Absence Of Acetate Research Articles

Gut microbiota-derived acetate attenuates lung injury induced by influenza infection via protecting airway tight junctions

BackgroundGut microbiota (GM) have been implicated as important regulators of gastrointestinal symptom which is commonly occurred along with respiratory influenza A virus (IAV) infection, suggesting the involvement of the gut-to-lung axis in a host’s response to IAV. IAV primarily destroys airway epithelium tight junctions (TJs) and consequently causes acute respiratory disease syndrome. It is known that GM and their metabolism produce an anti-influenza effect, but their role in IAV-induced airway epithelial integrity remains unknown.MethodsA mouse model of IAV infection was established. GM were analyzed using 16S rRNA gene sequencing, and short-chain fatty acids (SCFAs) levels were measured. GM depletion and fecal microbiota transplantation (FMT) were conducted to validate the role of GM in IAV infection. A pair-feeding experiment was conducted to reveal whether IAV-induced GM dysbiosis is attributed to impaired food intake. Furthermore, human bronchial epithelial (HBE) cells were cocultured with IAV in the presence or absence of acetate. TJs function was analyzed by paracellular permeability and transepithelial electronic resistance (TEER). The mechanism of how acetate affects TJs integrity was evaluated in HBE cells transfected with G protein-coupled receptor 43 (GPR43) short hairpin RNA (shRNA).ResultsIAV-infected mice exhibited lower relative abundance of acetate-producing bacteria (Bacteroides, Bifidobacterium, and Akkermansia) and decreased acetate levels in gut and serum. These changes were partly caused by a decrease in food consumption (due to anorexia). GM depletion exacerbated and FMT restored IAV-induced lung inflammatory injury. IAV infection suppressed expressions of TJs (occludin, ZO-1) leading to disrupted airway epithelial barrier function as evidenced by decreased TEER and increased permeability. Acetate pretreatment activated GPR43, partially restored IAV-induced airway epithelial barrier function, and reduced inflammatory cytokines levels (TNF-α, IL-6, and IL-1β). Such protective effects of acetate were absent in HBE cells transfected with GPR43 shRNA. Acetate and GPR43 improved TJs in an AMP-activated protein kinase (AMPK)-dependent manner.ConclusionCollectively, our results demonstrated that GM protected airway TJs by modulating GPR43-AMPK signaling in IAV-induced lung injury. Therefore, improving GM dysbiosis may be a potential therapeutic target for patients with IAV infection.

Co-cultivation of Saccharomyces cerevisiae strains combines advantages of different metabolic engineering strategies for improved ethanol yield

Glycerol is the major organic byproduct of industrial ethanol production with the yeast Saccharomyces cerevisiae. Improved ethanol yields have been achieved with engineered S. cerevisiae strains in which heterologous pathways replace glycerol formation as the predominant mechanism for anaerobic re-oxidation of surplus NADH generated in biosynthetic reactions. Functional expression of heterologous phosphoribulokinase (PRK) and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) genes enables yeast cells to couple a net oxidation of NADH to the conversion of glucose to ethanol. In another strategy, NADH-dependent reduction of exogenous acetate to ethanol is enabled by introduction of a heterologous acetylating acetaldehyde dehydrogenase (A-ALD). This study explores potential advantages of co-cultivating engineered PRK-RuBisCO-based and A-ALD-based strains in anaerobic bioreactor batch cultures. Co-cultivation of these strains, which in monocultures showed reduced glycerol yields and improved ethanol yields, strongly reduced the formation of acetaldehyde and acetate, two byproducts that were formed in anaerobic monocultures of a PRK-RuBisCO-based strain. In addition, co-cultures on medium with low acetate-to-glucose ratios that mimicked those in industrial feedstocks completely removed acetate from the medium. Kinetics of co-cultivation processes and glycerol production could be optimized by tuning the relative inoculum sizes of the two strains. Co-cultivation of a PRK-RuBisCO strain with a Δgpd1 Δgpd2 A-ALD strain, which was unable to grow in the absence of acetate and evolved for faster anaerobic growth in acetate-supplemented batch cultures, further reduced glycerol formation but led to extended fermentation times. These results demonstrate the potential of using defined consortia of engineered S. cerevisiae strains for high-yield, minimal-waste ethanol production.

