Experimental borosilicate bioactive glasses: Pulp cells cytocompatibility and mechanical characterization.

AIM: To assess in vitro the effect of two novel phase separated borosilicate glasses (PSBS) in the system SiO2 -B2 O3 -K2 O-CaO-Al2 O3 on dental pulp cells; and to compare their bioactivity and mechanical properties to a conventional fluoroaluminosilicate glass ionomer cement namely FUJI IX. METHODOLOGY: The cytocompatibility assessment of the two novel borosilicate glasses, one without alumina (PSBS8) and one containing alumina (PSBS16), was performed on cultured primary human pulp cells. Alamar blue assay was used to assess cell metabolic activity and cell morphology was evaluated by confocal imaging. The bioactivity in Stimulated Body Fluid was also evaluated after 1 and 3 weeks of immersion using SEM-EDX analysis. Vickers microhardness and flexural strength were assessed after incorporating the glass particles into a commercial glass ionomer cement (GIC) liquid containing both polyacrylic and polybasic carboxylic acid. RESULTS: The data revealed that the two borosilicate glasses enhanced cell viability ratios at all-time points in both direct and indirect contact assays. After 3 days of contact, PSBS8 without alumina showed higher viability rate (152%) compared to the PSBS16 containing alumina (145%) and the conventional glass ionomer particles (117%). EDX analysis confirmed an initial Ca/P ratio of 2.1 for 45S5K and 2.08 for PSBS8 without alumina after 3 weeks of immersion. The cement prepared using PSBS8 showed significantly higher Vickers hardness values (p = .001) than that prepared using PSBS16 (46.6 vs. 36.7 MPa). After 24 h of maturation, PSBS8 (without alumina) exhibited a flexural strength of 12.9 MPa compared to a value of 16.4 MPa for the commercial control. PSBS8 without alumina had a higher strength than PSBS16 with alumina, after 1 and 7 days of maturation (p = .001). CONCLUSIONS: The present in vitro results demonstrated that the borosilicate bioactive glass without alumina enhanced pulp cell viability, spreading and acellular bioactivity better than the conventional GIC and the experimental borosilicate glass containing alumina.

Synthesis, Characterization, Antibacterial Properties, and In Vitro Studies of Selenium and Strontium Co-Substituted Hydroxyapatite.

In this study, as a measure to enhance the antimicrobial activity of biomaterials, the selenium ions have been substituted into hydroxyapatite (HA) at different concentration levels. To balance the potential cytotoxic effects of selenite ions (SeO32-) in HA, strontium (Sr2+) was co-substituted at the same concentration. Selenium and strontium-substituted hydroxyapatites (Se-Sr-HA) at equal molar ratios of x Se/(Se + P) and x Sr/(Sr + Ca) at (x = 0, 0.01, 0.03, 0.05, 0.1, and 0.2) were synthesized via the wet precipitation route and sintered at 900 °C. The effect of the two-ion concentration on morphology, surface charge, composition, antibacterial ability, and cell viability were studied. X-ray diffraction verified the phase purity and confirmed the substitution of selenium and strontium ions. Acellular in vitro bioactivity tests revealed that Se-Sr-HA was highly bioactive compared to pure HA. Se-Sr-HA samples showed excellent antibacterial activity against both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus carnosus) bacterial strains. In vitro cell-material interaction, using human osteosarcoma cells MG-63 studied by WST-8 assay, showed that Se-HA has a cytotoxic effect; however, the co-substitution of strontium in Se-HA offsets the negative impact of selenium and enhanced the biological properties of HA. Hence, the prepared samples are a suitable choice for antibacterial coatings and bone filler applications.

The influence of experimental bioactive glasses on pulp cells behavior in vitro.

