ALBERT

Description: We study a model of lava flow to determine its thermal and dynamic characteristics from thermal measurements of the lava at its surface. Mathematically this problem is reduced to solving an inverse boundary problem. Namely, using known conditions at one part of the model boundary we determine the missing condition at the remaining part of the boundary. We develop a numerical approach to the mathematical problem in the case of steady-state flow. Assuming that the temperature and the heat flow are prescribed at the upper surface of the model domain, we determine the flow characteristics in the entire model domain using a variational (adjoint) method. We have performed computations of model examples and showed that in the case of smooth input data the lava temperature and the flow velocity can be reconstructed with a high accuracy. As expected, a noise imposed on the smooth input data results in a less accurate solution, but still acceptable below some noise level. Also we analyse the influence of optimization methods on the solution convergence rate. The proposed method for reconstruction of physical parameters of lava flows can also be applied to other problems in geophysical fluid flows.

Description: Studies of discrete volcanic explosions, that usually last less than 2 or 3 min, have suggested that the partitioning of seismic-acoustic energy is likely related to a range of physical mechanisms that depend on magma properties and other physical constraints such as the location of the fragmentation surface. In this paper, we explore the energy partition of a paroxysmal eruptive phase of Tungurahua volcano that lasted for over 4 hr, on 2006 July 14–15, using seismic-acoustic information recorded by stations on its flanks (near field). We find evidence of a linear scaling between seismic and acoustic energies, with time-dependent intensities, during the sustained explosive phase of the eruption. Furthermore, we argue that this scaling can be explained by two different processes: (1) the fragmentation region ultimately acts as the common source of energy producing both direct seismic waves, that travel through the volcanic edifice, and direct acoustic waves coming from a disturbed atmosphere above the summit; (2) the coupling of acoustic waves with the ground to cause seismic waves. Both processes are concurrent, however we have found that the first one is dominant for seismic records below 4 Hz. Here we use the linear scaling of intensities to construct seismic and acoustic indices, which, we argue, could be used to track an ongoing eruption. Thus, especially in strong paroxysms that can produce pyroclastic flows, the index correlation and their levels can be used as quantitative monitoring parameters to assess the volcanic hazard in real time. Additionally, we suggest from the linear scaling that the source type for both cases, seismic and acoustic, is dipolar and dominant in the near field.

Description: In this study, we isolated 15 endophytic fungi from five Sudanese medicinal plants. Each fungal endophytic strain was identified by sequencing of internal transcribed spacer (ITS) regions of rDNA. Ethyl acetate extracts were prepared from each endophyte cultivated in vitro and tested for their respective antibacterial activities and antiproliferative activities against human cancer cells. Antibacterial screening was carried out against two bacterial strains: Gram-negative Escherichia coli and Gram-positive methicillin-resistant Staphylococcus aureus , by the broth dilution method. Cell viability was evaluated by the MTT procedure after exposure of MCF7 breast cancer cells and HT29 or HCT116 human colon adenocarcinoma cells to each endophytic extract. Of interest, Byssochlamys spectabilis isolated from Euphorbia prostata showed cytotoxicity (IC 50 = 1.51 ± 0.2 μg mL –1 ) against MCF7 cells, but had a low effect against HT29 or HCT116 cells (IC 50 〉 20 μg mL –1 ). Cladosporium cladosporioides 2, isolated from Vernonia amygdalina leaves, showed antiproliferative activities against MCF7 cells (IC 50 = 10.5 ± 1.5 μg mL –1 ) only. On the other hand, B. spectabilis and Alternaria sp. extract had antibacterial activities against the S. aureus strain. The findings of this work revealed that endophytic fungi associated with medicinal plants from Sudan could be considered as an attractive source of new therapeutic compounds.

Description: Lying below Vatnajökull ice cap in Iceland, Bárðarbunga stratovolcano began experiencing wholesale caldera collapse in 2014 August 16, one of the largest such events recorded in the modern instrumental era. Simultaneous with this collapse is the initiation of a plate boundary rifting episode north of the caldera. Observations using the international constellation of radar satellites indicate rapid 50 cm d –1 subsidence of the glacier surface overlying the collapsing caldera and metre-scale crustal deformation in the active rift zone. Anomalous earthquakes around the rim of the caldera with highly nondouble-couple focal mechanisms provide a mechanical link to the dynamics of the collapsing magma chamber. A model of the collapse consistent with available geodetic and seismic observations suggests that the majority of the observed subsidence occurs aseismically via a deflating sill-like magma chamber.

Description: Deformation experiments on partially molten rocks in simple shear form melt bands at 20° to the shear plane instead of at the expected 45° principal compressive stress direction. These melt bands may play an important role in melt focusing in mid-ocean ridges. Such shallow bands are known to form for two-phase media under shear if strongly non-Newtonian power-law creep is employed for the solid phase, or anisotropy imposed. However laboratory experiments show that shallow bands occur regardless of creep mechanism, even in diffusion creep, which is nominally Newtonian. Here we propose that a couple of forms of two-phase damage allow for shallow melt bands even in diffusion creep.

Description: Resveratrol is a well-known triphenolic natural product present in red wine. For its contribution to human health, the demand for resveratrol as a food and nutrition supplement has increased significantly. In recent years, the rapid development of synthetic biology has promoted extensive work to increase the production of resveratrol in microbes. However, supplementation of expensive phenylpropanoic precursors was required in current engineered strains. Here, we first utilized the site-specific integration strategy to produce resveratrol in Escherichia coli . The genes tal , 4cl and sts were site-specific integrated into the loci of genes tyrR and trpED in the chromosome of E. coli BW25113 (DE3). The final strain was capable of producing 4.612 mg L –1 of resveratrol from glucose.

Description: While extensively studied in several model organisms, the role of small, non-coding RNAs in the stress response remains largely unexplored in Clostridium organisms. About 100 years after the first industrial Acetone–Butanol–Ethanol fermentation process, based on the Weizmann Clostridium acetobutylicum strain, strain tolerance to butanol remains a crucial factor limiting the economics of the process. Several studies have examined the response of this organism to metabolite stress, and several genes have been engaged to impart enhanced tolerance, but no sRNAs have yet been directly engaged in this task. We show that the two stress-responsive sRNAs, 6S and tmRNA, upon overexpression impart tolerance to butanol as assessed by viability assays under process-relevant conditions. 6S overexpression enhances cell densities as well as butanol titres. We discuss the likely mechanisms that these two sRNAs might engage in this tolerance phenotype. Our data support the continued exploration of sRNAs as a basis for engineering enhanced tolerance and enhanced solvent production, especially because sRNA-based strategies impose a minimal metabolic burden on the cells.

Description: Progressive deformation of upper mantle rocks via dislocation creep causes their constituent crystals to take on a non-random orientation distribution (crystallographic preferred orientation or CPO) whose observable signatures include shear-wave splitting and azimuthal dependence of surface wave speeds. Comparison of these signatures with mantle flow models thus allows mantle dynamics to be unraveled on global and regional scales. However, existing self-consistent models of CPO evolution are computationally expensive when used with 3-D and/or time-dependent convection models. Here we propose a new method, called ANPAR, which is based on an analytical parametrization of the crystallographic spin predicted by the second-order (SO) self-consistent theory. Our parametrization runs 2–6  x  10 4 times faster than the SO model and fits its predictions for CPO and crystallographic spin with a variance reduction 〉99 per cent. We illustrate the ANPAR model predictions for the deformation of olivine with three dominant slip systems, (010)[100], (001)[100] and (010)[001], for three uniform deformations (uniaxial compression, pure shear and simple shear) and for a corner-flow model of a spreading mid-ocean ridge.

Description: We investigate the capability of Time Reversal Mirror (TRM) algorithm to image local acoustic sources (〈3.5 km) associated with complex, sustained volcanic eruptions. Accurate source localization for volcano infrasound (low-frequency acoustic waves) is often challenging due to pronounced volcanic topography and emergent arrivals of infrasound signals. While the accuracy of the conventional approaches (e.g. triangulation and semblance method) can be severely compromised by the complex volcanic settings, a TRM-based method may have the potential to properly image acoustic sources by the use of full waveform information and numerical modelling of the time-reversed wavefield. We apply the TRM algorithm to a pyroclastic-laden eruption (sustained for ~60 s) at Santiaguito Volcano, Guatemala, and show that an ordinary TRM operation can undergo significant reduction of its focusing power due to strong topographic propagation effects (e.g. reflection and diffraction). We propose a weighted imaging condition to compensate for complicated transmission loss of the time-reversed wavefield and demonstrate that the presented condition significantly improves the focusing quality of TRM in the presence of complex topography. The consequent TRM source images exhibit remarkable agreement with the visual observation of the eruption implying that the TRM method with a proper imaging condition can be used to localize and track acoustic sources associated with complex volcanic eruptions.

Description: Ten indole alkaloids were obtained from the marine sponge-associated fungus Neosartorya siamensis KUFA 0017. We studied the antimicrobial properties of these and of three other compounds previously isolated from the soil fungus N. siamensis KUFC 6349. Only neofiscalin A showed antimicrobial activity against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE); with a minimum inhibitory concentration (MIC) of 8 μg mL –1 against both strains. Another compound, fiscalin C, presented synergistic activity against MRSA when combined with oxacillin, although alone showed no antibacterial effect. Moreover, neofiscalin A, when present at sub-MICs, hampered the ability of both MRSA and VRE strains to form a biofilm. Additionally, the biofilm inhibitory concentration values of neofiscalin A against the MRSA and VRE isolates were 96 and 80 μg mL –1 , respectively. At a concentration of 200 μg mL –1 , neofiscalin A was able to reduce the metabolic activity of the biofilms by ~50%. One important fact is that our results also showed that neofiscalin A had no cytotoxicity against a human brain capillary endothelial cell line.

