In vitro antioxidant and free-radical scavenging activities of polar leaf extracts of Vernonia amygdalina.

BACKGROUND: Plants are able to deliver a huge number of differing bioactive compounds which may supplement the requirements of the human body by acting as natural antioxidants. Antioxidants are mindful for the defense component of the life form against the pathologies related to the assault of free radicals. The main purpose of this study was to investigate the qualitative phytochemical composition of Vernonia amygdalina leaf extract and its antioxidant activity. METHOD: The powdered plant sample was successively extracted with aqueous, methanol and ethanol solvents using Soxhlet apparatus. The antioxidant activities of the crude leaf extract were determined using 1, 1- diphenyl-2-picryl hydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical, phosphomolybdate (PM) and hydrogen peroxide (H2O2) scavenging assay. All the examinations were drained triplicates and average values of each test were taken. RESULTS: Phytochemical investigation of the plant revealed that the three solvent extracts contained numerous bioactive compounds namely alkaloids, tannins, saponins, phenols, terpenoids, steroids, glycosides and sugars. The result showed that, the leaf extracts of V. amygdalina obtained from methanol extract exhibit the maximum antioxidant activity compared ethanol and aqueous extracts. The IC50 values of DPPH assay for the H2O, MeOH and EtOH extracts were 111.4, 94.92 and 94.83 µg/ml; of ABTS assay were 334.3, 179.8 and 256.9 µg/ml; of H2O2 assay were 141.6, 156 and 180.6 µg/ml, respectively. The maximum radical scavenging activity was obtained in DPPH assay while the lowest scavenging activity was obtained in ABTS assay method. The data obtained in the in vitro models clearly suggest that methanol extract has higher antioxidant activity due to a higher presence of phenolic constituents in the extract. CONCLUSION: This study revealed that V. amygdalina leaf has a noteworthy antioxidant and free radical scavenging activity mitigating the traditional use of the plant for different aliments.

The Effect of Non-Solvent Nature on the Rheological Properties of Cellulose Solution in Diluted Ionic Liquid and Performance of Nanofiltration Membranes.

The weak point of ionic liquids is their high viscosity, limiting the maximum polymer concentration in the forming solutions. A low-viscous co-solvent can reduce viscosity, but cellulose has none. This study demonstrates that dimethyl sulfoxide (DMSO), being non-solvent for cellulose, can act as a nominal co-solvent to improve its processing into a nanofiltration membrane by phase inversion. A study of the rheology of cellulose solutions in diluted ionic liquids ([EMIM]Ac, [EMIM]Cl, and [BMIM]Ac) containing up to 75% DMSO showed the possibility of decreasing the viscosity by up to 50 times while keeping the same cellulose concentration. Surprisingly, typical cellulose non-solvents (water, methanol, ethanol, and isopropanol) behave similarly, reducing the viscosity at low doses but causing structuring of the cellulose solution and its phase separation at high concentrations. According to laser interferometry, the nature of these non-solvents affects the mass transfer direction relative to the forming membrane and the substance interdiffusion rate, which increases by four-fold when passing from isopropanol to methanol or water. Examination of the nanofiltration characteristics of the obtained membranes showed that the dilution of ionic liquid enhances the rejection without changing the permeability, while the transition to alcohols increases the permeability while maintaining the rejection.

Turn air-captured CO2 with methanol into amino acid and pyruvate in an ATP/NAD(P)H-free chemoenzymatic system.

The use of gaseous and air-captured CO2 for technical biosynthesis is highly desired, but elusive so far due to several obstacles including high energy (ATP, NADPH) demand, low thermodynamic driving force and limited biosynthesis rate. Here, we present an ATP and NAD(P)H-free chemoenzymatic system for amino acid and pyruvate biosynthesis by coupling methanol with CO2. It relies on a re-engineered glycine cleavage system with the NAD(P)H-dependent L protein replaced by biocompatible chemical reduction of protein H with dithiothreitol. The latter provides a higher thermodynamic driving force, determines the reaction direction, and avoids protein polymerization of the rate-limiting enzyme carboxylase. Engineering of H protein to effectively release the lipoamide arm from a protected state further enhanced the system performance, achieving the synthesis of glycine, serine and pyruvate at g/L level from methanol and air-captured CO2. This work opens up the door for biosynthesis of amino acids and derived products from air.