OBJECTIVES: To assess in vitro the effect of experimental mesoporous BAG, on human dental pulp cells (hDPCs) behavior in terms of cytocompatiblity and bioactivity via mineralization potential. METHODS: Fine (FP) and large particles (LP) of a fixed BAG composition named 0NaMBG have been elaborated by a sol-gel process. In vitro assessment was achieved on cultured primary hDPCs using indirect contact. The effect of the concentration of 800µg/mL on cell metabolic activity and cytotoxicity were examined by performing Alamar blue and crystal violet assays. Alizarin Red staining was used to detect and quantify the formation of mineralized nodules and ALP activity was colourimetrically quantified. The expression of Odontogenic markers: DMP-1 and osteopontin (OPN) expression and cell morphology was evaluated by confocal microscopy. RESULTS: According to the Alamar blue and crystal violet assay, 0NaMBG samples were non-cytotoxic. Cells treated with 0NaMBG particles expressed higher metabolic activity than control cells, especially for LP. Both FP and LP significantly increased both extra and intra cellular ALP activity. hDPCs exhibited good cell spreading and adhesion in the presence of FP and LP extracts by confocal imaging. Further, Alizarin red S assay demonstrated more mineralization nodules and significant enhancement of the extracellular calcium deposition when cells were interfaced with both FP and LP compared to the control cells. Moreover, LP extracts enhanced the production and secretion of odontogenic markers: dentin matrix protein 1 (DMP-1) and osteopontin. SIGNIFICANCE: LP have a higher surface area and pore volume, which could explain their greater bioactivity in contact with pulp cells. The clinical relevance of these findings implicate that 0NaMBG could be used as fillers in dental therapeutic materials suitable for dentin and/or pulp tissues preservation.

Correction: Ga and Ce ion-doped phosphate glass fibres with antibacterial properties and their composite for wound healing applications.

Correction for 'Ga and Ce ion-doped phosphate glass fibres with antibacterial properties and their composite for wound healing applications' by Agata Lapa et al., J. Mater. Chem. B, 2019, DOI: .

Ga and Ce ion-doped phosphate glass fibres with antibacterial properties and their composite for wound healing applications.

Novel gallium/cerium-doped phosphate glass fibres (PGF) were successfully manufactured by the melt-quenching and melt-spinning process. The amorphous character of the materials produced was confirmed using X-ray powder diffraction (XRD), and the elemental composition was investigated with X-ray fluorescence confirming the presence of 2 mol% of Ga2O3 or CeO2. Fourier Transform Infrared Spectroscopy (FTIR) confirmed the presence of Q1 and Q2 structural phosphate species. Mechanical properties of the PGFs revealed tensile strength values of 428 ± 94 MPa and 379 ± 80 MPa, with elastic modulus values of 45 ± 4 GPa and 54 ± 9 GPa for Ce-PGF (diameter 25 µm) and Ga-PGF (diameter 18 µm), respectively. The influence of both dopants on the glass degradation properties was evaluated by tests in deionised water, which revealed a decreased dissolution rate for gallium-doped PGF in comparison to cerium-doped PGF. Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) measurements were used to explore ion release in cell culture medium, while ICP-mass spectrometry (ICP-MS) was used to measure ion release in deionised water. These techniques showed controlled release of therapeutic and antibacterial ions from the PGF. Antibacterial properties of Ce-PGF and Ga-PGF, based on turbidity measurements, were confirmed against Gram-positive bacteria. Moreover, Ce-doped phosphate glass fibres did not disturb the proliferation of human epidermal keratinocyte (HaCaT) cells or the mobility of mice embryonic fibroblasts (MEF). Applying an in vitro scratch assay showed full wound closure after 24 h of indirect incubation with Ga-PGF. Due to their superior processability as compared with Ga-PGFs, a fully degradable mesh based on Ce-PGF was designed and found to achieve high water uptake (up to 800%), suggesting its suitability for wound healing applications.

Sol-gel bioglasses in dental and periodontal regeneration: A systematic review.