Description: During unconventional protein secretion (UPS), proteins do not pass through the classical endoplasmic reticulum (ER)–Golgi-dependent pathway, but are transported to the cell membrane via alternative routes. One type of UPS is dependent on several autophagy-related (Atg) proteins in yeast and mammalian cells, but mechanisms for unconventional secretion are largely unknown for filamentous fungi. In this study, we investigated whether the autophagy machinery is used for UPS in the filamentous fungus Aspergillus niger . An aspartic protease, which we called PepN, was identified as being likely to be secreted unconventionally, as this protein is highly abundant in culture filtrates during carbon starvation while it lacks a conventional N-terminal secretion sequence. We analysed the presence of PepN in the culture filtrates of carbon starved wild-type, atg1 and atg8 deletion mutant strains by Western blot analysis and by secretome analysis using nanoLC-ESI-MS/MS (wild-type and atg8 deletion mutant). Besides the presence of carbohydrate-active enzymes and other types of proteases, PepN was abundantly found in culture filtrates of both wild-type and atg deletion strains, indicating that the secretion of PepN is independent of the autophagy machinery in A. niger and hence most likely occurs via a different mechanism.

Description: Actin-like MreB paralogs play important roles in cell shape maintenance, cell wall synthesis and the regulation of the D,L-endopeptidases, CwlO and LytE. The gram-positive bacteria, Bacillus amyloliquefaciens LL3, is a poly--glutamic acid (-PGA) producing strain that contains three MreB paralogs: MreB, Mbl and MreBH. In B. amyloliquefaciens , CwlO and LytE can degrade -PGA. In this study, we aimed to test the hypothesis that modulating transcript levels of MreB paralogs would alter the synthesis and degradation of -PGA. The results showed that overexpression or inhibition of MreB, Mbl or MreBH had distinct effects on cell morphology and the molecular weight of the -PGA products. In fermentation medium, cells of mreB inhibition mutant were 50.2% longer than LL3, and the -PGA titer increased by 55.7%. However, changing the expression level of mbl showed only slight effects on the morphology, -PGA molecular weight and titer. In the mreBH inhibition mutant, -PGA production and its molecular weight increased by 56.7% and 19.4%, respectively. These results confirmed our hypothesis that suppressing the expression of MreB paralogs might reduce -PGA degradation, and that improving the cell size could strengthen -PGA synthesis. This is the first report of enhanced -PGA production via suppression of actin-like MreB paralogs.

Description: Microbial production of acetone and butanol was one of the first large-scale industrial fermentation processes of global importance. During the first part of the 20th century, it was indeed the second largest fermentation process, superseded in importance only by the ethanol fermentation. After a rapid decline after the 1950s, acetone-butanol-ethanol (ABE) fermentation has recently gained renewed interest in the context of biorefinery approaches for the production of fuels and chemicals from renewable resources. The availability of new methods and knowledge opens many new doors for industrial microbiology, and a comprehensive view on this process is worthwhile due to the new interest. This thematic issue of FEMS Microbiology Letters, dedicated to the 100th anniversary of the first industrial exploitation of Chaim Weizmann's ABE fermentation process, covers the main aspects of old and new developments, thereby outlining a model development in biotechnology. All major aspects of industrial microbiology are exemplified by this single process. This includes new technologies, such as the latest developments in metabolic engineering, the exploitation of biodiversity and discoveries of new regulatory systems such as for microbial stress tolerance, as well as technological aspects, such as bio- and down-stream processing.

Description: The expansion of offshore renewable energy infrastructure and the need for trans-continental shelf power transmission require the use of submarine high-voltage (HV) cables. These cables have maximum operating surface temperatures of up to 70 °C and are typically buried 1–2 m beneath the seabed, within the wide range of substrates found on the continental shelf. However, the heat flow pattern and potential effects on the sedimentary environments around such anomalously high heat sources in the near-surface sediments are poorly understood. We present temperature measurements from a 2-D laboratory experiment representing a buried submarine HV cable, and identify the thermal regimes generated within typical unconsolidated shelf sediments—coarse silt, fine sand and very coarse sand. We used a large (2 x 2.5 m 2 ) tank filled with water-saturated spherical glass beads (ballotini) and instrumented with a buried heat source and 120 thermocouples to measure the time-dependent 2-D temperature distributions. The observed and corresponding Finite Element Method simulations of the steady state heat flow regimes and normalized radial temperature distributions were assessed. Our results show that the heat transfer and thus temperature fields generated from submarine HV cables buried within a range of sediments are highly variable. Coarse silts are shown to be purely conductive, producing temperature increases of 〉10 °C up to 40 cm from the source of 60 °C above ambient; fine sands demonstrate a transition from conductive to convective heat transfer between cf. 20 and 36 °C above ambient, with 〉10 °C heat increases occurring over a metre from the source of 55 °C above ambient; and very coarse sands exhibit dominantly convective heat transfer even at very low ( cf. 7 °C) operating temperatures and reaching temperatures of up to 18 °C above ambient at a metre from the source at surface temperatures of only 18 °C. These findings are important for the surrounding near-surface environments experiencing such high temperatures and may have significant implications for chemical and physical processes operating at the grain and subgrain scale; biological activity at both microfaunal and macrofaunal levels; and indeed the operational performance of the cables themselves, as convective heat transport would increase cable current ratings, something neglected in existing standards.

Description: Food-grade lactic acid bacteria, such as lactobacilli, represent good candidates for the development of mucosal vectors. Indeed, they are generally recognized as safe microorganisms and some strains display beneficial effects (probiotics). In this study, we described a new lactobacilli in vivo expression (LIVE) system for the production and delivery of therapeutic molecules at mucosal surfaces. The versatility and functionality of this system was successfully validated in several lactobacilli species; furthermore, we assessed in vivo LIVE system in two different mouse models of human pathologies: (i) a model of therapy against intestinal inflammation (inflammatory bowel diseases) and (ii) a model of vaccination against dental caries. We demonstrated that Lactobacillus gasseri expressing the anti-inflammatory cytokine IL-10 under LIVE system efficiently delivered the recombinant protein at mucosal surfaces and display anti-inflammatory effects. In the vaccination model against caries, LIVE system allowed the heterologous expression of Streptococcus mutans antigen GbpB by L. gasseri , leading to a stimulation of the host immune response.

Description: The microbial fermentation process is one of the sustainable and environment-friendly ways to produce 1-butanol and other bio-based chemicals. The success of the fermentation process greatly relies on the choice of bioreactors and the separation methods. In this review, the history and the performance of bioreactors for the acetone–butanol–ethanol (ABE) fermentation is discussed. The subject is then focused on in situ product recovery (ISPR) techniques, particularly for the integrated extraction-gas stripping. The usefulness of this promising hybrid ISPR device is acknowledged by its incorporation with batch, fed-batch and continuous processes to improve the performance of ABE fermentation.

Description: We present a strategy to thoroughly investigate the effects of prominent topography on the surface tilt due to a spherical pressure source. We use Etna's topography as a case of study and, for different source positions, we compare the tilt fields calculated through (i) a 3-D boundary element method and (ii) analytical half-space solutions. We systematically determine (i) the source positions leading to the strongest tilt misfits when numerical and analytical results are compared and (ii) the surface areas where the strongest distortions in the tilt field are most likely to be observed. We also demonstrate that, under critical circumstances, in terms of respective positions of pressure source and observation points, results of inversion procedures aimed at retrieving the source parameters can be misleading, if tilt data are analysed using models that do not account for topography.

Description: The agreement between shear wave velocities for the Earth's inner core observed from seismology with those derived from mineral physics is considerably worse than for any other region of the Earth. Furthermore, there is still debate as to the phase of iron present in the inner core, particularly when alloying with nickel and light elements is taken into account. To investigate the extent to which the mismatch between seismology and mineral physics is a function of either crystal structure and/or the amount of nickel present, we have used ab initio molecular dynamics simulations to calculate the elastic constants and seismic velocities ( V p and V s ) of face centred cubic (fcc) iron at Earth's inner core pressures (360 GPa) and at temperatures up to ~7000 K. We find that V p for fcc iron (fcc-Fe) is very similar to that for hexagonal close packed (hcp) iron at all temperatures. In contrast, V s for fcc-Fe is significantly higher than in hcp-Fe, with the difference increasing with increasing temperature; the difference between V s for the core (from seismology) and V s for fcc-Fe exceeds 40 per cent. These results are consistent with previous work at lower temperatures. We have also investigated the effect of 6.5 and 13 atm% Ni in fcc-Fe. We find that Ni only slightly reduces V p and V s (e.g. by 2 per cent in V s for 13 atm% Ni at 5500 K), and cannot account for the difference between the velocities observed in the core and those of pure fcc-Fe. We also tried to examine pre-melting behaviour in fcc-Fe, as reported in hcp-Fe by extending the study to very high temperatures (at which superheating may occur). However, we find that fcc-Fe spontaneously transforms to other hcp-like structures before melting; two hcp-like structures were found, both of hexagonal symmetry, which may most easily be regarded as being derived from an hcp crystal with stacking faults. That the structure did not transform to a true hcp phase is likely as a consequence of the limited size of the simulation box (108 atoms). At 360 GPa, in pure fcc-Fe, we find that the transition from fcc to the hcp-like structure occurs at 7000 K, whereas in the Ni bearing system, the transition occurs at higher temperature (7250 K). This reinforces previous work showing that fcc-Fe might transform to hcp-Fe just before melting, and that Ni tends to stabilize the fcc structure with respect to hcp.