Methanol fixation is the method of choice for droplet-based single-cell transcriptomics of neural cells.

The main critical step in single-cell transcriptomics is sample preparation. Several methods have been developed to preserve cells after dissociation to uncouple sample handling from library preparation. Yet, the suitability of these methods depends on the cell types to be processed. In this project, we perform a systematic comparison of preservation methods for droplet-based single-cell RNA-seq on neural and glial cells derived from induced pluripotent stem cells. Our results show that while DMSO provides the highest cell quality in terms of RNA molecules and genes detected per cell, it strongly affects the cellular composition and induces the expression of stress and apoptosis genes. In contrast, methanol fixed samples display a cellular composition similar to fresh samples and provide a good cell quality and little expression biases. Taken together, our results show that methanol fixation is the method of choice for performing droplet-based single-cell transcriptomics experiments on neural cell populations.

Investigation of composition, antioxidant, antimicrobial and cytotoxic characteristics from Juniperus sabina and Ferula communis extracts.

Plants have been one the most valuable sources of biologically active compounds. This study investigates the chemical composition, as well as the antioxidant, antimicrobial, and cytotoxic activities of methanolic and ethanolic extracts from Juniperus sabina and Ferula communis leaves, grown in Cyprus. Total phenolic and flavonoids content of methanol and ethanol extracts were quantified. Chemical constituents of the leaf extracts were analysed using gas chromatography/mass spectrometry (GC/MS). Mome inositol was the predominant component in the J. Sabina's extracts. The most dominant component in F. communis ethanolic extract was phytol, while in FCL methanolic extract 1,3,4,5 tetrahydroxycyclohexanecarboxylic acid. Antioxidant activities were evaluated by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical-scavenging ability. Antioxidant activity results revealed concentration dependent activity for methanolic and ethanolic extracts from the plant leaves. Antibacterial activity of plant extracts was tested against Gram-negative and Gram-positive bacteria using disk diffusion and minimal inhibitory concentration methods. Cytotoxic activity of plant extracts were evaluated on MCF-7 and MDA-MB-231 breast cancer cell lines, where they demonstrated their potential on the viability of both cell lines. The biological activity revealed by plants is due to the bioactive compounds found in the extracts. These bioactive components could be used as anticancer drug candidates.

Comparative analysis of phenolic compositions and biological activities of three endemic Teucrium L. (Lamiaceae) species from Turkey.

This study investigates the chemical compositions and biological activities of the methanol extracts of three endemic Teucrium species (T. ekimii, T. pestalozzae and T. semrae) collected from Turkey. Total phenolic and flavonoid contents were assessed spectrophotometrically. The total phenolic and flavonoid content in the T. ekimii methanolic extract were importantly higher than other both extracts. The polyphenolic components of the extracts were identified by liquid chromatography. Seven phenolic compounds were identified namely catechin, rutin, luteolin, apigenin chlorogenic acid, sinapic acid and rosmarinic acid. Antioxidant activities were determined by five in vitro assays namely phosphomolybdenum assay, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, ß-carotene bleaching assay, ferric ions reducing antioxidant power (FRAP) and cupric ions reducing antioxidant capacity (CUPRAC). The total antioxidant activity method exhibited that T. ekimii methanol extract exerted better antioxidant activity. The methanol extract of T. ekimii showed better antiradical scavenging activity as measured by DPPH assay. The antimicrobial capacities were determined by agar diffusion assay. Three endemic Teucrium species tested showed slight antibacterial activity only against Aeromonas hydrophila, Klebsiella pneumoniae and Streptococcus pneumoniae. The findings showed that three endemic Teucrium species may be utilized as natural sources of antioxidant and antimicrobial compounds in food and farmacy products.

1H NMR fecal metabolic phenotyping of periductal fibrosis- and cholangiocarcinoma-specific metabotypes defining perturbation in gut microbial-host co-metabolism.