Due to their osteoconductive and osteoinductive abilities, bioglasses (BGs) have attracted attention in tissue engineering, especially for mineralized tissue. The aim of this study is to review the current state of the art on the effects of BGs produced by sol-gel on cells for dental and periodontal regeneration. The study also discusses associated antibacterial properties. The research was performed by considering the Preferred Reporting Items for Systematic Reviews and the Meta-Analyses (PRISMA) statement. The research ranged 5 years' window time (from January, 01, 2012, to August, 31, 2017) and the relevant studies were identified based on the inclusion/exclusion criteria. A total of 45 articles were selected from 244 initial returns, plus seven further articles coming from other sources were selected for the same purpose. From this systematic study, it is revealed that only 13 of the 52 articles have proved both the ability of BGs to differentiate dental cells at genetic level and their ability of triggering cell-mediated mineralization, but only six of them showed, along with cells, the antibacterial properties of the glasses. This review shows that sol-gel BGs are not toxic, can sustain cell proliferation and differentiation at a genetic level, and can keep the bacterial population under control. Moreover, a standard methodology and an ideal material are suggested. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1210-1227, 2019.

Bioactivity evaluation of collagen-based scaffolds containing a series of Sr-doped melt-quench derived phosphate-based glasses.

Phosphate-based glasses have been attracting attention due to their possible medical applications arising from unique dissolution characteristics in the human body leading to the possibility of new tissue regeneration. In this study, the leaching kinetics of a series of melt-quenched Sr-doped phosphate glasses are presented. Regardless of the presence of Sr, all the glasses have an initial linear and sustained release of the ions followed by a plateau. To guarantee proper nutritional support to the growing tissue during regeneration and to mimic the 3-dimensional architecture of tissues, organic scaffold systems have been developed. However, their poor mechanical strength has limited their application. To overcome this problem, cross-linkers can be used although this then limits the solubility of the materials. To succeed in dealing with such a limitation, in this paper, by freeze-drying, the aforementioned soluble melt-quenched phosphate glasses were combined as powders with collagen fibres from bovine achilles tendon to make degradable scaffolds. The scaffolds were characterized by SEM, EDX and BET. Changes to the dissolution behaviour of the glasses arising from the presence of collagen interacting with the ions leached were reported. Furthermore, the ability of the scaffolds to induce hydroxyapatite (HA) formation was evaluated: one the elaborated scaffold could grow an HA-like layer after a week in SBF. Based on the results obtained, a possible application in restorative dentistry is proposed for one or more materials.

The influence of precursor addition order on the porosity of sol-gel bioactive glasses.

OBJECTIVE: The superior textural properties of sol-gel derived bioactive glasses compared to conventional melt quench glasses accounts for their accelerated bioactivity in vitro. Several studies have explored ways to improve the surface properties of sol-gel glasses in order to maximise their efficiency for bone and tooth regeneration. In this study, we investigated the effect of order of network modifying precursor addition on the textural properties of sol-gel derived bioactive glasses. METHODS: The effect of precursor addition order on the glass characteristics was assessed by switching the order of network modifying precursor (calcium acetate monohydrate and sodium acetate anhydrous) addition for a fixed composition of bioactive glass (75SiO2:5CaO:10Na2O:10P2O5). RESULTS: The results of this study showed that the order of precursor addition does influence the porosity of these glasses. For the glasses of a fixed composition and preparation conditions we achieved a doubling of surface area, a 1.5 times increase in pore volume and a 1.2 times decrease in pore size just by the mixing the network modifying precursors and adding them together in the sol-gel preparation. SIGNIFICANCE: This simple and straightforward route adaptation to the preparation of bioactive glasses would allow us to enhance the textural properties of existing and novel composition of bioactive glasses and thus accelerate their bioactivity.

Alcoholic liver disease on the intensive care unit - Outcomes and prognostication.

