A green reaction-based turn-off fluorescence sensor for determination of copper ions: DFT calculations, quenching mechanism, green chemistry metrics, and application in environmental samples.

When Cu(II) reacts with ascorbic acid (AA) to form Cu(I), Cu(I) can combine with eosin Y (EY) to form ionic associations, resulting in significant fluorescence quenching of the EY. Based on the turn-off of fluorescence in the chemosensor EY, a green reaction is proposed herein for the detection of Cu(II). The novel detection method for Cu(II) demonstrates simplicity, high sensitivity, and excellent selectivity, rendering it suitable for analyzing environmental samples. A static fluorescence quenching mechanism is validated through the Stern-Volmer relationship, and the thermodynamic parameters of the reaction are explored using a van 't Hoff plot. The reaction mechanism is investigated via fluorescence spectra, absorption spectra, and density-functional theory (DFT) calculations. The probe's green nature is confirmed by applying four green analytical chemistry metrics.

Fine sediment effects on seagrasses: A global review, quantitative synthesis and multi-stressor model.

This review collates research into fine sediment as a stressor of seagrass and emphasizes the multiple modes of action of this contaminant. The article is based on a bibliographic database search that identified 201 articles describing sediment impacts on seagrasses. Articles were classified by one of three non-exclusive modes of action: 1) light reduction; 2) smothering (burial), and 3) effects via rhizosphere physico-chemistry. Most citations (104) investigated multi-mode impacts of sediments, but the most frequently investigated single mode was light reduction (57 citations), followed by substrate rhizosphere chemistry (31) then smothering effects (6). Mud with high organic content is particularly problematic and smaller seagrasses are particularly vulnerable. Research gaps include polyphasic approaches, and studies of interactions between smothering, rhizosphere biogeochemistry and light climate. Identifying the thresholds of seagrass health indicators under mud stress should benefit coastal resource management, enabling improved decision-making and implementation of protective actions.

Nectary photosynthesis contributes to the production of manuka (Leptospermum scoparium) floral nectar.

Current models of floral nectar production do not include a contribution from photosynthesis by green nectary tissue, even though many species have green nectaries. Manuka (Leptospermum scoparium) floral nectaries are green, and in addition to sugars, their nectar contains dihydroxyacetone (DHA), the precursor of the antimicrobial agent in the honey. We investigated causes of variation in manuka floral nectar production, particularly the effect of light incident on the nectary. Flower gas exchange, chlorophyll fluorescence, and the effects on nectar of age, temperature, light, sucrose, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), pyridoxal phosphate, and 13 CO2 , were measured for attached and excised flowers. Flower age affected all nectar traits, whilst temperature affected total nectar sugar only. Increased light reduced floral CO2 efflux, increased nectar sugar production, and affected the ratio of DHA to other nectar sugars. DCMU, an inhibitor of photosystem II, reduced nectar sugar production. Pyridoxal phosphate, an inhibitor of the chloroplast envelope triose phosphate transporter, reduced nectar DHA content. Incubation of excised flowers with 13 CO2 in the light resulted in enrichment of nectar sugars, including DHA. Photosynthesis within green nectaries contributes to nectar sugars and influences nectar composition. Manuka nectar DHA arises from pools of triose phosphate that are modulated by nectary photosynthesis.

Composition and potential as a prebiotic functional food of a Giant Willow Aphid (Tuberolachnus salignus) honeydew honey produced in New Zealand.

The Giant Willow Aphid (Tuberolachnus salignus, GWA) is an invasive pest insect in New Zealand, which excretes honeydew. European honeybees collect this honeydew and make it into a type of honey that crystallizes in the comb, representing a significant loss to apiarists. This crystallization has been ascribed to high concentrations of oligosaccharides, particularly melezitose. In this research, the first carbohydrate profile of GWA honeydew honey, a sample of GWA honeydew honey was found to contain 37.8% total oligosaccharides of which 27.4% was melezitose, and 2.5% gluconic acid (higher than typical honeydew honeys); 41.2% monosaccharides (lower than typical honeydew honeys); and 0.054% salicylic acid (higher than previous estimates). Melezitose extracted from GWA honeydew honey was not significantly hydrolyzed in crude human-stomach and human-small-intestine simulations and may therefore meet the prebiotic criterion of human indigestibility.