Background: The liver fluke Opisthorchis viverrini (OV), which subsequently inhabits the biliary system and results in periductal fibrosis (PDF), is one of the primarily causes of cholangiocarcinoma (CCA), a bile duct cancer with an exceptionally high incidence in the northeast of Thailand and other Greater Mekong Subregion (GMS) countries. Insights in fecal metabolic changes associated with PDF and CCA are required for further molecular research related to gut health and potential diagnostic biological marker development. Methods: In this study, nuclear magnetic resonance (NMR) metabolomics was applied for fecal metabolic phenotyping from 55 fecal water samples across different study groups including normal bile duct, PDF and CCA groups. Results: By using NMR spectroscopy-based metabolomics, fecal metabolic profiles of patients with CCA or PDF and of individuals with normal bile duct have been established with a total of 40 identified metabolites. Further multivariate statistical analysis and hierarchical clustering heat map have demonstrated the PDF- and CCA-specific metabotypes through various altered metabolite groups including amino acids, alcohols, amines, anaerobic glycolytic metabolites, fatty acids, microbial metabolites, sugar, TCA cycle intermediates, tryptophan catabolism substrates, and pyrimidine metabolites. Compared to the normal bile duct group, PDF individuals showed the significantly elevated relative concentrations of fecal ethanol, glycine, tyrosine, and N-acetylglucosamine whereas CCA patients exhibited the remarkable fecal metabolic changes that can be evident through the increased relative concentrations of fecal uracil, succinate, and 5-aminopentanoate. The prominent fecal metabolic alterations between CCA and PDF were displayed by the reduction of relative concentration of methanol observed in CCA. The metabolic alterations associated with PDF and CCA progression have been proposed with the involvement of various metabolic pathways including TCA cycle, ethanol biogenesis, hexamine pathway, methanol biogenesis, pyrimidine metabolism, and lysine metabolism. Among them, ethanol, methanol, and lysine metabolism strongly reflect the association of gut-microbial host metabolic crosstalk in PDF and/or CCA patients. Conclusion: The PDF- and CCA-associated metabotypes have been investigated displaying their distinct fecal metabolic patterns compared to that of normal bile duct group. Our study also demonstrated that the perturbation in co-metabolism of host and gut bacteria has been involved from the early step since OV infection to CCA tumorigenesis.

A UHPLC-MS/MS method for the simultaneous determination of vancomycin, norvancomycin, meropenem, and moxalactam in human plasma and its clinical application.

We developed an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to determine four antibacterial drugs in human plasma for clinical usage. Samples were prepared using protein precipitation with methanol. Chromatographic separation was accomplished in 4.5 min on a BEH C18 column (2.1 × 50 mm, 1.7 µm) using a gradient elution of methanol and water (containing 7.71 g/L concentrated ammonium acetate, adjusted to pH 6.5 with acetic acid) at a flow rate of 0.4 mL/min. Positive electrospray was used for ionization. The method was linear in the concentration range 1-100 µg/mL for vancomycin, norvoncomycin, and meropenem; and 0.5-50 µg/mL for R-isomer of moxalactam and S-isomer of moxalactam. For all analytes, the intra- and inter-day accuracies and precisions were -8.47%-10.13% and less than 12%, respectively. The internal standard normalized recoveries and matrix effect were 62.72%-105.78% and 96.67%-114.20%, respectively. All analytes were stable at six storage conditions, with variations of less than 15.0%. The method was applied in three patients with central nervous system infection. The validated method might be useful for routine therapeutic drug monitoring and pharmacokinetic study.

Comparison between 5 extractions methods in either plasma or serum to determine the optimal extraction and matrix combination for human metabolomics.

BACKGROUND: Although metabolomics continues to expand in many domains of research, methodological issues such as sample type, extraction and analytical protocols have not been standardized, impeding proper comparison between studies and future research. METHODS: In the present study, five solvent-based and solid-phase extraction methods were investigated in both plasma and serum. All these extracts were analyzed using four liquid chromatography coupled with high resolution mass spectrometry (LC-MS) protocols, either in reversed or normal-phase and with both types of ionization. The performances of each method were compared according to putative metabolite coverage, method repeatability and also extraction parameters such as overlap, linearity and matrix effect; in both untargeted (global) and targeted approaches using fifty standard spiked analytes. RESULTS: Our results verified the broad specificity and outstanding accuracy of solvent precipitation, namely methanol and methanol/acetonitrile. We also reveal high orthogonality between methanol-based methods and SPE, providing the possibility of increased metabolome coverage, however we highlight that such potential benefits must be weighed against time constrains, sample consumption and the risk of low reproducibility of SPE method. Furthermore, we highlighted the careful consideration about matrix choice. Plasma showed the most suitable in this metabolomics approach combined with methanol-based methods. CONCLUSIONS: Our work proposes to facilitate rational design of protocols towards standardization of these approaches to improve the impact of metabolomics research.