The lifetime of the oxygen-evolving complex subunit PSBO depends on light intensity and carbon availability in Chlamydomonas.

PSBO is essential for the assembly of the oxygen-evolving complex in plants and green algae. Despite its importance, we lack essential information on its lifetime and how it depends on the environmental conditions. We have generated nitrate-inducible PSBO amiRNA lines in the green alga Chlamydomonas reinhardtii. Transgenic strains grew normally under non-inducing conditions, and their photosynthetic performance was comparable to the control strain. Upon induction of the PSBO amiRNA constructs, cell division halted. In acetate-containing medium, cellular PSBO protein levels decreased by 60% within 24 h in the dark, by 75% in moderate light, and in high light, the protein completely degraded. Consequently, the photosynthetic apparatus became strongly damaged, probably due to 'donor-side-induced photoinhibition', and cellular ultrastructure was also severely affected. However, in the absence of acetate during induction, PSBO was remarkably stable at all light intensities and less substantial changes occurred in photosynthesis. Our results demonstrate that the lifetime of PSBO strongly depends on the light intensity and carbon availability, and thus, on the metabolic status of the cells. We also confirm that PSBO is required for photosystem II stability in C. reinhardtii and demonstrate that its specific loss also entails substantial changes in cell morphology and cell cycle.

Acetate improves catalytic performance for rapid removal of Cr(VI) by sodium borohydride in aqueous environments

An efficient method of remediating Cr(VI) involves the conversion of Cr(VI) to Cr(III) using sodium borohydride (NaBH4). However, information regarding the role of organic substances in Cr(VI) remediation is limited. As such, in this study, we aimed to evaluate the role of common low-molecular-weight acetate in the rapid chemical reduction of Cr(VI) by NaBH4 and subsequent total chromium (TCr) removal. The results showed that acetate promoted the transformation of high-concentration Cr(VI) to Cr(III) by NaBH4. The presence of 0–4 mM acetate did not affect the subsequent precipitation of Cr(III). In the presence of 4 mM acetate, Cr(VI) conversion was 99.03%, and subsequent TCr removal was 95.87% within 15 min without adding precipitant. The addition of acetate reduced the requirement for a low initial pH to achieve high Cr(VI) reduction efficiency. In particular, in the presence of 10 mM acetate, the conversion of 100 mg/L Cr(VI) by NaBH4 at an initial pH of 4 was 99.26% compared with 36.89% in the absence of acetate. Regarding the promotion mechanism of acetate for the reduction of Cr(VI) by NaBH4, we did not observe direct electron transfer from acetate to Cr(VI) at the origin of the reduction, but the reduction of Cr(VI) was promoted by inhibiting the production of OH– through buffering. Cr in HCrO4- was the favorable site for nucleophilic reaction, which could be attacked by H- from BH4-. According to the results of qualitative and semiquantitative XPS, Fourier transform infrared spectroscopy, and elemental analyses of sediments, a small amount of Cr(VI) and approximately 3% of acetate was removed through precipitation adsorption by the precipitated Cr(III). This novel acetate–Cr(VI)–NaBH4 remediation technology, which is simple and easy to use, improves the removal of high-concentration Cr(VI) from wastewater.

Enhanced Acetate Tolerance and Recombinant Protein Accumulation in Escherichia coli by Transgenic Expression of a Heat Shock Protein from Carrot (Daucus carota L.).