Nanostructure of Gasification Charcoal (Biochar).

In this work, we investigate the molecular composition and nanostructure of gasification charcoal (biochar) by comparing it with heat-treated fullerene arc-soot. Using ultrahigh resolution Fourier transform ion-cyclotron resonance and laser desorption ionization time-of-flight mass spectrometry, Raman spectroscopy, and high resolution transmission electron microscopy we analyzed charcoal of low tar content obtained from gasification. Mass spectrometry revealed no magic number fullerenes such as C60 or C70 in the charcoal. The positive molecular ion m/ z 701, previously considered a graphitic part of the nanostructure, was found to be a breakdown product of pyrolysis and not part of the nanostructure. A higher mass distribution of ions similar to that found in thermally treated fullerene soot indicates that they share a nanostructure. Recent insights into the formation of all carbon fullerenes reveal that conditions in charcoal formation are not optimal for the formation of fullerenes, but instead, curved carbon structures coalesce into fulleroid-like structures. Microscopy and spectroscopy support such a stacked, fulleroid-like nanostructure, which was explored using reactive molecular dynamics simulations.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka honey: Part V - The rate determining step.

Monomer formation from dimeric DHA has previously been suggested as the rate-determining step in formation of methylglyoxal, the bioactive component in manuka honey. This step was studied by 1H NMR in DMSO­d6. First order reaction rate was 3.31 × 10-3 ±â€¯9.1 × 10-4 min-1. Upon titration with D2O, little change was observed until ∼15 mass% whereupon an exponential increase in rate occurred until indistinguishable from the rate observed in water. Acid or base caused rate accelerations. Theoretical modelling confirmed the existence of acid and base-catalysed mechanisms for dimer decomposition and the structures of two intermediates observed. In honey it is likely the base-catalysed decomposition predominates with water as catalyst but there is little rate acceleration at the levels of water present normally in honey however a small increase in the mass% of water in the honey could cause significant rate acceleration of dimer decomposition and hence formation of methylglyoxal.

Determination of Menaquinone-7 by a Simplified Reversed Phase- HPLC Method.

BACKGROUND: An efficient and accurate HPLC method was developed for the determination of menaquinone-7 (MK-7) in microbial fermentation using 2-propanol and n-hexane as extraction solvents as well as the eluent. METHODS: Extraction was carried out with 2-propanol and n-hexane (2:1, v/v) after enzymatic hydrolysis with 1% (w/v) lipase and ethanol water treatment prior to quantification in order to remove interfering lipids and denatured proteins. Chromatographic separation of MK-7 was accomplished isocratically on a C 18 Gemini column using a mobile phase mixture of 2- propanol: n-hexane (2:1, v/v) with a flow rate of 0.5 mL/min. UV detection was carried out from 200-400 nm and the chromatogram was extracted at a wavelength of 248 nm. A linear response was shown by the method with a coefficient of determination (R2) value of 0.9982. RESULTS: The recoveries of MK-7 were greater than 94% and the intra and inter day R.S.D values were less than 2%, demonstrating the accuracy of the method. The lower limit of detection (LOD) and the limit of quantification (LOQ) were 0.1 µg/mL and 0.29 µg/mL, respectively. CONCLUSION: The general usefulness of the described method is demonstrated by the application of this method in the analysis of MK-7 from Bacillus species. Under these conditions, the analysis of MK-7 was achieved in less than 8 minutes with a retention time of 7.19 ± 0.1 minutes.

High Level of Menaquinone-7 Production by Milking Menaquinone-7 with Biocompatible Organic Solvents.