Description: Viscoelastic behaviour of materials in nature is observed in post-event deformations due to seismic or volcanic activities. In this paper, by adopting the correspondence principle, we propose an inelastic model to predict first the Laplace-domain response of a transversely isotropic viscoelastic half-space due to a shear or tensile fault of polygonal shape. The displacement and stress fields in the time domain are then obtained using an efficient and accurate algorithm for the inverse Laplace transform. Numerical examples are presented to validate the proposed solution and to show the viscoelastic displacement and stress fields due to a strike-slip, dip-slip and tensile fault of rectangular shape. The obtained results indicate that both viscoelasticity and transverse isotropy play significant roles in the viscoelastic response of the half-space due to faults, which could be used as benchmarks for the future numerical analysis of realistic post-seismic or volcanic event.

Description: One of the unresolved questions concerning fault deformation is the degree and cause of localization of shear at depth beneath a fault. Geologic observations of exhumed shear zones indicate that while the motion is no longer planar, it can still be localized near the down-dip extension of the fault; however, the degree of localization is uncertain. We employ simple analytic and numerical models to investigate the structural form of distributed shear beneath a strike-slip fault, and the relative importance of the physical mechanisms that have the potential to localize a shear zone. For a purely depth dependent viscosity, = 0 exp (– z / z 0 ), we find that a shear zone develops with a half-width $\delta _w\sim \sqrt{z_0}$ for small z 0 at the base of the layer, where lengths are non-dimensionalized by the layer thickness ( d km). Including a non-linear stress–strain-rate relation ( $\dot{\epsilon }\propto \sigma ^n$ ) scales w by $1/\sqrt{n}$ , comparable to deformation length scales in thin viscous sheet calculations. We find that the primary control on the shear-zone width is the depth dependence of viscosity that arises from the temperature dependence of viscosity and the increase in temperature with depth. As this relationship is exponential, scaling relations give a dimensional half-width that scales approximately as $$ \tilde{\delta }_w\approx T_{\frac{1}{2}}\sqrt{\frac{Rd}{nQ\beta }}\text{ km}, $$ where $T_{\frac{1}{2}}$ (K) is the temperature at the midpoint of the layer, R (J mol –1 K –1 ) the gas constant, Q (J mol –1 ) the activation energy and β (K km –1 ) the geothermal gradient. This relation predicts the numerical results for the parameter range consistent with continental rheologies to within 2–5 per cent and shear-zone half-widths from 2 to 6 km. The inclusion of shear-stress heating reduces w by only an additional 5–25 per cent, depending on the initial width of the shear zone. While the width of the shear zone may not decrease significantly, local temperature increases from shear-stress heating range from 50 to 300 °C resulting in a reduction in viscosities beneath the fault of several orders of magnitude and a concomitant reduction in the stresses needed to drive the motion.

Description: The equation that relates pressure, temperature and volume and is described by parameters that are function of temperature at 1 bar (hereafter called thermal equation of state, TEOS), has practical computational advantages for petrological and geophysical applications over the equation that considers explicitly a thermal pressure. Some considerations that justify the use of the TEOS are discussed here. (1) The assumption that the parameters are function of temperature is perhaps better understood by looking at the Helmholtz energy function that is implicitly assumed in the case of an equation of state (EOS) derived from interatomic potentials. A test case shows that the Helmholtz energy related to the Vinet EOS and the Helmholtz energy from the Debye model are very similar. (2) The TEOS should be able to reproduce thermal expansion (α), isothermal bulk modulus ( K T ) and heat capacity ( C p and C v ) at high P , T computed from a lattice vibration model. The generalized Rydberg EOS applied to MgO is able to fit reasonably well the properties computed using Jacobs’ lattice dynamics formulation ( T range = 300–3000 K, P range = 1 bar–1500 kbar). (3) It is shown that in the case of MgO, the TEOS can be used quite successfully for extrapolation that goes beyond the P , T range of the measured/given data. Some physical constraints need to be applied to the derivation of the volume, bulk modulus and derivative of the bulk modulus with pressure at 1 bar. (4) The pressure dependence of the reference parameters in the TEOS that was inferred several decades ago is only apparent. A numerical computation demonstrates that the combined pressure effect in the terms defining the partial derivative of the reference V and K (and K') over temperature cancels out, making the reference parameters independent of pressure at any condition.

Description: The equation that relates pressure, temperature and volume and is described by parameters that are function of temperature at 1 bar (hereafter called thermal equation of state, TEOS), has practical computational advantages for petrological and geophysical applications over the equation that considers explicitly a thermal pressure. Some considerations that justify the use of the TEOS are discussed here. (1) The assumption that the parameters are function of temperature is perhaps better understood by looking at the Helmholtz energy function that is implicitly assumed in the case of an equation of state (EOS) derived from interatomic potentials. A test case shows that the Helmholtz energy related to the Vinet EOS and the Helmholtz energy from the Debye model are very similar. (2) The TEOS should be able to reproduce thermal expansion (α), isothermal bulk modulus ( K T ) and heat capacity ( C p and C v ) at high P , T computed from a lattice vibration model. The generalized Rydberg EOS applied to MgO is able to fit reasonably well the properties computed using Jacobs’ lattice dynamics formulation ( T range = 300–3000 K, P range = 1 bar–1500 kbar). (3) It is shown that in the case of MgO, the TEOS can be used quite successfully for extrapolation that goes beyond the P , T range of the measured/given data. Some physical constraints need to be applied to the derivation of the volume, bulk modulus and derivative of the bulk modulus with pressure at 1 bar. (4) The pressure dependence of the reference parameters in the TEOS that was inferred several decades ago is only apparent. A numerical computation demonstrates that the combined pressure effect in the terms defining the partial derivative of the reference V and K (and K') over temperature cancels out, making the reference parameters independent of pressure at any condition.

Description: Anaerobic Clostridium spp. is an important bioproduction microbial genus that can produce solvents and utilize a broad spectrum of substrates including cellulose and syngas. Genome-scale metabolic (GSM) models are increasingly being put forth for various clostridial strains to explore their respective metabolic capabilities and suitability for various bioconversions. In this study, we have selected representative GSM models for six different clostridia ( Clostridium acetobutylicum , C. beijerinckii , C. butyricum , C. cellulolyticum , C. ljungdahlii and C. thermocellum ) and performed a detailed model comparison contrasting their metabolic repertoire. We also discuss various applications of these GSM models to guide metabolic engineering interventions as well as assessing cellular physiology.

Description: Biobutanol is a next-generation liquid biofuel with properties akin to those of gasoline. There is a widespread effort to commercialize biobutanol production from agricultural residues, such as corn stover, which do not compete with human and animal foods. This pursuit is backed by extensive government mandates to expand alternative energy sources. This review provides an overview of research on biobutanol production using corn stover feedstock. Structural composition, pretreatment, sugar yield (following pretreatment and hydrolysis) and generation of lignocellulose-derived microbial inhibitory compounds (LDMICs) from corn stover are discussed. The review also discusses different Clostridium species and strains employed for biobutanol production from corn stover-derived sugars with respect to solvent yields, tolerance to LDMICs and in situ solvent recovery (integrated fermentation). Further, the economics of cellulosic biobutanol production are highlighted and compared to corn starch-derived ethanol and gasoline. As discussed herein, the economic competitiveness of biobutanol production from corn stover largely depends on feedstock processing and fermentation process design.

Description: Biological production in heterologous hosts is of interest for the production of the C4 alcohol (butanol) and other chemicals. However, some hurdles need to be overcome in order to achieve an economically viable process; these include avoiding the consumption of butanol and maintaining tolerance to this solvent during production. Pseudomonas putida is a potential host for solvent production; in order to further adapt P. putida to this role, we generated mini-Tn 5 mutant libraries in strain BIRD-1 that do not consume butanol. We analyzed the insertion site of the mini-Tn 5 in a mutant that was deficient in assimilation of butanol using arbitrary PCR followed by Sanger sequencing and found that the transposon was inserted in the malate synthase B gene. Here, we show that in a second round of mutagenesis a double mutant unable to take up butanol had an insertion in a gene coding for a multisensor hybrid histidine kinase. The genetic context of the histidine kinase sensor revealed the presence of a set of genes potentially involved in butanol assimilation; qRT-PCR analysis showed induction of this set of genes in the wild type and the malate synthase mutant but not in the double mutant.

Description: The heavy dependence on petroleum-derived fuel has raised concerns about energy sustainability and climate change, which have prompted researchers to explore fuel production from renewable sources. 1-Butanol and isobutanol are promising biofuels that have favorable properties and can also serve as solvents or chemical feedstocks. Microbial production of these alcohols provides great opportunities to access a wide spectrum of renewable resources. In recent years, research has improved the native 1-butanol production and has engineered isobutanol production in various organisms to explore metabolic diversity and a broad range of substrates. This review focuses on progress in metabolic engineering for the production of these two compounds using various resources.