In Escherichia coli (E. coli) culture, acetate accumulates as an undesirable by-product of aerobic fermentation on glucose and inhibits cell growth and recombinant protein production. We examined whether the heterologous expression of a eukaryotic heat shock protein (Hsp) can confer tolerance to acetate in E. coli. Transgenic cell lines (TCLs) heterologously expressing a small heat shock protein (sHsp) from carrot (Daucus carota L.), DcHsp17.7, were exposed to heat, sodium acetate, and alkaline conditions. The cell growth and cell viability were examined by measuring O.D.600 and colony-forming units (CFU), respectively. The His-tagged recombinant alcohol dehydrogenase (ADH) gene cloned in a pET11a expression vector was introduced into TCL1 and expressed by isopropyl β-D-1-thiogalactopyranoside treatment. After purifying using Ni-NTA affinity chromatography, its accumulation levels were examined using SDS-PAGE in the presence of acetate. TCLs constitutively expressing DcHsp17.7 showed improved growth, cell density, and cell viability under the stress conditions of heat, acetate, and alkaline compared to an empty vector control line. In acetate stress conditions, TCL1 accumulated more cellular proteins (approximately 130%) than the control. The recombinant ADH accumulated to a higher level in TCL1 (2.2-fold at 16 °C) than the control. The addition of acetate reduced the recombinant ADH level by 70% in the control when compared with the absence of acetate. In contrast, recombinant ADH accumulation was not affected by acetate in TCL1. In the presence of acetate, TCL1 accumulated 6.4-fold more recombinant ADH than did the control. Furthermore, recombinant ADH produced in TCL1 showed 1.5-fold higher enzyme activity than that produced in the control in the presence or absence of acetate. Our study showed that heterologously expressed DcHsp17.7 from carrot can alleviate the negative effects of acetate on E. coli.

Acetate Activates Lactobacillus Bacteriocin Synthesis by Controlling Quorum Sensing.

Bacteriocins are useful for controlling the composition of microorganisms in fermented food. Bacteriocin synthesis is regulated by quorum sensing mediated by autoinducing peptides. In addition, short-chain fatty acids, especially acetic acid, reportedly regulate bacteriocin synthesis. Five histidine kinases that regulated the synthesis of bacteriocins were selected to verify their interactions with acetate. Acetate activated the kinase activity of PlnB, SppK, and HpK3 in vitro and increased the yield of their cognate bacteriocins plantaricin EF, sakacin A, and rhamnosin B in vivo. The antimicrobial activity against Staphylococcus aureus of the fermentation supernatants of Lactobacillus plantarum, Lactobacillus sakei, and Lactobacillus rhamnosus with addition of acetate increased to 298%, 198%, and 289%, respectively, compared with that in the absence of acetate. Our study elucidated the activation activity of acetate in bacteriocin synthesis, and it might provide a potential strategy to increase the production of bacteriocin produced by Lactobacillus. IMPORTANCE Bacteriocins produced by lactic acid bacteria (LAB) are particularly useful in food preservation and food safety. Bacteriocins might increase bacterial competitive advantage against the indigenous microbiota of the intestines; at the same time, bacteriocins could limit the growth of undesired microorganisms in yogurt and other dairy products. This study confirmed that three kinds of histidine kinases were activated by acetate and upregulated bacteriocin synthesis both in vitro and in vivo. The increasing yield of bacteriocins reduced the number of pathogens and increased the number of probiotics in milk. Bacteriocin synthesis activation by acetate may have a broad application in the preservation of dairy products and forage silage.

Water-soluble bitumen degradation products can fuel nitrate reduction from non-radioactive bituminized waste

