The reflection spectra of the GaAs/metals interface were calculated according to the Fresnel formula. The mean reflectance of 25 nm Cr, Ni and Ti are over 80% from 0.3 to 1.5 THz. The results show that ultra-thin metal films can be used for reflectors as well as the electrodes in the THz band.

The Waiting Room Assessment to Virtual Emergency Department pathway: Initiating video-based telemedicine in the pediatric emergency department.

The COVID-19 pandemic has presented pediatric emergency departments with unique challenges, resulting in a heightened demand for adapted clinical pathways. In response to this need, the Montreal Children's Hospital pediatric emergency department introduced the WAVE (Waiting Room Assessment to Virtual Emergency Department) pathway, a video-based telemedicine pathway for selected non-critical patients, aiming to reduce safety issues related to emergency department overcrowding, while providing timely care to all children presenting and registering at our emergency department. The objective of the WAVE pilot phase was to evaluate the feasibility and acceptability of telemedicine in our pediatric emergency department, which was previously unfamiliar with this mode of care delivery. During the six-week, three-evening per week deployment, we conducted 18 five-hour telemedicine shifts. In total, 27 patients participated in the WAVE pathway. Results from this pilot phase met four of five a priori feasibility and acceptability criteria. Overall, participating families were satisfied with this novel care pathway and reported no disruptive technological barriers.

Comprehensive binding analysis of polybrominated diphenyl ethers and aryl hydrocarbon receptor via an integrated molecular modeling approach.

Polybrominated diphenyl ethers (PBDEs) are often suspected to activate the signal transduction pathway of aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, for the induction of toxicity. Hence, the binding property of PBDEs with AhR is assumed to be associated with the ligand-dependent activation of AhR that may introduce many drug-metabolizing enzymes of genes encoding. However, the binding mechanism and the structural effect of PBDEs on their binding properties of AhR still need to be unraveled for toxicology research. A comprehensive study of the PBDEs-AhR binding mechanism was investigated using an integrated molecular modeling approach with two-dimensional quantitative structure-activity relationships (2D-QSAR), three-dimensional QSAR (3D-QSAR), and molecular docking simulation. Molecular docking revealed the differences in binding domains among 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-AhR complex and two PBDE-AhR complexes. A 2D-QSAR model was developed to analyze the overall structural effects of PBDEs on the binding affinity of AhR. It provided an insight into major physico-chemical properties by multiple linear regression based on genetic algorithm with reasonable results. The 3D-QSAR modeling discovered the detailed interaction features of binding sites, configurations and interaction fields of AhR with different PBDE ligands. This study demonstrated that the descriptors of Smin69 and MoRSEC15 were related to electronic properties and had a great effect on the relative binding affinities. The position of Br substitutions exhibited a significant influence on the interactions between AhR and PBDEs, including halogen interaction, π-S interaction, π-π stacking interaction, and hydrophobic effect. This integrated molecular modeling approach provided a comprehensive analysis of the structural effects of PBDEs on their binding properties with AhR at molecular level.

Evaluation of structurally different brominated flame retardants interacting with the transthyretin and their toxicity on HepG2 cells.

Brominated flame retardants (BFRs) are found at quantifiable levels in both humans and wildlife and may potentially cause a health risk. For BFRs and their derivatives, limited information regarding the relationship among the structure, binding affinity to the target protein and toxicity is currently available. In the present work, representative BFRs with different hydroxyl- or bromo-substituents, namely 2, 2', 4, 4'-tetrabromodiphenyl ether (BDE-47), 3-hydroxy-2, 2', 4, 4'-tetrabromodiphenyl ether (3-OH-BDE-47) and tetrabromobisphenol A (TBBPA), were selected to investigate the interactions with transthyretin (TTR) by electrospray ionization mass spectrometry (ESI-MS) and cytotoxicity on HepG2 cells. It was noted that BDE-47 had a weak binding affinity to TTR, while 3-OH-BDE-47 and TBBPA had a stronger binding affinity than BDE-47 and thyroxine (T4). Hence, 3-OH-BDE-47 and TBBPA could affect the binding of TTR with its native ligand T4 by competitive binding to TTR, even at equal concentrations, which might be associated with BFR toxicity of endocrine disruption. Negative cooperativity was found for 3-OH-BDE-47 and TBBPA binding to TTR, similar to T4 with a well-established negatively cooperative binding mechanism. The tendency of toxic effects on HepG2 cells for these three BFRs was, 3-OH-BDE-47 > TBBPA > BDE-47, and this order was in good agreement with the binding ability explored by ESI-MS experiments and molecular docking simulation. The observations obtained by this study demonstrate that the binding properties of these BFRs to TTR and their cytotoxicity are correlated with structure differentials and functional substituents.