Description: Biobased production of butanol promises a more sustainable route for industrial production. However, butanol toxicity remains a barrier for achieving high product titers. Investigation into butanol stress has shed some light on its modes of toxicity. Unfortunately, there still remain significant shortfalls in our understanding of the complex interactions of butanol with cells. To address this knowledge gap, a diverse range of tools have been employed to gain a better understanding of the adverse effects of butanol on the cell. These findings have lead to the identification of possible molecular mechanisms associated with butanol tolerance, which can be harnessed for future strain development efforts. This review focuses on recent efforts to address the toxicity of butanol in microbial producers and offers some perspectives on the future direction of this research sector.

Description: Biobutanol outperforms bioethanol as an advanced biofuel, but is not economically competitive in terms of its titer, yield and productivity associated with feedstocks and energy cost. In this work, the synergistic effect of calcium and zinc was investigated in the acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum using glucose, xylose and glucose/xylose mixtures as carbon source(s). Significant improvements associated with enhanced glucose/xylose utilization, cell growth, acids re-assimilation and butanol biosynthesis were achieved. Especially, the maximum butanol and ABE production of 16.1 and 25.9 g L –1 were achieved from 69.3 g L –1 glucose with butanol/ABE productivities of 0.40 and 0.65 g L –1 h –1 compared to those of 11.7 and 19.4 g/L with 0.18 and 0.30 g L –1 h –1 obtained in the control respectively without any supplement. More importantly, zinc was significantly involved in the butanol tolerance based on the improved xylose utilization under various butanol-shock conditions (2, 4, 6, 8 and 10 g L –1 butanol). Under the same conditions, calcium and zinc co-supplementation led to the best xylose utilization and butanol production. These results suggested that calcium and zinc could play synergistic roles improving ABE fermentation by C. acetobutylicum .

Description: The presence of calcite in and near faults, as the dominant material, cement, or vein fill, indicates that the mechanical behaviour of carbonate-dominated material likely plays an important role in shallow- and mid-crustal faulting. To better understand the behaviour of calcite, under loading conditions relevant to earthquake nucleation, we sheared powdered gouge of Carrara Marble, 〉98 per cent CaCO 3 , at constant normal stresses between 1 and 100 MPa under water-saturated conditions at room temperature. We performed slide-hold-slide tests, 1–3000 s, to measure the amount of static frictional strengthening and creep relaxation, and velocity-stepping tests, 0.1–1000 μm s –1 , to evaluate frictional stability. We observe that the rates of frictional strengthening and creep relaxation decrease with increasing normal stress and diverge as shear velocity is increased from 1 to 3000 μm s –1 during slide-hold-slide experiments. We also observe complex frictional stability behaviour that depends on both normal stress and shearing velocity. At normal stresses less than 20 MPa, we observe predominantly velocity-neutral friction behaviour. Above 20 MPa, we observe strong velocity-strengthening frictional behaviour at low velocities, which then evolves towards velocity-weakening friction behaviour at high velocities. Microstructural analyses of recovered samples highlight a variety of deformation mechanisms including grain size reduction and localization, folding of calcite grains and fluid-assisted diffusion mass transfer processes promoting the development of calcite nanograins in the highly deformed portions of the experimental fault. Our combined analyses indicate that calcite fault gouge transitions from brittle to semi-brittle behaviour at high normal stress and slow sliding velocities. This transition has important implications for earthquake nucleation and propagation on faults in carbonate-dominated lithologies.

Description: Pseudomonas putida KT2440 is a saprophytic, environmental microorganism that plays important roles in the biodegradation of environmental toxic compounds and production of polymers, chemicals and secondary metabolites. Gene deletion of KT2440 usually involves cloning of the flanking homologous fragments of the gene of interest into a suicide vector followed by transferring into KT2440 via triparental conjugation. Selection and counterselection steps are then employed to generate gene deletion mutant. However, these methods are tedious and are not suitable for the manipulation of multiple genes simultaneously. Herein, a two-step, markerless gene deletion method is presented. First, homologous armsflanked loxP-neo-loxP was knocked-in to replace the gene of interest, then the kanamycin resistance marker is removed by Cre recombinase catalyzed site-specific recombination. Both two-plasmid and one-plasmid gene systems were established. MekR/P mekA regulated gene expression system was found to be suitable for tight Cre expression in one-plasmid deletion system. The straightforward, time saving and highly efficient markerless gene deletion strategy has the potential to facilitate the genetics and functional genomics study of P. putida KT2440.

Description: Flow cytometry, in combination with fluorescent staining, was used to evaluate population heterogeneity in acetone-butanol–ethanol fermentation that was carried out with type strain Clostridium beijerinckii NCIMB 8052 and non-type C. pasteurianum NRRL B-598. A combination of propidium iodide (PI) and carboxyfluorescein diacetate (CFDA), PI plus Syto-9 and bis-oxonol (BOX) alone were employed to distinguish between active and damaged cells together with simultaneous detection of spores. These strategies provided valuable information on the physiological state of clostridia. CFDA and PI staining gave the best separation of four distinct subpopulations of enzymatically active cells, doubly stained cells, damaged cells and spores. Proportional representation of cells in particular sub-regions correlated with growth characteristics, fermentation parameters such as substrate consumption and product formation in both species under different cultivation conditions.

Description: A knowledge of subsurface temperatures in sedimentary basins, fault zones, volcanic environments and polar ice sheets is of interest for a wide variety of geophysical applications. However, the process of drilling deep boreholes in these environments to provide access for temperature and other measurements invariably disturbs the temperature field around a newly created borehole. Although this disturbance dissipates over time, most temperature measurements are made while the temperature field is still disturbed. Thus, the measurements must be ‘corrected’ for the drilling-disturbance effect if the undisturbed temperature field is to be determined. This paper provides compact analytical solutions for the thermal drilling disturbance based on 1-D (radial) and 2-D (radial and depth) Green's functions (GFs) in cylindrical coordinates. Solutions are developed for three types of boundary conditions (BCs) at the borehole wall: (1) prescribed temperature, (2) prescribed heat flux and (3) a prescribed convective condition. The BC at the borehole wall is allowed to vary both with depth and time. Inclusion of the depth dimension in the 2-D solution allows vertical heat-transfer effects to be quantified in situations where they are potentially important, that is, near the earth's surface, at the bottom of a well and when considering finite-drilling rates. The 2-D solution also includes a radial- and time-dependent BC at the earth's surface to assess the impact of drilling-related infrastructure (drilling pads, mud pits, permanent shelters) on the subsurface temperature field. Latent-heat effects due to the melting and subsequent refreezing of interstitial ice while drilling a borehole through ice-rich permafrost can be included in the GF solution as a moving-plane heat source (or sink) located at the solid–liquid interface. Synthetic examples are provided illustrating the 1-D and 2-D GF solutions. The flexibility of the approach allows the investigation of thermal drilling effects in rock or ice for a wide variety of drilling technologies. Numerical values for the required radial GFs G R are available through the Advanced Cooperative Arctic Data and Information Service at doi:10.5065/D64F1NS6.

Description: Simultaneous saccharification and fermentation (SSF) process was applied for biobutanol production by Clostridium saccharobutylicum DSM 13864 from corn stover (CS). The key influential factors in SSF process, including corn steep liquor concentration, dry biomass and enzyme loading, SSF temperature, inoculation size and pre-hydrolysis time were optimized. In 5-L bioreactor with SSF process, butanol titer and productivity of 12.3 g/L and 0.257 g/L/h were achieved at 48 h, which were 20.6% and 21.2% higher than those in separate hydrolysis and fermentation (SHF), respectively. The butanol yield reached 0.175 g/g pretreated CS in SSF, representing 50.9% increase than that in SHF (0.116 g/g pretreated CS). This study proves the feasibility of efficient and economic production of biobutanol from CS by SSF.

Description: The elastic properties of homogeneous, isotropic materials are well constrained. However, in heterogeneous and evolving materials, these essential properties are less well-explored. During sintering of volcanic ash particles by viscous processes as well as during compaction and cementation of sediments, microstructure and porosity undergo changes that affect bulk dynamic elastic properties. Here using a model system of glass particles as an analogue for initially granular rock-forming materials, we have determined porosity and P -wave velocity during densification. Using these results, we test models for the kinetics of densification and the resultant evolution of the elastic properties to derive a quantitative description of the coupling between the kinetics of isotropic densification and the evolving dynamic elastic moduli. We demonstrate the power of the resultant model on a wide range of data for non-coherent sediments as well as sedimentary and volcanic rocks. We propose that such constraints be viewed as an essential ingredient of time-dependent models for the deformation of evolving materials in volcanoes and sedimentary basins.