An important fraction of the currently stored volume of long-lived intermediate level radioactive waste in Belgium is immobilized as Eurobitum. This type of waste typically contains large amounts of NaNO3 homogeneously dispersed in a hard bituminous matrix. Geological disposal of Eurobitum in a water-saturated sedimentary formation such as Boom Clay will result in the leaching of high concentrations of NaNO3 in the Boom Clay formation. This could cause a geochemical perturbation of the surrounding clay, possibly affecting some of the favorable characteristics of the host formation such as its hydraulic conductivity, sorption potential or the redox conditions. The latter might result in a decrease of its reducing capacity and an increase in the mobility of redox-sensitive radionuclides. Microbial nitrate reduction is a well-known process, which could be stimulated by the chemical and radiolytical water-soluble organic bitumen degradation products. The present study carried out different series of long-term anoxic batch experiments to investigate the ability of the microbial community of Boom Clay borehole water to reduce nitrate, leaching from thermally aged non-radioactive Eurobitum in the presence or absence of acetate, formate and oxalate, being part of the most important organic bitumen degradation products. Obtained results indicate that all three organic bitumen degradation products can be used as electron donor to fuel microbial nitrate reduction, albeit with a different efficiency. The highest nitrate reduction rate was observed in the presence of acetate, while oxalate was the least efficient electron donor for nitrate reduction. Moreover, organic compounds that leached from the Eurobitum during the course of the experiment were used as electron donor for microbial nitrate reduction in all conditions. Furthermore, calcium oxalate crystals were observed, indicating that if oxalate is present, it will probably be less bioavailable compared to other organic compounds.

Propionate, together with triple antibiotics, inhibits the growth of Enterococci.

Enterococci are Gram-positive facultative anaerobic bacteria that colonize the oral cavity and gastrointestinal tract. Enterococcal infections, mainly caused by Enterococcus faecalis and Enterococcus faecium, include apical periodontitis, endocarditis, and bloodstream infections. Recently, vancomycinresistant Enterococci are considered major pathogens that are common but difficult to treat, especially in nosocomial settings. Moreover, E. faecalis is closely associated with recurrent endodontic infections and failed endodontic treatment. In this study, we investigated the effects of short-chain fatty acids (SCFAs), acetate, propionate, and butyrate, which are metabolites fermented by gut microbiota, on the growth of Enterococci. Enterococci were cultured in the presence or absence of acetate, propionate, or butyrate, and the optical density at 600 nm was measured to determine bacterial growth. The minimum inhibitory concentration/minimum bactericidal concentration test was conducted. Bacteria were treated with a SCFA, together with clinically used endodontic treatment methods such as triple antibiotics (metronidazole, minocycline, and ciprofloxacin) and chlorhexidine gluconate (CHX) to determine the effects of combination treatment. Of the SCFAs, propionate had a bacteriostatic effect, inhibiting the growth of E. faecalis in a dose-dependent manner and also that of clinical strains of E. faecalis isolated from dental plaques. Meanwhile, acetate and butyrate had minimal effects on E. faecalis growth. Moreover, propionate inhibited the growth of other Enterococci including E. faecium. In addition, combination treatment of propionate and triple antibiotics led to further growth inhibition, whereas no cooperative effect was observed at propionate plus CHX. These results indicate that propionate attenuates the growth of Enterococci, suggesting propionate as a potential agent to control Enterococcal infections, especially when combined with triple antibiotics.

Direct measurement of dissolved dinitrogen to refine reactive modelling of denitrification in agricultural soils

Nitrogen fertilizers used in agriculture often cause nitrate leaching towards shallow groundwater, especially in lowland areas where soil permeability, ploughing, clay content, and the flat topography minimizes surface runoff. The introduction of good agricultural practices to reduce the nitrate amount entering the groundwater system is crucial to ameliorate the kinetic control on nitrate denitrification capacity. With this aim, a series of anaerobic mesocosms, consisting of loamy and clay soils and nitrate rich water, were modelled using acetate and natural organic matter as electron donors. Acetate was chosen because it is the main intermediate in many biodegradation pathways of organic compounds, and hence it is a suitable carbon source for denitrification. To account for the spatial variability of soil parameters, the experiments were performed in triplicates. The geochemical code PHREEQC(3) was used to simulate kinetic denitrification, and equilibrium reactions of gas and mineral phases. The reactive modelling results highlighted a rapid acetate and nitrate degradation rate, a rapid production of dissolved inorganic carbon and dinitrogen, and a steady concentration of dissolved iron and sulphate, suggesting that the main pathway of nitrate attenuation is through denitrification; concomitantly excluding the occurrence of other processes leading to nitrate consumption. In the absence of acetate, the loamy soil, poor of natural organic matter, did not allow to complete the denitrification process.This modelling study investigates in detail the relationship between the denitrification process in natural soils, with excess and in limitation of organic substrates, and the occurrence and fate of dissolved dinitrogen analysed with a high precision membrane inlet mass spectrometer. Results demonstrate that modelling nitrate degradation processes as a whole, using geochemical datasets and codes, will improve the estimates of agricultural landscapes denitrification and support better nitrogen management, especially in lowland environments.