[An LC-MS/MS Analytical Method for Moenomycin A in Livestock Products].

A simple and sensitive method for the determination of moenomycin A residues in livestock products using LC-MS/MS was developed. Moenomycin A, a residual definition of flavophospholipol, was extracted from samples with a mixture of ammonium hydroxide and methanol (1 : 9, v/v) preheated at 50℃. The crude extracted solutions were evaporated and purified by liquid-liquid partitioning between a mixture of ammonium hydroxide, methanol and water (1 : 60 : 40, v/v/v) and ethyl acetate. The alkaline layer was taken, and cleaned up using a strong anion exchange (InertSep SAX) solid phase extraction cartridge. The LC separation was performed on an Inertsil C8 column with liner gradient elution using 0.3 vol% formic acid and acetonitrile containing 0.3 vol% formic acid. Moenomycin A was detected using tandem mass spectrometry with negative ion electrospray ionization. Recovery tests were conducted using three porcine samples (muscle, fat and liver) and chicken eggs. Samples were spiked with moenomycin A at 0.01 mg/kg and at the Japanese Maximum Residue Limits (MRLs) established for each sample. The trueness ranged from 79 to 93% and precision ranged from 0.5 to 2.8%. The limit of quantification (S/N≥10) of the developed method is 0.01 mg/kg. The developed method would thus be very useful for regulatory monitoring of flavophospholipol in livestock products.

Synthesis and characterization of magnetic bifunctional nano-catalyst for the production of biodiesel from Madhuca indica oil.

The reusable magnetic multimetal nano-catalyst (Fe3O4.Cs2O) was synthesized using co-precipitation and incipient wetness impregnation methods. It was used to esterify and transesterify Madhuca indica (M. indica) oil to produce biodiesel with methanol. The prepared catalyst, caesium oxide doped on the nano-magnetite core, was characterized using field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). Further, the activity of the catalyst was investigated by subjecting it to a biodiesel reaction. To maximize biodiesel conversion, studies were carried out by varying the process variables like catalyst concentration, methanol-to-oil molar ratio, reaction temperature, and reaction time. A maximum conversion of 97.4% was obtained at the holding conditions of 18:1 methanol-to-oil ratio, 7 wt% catalyst loading, 65 °C reaction temperature, and 300 min reaction time. Moreover, the catalyst recyclability study showed that it could be recycled up to 12 cycles with a conversion of 90% and above. The biodiesel's fuel properties were analysed and found to be within the limits of ASTM D6751 standard.

Understanding the Chemical Composition and Biological Activities of Different Extracts of Secamone afzelii Leaves: A Potential Source of Bioactive Compounds for the Food Industry.

Secamone afzelii (Roem. & Schult.) K. Schum (family Asclepiadaceae) is a creeping woody climber used to treat ailments in many traditional medicine systems. The present study aims to examine the antioxidant and enzyme inhibition activities of S. afzelii leaf using different compositions of methanol-water mixture as an extraction solvent. The extracts were characterized by HPLC-ESI-MSn in terms of chemical compounds. The in silico results show that compound 23 (quercitrin) has the higher docking scores among the selected substances and the MD simulation revealed that the interactions with the enzymatic pocket are stable over the simulation time and strongly involve the tyrosinase catalytic Cu atoms. All together the results showed that both 80% and 100% methanolic extracts contained significantly (p < 0.05) the highest total phenolics content while the highest content of total flavonoids was significantly (p < 0.05) extracted by 100% methanol. About 26 compounds were tentatively identified by HPLC-ESI-MSn and 6 of them were quantified using standards. Results showed that the extracts were rich in flavonoids with a relatively high abundance of two kaempferol glycosides comprising 60% of quantified compounds. The 100% and 80% methanol extracts recorded significantly (p < 0.05) the highest total antioxidant, DPPH and ABTS activity as well as tyrosinase and ⍺-amylase inhibitory activities. The best significant (p < 0.05) cholinesterase inhibitory activity and reducing capacity of Fe+++ and Cu++ was recorded from the 80% methanolic extract while 100% ethanolic extract gave the highest significant (p < 0.05) butyrylcholinesterase inhibitory activity. The best glucosidase activity was observed in the 50% and 80% methanolic extracts. Although the water extract displayed the least total phenolics and flavonoids content and consequently the lowest antioxidant and enzyme inhibition activity, it displayed significantly (p < 0.05) the highest chelating power. In conclusion, these results demonstrated the richness of S. afzelii leaf as a potential source of bioactive compounds for the food industry, for the preparation of food supplements and functional foods.