Description: Tarim Craton is a Precambrian block situated in northwest China, just north of the Tibetan Plateau, where a large sedimentary basin with abundant hydrocarbon potential has developed. Accurate heat flow data for Tarim is vital for understanding the lithospheric evolution and hydrocarbon generation in this area; however, there were unavailable until now, due to a lack of high quality steady-state temperature logging data. Here, we report 10 new heat flow values derived from steady-state temperature logging and measured thermal conductivity data. New heat flow values range from 40.1 to 49.4 mW m –2 , with a mean of 43.1 ± 3.0 mW m –2 . In addition, radiogenic heat production from the sediments accounts for 20 per cent of the observed surface heat flow, whilst the mantle heat flow is estimated to be as low as 6–15 mW m –2 ; this indicates a dominant contribution from crustal heat, to the observed heat flow. The average heat flow and crustal temperature in the Tarim Craton are markedly lower than those in the Tibetan Plateau, whilst the calculated rheological strength of the lithosphere, beneath Tarim, is sufficiently large to resist the elevation-induced gravitational potential energy difference between Tarim and Tibet. This inherited thermal and rheological contrast, between the craton and Plateau, can be traced back to before the India–Asia collision; this accounts for the differential active deformation pattern in the Tarim Craton and adjacent areas.

Description: We present a study of the seismic velocity variations that occurred in the structure before the large 2010 eruption of Merapi volcano. For the first time to our knowledge, the technique of coda wave interferometry is applied to both families of similar events (multiplets) and to correlation functions of seismic noise. About half of the seismic events recorded at the summit stations belong to one of the ten multiplets identified, including 120 similar events that occurred in the last 20 hr preceding the eruption onset. Daily noise cross-correlation functions (NCF) were calculated for the six pairs of short-period stations available. Using the stretching method, we estimate time-series of apparent velocity variation (AVV) for each multiplet and each pair of stations. No significant velocity change is detected until September 2010. From 10 October to the beginning of the eruption on 26 October, a complex pattern of AVV is observed with amplitude of up to ±1.5 per cent. Velocity decrease is first observed from families of deep events and then from shallow earthquakes. In the same period, AVV with different signs and chronologies are estimated from NCF calculated for various station pairs. The location in the horizontal plane of the velocity perturbations related with the AVV obtained from NCF is estimated by using an approach based on the radiative transfer approximation. Although their spatial resolution is limited, the resulting maps display velocity decrease in the upper part of the edifice in the period 12–25 October. After the eruption onset, the pattern of velocity perturbations is significantly modified with respect to the previous one. We interpret these velocity variations in the framework of a scenario of magmatic intrusion that integrates most observations. The perturbation of the stress field associated with the magma migration can induce both decrease and increase of the seismic velocity of rocks. Thus the detected AVVs can be considered as precursors of volcanic eruptions in andesitic volcanoes, without taking their sign into account.

Description: We previously reported that the β-1,4-Mannanase ( manB ) gene from Bacillus pumilus functions as a good reporter gene in Lactobacillus casei . Two vectors were constructed. One carries the signal peptide of secretion protein Usp45 (SP Usp45 ) from Lactococcus lactis (pELSH), and the other carries the full-length S-layer protein, SlpA, from L. acidophilus (pELWH). In this work, another vector, pELSPH, was constructed to include the signal peptide of protein SlpA (SP SlpA ), and the capacity of all three vectors to drive expression of the manB gene in L. casei was evaluated. The results showed that SP Usp45 is functionally recognized and processed by the L. casei secretion machinery. The SP Usp45 -mediated secretion efficiency was ~87%, and SP SlpA drove the export of secreted ManB with ~80% efficiency. SP SlpA secretion was highly efficient, and expressed SlpA was anchored to the cell wall by an unknown secretion mechanism. Full-length SlpA drove the cell wall-anchored expression of an SlpA-ManB fusion protein but at a much lower level than that of protein SlpA.

Description: The porin MspA of Mycobacterium smegmatis is a biological nanopore used for DNA sequencing. The octameric MspA pore can be isolated from M. smegmatis in milligram quantities, is extremely stable against denaturation and rapidly inserts into lipid membranes. Here, we show that MspA pores composed of different Msp subunits are formed in M. smegmatis and that hetero-oligomers of different Msp monomers increase the heterogeneity of MspA pores designed for DNA sequencing. To improve the quality of preparations of mutant MspA proteins, all four msp genes were deleted from the M. smegmatis genome after insertion of an inducible porin gene from M. tuberculosis. In the msp quadruple mutant M. smegmatis ML712 no Msp porins were detected and mutant MspA proteins were produced at wild-type levels. Lipid bilayer experiments demonstrated that MspA pores isolated from ML712 formed functional channels and had a narrower conductance distribution than pores purified from M. smegmatis with background msp expression. Thus, the M. smegmatis msp quadruple mutant improves the homogeneity of MspA pores designed for DNA sequencing and might also facilitate the identification and functional characterization of other mycobacterial pore proteins.

Description: Mostly, butanol is formed as a product by saccharolytic anaerobes, employing the so-called ABE fermentation (for acetone–butanol–ethanol). However, this alcohol can also be produced from gaseous substrates such as syn(thesis) gas (major components are carbon monoxide and hydrogen) by autotrophic acetogens. In view of economic considerations, a biotechnological process based on cheap and abundant gases such as CO and CO 2 as a carbon source is preferable to more expensive sugar or starch fermentation. In addition, any conflict for use of substrates that can also serve as human nutrition is avoided. Natural formation of butanol has been found with, e.g. Clostridium carboxidivorans , while metabolic engineering for butanol production was successful using, e.g. C. ljungdahlii . Production of butanol from CO 2 under photoautotrophic conditions was also possible by recombinant DNA construction of a respective cyanobacterial Synechococcus sp. PCC 7942 strain.

Description: The influx of groundwater into hot kimberlite deposits results in the reaction of water with olivine-rich rocks. The products of the reaction are serpentine and release of latent heat. The rise of temperature due to the heat release increases the rate of the reaction. Under certain conditions, this self-speeding up of the reaction can result in instabilities associated with a significantly higher final serpentinization in slightly warmer regions of the kimberlite deposit. We conduct linear stability analysis of serpentinization in an isolated volume of porous kimberlitic rocks saturated with water and an inert gas. There is a counteracting interplay between the heat release tending to destabilize the uniform distribution of parameters and the heat conduction tending to stabilize it by smoothing out temperature perturbations. We determine the critical spatial scale separating the parameters where one phenomenon dominates over another. The perturbations of longer-than-critical length grow, whereas the perturbations of shorter-than-critical length fade. The analytical results of the linear stability analysis are supported by direct numerical simulations using a full nonlinear model.

Description: Lichen-forming fungi and extracts derived from them have been used as alternative medicine sources for millennia and recently there has been a renewed interest in their known bioactive properties for anticancer agents, cosmetics and antibiotics. Although lichen-forming fungus-derived compounds are biologically and commercially valuable, few studies have been performed to determine their modes of action. This study used chemical-genetic and chemogenomic high-throughput analyses to gain insight into the modes of action of Caloplaca flavoruscens extracts. High-throughput screening of 575 lichen extracts was performed and 39 extracts were identified which inhibited yeast growth. A C. flavoruscens extract was selected as a promising antifungal and was subjected to genome-wide haploinsufficiency profiling and homozygous profiling assays. These screens revealed that yeast deletion strains lacking Rsc8, Pro1 and Toa2 were sensitive to three concentrations (IC 25.5 , IC 25 and IC 50 , respectively) of C. flavoruscens extract. Gene-enrichment analysis of the data showed that C. flavoruscens extracts appear to perturb transcription and chromatin remodeling.

Description: Plant lectins, which are proteins/glycoproteins present in a wide range of vegetables, fruits, cereals and beans, are resistant to digestive enzymes and food cooking temperatures. They bind reversibly to specific glycosidic residues expressed on the membrane of intestinal epithelial cells (IEC) and cause anti-nutritional effects in humans and animals. Soybean lectin (SBA) has been detected in poultry diets, and its ability to bind to the intestinal epithelium has been reported. The development of new methods for removing SBA from feeds or to prevent interaction with the intestinal mucosa is of interest. In this study, the in vitro cytotoxicity of SBA on IEC of chicks was demonstrated for the first time. The LD 50 , assessed after 2 h exposure of IEC to SBA, was 6.13 μg mL –1 . The ability of Bifidobacterium infantis CRL1395 to bind SBA on the bacterial envelope was confirmed, and prevention of IEC cytotoxicity by lectin removal was demonstrated. Safety of B. infantis CRL1395, resistance to gastrointestinal stress and adhesion were also determined. It was concluded that the early administration of B. infantis CRL1395 to chicks would effectively reduce the toxicity of SBA. Besides, it would favour the colonization of the gut with a beneficial microbiota.

Description: Inelastic deformation can either occur with dilatancy or compaction, implying differences in porosity changes, failure and petrophysical properties. In this study, the roles of water as a pore fluid, and of temperature, on the deformation and failure of a micritic limestone (white Tavel limestone, porosity 14.7 per cent) were investigated under triaxial stresses. For each sample, a hydrostatic load was applied up to the desired confining pressure (from 0 up to 85 MPa) at either room temperature or at 70 °C. Two pore fluid conditions were investigated at room temperature: dry and water saturated. The samples were deformed up to failure at a constant strain rate of ~10 –5 s –1 . The experiments were coupled with ultrasonic wave velocity surveys to monitor crack densities. The linear trend between the axial crack density and the relative volumetric strain beyond the onset of dilatancy suggests that cracks propagate at constant aspect ratio. The decrease of ultrasonic wave velocities beyond the onset of inelastic compaction in the semi-brittle regime indicates the ongoing interplay of shear-enhanced compaction and crack development. Water has a weakening effect on the onset of dilatancy in the brittle regime, but no measurable influence on the peak strength. Temperature lowers the confining pressure at which the brittle–semi-brittle transition is observed but does not change the stress states at the onset of inelastic compaction and at the post-yield onset of dilatancy.