Degradation of 2,4,6-trichlorophenol and determination of bacterial community structure by micro-electrical stimulation with or without external organic carbon source

This study aimed to investigate the degradation efficiency of 2,4,6-trichlorophenol through a batch of potentiostatic experiments (0.2 V vs. Ag/AgCl). Efficiencies in the presence and absence of acetate and glucose were compared through open-circuit reference experiments. Significant differences in degradation efficiency were observed in six reactors. The highest and lowest degradation efficiencies were observed in the closed-circuit reactor fed with glucose and in the open-circuit reactor, respectively. This finding was due to the enhanced bacterial metabolism caused by the application of micro-electrical field and degradable organics as co-substrates. The different treatment efficiencies were also caused by the distinct bacterial communities. The composition of bacterial community was affected by adding different organics as co-substrates. At the phylum level, the most dominant bacteria in the reactor with the added acetate and glucose were Proteobacteria and Firmicutes, respectively.

Synthesis, Structures, and Reactivity of Single and Double Cyclometalated Complexes Formed by Reactions of [Cp*MCl2]2 (M = Ir and Rh) with Dinaphthyl Phosphines.

Reactions of two dinaphthyl phosphines with [Cp*IrCl2]2 have been carried out. In the case of di(α-naphthyl)phenylphosphine (1a), a simple P-coordinated neutral adduct 2a is obtained. However, tert-butyldi(α-naphthyl)phenylphosphine (1b) is cyclometalated to form [Cp*IrCl(P^C)] (3b). Complexes 2a and 3a undergo further cyclometalation to give the corresponding double cyclometalated complexes [Cp*Ir(C^P^C)] (4a,b) upon heating. In the presence of sodium acetate, reactions of 1a,b with [Cp*IrCl2]2 directly afford the final double cyclometalated complexes (4a,b). In the absence of acetate, [Cp*RhCl2]2 shows no reaction with 1a,b, whereas with acetate the reactions form the corresponding single cyclometalated complexes [Cp*RhCl(P^C)] (5a,b), which react with tBuOK to form the corresponding rhodium hydride complexes (6a,b). Treatment of 4a with CuCl2 or I2 leads to opening of two Ir–C σ bonds to yield the corresponding P-coordinated iridium dihalide (7 or 8) by means of an intramolecular C–C coupling reaction. A new chiral phosphine (11) is formed by the ligand-exchange reaction of 8 with PMe3. Reactions of the single cycloiridated complex 3b with terminal aromatic alkynes result in the corresponding five- and six-membered doubly cycloiridated complex 12 and/or η2-alkene coordinated complexes 13–15; the latter discloses that the electronic effect of terminal alkynes affects the regioselectivity. While the single cyclorhodated complex 5b reacts with terminal aromatic alkynes to form the corresponding six-membered cyclometalated complexes 16a–c by vinylidene rearrangement/1,1-insertion. Plausible pathways for formation of insertion products 13–16 were proposed. Molecular structures of twelve new complexes were determined by X-ray diffraction.

Novel quantitative insights into carbon sources for synthesis of poly hydroxybutyrate in Synechocystis PCC 6803.