BACKGROUND: The effect of milking menaquinone-7 (MK-7) with biocompatible organic solvents on MK-7 production in B. subtilis fermentation was studied. Periodic milking of MK-7 from the fermentation medium by n-hexane significantly enhanced the total MK-7 production ~ 1.7 fold at the end of 72 hours of fermentation (p < 0.05) as compared to the control medium. METHODS: Milking of MK-7 with a mixture of n-hexane phase modified with n-butanol was also explored. Although milking of MK-7 by a mixture of n-hexane and n-butanol (1:2, v/v), was found to be appropriate in terms of high extraction capacity, no significant increase in total MK-7 concentration was observed. Biocompatibility between the extraction solvents and B. subtilis was also examined. RESULTS AND CONCLUSION: The results indicated that the mixture of n-hexane and n-butanol exhibited some detrimental effects. However, n-hexane alone exhibited delayed toxicity starting at 84 hours of periodic milking and could, therefore, be considered as the most promising organic solvent for milking MK-7 in B. subtilis fermentation while enhancing the productivity of the system.

Influence of genotype, floral stage, and water stress on floral nectar yield and composition of manuka (Leptospermum scoparium).

Background and Aims: Floral nectar can be variable in composition, influencing pollinator behaviour and the composition of honey derived from it. The non-peroxide antibacterial activity of manuka (Leptospermum scoparium, Myrtaceae) honey results from the chemical conversion of the triose sugar dihydroxyacetone (DHA), after DHA accumulates for an unknown reason in the nectar. This study examined variation in nectar DHA, glucose, fructose and sucrose content with floral stage of development, between manuka genotypes with differing flower morphology, and in response to water stress. Methods: Six manuka genotypes were grown without nectar-feeding insects. Stages of flower development were defined, nectar was harvested and its composition was compared between stages and genotypes, and with floral morphology. Water stress was imposed and its effect on nectar composition was examined. Key Results: Nectar was present from soon after flower opening until the end of petal abscission, with the quantity of accumulated nectar sugars rising, then stabilizing or falling, indicating nectar secretion followed by reabsorption in some genotypes. The quantity of DHA, the ratio of DHA to other nectar sugars and the fructose to glucose ratio also varied with stage of development, indicating differences in rates of production and reabsorption between nectar components. Nectar composition and yield per flower also differed between genotypes, although neither was positively related to nectary area or stomatal density. Drying soil had no effect on nectar composition or yield, but variation in nectar yield was correlated with temperature prior to nectar sampling. Conclusions: Manuka nectar yield and composition are strongly influenced by plant genotype, flower age and the environment. There were clear stoichiometric relationships between glucose, fructose and sucrose per flower, but DHA per flower was only weakly correlated with the amount of other sugars, suggesting that accumulation of the triose sugar is indirectly coupled to secretion of the larger sugars by the nectary parenchyma.

Magnetic immobilization of bacteria using iron oxide nanoparticles.

Bacterial cell immobilization is a novel technique used in many areas of biosciences and biotechnology. Iron oxide nanoparticles have attracted much attention in bacterial cell immobilization due to their unique properties such as superparamagnetism, large surface area to volume ratio, biocompatibility and easy separation methodology. Adhesion is the basis behind many immobilization techniques and various types of interactions determine bacterial adhesion. Efficiency of bacterial cell immobilization using iron oxide nanoparticles (IONs) generally depends on the physicochemical properties of the IONs and surface properties of bacterial cells as well as environmental/culture conditions. Bacteria exhibit various metabolic responses upon interaction with IONs, and the potential applications of iron oxide nanoparticles in bacterial cell immobilization will be discussed in this work.

Impact of 3-Aminopropyltriethoxysilane-Coated Iron Oxide Nanoparticles on Menaquinone-7 Production Using B. subtilis.

One of the major issues associated with industrial production of menaquinone-7 (MK-7) is the low fermentation yield. In this study, we investigated the effect of iron oxide nanoparticles coated with 3-aminopropyltriethoxysilane (IONs@APTES) on the production of MK-7 using B. subtilis (ATCC 6633). Decoration of B. subtilis cells with IONs@APTES significantly enhanced both MK-7 production and yield. An approximately two-fold increase in MK-7 production (41 mg/L) was observed in the presence of 500 µg/mL IONs@APTES, as compared to MK-7 production using untreated bacteria (22 mg/L). This paper, therefore, illustrates the immense biotechnological potential of IONs@APTES in increasing MK-7 concentration using B. subtilis, and its future role in bioprocess engineering.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka honey: Part IV - Formation of HMF.