Microwave-Assisted Extraction Optimization and Effect of Drying Temperature on Catechins, Procyanidins and Theobromine in Cocoa Beans.

Cocoa beans (Theobroma cacao L.) are an important source of polyphenols. Nevertheless, the content of these compounds is influenced by post-harvest processes. In this sense, the concentration of polyphenols can decrease by more than 50% during drying. In this study, the process of procyanidins extraction was optimized and the stability of catechins, procyanidins, and theobromine to different drying temperatures was evaluated. First, the effectiveness of methanol, ethanol, acetone, and water as extract solvents was determined. A Box-Behnken design and response surface methodology were used to optimize the Microwave-Assisted Extraction (MAE) process. The ratios of methanol-water, time, and temperature of extraction were selected as independent variables, whereas the concentration of procyanidins was used as a response variable. Concerning the drying, the samples were dried using five temperatures, and a sample freeze-dried was used as a control. The quantitative analyses were carried out by HPLC-DAD-ESI-IT-MS. The optimal MAE conditions were 67 °C, 56 min, and 73% methanol. Regarding the drying, the maximum contents of procyanidins were obtained at 40 °C. To our knowledge, this is the first time that the stability of dimers, trimers, and tetramers of procyanidins on drying temperature was evaluated. In conclusion, drying at 40 °C presented better results than the freeze-drying method.

Chemical Characterisation, Antidiabetic, Antibacterial, and In Silico Studies for Different Extracts of Haloxylon stocksii (Boiss.) Benth: A Promising Halophyte.

The objective of the study is to evaluate the chemical characterisation, and biological and in silico potential of Haloxylon stocksii (Boiss.) Benth, an important halophyte commonly used in traditional medicine. The research focuses on the roots and aerial parts of the plant and extracts them using two solvents: methanol and dichloromethane. Chemical characterisation of the extracts was carried out using total phenolic contents quantification, GC-MS analysis, and LC-MS screening. The results exhibited that the aerial parts of the plant have significantly higher total phenolic content than the roots. The GC-MS and LC-MS analysis of the plant extracts revealed the identification of 18 bioactive compounds in each. The biological evaluation was performed using antioxidant, antibacterial, and in vitro antidiabetic assays. The results exhibited that the aerial parts of the plant have higher antioxidant and in vitro antidiabetic activity than the roots. Additionally, the aerial parts of the plant were most effective against Gram-positive bacteria. Molecular docking was done to evaluate the binding affinity (BA) of the bioactive compounds characterised by GC-MS with diabetic enzymes used in the in vitro assay. The results showed that the BA of γ-sitosterol was better than that of acarbose, which is used as a standard in the in vitro assay. Overall, this study suggests that the extract from aerial parts of H. stocksii using methanol as a solvent have better potential as a new medicinal plant and can provide a new aspect to develop more potent medications. The research findings contribute to the scientific data of the medicinal properties of Haloxylon stocksii and provide a basis for further evaluation of its potential as a natural remedy.

Ex Vivo Antiplatelet and Thrombolytic Activity of Bioactive Fractions from the New-Fangled Stem Buds of Ficus religiosa L. with Simultaneous GC-MS Examination.