BACKGROUND: Hospital admissions with decompensated chronic alcoholic liver disease have been increasing, leading to increased pressure on intensive care unit services. We aimed to determine the outcome and prognostic factors for patients with alcoholic liver disease requiring admission to intensive care unit. METHODS: This was a retrospective study over 5 years (January 2006-December 2010) of all intensive care unit admissions with alcoholic liver disease to either of the two Leeds Teaching Hospitals NHS Trust general intensive care units. A detailed case note review was conducted based on a pre-established proforma. Eighty-two patients included. Primary outcome was hospital mortality. RESULTS: The overall intensive care unit and hospital mortality were 46% and 67%, respectively. Hospital mortality in patients successfully discharged from intensive care unit with the intent of recovery remained high at 21%. Variceal bleed was the only indicator that had a mortality <60%. Factors which suggested a poor outcome included sepsis (86% mortality) and hepato-renal syndrome (86% mortality). A Sequential Organ Failure Assessment score of greater than 10 on intensive care unit admission was associated with 97% hospital mortality. Sequential Organ Failure Assessment score increased from a mean of 10.9-12.5 in those that did not survive hospital. Patients with first alcoholic liver disease related admission had poorer outcomes. CONCLUSION: These results are similar to previous studies with no significant improvement in outcomes. Alcoholic liver disease is not a contra-indication to intensive care unit admission but assessment of the individual patient is required. The most appropriate objective factors to guide prognostication are the presenting intensive care unit diagnosis and Sequential Organ Failure Assessment score. First presentation of alcoholic liver disease is not a positive prognostic indicator.

Dirigent proteins in plants: modulating cell wall metabolism during abiotic and biotic stress exposure.

Dirigent (DIR) proteins were found to mediate regio- and stereoselectivity of bimolecular phenoxy radical coupling during lignan biosynthesis. Here we summarize the current knowledge of the importance of DIR proteins in lignan and lignin biosynthesis and highlight their possible importance in plant development. We focus on the still rather enigmatic Arabidopsis DIR gene family, discussing the few members with known functional importance. We comment on recent discoveries describing the detailed structure of two DIR proteins with implications in the mechanism of DIR-mediated catalysis. Further, we summarize the ample evidence for stress-induced dirigent gene expression, suggesting the role of DIRs in adaptive responses. In the second part of our work, we present a preliminary bioinformatics-based characterization of the AtDIR family. The phylogenetic analysis of AtDIRs complemented by comparison with DIR proteins of mostly known function from other species allowed us to suggest possible roles for several members of this family and identify interesting AtDIR targets for further study. Finally, based on the available metadata and our in silico analysis of AtDIR promoters, we hypothesize about the existence of specific transcriptional controls for individual AtDIR genes and implicate them in various stress responses, hormonal regulations, and developmental processes.

Bioactive glass for dentin remineralization: A systematic review.

BACKGROUND AND OBJECTIVES: Strategies to achieve dentin remineralization is at present an important target of restorative dentistry. Remineralization of dentin by a bioactive material is complete only when the tissue regains its functionality. This is achieved when there is adequate apatite formation which most importantly translates into improved mechanical properties of dentin as a result of intrafibrillar mineralization. Bioactive glass (BAG) is a well-known implant material for bone regeneration and is proven to have excellent ability of apatite formation. Hence, recent studies have proposed BAGs as one of the most desired materials for remineralization of dentin. Therefore the aim of this systematic review was to scope the evidence of bioactive glass to remineralize dentin. METHODS: The following research question was formulated: "Is there strong evidence for bioactive glass to remineralize dentin?" Three databases (Web of science, PubMed and Science direct) were scanned independently following PRISMA guidelines. Inclusion and exclusion criteria were set to identify relevant articles based on title and abstract screening. Finally, potentially relevant articles were downloaded and the full text was scrutinized to select the articles included in this review. RESULTS: The first phase of search returned 303 articles. A total of 19 papers with full text were scrutinized for inclusion, of which 3 papers were chosen for the final synthesis. All three studies confirm that BAG treatment leads to enhanced apatite formation in dentin. Only 1 of the 3 studies has reported the mechanical properties of dentin after BAG treatment and it revealed that the Young's modulus and flexural bend strength of BAG treated dentin were much lower than natural dentin even though they had similar apatite content. CONCLUSIONS: This review highlights the importance of assessing the mechanical properties of dentin alongside to the newly formed apatite content in order to prove BAGs efficiency to remineralize this tissue. Though studies have confirmed that BAGs stimulate excellent apatite formation in dentin, it should be concluded that there isn't sufficient evidence for bioactive glass to effectively remineralize this tissue as the mechanical properties of the BAG treated dentin haven't been well explored.