During a study of the conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) in maturing New Zealand manuka honey, the kinetics of formation of 5-(hydroxymethyl)furfural (HMF) was studied at temperatures from 4 to 37°C. Formation of HMF was first-order during an induction period and zero-order thereafter indicating that the mechanism includes the formation of certain critical intermediates and that these require time to build up; the duration of the induction period depended primarily upon temperature. The zero-order rate constant at 37°C was the same for manuka honey and clover honey doped with 2000 or 10,000mg/kg DHA and for artificial honey with 2000mg/kg of DHA and either alanine or proline and alanine added. Zero-order rate constants for artificial honey with added amino acids were less than for a control without amino acids. A simulation was created to predict the formation of HMF over time at 37°C in manuka honey.

Iron oxide nanoparticles in modern microbiology and biotechnology.

Iron oxide nanoparticles (IONs) are one of the most developed and used nanomaterials in biotechnology and microbiology. These particles have unique physicochemical properties, which make them unique among nanomaterials. Therefore, many experiments have been conducted to develop facile synthesis methods for these particles and to make them biocompatible. Various effects of IONs on microorganisms have been reported. Depending on the microbial strain and nanoparticle (NP) concentration, IONs can stimulate or inhibit microbial growth. Due to the superparamagnetic properties of IONs, these NPs have used as nano sources of heat for hyperthermia in infected tissues. Antibiotic-loaded IONs are used for targeted delivery of chemical therapy direct to the infected organ and IONs have been used as a dirigible carrier for more potent antimicrobial nanomaterials such as silver nanoparticles. Magnetic NPs have been used for specific separation of pathogen and non-pathogen bacterial strains. Very recently, IONs were used as a novel tool for magnetic immobilization of microbial cells and process intensification in a biotechnological process. This review provides an overview of application of IONs in different microbial processes. Recommendations are also given for areas of future research.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka honey: Part I--Honey systems.

The kinetics of conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) were investigated in manuka honeys and DHA-doped clover honeys stored between 4 and 37°C. Both the disappearance of DHA and appearance of MGO were confirmed as overall, first order reactions, albeit probably composites of multiple reactions. Increasing the storage temperature accelerated the rate of DHA loss and the initial rate of formation of MGO, but better conversion efficiency was observed at lower temperature. At 37°C, more MGO was lost at later times in manuka honey compared to DHA-doped-clover honey. Thirty-seven New Zealand manuka honeys and four clover honeys were analysed for various chemical and physical properties; comparison of rate constants and these parameters identified some positive correlations.

Kinetics of the conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka honey: Part II--Model systems.

The irreversible dehydration reaction of dihydroxyacetone (DHA) to methylglyoxal (MGO) in a honey model system has been examined to investigate the influence of added perturbant species on the reaction rate. The secondary amino acid proline, primary amino acids (alanine, lysine and serine), and iron, or combinations of these perturbants, were added to artificial honey with either DHA or MGO and stored at 20, 27 and 37°C. These systems were monitored over time. A 1:1 conversion of DHA to MGO was not observed in any system studied, including the control system with no added perturbants. Addition of proline to the matrix increased consumption of DHA but did not produce any more MGO than the control sample. Lysine and serine behaved similarly. Alanine enhanced the conversion of DHA to MGO and had the best efficiency of conversion of DHA to MGO for the amino acids studied. An iron II salt enhanced the conversion of DHA to MGO, even in the presence of proline.

Kinetics of conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka honey: Part III--A model to simulate the conversion.

A kinetic model for the conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) in honey is proposed; a building block approach was used to create the model. Artificial honeys doped with DHA and individual perturbants were fitted first, then multiple perturbants (alanine, proline and iron, and combinations of these) were fitted before comparing the simulation to real honey samples (doped clover and manuka honey). The main responses in the prediction model were DHA, MGO, proline, primary amino acids, acidity, 3-phenyllactic acid and 4-methoxyphenyllactic acid. Three temperatures (20, 27 and 37°C) were studied and the conversion of DHA to MGO was monitored over at least 1year. Differences in the conversion between clover doped with DHA and manuka honey were observed. The simulation fitted well for the honeys tested.