Different parts of Ficus religiosa are the common components of various traditional formulations for the treatment of several blood disorders. The new-fangled stem buds' powder was extracted with 80% ethanol and successively fractionated by chloroform and methanol. Chloroform and methanol fractions of Ficus religiosa (CFFR and MFFR) were tested for antiplatelet, antithrombotic, thrombolytic, and antioxidant activity in ex vivo mode. The MFFR was particularly investigated for GC-MS and toxicity. The antiplatelet activity of the CFFR, MFFR, and standard drug aspirin at 50 µg/mL was 54.32%, 86.61%, and 87.57%, and a significant delay in clot formation was noted. CFFR at different concentrations did not show a significant effect on the delay of clot formation, antiplatelet, and free radical scavenging activity. The most possible marker compounds for antiplatelet and antioxidant activity identified by GC-MS in the MFFR are salicylate derivatives aromatic compounds such as benzeneacetaldehyde (7), phenylmalonic acid (13), and Salicylic acid (14), as well as Benzamides derivatives such as carbobenzyloxy-dl-norvaline (17), 3-acetoxy-2(1H)-pyridone (16), and 3-benzylhexahydropyrrolo [1,2-a] pyrazine-1,4-dione (35). A toxicity study of MFFR did not show any physical indications of toxicity and mortality up to 1500 mg/kg body weight and nontoxic up to 1000 mg/kg, which is promising for the treatment of atherothrombotic diseases.

Phytochemical Analysis and Profiling of Antioxidants and Anticancer Compounds from Tephrosia purpurea (L.) subsp. apollinea Family Fabaceae.

Tephorosia purpurea subsp. apollinea was extracted with methanol and n-hexane to obtain sub-fractions. The chemical compounds identified with GC-MS and HPLC in T. purpurea subsp. apollinea extracts showed antioxidant and anticancer properties. The antioxidant and anticancer activities were investigated using DDPH and ABTS assays, and MTT assay, respectively. Stigmasta-5,24(28)-dien-3-ol, (3 ß,24Z)-, 9,12,15-octadecatrienoic acid methyl ester, phytol, chlorogenic acid, and quercetin were the major chemical compounds detected in T. purpurea subsp. apollinea. These compounds possessed antioxidant and anticancer properties. The methanol extract showed antioxidant properties with DDPH and ABTS radical scavenging of 84% and 94%, respectively, relative to ascorbic acid and trolox. The anticancer effects of T. purpurea subsp. apollinea against the cancer cell lines MCF7 (IC50 = 102.8 ± 0.6 µg/mL), MG63 (IC50 = 118.3 ± 2.5 µg/mL), T47D (IC50 = 114.7 ± 1.0 µg/mL), HeLa (IC50 = 196.3 ± 2.3 µg/mL), and PC3 (IC50 = 117.7 ± 1.1 µg/mL) were greater than its anticancer effects against U379 (IC50 = 248.4 ± 7.5 µg/mL). However, it had no adverse effects on the normal cells (WI38) (IC50 = 242.9 ± 1.8 µg/mL). Therefore, the major active constituents presented in T. purpurea subsp. apollinea can be isolated and studied for their potential antioxidant and anticancer effects against breast, cervical, and prostate cancers and osteosarcoma.

Impact of instrumentation reliability on mainstream suspended partial denitrification-anammox (PdNA).

This study successfully revealed the importance of probe reliability and sensitivity with ion sensitive electrode (ISE) probes on achieving high partial denitrification (PdN) efficiency; and decreasing carbon overdosing events that cause the decline of microbial populations and performance of PdNA. In a mainstream integrated hybrid granule-floc system, an average PdN efficiency of 76% was achieved with acetate as the carbon source. Thauera was identified as the dominant PdN species; its presence in the system was analogous to instrumentation reliability and PdN selection and was not a consequence of bioaugmentation. Up to 27-121 mg total inorganic nitrogen/L/d, an equivalent of 18-48% of the overall total inorganic nitrogen removed, was achieved through the PdNA pathway. Candidatus Brocadia was the main anoxic ammonium oxidizing bacteria species that was seeded from sidestream and enriched and retained in the mainstream system with observed growth rates of 0.04-0.13 day-1 . Moreover, there was no direct negative impact of methanol's use for post-polishing on anoxic ammonium oxidizing bacteria activity and growth. PRACTITIONER POINTS: Stress testing with ISE sensors revealed the importance of probe reliability and sensitivity on PdN selection and PdNA performance. Up to 121 mg TIN/L/d was achieved via PdNA in a mainstream suspended hybrid granule-floc partial denitrification-anammox (PdNA) system. Candidatus Brocadia was the dominant AnAOB species with observed growth rates of 0.04-0.13 day-1. There was no direct negative impact of methanol's use for post-polishing on AnAOB activity and growth.