Hormonal regulation of secondary cell wall formation.

Secondary cell walls (SCWs) have critical functional importance but also constitute a high proportion of the plant biomass and have high application potential. This is true mainly for the lignocellulosic constituents of the SCWs in xylem vessels and fibres, which form a structured layer between the plasma membrane and the primary cell wall (PCW). Specific patterning of the SCW thickenings contributes to the mechanical properties of the different xylem cell types, providing the plant with mechanical support and facilitating the transport of solutes via vessels. In the last decade, our knowledge of the basic molecular mechanisms controlling SCW formation has increased substantially. Several members of the multi-layered regulatory cascade participating in the initiation and transcriptional regulation of SCW formation have been described, and the first cellular components determining the pattern of SCW at the subcellular resolution are being uncovered. The essential regulatory role of phytohormones in xylem development is well known and the molecular mechanisms that link hormonal signals to SCW formation are emerging. Here, we review recent knowledge about the role of individual plant hormones and hormonal crosstalk in the control over the regulatory cascades guiding SCW formation and patterning. Based on the analogy between many of the mechanisms operating during PCW and SCW formation, recently identified mechanisms underlying the hormonal control of PCW remodelling are discussed as potentially novel mechanisms mediating hormonal regulatory inputs in SCW formation.

The multifaceted dissociation chemistry of anionic aggregates containing functionalised amines and CO2.

Specific products and their relative abundances observed in gas-phase collision-induced dissociation experiments for [amine-H + CO2 + H2O](-) reflect the behaviour of the bulk aqueous amines in the presence of CO2.

2-Carboxy-D-arabinitol 1-phosphate (CA1P) phosphatase: evidence for a wider role in plant Rubisco regulation.

The genes for CA1Pase (2-carboxy-D-arabinitol-1-bisphosphate phosphatase) from French bean, wheat, Arabidopsis and tobacco were identified and cloned. The deduced protein sequence included an N-terminal motif identical with the PGM (phosphoglycerate mutase) active site sequence [LIVM]-x-R-H-G-[EQ]-x-x-[WN]. The corresponding gene from wheat coded for an enzyme with the properties published for CA1Pase. The expressed protein lacked PGM activity but rapidly dephosphorylated 2,3-DPG (2,3-diphosphoglycerate) to 2-phosphoglycerate. DTT (dithiothreitol) activation and GSSG inactivation of this enzyme was pH-sensitive, the greatest difference being apparent at pH 8. The presence of the expressed protein during in vitro measurement of Rubisco (ribulose-1,5-bisphosphate carboxylase/oxygenase) activity prevented a progressive decline in Rubisco turnover. This was due to the removal of an inhibitory bisphosphate that was present in the RuBP (ribulose-1,5-bisphosphate) preparation, and was found to be PDBP (D-glycero-2,3-pentodiulose-1,5-bisphosphate). The substrate specificity of the expressed protein indicates a role for CA1Pase in the removal of 'misfire' products of Rubisco.

Tandem mass spectrometry measurement of the collision products of carbamate anions derived from CO2 capture sorbents: paving the way for accurate quantitation.