Many freshwater cyanobacteria accumulate polyhydroxybutyrate (PHB) under nitrogen or phosphorus deprivation. While prior literature has shed lights on transcriptomic and metabolomic changes in the model cyanobacterium Synechocystis PCC 6803 cells, the quantitative contributions of the newly fixed carbon following nitrogen deprivation or the externally added acetate to PHB synthesis are not clear. Similarly, it is not clear how photomixotrophy affects precursor contributions. In this study, we show that (i) the pre-growth mode (photoautotrophic or photomixotrophic), while significantly impacting glycogen levels, does not have any significant effect on PHB levels, (ii) the carbon fixed following nitrogen deprivation contributes 26% of C for PHB synthesis in photoautotrophically pre-grown cells and its contribution to the PHB synthesis goes down with the addition of acetate at the resuspension phase or with photomixotrophic pre-growth, (iii) the acetate added at the start of nitrogen deprivation, doubles the intracellular PHB levels and contributes 44-48% to PHB synthesis and this value is not greatly affected by how the cells were pre-grown. Indirectly, the labeling studies also show that the intracellular C recycling is the most important source of precursors for PHB synthesis, contributing about 74-87% of the C for PHB synthesis in the absence of acetate. The addition of acetate significantly reduces its contribution. In photoautotrophic pre-growth followed by acetate addition under nitrogen starvation, the contribution of intracellular C reduces to about 34%. Thus, our study provides several novel quantitative insights on how prior nutritional status affects the precursor contributions for PHB synthesis.

When Glass and Metal Corrode Together, V: Sodium Copper Formate

ABSTRACTA sodium copper formate, first described in 2002, can also form on copper alloys with sodium originating from soda glass hydrolysis. The frequent occurrence (50% of 250 cases of glass-induced metal corrosion) is due to the presence of formaldehyde emissions in storage and its direct reaction to formate in the alkaline surface films. The compound can be produced without the presence of acetate when chalconatronite or metal coupons immersed in soda solution are exposed to formaldehyde vapours. The crystal structure, derived from X-ray powder diffraction, yielded the sum formula Cu4Na4O(HCOO)8(OH)2·4H2O. Except for the absence of acetate in the structure, this is in good agreement with the 2002 publication. To assess which kind of combined glass/metal objects are affected, a number of large museum collections were surveyed. Sodium copper formate was detected as a corrosion product using mainly micro-Raman spectroscopy, for instance, on 18 painted Limoges enamels, eight glass vessels with metal mountings, glass beads on metal wire from 11 bridal crowns, nine Christmas tree balls with wire decoration, 40 pieces of jewellery with glass gems, three daguerreotypes, and one miniature with cover glasses.

Autocatalytic and Non-Catalytic Oxidation of Manganese(II) on a Platinum Electrode in Acetic Buffer

Abstract The influence of acetate on Mn2+ oxidation by a platinum electrode has been investigated in aqueous solution with cyclic voltammetry. It is found that acetate at concentrations below 0.5 M gives rise to autocatalytic oxidation of Mn2+ with MnO2 serving as the catalyst, while in 0.5–1 M acetate (as well as in the absence of acetate) Mn2+ is oxidized non-catalytically. The autocatalytic oxidation is revealed by a separate voltammetric wave which has a much sharper rise and a lower potential compared to non-catalytic waves, by a sigmoid kinetics during chronoamperometry and by an enhanced Mn2+ uptake with electrodeposited MnO2. The increase in temperature and Mn2+ concentration promotes autocatalysis while acidification and the increase in sweep rate inhibits it that is thought to be due to the acceleration or delay of MnO2 formation from Mn3+. It is suggested that acetate acts in two ways: (1) thermodynamically stabilizes Mn3+ by complexation to enable the autocatalytic reaction Mn2++MnO2+OAc−+H+↔Mn3+−OAc+ MnIIIOOH and (2) delays the formation of MnO2 (presumably on the stage of Mn(III) crystallization) which leads to the inhibition of autocatalysis at high acetate concentrations.

PSR1 Is a Global Transcriptional Regulator of Phosphorus Deficiency Responses and Carbon Storage Metabolism in Chlamydomonas reinhardtii.