Dopant-enriched nitrogen gas to boost ionization of glycoproteins analyzed with native liquid chromatography coupled to nano-electrospray ionization.

Proteins carry a plethora of post-translational modifications (PTMs), such as glycosylation or phosphorylation, which may affect stability and activity. Analytical strategies are needed to investigate these PTMs in their native state to determine the link between structure and function. The coupling of native separation techniques with mass spectrometry (MS) has emerged as a powerful tool for in-depth protein characterization. Yet obtaining high ionization efficiency still can be challenging. Here, we explored the potential of dopant-enriched nitrogen (DEN) gas to improve nano-electrospray ionization (nano-ESI)-MS of native proteins after anion exchange chromatography. The dopant gas was enriched with different dopants (acetonitrile, methanol, and isopropanol) and the effects were compared with the use of solely nitrogen gas for six proteins covering a wide range of physicochemical properties. The use of DEN gas resulted generally in lower charge states, independent of the selected dopant. Moreover, less adduct formation was observed, particularly for the acetonitrile-enriched nitrogen gas. Importantly, striking differences in MS signal intensity and spectral quality were observed for extensively glycosylated proteins, where isopropanol- and methanol-enriched nitrogen appeared to be most beneficial. Altogether, the use of DEN gas improved nano-ESI of native glycoproteins and increased spectral quality for highly glycosylated proteins that normally suffer from low ionization efficiency.

Moorella caeni sp. nov., isolated from thermophilic anaerobic sludge from a methanol-fed reactor.

Strain AMPT has been previously suggested as a strain of the species Moorella thermoacetica Jiang et al. 2009 (based on the high 16S rRNA gene identity, 98.3 %). However, genome-based phylogenetic analysis of strain AMPT reveals that this bacterium is in fact a novel species of the genus Moorella. Genome relatedness indices between strain AMPT and Moorella thermoacetica DSM 521T were below the minimum threshold values required to consider them members of the same species (digital DNA-DNA hybridization, 52.2 % (<70%); average nucleotide identity, 93.2 % (<95%)). Based on phylogenetic and phenotypic results we recommend that strain AMPT (DSM 21394T=JCM 35360T) should be classified as representing new species, for which we propose the name Moorella caeni sp. nov.

Enantiomeric Separation and Degradation of Benoxacor Enantiomers in Horticultural Soil by Normal-Phase and Reversed-Phase High Performance Liquid Chromatography.

The separation of benoxacor enantiomers on six commercial chiral columns was investigated by high-performance liquid chromatography (HPLC) under normal-phase and reversed-phase conditions. The mobile phases included hexane/ethanol, hexane/isopropanol, acetonitrile/water, and methanol/water. The effects of the chiral stationary phases (CSPs), temperature, and mobile phase composition and ratio on the separation of benoxacor enantiomers were examined. Under normal-phase conditions, the two benoxacor enantiomers were completely separated on Chiralpak AD, Chiralpak IC, Lux Cellulose-1, and Lux Cellulose-3 columns and partially separated on a Lux Cellulose-2 column. Under reversed-phase conditions, benoxacor enantiomers were completely separated on a Lux Cellulose-3 column and partially separated on Chiralpak IC and Lux Cellulose-1 columns. Normal-phase HPLC performed better than reversed-phase HPLC for the separation of benoxacor enantiomers. As the column temperature increased from 10 °C to 4 °C, the enthalpy (ΔH) and entropy (ΔS) results indicated that the resolution was strongly affected by the temperature and that the lowest temperature did not always produce the best resolution. An optimized separation method on the Lux Cellulose-3 column was used to investigate the stability of benoxacor enantiomers in solvents and the degradation of benoxacor enantiomers in three types of horticultural soil. Benoxacor enantiomers were stable, and degradation or racemization were not observed in methanol, ethanol, isopropanol, acetonitrile, hexane, or water (pH = 4.0, 7.0, and 9.0). In three horticultural soils, the degradation rate of S-benoxacor was faster than that of R-benoxacor, resulting in soil enrichment with R-benoxacor. The results of this study will help to improve the risk assessment of enantiomer levels of benoxacor in the environment.