The reaction between CO(2) and aqueous amines to produce a charged carbamate product plays a crucial role in post-combustion capture chemistry when primary and secondary amines are used. In this paper, we report the low energy negative-ion CID results for several anionic carbamates derived from primary and secondary amines commonly used as post-combustion capture solvents. The study was performed using the modern equivalent of a triple quadrupole instrument equipped with a T-wave collision cell. Deuterium labeling of 2-aminoethanol (1,1,2,2,-d(4)-2-aminoethanol) and computations at the M06-2X/6-311++G(d,p) level were used to confirm the identity of the fragmentation products for 2-hydroxyethylcarbamate (derived from 2-aminoethanol), in particular the ions CN(-), NCO(-) and facile neutral losses of CO(2) and water; there is precedent for the latter in condensed phase isocyanate chemistry. The fragmentations of 2-hydroxyethylcarbamate were generalized for carbamate anions derived from other capture amines, including ethylenediamine, diethanolamine, and piperazine. We also report unequivocal evidence for the existence of carbamate anions derived from sterically hindered amines (Tris(2-hydroxymethyl)aminomethane and 2-methyl-2-aminopropanol). For the suite of carbamates investigated, diagnostic losses include the decarboxylation product (-CO(2), 44 mass units), loss of 46 mass units and the fragments NCO(-) (m/z 42) and CN(-) (m/z 26). We also report low energy CID results for the dicarbamate dianion ((-)O(2)CNHC(2)H(4)NHCO(2)(-)) commonly encountered in CO(2) capture solution utilizing ethylenediamine. Finally, we demonstrate a promising ion chromatography-MS based procedure for the separation and quantitation of aqueous anionic carbamates, which is based on the reported CID findings. The availability of accurate quantitation methods for ionic CO(2) capture products could lead to dynamic operational tuning of CO(2) capture-plants and, thus, cost-savings via real-time manipulation of solvent regeneration energies.

Extensin network formation in Vitis vinifera callus cells is an essential and causal event in rapid and H(2)O(2)-induced reduction in primary cell wall hydration.

BACKGROUND: Extensin deposition is considered important for the correct assembly and biophysical properties of primary cell walls, with consequences to plant resistance to pathogens, tissue morphology, cell adhesion and extension growth. However, evidence for a direct and causal role for the extensin network formation in changes to cell wall properties has been lacking. RESULTS: Hydrogen peroxide treatment of grapevine (Vitis vinifera cv. Touriga) callus cell walls was seen to induce a marked reduction in their hydration and thickness. An analysis of matrix proteins demonstrated this occurs with the insolubilisation of an abundant protein, GvP1, which displays a primary structure and post-translational modifications typical of dicotyledon extensins. The hydration of callus cell walls free from saline-soluble proteins did not change in response to H(2)O(2), but fully regained this capacity after addition of extensin-rich saline extracts. To assay the specific contribution of GvP1 cross-linking and other wall matrix proteins to the reduction in hydration, GvP1 levels in cell walls were manipulated in vitro by binding selected fractions of extracellular proteins and their effect on wall hydration during H(2)O(2) incubation assayed. CONCLUSIONS: This approach allowed us to conclude that a peroxidase-mediated formation of a covalently linked network of GvP1 is essential and causal in the reduction of grapevine callus wall hydration in response to H(2)O(2). Importantly, this approach also indicated that extensin network effects on hydration was only partially irreversible and remained sensitive to changes in matrix charge. We discuss this mechanism and the importance of these changes to primary wall properties in the light of extensin distribution in dicotyledons.

Association of hemicellulose- and pectin-modifying gene expression with Eucalyptus globulus secondary growth.

Wood properties are ultimately related to the morphology and biophysical properties of the xylem cell wall. Although the cellulose and lignin biosynthetic pathways have been extensively studied, modifications of other wall matrix components during secondary growth have attracted relatively less attention. In this work, thirty-eight new Eucalyptus cDNAs encoding cell wall-modifying proteins from nine candidate families that act on the cellulose-hemicellulose and pectin networks were cloned and their gene expression was investigated throughout the developing stem. Semi-quantitative RT-PCR revealed distinct, gene-specific transcription patterns for each clone, allowing the identification of genes up-regulated in xylem or phloem of stem regions undergoing secondary growth. Some genes, namely an endo-1,4-beta-glucanase, one mannan-hydrolase and three pectin methylesterases showed transcription in juvenile and also in mature stages of wood development. The patterns of gene expression using samples from tension and opposite wood disclosed a general trend for up-regulation in tension wood and/or down-regulation in opposite wood. Localised gene expression of two selected representative clones, EGl-XTH1 and EGl-XTH4, obtained through in situ hybridization confirms the RT-PCR results and association with secondary xylem formation. Likewise, immunolocalisation studies with the anti-pectin antibody (JIM5) also supported the idea that the development of tissue-specific pectin characteristics is important during secondary growth. These results emphasize an involvement of hemicellulose and pectin biochemistry in wood formation, suggesting that the controlled and localised modification of these polysaccharides may define cell properties and architecture and thus, contribute to determining different biophysical characteristics of Eucalyptus wood.