Many eukaryotic microalgae modify their metabolism in response to nutrient stresses such as phosphorus (P) starvation, which substantially induces storage metabolite biosynthesis, but the genetic mechanisms regulating this response are poorly understood. Here, we show that P starvation-induced lipid and starch accumulation is inhibited in a Chlamydomonas reinhardtii mutant lacking the transcription factor Pi Starvation Response1 (PSR1). Transcriptomic analysis identified specific metabolism transcripts that are induced by P starvation but misregulated in the psr1 mutant. These include transcripts for starch and triacylglycerol synthesis but also transcripts for photosynthesis-, redox-, and stress signaling-related proteins. To further examine the role of PSR1 in regulating lipid and starch metabolism, PSR1 complementation lines in the psr1 strain and PSR1 overexpression lines in a cell wall-deficient strain were generated. PSR1 expression in the psr1 lines was shown to be functional due to rescue of the psr1 phenotype. PSR1 overexpression lines exhibited increased starch content and number of starch granules per cell, which correlated with a higher expression of specific starch metabolism genes but reduced neutral lipid content. Furthermore, this phenotype was consistent in the presence and absence of acetate. Together, these results identify a key transcriptional regulator in global metabolism and demonstrate transcriptional engineering in microalgae to modulate starch biosynthesis.

EXTRACTION OF NI AND CO VIA D2EHPA IN THE PRESENCE OF ACETATE ION

The effect of acetate ion introduced to aqueous solutions was investigated for the extraction/separation of Ni and Co from a dilute sulfate media. The separation of Co and Ni was improved by introducing acetate to aqueous media. By the use of the slope analysis method, the stoichiometric coefficient of the extractant was found to be 4 for cobalt and 3.5 or 4 for nickel depending on the acetate concentration. Fourier Transform Infrared Spectroscopy (FT-IR) was utilized to examine the probablemetal-acetate-D2EHPA complexes. Increasing the level of acetate, up to 0.4 M showed a relatively significant effect on the extraction curve of Ni, but there was no influence in the Co extraction curve. At the acetate level of 0.6 M, the pH difference at 70 extraction percentage reached 0.55 in compared to 0.2in the absence of acetate.

Genuine open form of the pentameric ligand-gated ion channel GLIC.

Pentameric ligand-gated ion channels (pLGICs) mediate fast chemical neurotransmission of nerve signalling in the central and peripheral nervous systems. GLIC is a bacterial homologue of eukaryotic pLGIC, the X-ray structure of which has been determined in three different conformations. GLIC is thus widely used as a model to study the activation and the allosteric transition of this family of receptors. The recently solved high-resolution structure of GLIC (2.4 Å resolution) in the active state revealed two bound acetate molecules in the extracellular domain (ECD). Here, it is shown that these two acetates exactly overlap with known sites of pharmacological importance in pLGICs, and their potential influence on the structure of the open state is studied in detail. Firstly, experimental evidence is presented for the correct assignment of these acetate molecules by using the anomalous dispersion signal of bromoacetate. Secondly, the crystal structure of GLIC in the absence of acetate was solved and it is shown that acetate binding induces local conformational changes that occur in strategic sites of the ECD. It is expected that this acetate-free structure will be useful in future computational studies of the gating transition in GLIC and other pLGICs.

