ALBERT

Description: Although nitric oxide (NO) is an important signaling molecule in bacteria and higher organisms, excessive intracellular NO is highly reactive and dangerous. Therefore, living cells need strict regulation systems for cellular NO homeostasis. Recently, we discovered that Streptomyces coelicolor A3(2) retains the nitrogen oxide cycle (NO 3 – -〉NO 2 – -〉NO-〉NO 3 – ) and nitrite removal system. The nitrogen oxide cycle regulates cellular NO levels, thereby controlling secondary metabolism initiation (red-pigmented antibiotic, RED production) and morphological differentiation. Nitrite induces gene expression in neighboring cells, suggesting another role for this cycle as a producer of transmittable intercellular communication molecules. Here, we demonstrated that ammonium-producing nitrite reductase (NirBD) is involved in regulating NO homeostasis in S. coelicolor A3(2). NirBD was constitutively produced in culture independently of GlnR, a known transcriptional factor. NirBD cleared the accumulated nitrite from the medium. Nir deletion mutants showed increased NO-dependent gene expression at later culture stages, whereas the wild-type M145 showed decreased expression, suggesting that high NO concentration was maintained in the mutant. Moreover, the nir deletion mutant produced more RED than that produced by the wild-type M145. These results suggest that NO 2 – removal by NirBD is important to regulate NO homeostasis and to complete NO signaling in S. coelicolor .

Description: Earlier, vitamin C was demonstrated to sterilize Mycobacterium tuberculosis culture via Fenton's reaction at high concentration. It alters the regulatory pathways associated with stress response and dormancy. Since (p)ppGpp is considered to be the master regulator of stress response and is responsible for bacterial survival under stress, we tested the effect of vitamin C on the formation of (p)ppGpp. In vivo estimation of (p)ppGpp showed a decrease in (p)ppGpp levels in vitamin C-treated M. smegmatis cells in comparison to the untreated cells. Furthermore, in vitro (p)ppGpp synthesis using Rel MSM enzyme was conducted in order to confirm the specificity of the inhibition in the presence of variable concentrations of vitamin C. We observed that vitamin C at high concentration can inhibit the synthesis of (p)ppGpp. We illustrated binding of vitamin C to Rel MSM by isothermal titration calorimetry. Enzyme kinetics was followed where K 0.5 was found to be increased with the concomitant reduction of V max value suggesting mixed inhibition. Both long-term survival and biofilm formation were inhibited by vitamin C. The experiments suggest that vitamin C has the potential to be developed as the inhibitor of (p)ppGpp synthesis and stress response, at least in the concentration range used here.

Description: The culturability of Escherichia coli , Ralstonia eutropha and Bacillus subtilis after incubation in phosphate-buffered saline at either 5°C or 30°C was determined. The culturability of B. subtilis showed little dependence on temperature. The culturability of E. coli rapidly decreased at 30°C but remained almost constant at 5°C. In contrast, the culturability of R. eutropha decreased by three orders of magnitude at 5°C within 24 h but only moderately decreased (one order of magnitude) at 30°C. Remarkably, prolonged incubation of R. eutropha at 30°C resulted in a full recovery of colony forming units in contrast to only a partial recovery at 5°C. Ralstonia eutropha cells at 30°C remained culturable for 3 weeks while culturability at 5°C constantly decreased. The effect of temperature was significantly stronger in a polyhydroxybutyrate-negative mutant. Our data show that accumulated polyhydroxybutyrate has a cold-protective function and can prevent R. eutropha entering the viable but not culturable state.

Description: This is the second of three papers investigating properties of titanohematite-bearing quartzo-feldspathic rocks that create a significant remanent magnetic anomaly in the Modum District, South Norway. The first paper provided initial magnetic results, mineralogical characterization and evidence for the presence of lamellar magnetism. In this paper, knowledge of lamellar magnetic properties is explored through experiments where ilmenite lamellae were magnetized below 57 K, and interact magnetically along interfaces with the titanohematite host. Samples with known NRM directions were placed in specific orientations in an MPMS then cooled in zero field to 5 K, where hysteresis loops were measured in fields up to 5 Tesla. This assured that results were ultimately related to the natural lamellar magnetism produced during cooling ~1 billion years ago. In a second set of experiments the same oriented samples, were subjected to a +5 Tesla (T) field then field cooled to 5 K before hysteresis experiments. The first experiments consistently produced asymmetric shifted hysteresis loops with two loop separations, one in a positive field and one in a negative field. Without exception, when the NRM was oriented toward the negative field end of the MPMS, the bimodal loop showed a dominant loop separation in a positive field. By contrast, when the NRM was oriented toward the positive field end of the MPMS, the bimodal loop showed a dominant loop separation in a negative field. Both observations are consistent with antiferromagnetic coupling between the hard magnetization of ilmenite and the more easily shifted lamellar magnetism of the hematite. The bimodal nature of the loops indicates that the NRMs are vector sums of natural lamellar moments, which are oriented both positively and negatively, and that these opposite moments control the orientations of ilmenite magnetizations when cooling through 57 K. Here, extreme exchange biases up to 1.68 T were measured. The second set of experiments produced asymmetric shifted hysteresis loops with one opening always in the negative field. These observations indicate that the +5 T field applied at room temperature rotated the hematite lamellar magnetism in a positive direction, so that upon cooling all the ilmenite lamellae acquired negative magnetic moments, thus causing unimodal negatively shifted loops. Here, the largest exchange bias among the unimodal loops was only 0.7 T. These results will be used in paper III to build a better understanding of lamellar magnetism at the atomic layer scale.