Description: Fermentation-based production of butyric acid is robust and efficient. Modern catalytic technologies make it possible to convert butyric acid to important fine chemicals and biofuels. Here, current chemocatalytic and biocatalytic conversion methods are reviewed with a focus on upgrading butyric acid to 1-butanol or butyl-butyrate. Supported Ruthenium- and Platinum-based catalyst and lipase exhibit important activities which can pave the way for more sustainable process concepts for the production of green fuels and chemicals.

Description: Clostridium acetobutylicum is an industrially important Gram-positive organism, which is capable of producing economically important chemicals in the ABE (Acetone, Butanol and Ethanol) fermentation process. Renewed interests in the ABE process necessitate the availability of additional genetics tools to facilitate the derivation of a greater understanding of the underlying metabolic and regulatory control processes in operation through forward genetic strategies. In this study, a xylose inducible, mariner -based, transposon system was developed and shown to allow high-efficient random mutagenesis in the model strain ATCC 824. Of the thiamphenicol resistant colonies obtained, 91.9% were shown to be due to successful transposition of the catP- based mini-transposon element. Phenotypic screening of 200 transposon clones revealed a sporulation-defective clone with an insertion in spo0A , thereby demonstrating that this inducible transposon system can be used for forward genetic studies in C. acetobutylicum .

Description: Bacteria are becoming increasingly resistant to currently used antibiotics. At the same time, little progress has been made in discovering new antibacterial drugs to combat resistant organisms. History teaches us that ‘high tech’ target-based complex methods are not synonymous with success and a return to simple, systematic screening of natural products against bacteria from traditional and novel resources holds our greatest hope of success.

Description: The economic upturn of the past 200 years would not have been conceivable without fossil resources such as coal and oil. However, the fossil-based economy increasingly reaches its limits and displays contradictions. Bioeconomy, strategically combining economy and ecology willing to make biobased and sustainable growth possible, is promising to make a significant contribution towards solving these issues. In this context, microbial bioconversions are promising to support partially the increasing need for materials and fuels starting from fresh, preferably waste, biomass. Butanol is a very attractive molecule finding applications both as a chemical platform and as a fuel. Today it principally derives from petroleum, but it also represents the final product of microbial catabolic pathways. Because of the need to maximize yield, titer and productivity to make the production competitive and viable, the challenge is to transform a robustly regulated metabolic network into the principal cellular activity. However, this goal can only be accomplished by a profound understanding of the cellular physiology, survival strategy and sensing/signalling cascades. Here, we shortly review on the natural cellular pathways and circumstances that lead to n -butanol accumulation, its physiological consequences that might not match industrial needs and on possible solutions for circumventing these natural constraints.

Description: Butanol is a very interesting substance both for the chemical industry and as a biofuel. The classical distillation process for the removal of butanol is far too energy demanding, at a factor of 220% of the energy content of butanol. Alternative separation processes studied are hybrid processes of gas-stripping, liquid–liquid extraction and pervaporation with distillation and a novel adsorption/drying/desorption hybrid process. Compared with the energy content of butanol, the resulting energy demand for butanol separation and concentration of optimized hybrid processes is 11%–22% for pervaporation/distillation and 11%–17% for liquid–liquid extraction/distillation. For a novel adsorption/drying/desorption process, the energy demand is 9.4%. But all downstream process options need further proof of industrial applicability.

Description: Acetone–butanol–ethanol (ABE) fermentation is a metabolic process of clostridia that produces bio-based solvents including butanol. It is enabled by an underlying metabolic reaction network and modulated by cellular gene regulation and environmental cues. Mathematical modeling has served as a valuable strategy to facilitate the understanding, characterization and optimization of this process. In this review, we highlight recent advances in system-level, quantitative modeling of ABE fermentation. We begin with an overview of integrative processes underlying the fermentation. Next we survey modeling efforts including early simple models, models with a systematic metabolic description, and those incorporating metabolism through simple gene regulation. Particular focus is given to a recent system-level model that integrates the metabolic reactions, gene regulation and environmental cues. We conclude by discussing the remaining challenges and future directions towards predictive understanding of ABE fermentation.

Description: Cry proteins are pesticidal toxins produced by the bacterium Bacillus thuringiensis (Bt), which aggregate in sporulating cells to form a crystal. Except in a relatively few cases, these crystals are located outside the exosporium that surrounds the spore. Bt2-56 is a strain of Bt that has the relatively uncommon characteristic of locating its Cry protein-containing crystal within the exosporium, and in association with a long, multifiber filament. With the ultimate goal of both understanding and manipulating the localization of Cry proteins within the exosporium, we sought to identify the genes coding for the exosporium-localized Cry proteins in Bt2-56. Herein we show (i) that five cry-like genes are present in the genome of Bt2-56, (ii) that two pairs of these genes show organizational similarity to a relatively uncommon gene configuration that coexpress a cry gene along with a gene whose product aids crystal formation and (iii) that when one of these two gene pairs (cry21A-cdA) is expressed in an acrystalliferous strain of Bt, crystals are formed that localize within the exosporium. In Bt ssp. finitimus , the only other strain in which crystal localization has been studied, a Cry protein needed expression of two non-cry ORFs in order to localize within the exosporium, indicating that there are some mechanistic differences for crystal localization between Bt ssp. finitimus and Bt2-56.

Description: The commercialization of the n -butanol bioprocess is largely dependent on the price of feedstocks. Renewable cellulose appears to be an appealing feedstock. The microbial production of n -butanol still remains challenging because of the limited availability of intracellular NADH. To address this issue, an Escherichia coli strain carrying the clostridial CoA-dependent pathway was supplied with heterologous formate dehydrogenase. With the cellulose hydrolysate of rice straw, this single strain produced cellulosic biobutanol with a production yield at 35% of the theoretical and a productivity of 0.093 g L –1  h –1 . In an alternative method, the production involved a co-culture system consisting of two separate strains provided with the full CoA-dependent pathway. This system achieved a production yield and productivity reaching 62.8% of the theoretical and 0.163 g L –1  h –1 , respectively. The result indicates that the E. coli co-culture system is technically promising for the production of cellulosic biobutanol.

Description: Remote sensing thermal data of active lava flows allow the evaluation of effusion rates. This is made possible by a simple formula relating the lava effusion rate to the heat flux radiated per unit time from the surface of the flow. Due to the assumptions of the model, this formula implies that heat flux, surface temperature and lava temperature vary as a function of the flow thickness. These relationships, never verified or validated before, have been used by several authors as a proof of the weakness of the model. Here, multispectral infrared and visible imaging spectrometer (MIVIS) high spatial resolution (5–10 m) thermal data acquired during Etna's 2001 eruption were used to investigate downflow heat flux variations in the lava flow emitted from a vent located at 2100 m a.s.l. A high correlation between the downflow heat flux and the lava flow thickness (measured from a pre-existing digital elevation model) was found. Topography beneath the flow appears to play an important role both in lava emplacement mechanisms and flow dynamics. MIVIS-derived downflow effusion rates are consistent with the law of conservation of mass assessing the reliability of remote sensing techniques.

Description: Hepatitis E virus (HEV) is the pathogen causing hepatitis E (HE). It arouses global public health concern since it is a zoonotic disease. The objective of this letter is to report a cost-effective internal control prepared for monitoring procedures of HEV reverse transcriptase (RT)-PCR detection. A selected conserved HEV RNA fragment was integrated into the downstream of the truncated MS2 bacteriophage genome based on Armored RNA technology. The resulting MS2-HEV gene harbored by the pET-28b-MS2-HEV plasmid was transformed into E. coli BL21(DE3) for expression analysis by SDS-PAGE. The expression products were purified and concentrated by ultrasonication and ultrafiltration separation. The morphology and stability properties of the virus-like particles (VLPs) were evaluated by electron microscopy scanning and nuclease challenges, respectively. SDS-PAGE results showed that the constructed MS2-HEV gene expressed efficiently and the purity of the VLPs was highly consistent with the result in electron microscopy. Stability evaluation results demonstrated that the prepared VLPs exhibited strong resistance to DNase I and RNase A attacks and also performed long-lasting protection of coated HEV RNA for at least 4 months at –20°C. These data revealed that the prepared VLPs meet the basic requirements of use as internal control material in the HEV RNA amplification assay.

Description: Epigenetics is the study of heritable changes in gene expression without concomitant changes in DNA sequence. Due to its relevance in development, differentiation and human health, epigenetics has recently become an emerging area of science with regard to eukaryotic organisms and has shown enormous potential in synthetic biology. However, significant examples of epigenetic regulation in bacterial synthetic biology have not yet been reported. In the current study, we present the first model of such an epigenetic circuit. We termed the circuit the alternator circuit because parental cells carrying this circuit and their progeny alternate between distinct and heritable cellular fates without undergoing changes in genome sequence. Furthermore, we demonstrated that the alternator circuit exhibits hysteresis because its output depends not only on its present state but also on its previous states.