THE ROLE OF G-PROTEIN COUPLED RECEPTOR 43 IN HIGH GLUCOSE-INDUCED OXIDATIVE STRESS IN HUMAN BLOOD VESSELS

Hyperglycaemia is a hallmark of diabetes and has been found to activate a reactive oxygen species (ROS)-generating enzyme, NADPH oxidase 2 (Nox2), which is involved in inflammation and diabetic cardiovascular complications. G-protein coupled receptor 43 (GPCR43) has been reported recently to be expressed in various tissues; however, its role in the human vascular system remains unknown. Acetate is agonist of GPCR43. In this study we investigated the expression and redox-signalling pathway of GPCR43 in human saphenous vein sections in a cultured model of high glucose-induced Nox2 activation and vessel dysfunction. Vessels were cultured in normal (5 mM, control) or high glucose (30 mM) medium for 24 hours in the presence or absence of acetate (5 mM) to activate GPCR43. The morphological and structural integrities of the vessels were confirmed by histological examination (H & E staining). GPCR43 was detected throughout the entire vessel wall including the endothelium, smooth muscles and adventitia. Compared to control vessels, high glucose increased (33%±10%) the expression of Nox2 in the endothelium as evaluated using immunofluorescence quantification. Increased Nox2 expression was accompanied by increased ROS production, as detected by NADPH-dependent lucigenin-chemiluminescence, and an increase in the phosphorylation of redox-sensitive signalling pathways i.e. ERK1/2 (77%±18%) and JNK (99%±23%). Acetate has no significant effect on GPCR43 expression, but prevented significantly the high glucose-induced Nox2 activation and ERK1/2 and JNK phosphorylation. In conclusion, GPCR43 is highly expressed in human vasculature and may have a therapeutic potential to reduce high glucose-induced Nox2 activation and oxidative damage of vascular function in diabetes.

The role of acetogens in microbially influenced corrosion of steel.

Microbially influenced corrosion (MIC) of iron (Fe0) by sulfate-reducing bacteria (SRB) has been studied extensively. Through a mechanism, that is still poorly understood, electrons or hydrogen (H2) molecules are removed from the metal surface and used as electron donor for sulfate reduction. The resulting ferrous ions precipitate in part with the sulfide produced, forming characteristic black iron sulfide. Hydrogenotrophic methanogens can also contribute to MIC. Incubation of pipeline water samples, containing bicarbonate and some sulfate, in serum bottles with steel coupons and a headspace of 10% (vol/vol) CO2 and 90% N2, indicated formation of acetate and methane. Incubation of these samples in serum bottles, containing medium with coupons and bicarbonate but no sulfate, also indicated that formation of acetate preceded the formation of methane. Microbial community analyses of these enrichments indicated the presence of Acetobacterium, as well as of hydrogenotrophic and acetotrophic methanogens. The formation of acetate by homoacetogens, such as Acetobacterium woodii from H2 (or Fe0) and CO2, is potentially important, because acetate is a required carbon source for many SRB growing with H2 and sulfate. A consortium of the SRB Desulfovibrio vulgaris Hildenborough and A. woodii was able to grow in defined medium with H2, CO2, and sulfate, because A. woodii provides the acetate, needed by D. vulgaris under these conditions. Likewise, general corrosion rates of metal coupons incubated with D. vulgaris in the presence of acetate or in the presence of A. woodii were higher than in the absence of acetate or A. woodii, respectively. An extended MIC model capturing these results is presented.

Enrichment of amino acid-oxidizing, acetate-reducing bacteria

In anaerobic condition, amino acids are oxidatively deaminated, and decarboxylated, resulting in the production of volatile fatty acids. In this process, excess electrons are produced and their consumption is necessary for the accomplishment of amino acid degradation. In this study, we anaerobically constructed leucine-degrading enrichment cultures from three different environmental samples (compost, excess sludge, and rice field soil) in order to investigate the diversity of electron-consuming reaction coupled to amino acid oxidation. Constructed enrichment cultures oxidized leucine to isovalerate and their activities were strongly dependent on acetate. Analysis of volatile fatty acids (VFAs) profiles and community structure analysis during batch culture of each enrichment indicated that Clostridium cluster I coupled leucine oxidation to acetate reduction in the enrichment from the compost and the rice field soil. In these cases, acetate was reduced to butyrate. On the other hand, Clostridium cluster XIVb coupled leucine oxidation to acetate reduction in the enrichment from the excess sludge. In this case, acetate was reduced to propionate. To our surprise, the enrichment from rice field soil oxidized leucine even in the absence of acetate and produced butyrate. The enrichment would couple leucine oxidation to reductive butyrate synthesis from CO2. The coupling reaction would be achieved based on trophic link between hydrogenotrophic acetogenic bacteria and acetate-reducing bacteria by sequential reduction of CO2 and acetate. Our study suggests anaerobic degradation of amino acids is achieved yet-to-be described reactions.