Isolation of maltol glucoside from the floral nectar of New Zealand manuka (Leptospermum scoparium).

Maltol glucoside (3-(ß-D-glucopyranosyloxy)-2-methyl-4H-pyran-4-one), 1, was isolated from a preparation of the floral nectar from the New Zealand manuka tree (Leptospermum scoparium). 1 eluted just after dihydroxyacetone in HPLC of underivatized nectar and showed a UV absorbance maximum of 258 nm. The structure of 1 was confirmed by NMR and high resolution mass spectrometry.

Regional, annual, and individual variations in the dihydroxyacetone content of the nectar of maÌ nuka (Leptospermum scoparium) in New Zealand.

A method was designed and validated for the analysis of dihydroxyacetone in the floral nectar of ma̅nuka (Leptospermum scoparium). The method was applied to samples collected from different regions of the North Island and the Nelson region of the upper South Island of New Zealand during the period 2009-2012 as well as to nectar samples from some Australian Leptospermum species. The ratio of dihydroxyacetone to total sugar (DHA/Tsugar) was classified as low (<0.001 mg/mg), moderate (0.001-0.002 mg/mg), or high (>0.002 mg/mg). Inter- and intraregional variation were observed as well as interannual variation with variation from low to high classification occurring within one region and from low to moderate between years. Australian species also demonstrated elevated levels of dihydroxyacetone in the nectar. Some garden cultivars were shown to produce very high nectar DHA/Tsugar, and a survey of cultivars was undertaken; cultivars with single-flowered red or pink flowers were the most common producers of very high nectar DHA/Tsugar.

Effect of high pressure processing on the conversion of dihydroxyacetone to methylglyoxal in New Zealand manuka (Leptospermum scoparium) honey and models thereof.

The effect of high pressure processing (HPP) on the conversion of dihydroxyacetone (DHA) to methylglyoxal (MGO) was examined in New Zealand manuka honey and models thereof. The objective was to confirm that previously reported increases of MGO with HPP treatment originated from conversion of DHA. RP-HPLC was used to quantify DHA, MGO and hydroxymethylfurfural (HMF) after derivatisation with O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) or (in the case of MGO) separately with o-phenylenediamine (OPD). Fresh and stored manuka honey, clover honey with DHA added and artificial 26 honey with DHA added were subjected to nine different pressures and holding times and compared to untreated samples. There was no consistent trend of decrease in DHA or increase in MGO for any of the samples with any treatment. Samples showed random change generally within 5-10% of an untreated sample for MGO, DHA and HMF. HPP does not accelerate the conversion of DHA to MGO in honey.

The unique manuka effect: why New Zealand manuka honey fails the AOAC 998.12 C-4 sugar method.

Conversion of dihydroxyacteone (DHA) to methylglyoxal (MGO) has been shown to be the key mechanism for the growth in "apparent" C-4 sugar content in nonperoxide activity (NPA) manuka honey. This reaction is enhanced by heating and storage time and is demonstrated for the first time in clover honey adulterated with DHA purchased from a chemical supplier and in manuka honey containing naturally occurring DHA and MGO. After heating at 37 °C for 83 days, pure clover honey with no added DHA has the same apparent C-4 sugar content as at t = 0 days. The same clover honey adulterated with synthetic DHA added at t = 0 days and heated at 37 °C over the same time scale shows a change in apparent C-4 sugars from 2.8 to 5.0%. Four NPA manuka honeys heated over longer periods show an increase in apparent C-4 sugars of up to 280% after 241 days. This study strongly suggests that a protein fractionation effect occurs in the conversion of DHA to MGO in higher NPA manuka honey, rendering the remaining δ(13)C protein value more negative and falsely indicating C-4 sugar addition when using the AOAC 998.12 method.