Description: Bacteriophages produce endolysins (peptidoglycan hydrolases) to lyse the host cell from within and release nascent bacteriophage particles. Recombinant endolysins can lyse Gram-positive bacteria when added exogenously. As a potential alternative antimicrobial, we cloned and expressed the enterococcal VD13 bacteriophage endolysin. VD13 endolysin has a CHAP catalytic domain with 92% identity with the bacteriophage IME-EF1 endolysin. The predicted size of VD13 endolysin is ~27 kDa as verified by SDS-PAGE. The VD13 endolysin lyses Enterococcus faecalis strains, but not E. faecium or other non-enterococci. VD13 endolysin has activity from pH 4 to pH 8, with peak activity at pH 5, and exhibits greater activity in the presence of calcium. Optimum activity at pH 5 occurs in the absence of NaCl. VD13 endolysin, in ammonium acetate (C 2 H 3 O 2 NH 4 ) calcium chloride (CaCl 2 ) buffer pH 5, is stimulated to higher activity upon heating at temperatures up to 65°C for 30 min, whereas activity is lost upon heating to 42°C, in pH 7 buffer.

Description: Cultivation in a bioreactor of immobilized deep-sea hydrothermal microbial community was tested in order to assess the stability and reactivity of this new system. A community composed of eight hydrothermal strains was entrapped in a polymer matrix that was used to inoculate a continuous culture in a gas-lift bioreactor. The continuous culture was performed for 41 days at successively 60°C, 55°C, 60°C, 85°C and 60°C, at pH 6.5, in anaerobic condition and constant dilution rate. Oxic stress and pH variations were tested at the beginning of the incubation. Despite these detrimental conditions, three strains including two strict anaerobes were maintained in the bioreactor. High cell concentrations (3 x 10 8 cells mL –1 ) and high ATP contents were measured in both liquid fractions and beads. Cloning-sequencing and qPCR revealed that Bacillus sp. dominated at the early stage, and was later replaced by Thermotoga maritima and Thermococcus sp. Acetate, formate and propionate concentrations varied simultaneously in the liquid fractions. These results demonstrate that these immobilized cells were reactive to culture conditions. They were protected inside the beads during the stress period and released in the liquid fraction when conditions were more favorable. This confirms the advantage of immobilization that highlights the resilience capacity of certain hydrothermal microorganisms after a stress period.

Description: To study the viability of a gyrA S83 stop mutation found in an Escherichia coli J53 ciprofloxacin-resistant strain (J53 CipR27), a pBR322 derivative was constructed with a TAG mutation in the bla gene knocking out ampicillin resistance. Ampicillin resistance was restored, suggesting that the strain contains tRNA suppressor activity able to suppress the UAG codon gyrA and allow viability. The method was applied to 22 unique clinical E. coli isolates, and all were found to have low-level suppressor activity.

Description: Enterotoxigenic Escherichia coli (ETEC) bacteria producing heat-stable toxin (STa) and/or heat-labile toxin (LT) are among top causes of children's diarrhea and travelers’ diarrhea. Currently no vaccines are available for ETEC associated diarrhea. A major challenge in developing ETEC vaccines is the inability to stimulate protective antibodies against the key STa toxin that is potently toxic and also poorly immunogenic. A recent study suggested toxoid fusion 3xSTa N12S -dmLT, which consists of a monomer LT toxoid (LT R192G/L211A ) and three copies of STa toxoid STa N12S , may represent an optimal immunogen inducing neutralizing antibodies against STa toxin [IAI 2014, 82(5):1823-32]. In this study, we immunized mice with this fusion protein following a different parenteral route and using different adjuvants to further characterize immunogenicity of this toxoid fusion. Data from this study showed that 3xSTa N12S -dmLT toxoid fusion induced neutralizing anti-STa antibodies in the mice following subcutaneous immunization, as effectively as in the mice under intraperitoneal route. Data also indicated that double mutant LT (dmLT) can be an effective adjuvant for this toxoid fusion in mice subcutaneous immunization. Results from this study affirmed that toxoid fusion 3xSTa N12S -dmLT induces neutralizing antibodies against STa toxin, suggesting this toxoid fusion is potentially a promising immunogen for ETEC vaccine development.

Description: We performed complex conductivity measurements on 28 core samples from the hole drilled for the Humu'ula Groundwater Research Project (Hawai'i Island, HI, USA). The complex conductivity measurements were performed at 4 different pore water conductivities (0.07, 0.5, 1.0 or 2.0, and 10 S m –1 prepared with NaCl) over the frequency range 1 mHz to 45 kHz at 22 ± 1 °C. The in-phase conductivity data are plotted against the pore water conductivity to determine, sample by sample, the intrinsic formation factor and the surface conductivity. The intrinsic formation factor is related to porosity by Archie's law with an average value of the cementation exponent m of 2.45, indicating that only a small fraction of the connected pore space controls the transport properties. Both the surface and quadrature conductivities are found to be linearly related to the cation exchange capacity of the material, which was measured with the cobalt hexamine chloride method. Surface and quadrature conductivities are found to be proportional to each other like for sedimentary siliclastic rocks. A Stern layer polarization model is used to explain these experimental results. Despite the fact that the samples contain some magnetite (up to 5 per cent wt.), we were not able to identify the effect of this mineral on the complex conductivity spectra. These results are very encouraging in showing that galvanometric induced polarization measurements can be used in volcanic areas to separate the bulk from the surface conductivity and therefore to define some alteration attributes. Such a goal cannot be achieved with resistivity alone.

Description: We investigate the relationship between complex conductivity spectra and both permeability and pore mean size and distribution of 22 core samples (21 volcanic rocks and 1 clayey sandstone). The volcanic core samples were extracted from a wellbore drilled for the Humu‘ula Groundwater Research Project in the Humu‘ula saddle region between Mauna Kea and Mauna Loa volcanoes (Hawaii). The quadrature conductivity spectra of volcanic rocks exhibit a subtle, but generally detectable, relaxation frequency in the range 0.3 Hz to 45 kHz similar to the relaxation frequency observed for clayey sandstones. We find a fair relationship between this relaxation frequency and the pore size determined by mercury porosimetry. Combined with the intrinsic formation factor of the core samples, the relaxation frequency can be used as an indicator of the permeability of the material. The predicted values of the permeability are grossly consistent with the permeability values to air (in the range 0.001–100 mD) within two orders of magnitude. The measured permeability values are highly correlated to the peak of the pore size distribution determined from mercury porosimetry divided by the intrinsic formation factor. By fitting the complex conductivity spectra with the pore size distribution, we obtain the normalized chargeability of the core samples, which is, in turn, highly correlated to the measured cation exchange capacity.

Description: Carocin D is a bacteriocin produced by Pectobacterium carotovorum subsp. carotovorum Pcc21. Carocin D inhibits the growth of P . carotovorum subsp. carotovorum and closely related strains. Pectobacterium carotovorum subsp. carotovorum is a causative bacterium for soft rot disease and leads to severe economic losses. Bacteriocins recognize and interact with a specific membrane protein of target bacteria as a receptor. To identify the receptor responsible for carocin D recognition, mutants that underwent a phenotypic change from carocin D sensitivity to carocin D insensitivity were screened. Based on Tn 5 insertions, carocin D sensitivity was dependent on expression of the outer membrane protein OmpF. The insensitivity of the mutant (Pcc3MR) to carocin D was complemented with ompF from carocin D-sensitive strains, not from carocin D-resistant strains. The selectivity between sensitive and resistant strains could be attributed to variation in OmpFs in the cell-surface-exposed regions. Based on sequence analysis and complementation assays, it appears that carocin D uses OmpF as a receptor and is translocated by the TonB system. According to previously reported translocation mechanisms of colicins, OmpF works along with the TolA system rather than the TonB system. Therefore, the current findings suggest that carocin D is imported by a unique colicin-like bacteriocin translocation system.

Description: The Puyehue-Cordón Caulle Volcanic Complex (PCCVC) is one of the best examples of tectonic control on volcanism at the Southern Volcanic Zone of the Andes (southern Chile). The PCCVC comprises several volcanic centres that erupted dominantly SiO 2 -rich magmas at the intersection of the trench-parallel Liquiñe-Ofqui Fault Zone (LOFZ) and an inherited NW–SE basement structure. The PCCVC began an explosive and later effusive eruption on 2011 June 4 causing decimetre- to metre-scale surface deformation that was observed by a series of Envisat ASAR satellite scenes. We modelled this data and complemented it with time-series of two continuous GPS stations and seismicity recorded by a local network. Deformation during the first 3 days of the eruption can be modelled either by two point sources aligned with the NW–SE Cordón Caulle graben or by a closing dyke with a significant component of left-lateral motion along the graben. These models are discussed with respect to their implications on the estimated rheology and the eruption mechanism. GPS observations near the volcanic complex reveal an additional, more localised effect related to the LOFZ in the south of the complex. Coeruptive deformation at the main geological structures of the PCCVC is further supported by relocated seismicity, which is concentrated along the Cordón Caulle graben and to the western side of the LOFZ.