Characterization of in vitro metabolism of focal adhesion kinase inhibitors by LC/MS/MS.

Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, is critically involved in cell migration, spreading and proliferation at the early step of various cancers. Small molecule inhibitors of FAK are effective to inhibit its activation in the process of tumor formation in cell. To better understand biotransformation of FAK inhibitors, this work has investigated in vitro phase I metabolism of inhibitors (namely PF-573228, PF-562271 and PF-03814735) by rat liver microsomes model. Using liquid chromatography - quadrupole time of flight mass spectrometry and tandem mass spectrometry (LC/Q-TOF/MS and MS/MS), three metabolites of PF-573228 and PF-562271 were observed and characterized, respectively. These in vitro metabolites were reported for the first time. The structures and fragmentation patterns of these metabolites were elucidated, and phase I metabolic pathways for FAK inhibitors were proposed. The main metabolic pathways of PF-573228 were hydroxylation, dehydrogenation and N-dealkylation. For PF-562271, they were hydroxylation and dehydrogenation. Hydroxylation was observed as the primary metabolism for PF-0381473.

Nitrogen-Doped Carbon-Supported Nickel Nanoparticles: A Robust Catalyst to Bridge the Hydrogenation of Nitriles and the Reductive Amination of Carbonyl Compounds for the Synthesis of Primary Amines.

An efficient method was developed for the synthesis of primary amines either from the hydrogenation of nitriles or reductive amination of carbonyl compounds. The reactions were catalyzed by nitrogen-doped mesoporous carbon (MC)-supported nickel nanoparticles (abbreviated as MC/Ni). The MC/Ni catalyst demonstrated high catalytic activity for the hydrogenation of nitriles into primary amines in high yields (81.9-99 %) under mild reaction conditions (80 °C and 2.5 bar H2 ). The MC/Ni catalyst also promoted the reductive amination of carbonyl compounds for the synthesis of primary amines at 80 °C and 1 bar H2 . The hydrogenation of nitriles and the reductive amination proceeded through the same intermediates for the generation of the primary amines. To the best of our knowledge, no other heterogeneous non-noble metal catalysts have been reported for the synthesis of primary amines under mild conditions, both from the hydrogenation of nitriles and reductive amination.

Quantitative-Profiling Method of Serum Steroid Hormones by Hydroxylamine-Derivatization HPLC-MS.

A sensitive and rapid high performance liquid chromatography-mass spectrometry (HPLC-MS) method was developed and validated for simultaneous quantification of ten steroid hormones, including estrogens, androgens, progesterones, and corticosteroids four classes of steroids. The following ten steroid hormones were analyzed: progesterone, 21-deoxycortisol, estrone, 4-androstenedione, testosterone, dihydro-testosterone, androstenone, dehydroepiandrosterone, corticosterone and cortisone. Stable deuterated isotopes were used as internal standards for quantification. Sample preparation with and without derivatization were performed after liquid-liquid extraction, and the corresponding results were compared according to sensitivity and selectivity. Hydroxylamine derivatization was found to improve the ionization efficiency of the analytes for electrospray ionization MS analysis. The gradient of mobile phase and experimental parameters for HPLC separation were optimized. The lower limits of quantification were in the range of 0.05-5 ng mL-1 with wide linear range for the ten steroid hormones. The intra-day precision < 11.1% and recovery of 84.5-120% with negligible matrix effect were achieved, where within the acceptance limits of the FDA guideline. Total HPLC-MS analysis time was 6 min. This method enables simultaneous quantification of steroids in human serum. It will be helpful for the serum steroid profiling in order to understand various endocrinology diseases.

Study on the Structural Effect of Maltoligosaccharides on Cytochrome c Complexes Stabilities by Native Mass Spectrometry.