A proteomic analysis of the wound response in Medicago leaves reveals the early activation of a ROS-sensitive signal pathway.

Wounded Medicago truncatula leaves produce a burst of O(2)(-) (phase I) between 1 and 15 min, then of O(2)(-) and H(2)O(2) (phase II) between 1 and 3 h. Our previous results suggest reactive oxygen species (ROS) may provide signals to mobilise early (6 h), apoplastic, wound-responsive proteins (WRPs). 2DE and MALDI-TOF/TOF were used to analyse how the suppression of ROS production at different time points by diphenyleneiodonium (DPI), affects the expression of WRPs. Rapid (≤3 min) DPI inhibition of phase I O(2)(-) production suppressed the differential regulation of 7 out of 19 WRPs, which were consequently classified as ROS-dependent WRPs. DPI inhibition of only phase II ROS production failed to suppress the wound regulation of 18 out of 19 WRPs, but led to the altered expression of 1 ROS-dependent WRP and 2 non-WRPs (Group B). The data indicates Group B proteins are alternatively targeted via the modulation of phase II ROS production. This reinforces an important role for phase I O(2)(-) signalling in the early wound response, but indicates that this response is partly regulated by phase II of the oxidative burst. This data provides an informed basis for further proteomic studies aimed at identifying early activated O(2)(-) signalling components in wounded Medicago.

N-Nitrosopiperazines form at high pH in post-combustion capture solutions containing piperazine: a low-energy collisional behaviour study.

During the process of exploring aqueous piperazine chemistry under simulated flue-gas scrubbing conditions, positive-ion electrospray ionisation mass spectrometric (ESI-MS) analyses of the resulting reaction mixtures in a triple quadrupole system revealed the presence of peaks at m/z 116 and 145, the putative N-nitroso derivatives of piperazine. Confirmation of the presence of these species in the reaction mixtures was achieved using collision-induced dissociation experiments. A purchased standard, together with in-house synthesised N-nitrosopiperazine standards (including N-nitroso derivatives derived from deuterium-labelled precursor materials), were used for this purpose. Across a small range of collision energies, large fluctuations in the abundance of the two major product ions of protonated N-nitrosopiperazine, m/z 86 and 85, were observed. Using B3LYP/6-311 + +G(d,p) computations, the potential energy surface was determined for loss of NO and [H,N,O]. At an activation energy slightly in excess of 1 eV, intramolecular isomerisation precedes loss of NO (m/z 86) via a 4,1 H-shift, and at activation energies between 2.1-2.3 eV, consecutive loss of NO and atomic hydrogen competes with the direct loss of nitrosyl hydride (m/z 85). It is recommended that any multiple reaction monitoring method for quantifying N-nitrosopiperazines at low collision energies use the sum of both transitions (m/z 116 ← 85, m/z 116 ← 86) to avoid errors that could be introduced by subtle changes in ES source conditions or collision voltages. This approach is adopted in an HPLC/MS/MS method used to monitor the degradation of N-nitrosopiperazine exposed to (i) broad-band UV light and (ii) heat typical of an amine regeneration (stripper) tower. The results reveal that aqueous N-nitrosopiperazine is thermally stable at 150°C but will degrade slowly upon exposure to UV light.