Description: In this study, equations are developed that predict for synthetic sedimentary rocks (clastics, carbonates and evapourates) thermal properties comprising thermal conductivity, specific heat capacity and thermal diffusivity. The rock groups are composed of mineral assemblages with variable contents of 15 major rock-forming minerals and porosities of 0–30 per cent. Petrophysical properties and their well-logging-tool-characteristic readings were assigned to these rock-forming minerals and to pore-filling fluids. Relationships are explored between each thermal property and other petrophysical properties (density, sonic interval transit time, hydrogen index, volume fraction of shale and photoelectric absorption index) using multivariate statistics. The application of these relations allows computing continuous borehole profiles for each rock thermal property. The uncertainties in the prediction of each property vary depending on the selected well-log combination. Best prediction is in the range of 2–8 per cent for the specific heat capacity, of 5–10 per cent for the thermal conductivity, and of 8–15 for the thermal diffusivity, respectively. Well-log derived thermal conductivity is validated by laboratory data measured on cores from deep boreholes of the Danish Basin, the North German Basin, and the Molasse Basin. Additional validation of thermal conductivity was performed by comparing predicted and measured temperature logs. The maximum deviation between these logs is 〈3 °C. The thermal-conductivity calculation allowed an evaluation of the depth range in which the palaeoclimatic effect on the subsurface temperature field can be observed in the North German Basin. This effect reduces the surface heat-flow density by 25 mW m –2 .

Description: Coal fires are severe hazards to environment, health and safety throughout the world. Efficient and economical extinguishing of these fires requires that the extent of the subsurface coal fires should be delineated. Electrical and electromagnetic methods have been used to detect coal fires in recent years. However, the resistivity change of coal-bearing rocks at high temperature is rarely investigated. The resistivity characteristics of coal fires at different temperatures and depths are seldomly researched as well. In this paper, we present the results of measurements of several coal-bearing rocks’ resistivity and permeability under high temperature. Two major causes for the change in resistivity with increasing temperature are recognized, there are the increase of charge carriers and thermal fracturing, of which the first one is probably the dominant cause. A set of 2-D simulations is carried out to compare the relation of resolution and efficiency of coal fires detection to temperature and depth when adopting the electrical resistance tomography. The simulation results show that the resolution and efficiency decrease with the decrease of temperature and the increase of depth. Finally, the electrical resistance tomography is used to delineate coal fires in the Anjialing Open Pit Mine. Most low-resistivity regions are verified as coal-fire areas according to the long-term monitoring of borehole temperature. The results indicate that the electrical resistance tomography can be used as a tool for the detection of coal fires.

Description: Pseudomonas putida KT2440 is a saprophytic and generally recognized as safe microorganism that plays important roles in the biodegradation and production of value-added chemicals. Chromosomal gene deletion of P. putida KT2440 usually involves time-consuming gene cloning, conjugal transfer and counterselection. Recently, we developed a P. putida KT2440 markerless gene deletion method based on recombineering and Cre/ loxP site-specific recombination. PCR-based Red recombineering circumvents the tedious cloning steps and is more amenable to high-throughput manipulation. Here we report an improved scarless gene deletion strategy based on recombineering and intron-encoded homing endonuclease I-SceI-mediated double-strand break repair. Sixteen drug exporter gene(s) were deleted and the minimal inhibition concentrations of the mutants to a variety of antibiotics were determined. The robustness of the procedure was also demonstrated by sequential deletion of five large genomic regions. Up to 59% recombination efficiency was achieved for a 54.8 kb deletion, and the efficiency of RecA-mediated double-strand break repair, which was boosted by Red recombinase, was nearly 100%. The strain with a 3.76% genome reduction showed an improved growth rate and transformation efficiency. The straightforward, time-saving and highly efficient scarless deletion approach has the potential to facilitate the genetic study, and biotechnological and environmental applications of P. putida KT2440.

Description: Human health has been seriously endangered by highly prevalent salmonellosis and multidrug-resistant Salmonella strains. Current vaccines suffer from variable immune-protective effects, so more effective ones are needed to control Salmonella infection . Bacterial ghosts have been produced by the expression of lysis gene E from bacteriophage PhiX174 and can be filled with considerable exogenous substances such as DNA or drugs as a novel platform. In this study, Salmonella enteritidis (SE) ghosts were developed and loaded with Neisseria gonorrhoeae porin B (porB) to construct a novel inactive vaccine. Our new studies show that SE ghosts loaded with porB displayed increased production of pro-inflammatory cytokines (IL-1β, IL-6, IL-10 and IL-12p70) in bone marrow-derived dendritic cells (BMDCs), and elicited significantly higher specific systemic and mucosal immune responses to Salmonella than SE ghosts alone. In addition, the novel porB-loaded ghosts conferred higher protective effects on virulent Salmonella challenge. For the first time, we demonstrate that N. gonorrhoeae porB, as a novel adjuvant, can increase the immunogenicity of SE ghosts. Our studies suggested that Salmonella enteritidis ghosts loaded with Neisseria gonorrhoeae porin B might be a useful mucosal Salmonella vaccine candidate for practical use in the future.

Description: Accurate information on thermal conductivity and thermal diffusivity of materials is of central importance in relation to geoscience and engineering problems involving the transfer of heat. Several methods, including the classical divided bar technique, are available for laboratory measurements of thermal conductivity, but much fewer for thermal diffusivity. We have generalized the divided bar technique to the transient case in which thermal conductivity, volumetric heat capacity and thereby also thermal diffusivity are measured simultaneously. As the density of samples is easily determined independently, specific heat capacity can also be determined. The finite element formulation provides a flexible forward solution for heat transfer across the bar, and thermal properties are estimated by inverse Monte Carlo modelling. This methodology enables a proper quantification of experimental uncertainties on measured thermal properties and information on their origin. The developed methodology was applied to various materials, including a standard ceramic material and different rock samples, and measuring results were compared with results applying traditional steady-state divided bar and an independent line-source method. All measurements show highly consistent results and with excellent reproducibility and high accuracy. For conductivity the obtained uncertainty is typically 1–3 per cent, and for diffusivity uncertainty may be reduced to about 3–5 per cent. The main uncertainty originates from the presence of thermal contact resistance associated with the internal interfaces in the bar. These are not resolved during inversion and it is imperative that they are minimized. The proposed procedure is simple and may quite easily be implemented to the many steady-state divided bar systems in operation. A thermally controlled bath, as applied here, may not be needed. Simpler systems, such as applying temperature-controlled water directly from a tap, may also be applied.

Description: The food-grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L. lactis to secrete murine IL-18. The mature murine IL-18 gene was inserted downstream of a nisin promoter in pNZ8149 plasmid and the construct was used to transform L. lactis NZ3900 . The transformants were selected on Elliker agar and confirmed by restriction enzyme digestion and sequencing. The expression and secretion of IL-18 protein was verified by SDS-PAGE, western blotting and ELISA. The biological activity of recombinant IL-18 was determined by its ability to induce interferon (IFN)- production in L. lactis co-cultured with murine splenic T cells. The amounts of IL-18 in bacterial lysates and supernatants were 3–4 μg mL –1 and 0.6–0.7 ng mL –1 , respectively. The successfully generated L. lactis strain that expressed biologically active murine IL-18 can be used to evaluate the possible therapeutic effects of IL-18 on mucosal surfaces.

Description: It is important to know the electrokinetic properties of crustal rocks for interpreting the conductivity mechanisms and seismoelectric phenomena during earthquakes and seismoelectric well logging. In this study, electrokinetic experiments are conducted using a special core-holder by employing an AC lock-in technique. A series of experiments are conducted on 10 sandstone samples to measure the streaming potentials and streaming currents, and the experiments on each sample are done at six different salinities. The streaming potential coefficient and streaming current coefficient are calculated from the measured streaming potentials and streaming currents. The experimental results show that streaming potential coefficient and streaming current coefficient decrease as the salinity increases. The dependence of these two coefficients on permeability and pore radius are analysed and compared with previous works. At low salinities, the streaming potential coefficient and streaming current coefficient increase with the increasing permeability and pore radius. At high salinities, the streaming potential coefficient (streaming current coefficient) almost share a same value for 10 different samples. This conclusion indicates that the differences of rock parameters can only be well recognized at lower salinities, and the electrokinetic signals are invalid at high salinities, which offers a restrictive condition for using the amplitude of electrokinetic signals to estimate rock parameters. The zeta-potential have also been estimated through combined measurements of streaming potential and streaming current. The surface conductivity and its contribution to electrokinetic effects are determined from a comparison of zeta-potentials by two different methods, and then the validation of the Helmholz–Smoluchowski equation for a capillary tube is tested in rocks. We also compare our date with theoretical and experimental works, and set up an expression about the relationship between zeta potential and salinity, which fits the experimental data well.

Description: We used ab initio molecular dynamics to calculate the elastic constants of MgSiO 3 , FeSiO 3 and (Fe 0.5 Mg 0.5 )SiO 3 perovskite under lower-most mantle conditions (136 GPa, 2000–4000 K). At 0 K, V p and V s for FeSiO 3 perovskite are 12 and 18 per cent lower, respectively than those for MgSiO 3 , in agreement with previous work. The difference in velocity, however, increases with temperature for high spin (but not low spin) states and at 4000 K, V p and V s for the end-member FeSiO 3 are, respectively, 19 and 34 per cent lower than those of MgSiO 3 . We find that (dln V s /dln V p ) T,P of Fe x Mg 1- x O 3 remains below 2 for all relevant iron concentrations and lower than the value of 3 seen in most ultralow velocity zones (ULVZ). Moreover, we are unable to match simultaneously the observed density increase and shear wave velocity reduction shown by ULVZs by increasing the ferrous iron content in perovskite except for the largest density increases and the smallest shear wave decreases observed in ULVZs. Assuming that current seismic estimates of ULVZ properties are accurate, our results suggest that ferrous iron enrichment alone cannot explain ULVZs.