Noncovalent interactions between ligands and targeting proteins are essential for understanding molecular mechanisms of proteins. In this work, we investigated the interaction of Cytochrome c (Cyt c) with maltoligosaccharides, namely maltose (Mal II), maltotriose (Mal III), maltotetraose (Mal IV), maltopentaose (Mal V), maltohexaose (Mal VI) and maltoheptaose (Mal VII). Using electrospray ionization mass spetrometry (ESI-MS) assay, the 1:1 and 1:2 complexes formed by Cyt c with maltoligosaccharide ligand were observed. The corresponding association constants were calculated according to the deconvoluted spectra. The order of the relative binding affinities of the selected oligosaccharides with Cyt c were as Mal III > Mal IV > Mal II > Mal V > Mal VI > Mal VII. The results indicated that the stability of noncovalent protein complexes was intimately correlated to the molecular structure of bound ligand. The relevant functional groups that could form H-bonds, electrostatic or hydrophobic forces with protein's amino residues played an important role for the stability of protein complexes. In addition, the steric structure of ligand was also critical for an appropriate interaction with the binding pocket of proteins.

Nitrogen-Doped Carbon Materials for the Metal-Free Reduction of Nitro Compounds.

In this study, nitrogen-doped carbon materials (labeled as NC-T) were easily prepared by thermal treatment of the melamine-chitosan composite. NC-T catalysts demonstrated good activity toward the reduction of nitro compounds by using hydrazine hydrate (N2H4·H2O) as the reductant. The activity of NC-T enhanced with the rise of the pyrolysis temperature because of the enhanced ratio of graphitic-type nitrogen, which might be the active sites for the reduction of nitro compounds. NC-950 showed high activity for the selective reduction of nitro compounds in hexane to produce amines at yields from 87.5 to 100% at 90 °C using 2 equiv of N2H4·H2O. The NC-950 catalyst demonstrated comparable or even higher catalytic activity in comparison with the reported metal catalysts. The hydrogenation of nitro compounds with N2H4·H2O proceeded via the direct way involving hydroxylamine as the reaction intermediate.

Interactions of perfluorooctanoic acid and perfluorooctanesulfonic acid with serum albumins by native mass spectrometry, fluorescence and molecular docking.

The binding information of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) with bovine and human serum albumins was investigated and characterized in details by using a combination method of electrospray ionization mass spectrometry (ESI-MS), fluorescence, circular dichroism (CD) and molecular docking (MD). The ESI-MS analysis revealed that maximally eight PFOA or PFOS molecules could bind to serum albumins at high mole ratios of PFOA/PFOS. Association constants were measured by ESI-MS and suggested that PFOS had a better binding affinity than PFOA. PFOA and PFOS were likely to bind with serum albumins in more than one pocket. The CD data demonstrated that binding of PFOA and PFOS could change the conformation of serum albumins with decreasing α-helix content, which may affect the protein physiological function. The phenomenon of protein fluorescence quenching by the binding of PFOA and PFOS indicated that the hydrophobic pocket proximate to Trp 214 in human serum albumin might be one of the dominated binding sites. This assumption was further confirmed by MD simulation. Consistent to ESI-MS observation, MD results also displayed a stronger binding affinity of PFOS than PFOA according to the calculated binding free energy, which is probably ascribed to one more hydrogen bond formed in the PFOS-bound protein complexes.

Temporal and spatial impact of Spartina alterniflora invasion on methanogens community in Chongming Island, China.

Methane production by methanogens in wetland is recognized as a significant contributor to global warming. Spartina alterniflora (S. alterniflora), which is an invasion plant in China's wetland, was reported to have enormous effects on methane production. But studies on shifts in the methanogen community in response to S. alterniflora invasion at temporal and spatial scales in the initial invasion years are rare. Sediments derived from the invasive species S. alterniflora and the native species Phragmites australis (P. australis) in pairwise sites and an invasion chronosequence patch (4 years) were analyzed to investigate the abundance and community structure of methanogens using quantitative real-time PCR (qPCR) and Denaturing gradient gel electrophoresis (DGGE) cloning of the methyl-coenzyme M reductase A (mcrA) gene. For the pairwise sites, the abundance of methanogens in S. alterniflora soils was lower than that of P. australis soils. For the chronosequence patch, the abundance and diversity of methanogens was highest in the soil subjected to two years invasion, in which we detected some rare groups including Methanocellales and Methanococcales. These results indicated a priming effect at the initial invasion stages of S. alterniflora for microorganisms in the soil, which was also supported by the diverse root exudates. The shifts of methanogen communities after S. alterniflora invasion were due to changes in pH, salinity and sulfate. The results indicate that root exudates from S. alterniflora have a priming effect on methanogens in the initial years after invasion, and the predominate methylotrophic groups (Methanosarcinales) may adapt to the availability of diverse substrates and reflects the potential for high methane production after invasion by S. alterniflora.

Genomic analysis of para-Bombay individuals in south-eastern China: the possibility of linkage and disequilibrium between FUT1 and FUT2.

BACKGROUND: The para-Bombay phenotype results from a variety of mutations in the α-(1,2)-fucosyltransferase gene (FUT1). We investigated samples from seven Chinese probands serologically typed as having the para-Bombay phenotype. MATERIALS AND METHODS: The para-Bombay phenotype was identified by standard serological methods. Genetic mutations of FUT1 and FUT2 genes were analysed by DNA sequencing. Heterozygous mutations of FUT1 were identified by TOPO cloning sequencing. Blood samples from 331 randomly-selected Chinese donors were analysed with the SNaPshot system to distinguish five known mutations (Se C357T, A385T, G428A, G716A and FUT1 880delTT) in the FUT1 and FUT2 genes. The genetic characteristics of all para-Bombay probands identified in the Fujian Blood Centre, including those in the present study, were also summarised. RESULTS: Three FUT1 genotypes, h1/h1 (5 individuals), h1/h6 (1 individual) and h3/h2 (1 individual), and three FUT2 genotypes, Se(357)/Se(357) (5 individuals), Se(357)/Se(357, 385) (1 individual) and Se(357)/Se(357, 716) (1 individual) were observed in seven para-Bombay probands. Among 331 donors, only one individual carried the G716A and 880delTT mutations in heterozygosity; this subjects FUT1 and FUT2 genotypes were H/h2 and Se(357)/Se(357, 716), respectively. CONCLUSION: The review of all para-Bombay probands identified in the Fujian Blood Centre showed that h1 and h2 are the predominant non-functional FUT1 alleles in Fujian para-Bombay individuals. Our data confirm the hypothesis that the h2 allele is linked to Se(357, 716), and the concurrence of unique FUT1 and FUT2 mutations is geographically specific.

[Determination of available as (III) and as (V) in soils by flow injection hydride generation atomic fluorescence spectrometry].

A method for the determination of available As (III) and As (V) in soils by flow injection hydride generation atomic fluorescence spectrometry (FI-HG-AFS) was presented. As the concentration of As (III) in extract was determined by HG-AFS in the medium of 0.1 mol.L-1 citric acid solution and the total arsenic was determined after FI on-line reduction of As (V) by L-cysteine, the concentration of As (V) was calculated by the difference. The manifolds for FI on-line reduction of As (V) were designed. The chemical conditions and the operating parameters were optimized. The detection limits (3 sigma) of As (III) and As (V) are 0.11 and 0.07 ng.mL-1, respectively, and the relative standard deviation is 1.43 for 7 determinations of 10 ng.mL-1 As (III) solution. The developed method was applied to the determination of available As (III) and As (V) in real soils and the recovery was 82%-104%.

Morphological control and evolution of octahedral and truncated trisoctahedral Pt-Au alloy nanocrystals under microwave irradiation.

Uniform and well-defined truncated trisoctahedral and octahedral Pt-Au alloy nanocrystals were fabricated by co-reducing H(2)PtC(l6)-HAuC(l4) with tetraethylene glycol (TEG) under microwave irradiation for only 140 s. Iodide ions were critical to the morphological control and evolution of Pt-Au alloy nanostructures. The as-prepared Pt-Au alloy nanocrystals exhibited efficient electrocatalytic activities.

[Effects of filamentous macroalgae on the methane emission from urban river: a review].

The global warming caused by greenhouse gases emission has raised serious concerns. Recent studies found that the carbon dioxide (CO2) and methane (CH4) emissions from river ecosystem can partly offset the carbon sequestration by terrestrial ecosystem, leading to a rethink of the effects of river ecosystem on the global carbon balance and greenhouse gases emission inventory. As an important primary producer in urban river ecosystem, filamentous macroalgae can deeply affect the carbon cycle process of river system through changing the abiotic and biotic factors in the interface of water-sediment. This paper reviewed the effects of filamentous macroalgae on the CH4 emission from urban river system from the aspects of 1) the effects of urbanization on the river ecosystem and its CH4 emission flux, 2) the effects of filamentous macroalgae on the CH4 generation and emission process in natural river systems, and 3) the effects of filamentous macroalgae on the primary productivity and CH4 emission process in urban river systems. The current problems and future directions in related researches were discussed and prospected.