Description: The temperature dependence of magnetic susceptibility ( –T curves) and magnetization ( M–T curves) has been used as a routine rock magnetic tool to characterize the magnetic mineralogy and magnetic granulometry of Chinese loess/palaeosols. However, palaeoclimatic interpretation of these thermomagnetic analyses remains controversial. In the present study, total organic carbon (TOC), thermomagnetic and low-temperature magnetic experiments on Mid-Late Pleistocene loess/palaeosols in Central Asia and the Chinese Loess Plateau (CLP) have been conducted. We found that the M ( T ) cooling curves at room temperature were mostly lower than the corresponding heating curves, whereas for the ( T ) analyses the cooling curves at room temperature were always much higher than the heating curves. Low-temperature magnetic measurements demonstrated that a large amount of superparamagnetic ferrimagnetic particles were produced during the thermal treatment and resulted in the aforementioned differences. This finding further indicated that the use of the M–T curves to estimate the relative content of maghemite in the loess/palaeosols from the CLP was problematic. In addition, a positive correlation exists between the TOC and the frequency-dependent susceptibility ( FD ) in the CLP, suggesting that stronger pedogenesis would result in the simultaneous increase in the content of both maghemite and organic matter. Consequently, the parameters 1 (representing the relative content of pedogenic maghemite), 2 ([ ph – ] + 1 ) and ph (related to the organic matter concentration), which can be calculated from the –T analyses, can potentially be used as new indicators of pedogenesis and palaeoclimate in Central Asia and the CLP.

Description: Pseudomonas plecoglossicida is a facultative fish pathogen. Recent studies showed that P. plecoglossicida infection in fish was associated with temperature. The aim of this study was to compare the secretomes of P. plecoglossicida cultured in vitro at representative temperatures for pathogenic (20°C) and less pathogenic (30°C) phenotypes. Thirteen proteins in the culture supernatants of P. plecoglossicida showed significant difference in abundance at 20 vs. 30°C. Four proteins were strongly increased at 20°C, including two hemolysin co-regulated proteins (Hcp) that are part of the bacterial type VI secretion system (T6SS), flagellin and an unknown protein. Immunoblot analysis verified the induced secretion of Hcps at 20°C. Furthermore, the upregulation of Hcps at 20°C was confirmed at transcriptional level by RT-qPCR analysis, which also demonstrated the induction of expression of other T6SS-related genes at 20°C. Taken together, we demonstrate the presence of two functionally active T6SS proteins in fish pathogenic P. plecoglossicida strains, as evidenced by the secretion of the T6SS substrate Hcp, the production of which were found to be controlled by temperature. Our findings also support efforts to develop vaccines targeting secreted virulence factors as prophylactic strategies for diseases in fish caused by P. plecoglossicida .

Description: Previous magnetotelluric (MT) studies of the high-temperature Coso geothermal system in California identified a subvertical feature of low resistivity (2–5 Ohm m) and appreciable lateral extent (〉1 km) in the producing zone of the East Flank field. However, these models could not reproduce gross 3-D effects in the recorded data. We perform 3-D full-tensor inversion and retrieve a resistivity model that out-performs previous 2-D and 3-D off-diagonal models in terms of its fit to the complete 3-D MT data set as well as the degree of modelling bias. Inclusion of secondary Z xx and Z yy data components leads to a robust east-dip (60) to the previously identified conductive East Flank reservoir feature, which correlates strongly with recently mapped surface faults, downhole well temperatures, 3-D seismic reflection data, and local microseismicity. We perform synthetic forward modelling to test the best-fit dip of this conductor using the response at a nearby MT station. We interpret the dipping conductor as a fractured and fluidized compartment, which is structurally controlled by an unmapped blind East Flank fault zone.

Description: The magnetic susceptibility measured in alternating field can in general be resolved into a component that is in-phase with the applied field and a component that is out-of-phase. While in non-conductive diamagnetic, paramagnetic and many ferromagnetic materials the phase is effectively zero, in some ferromagnetic minerals, such as pyrrhotite, hematite, titanomagnetite or small magnetically viscous grains of magnetite, it is clearly non-zero. The anisotropy of out-of-phase susceptibility (opAMS) can then be used as a tool for the direct determination of the magnetic subfabrics of the minerals with non-zero phase. The error in determination of out-of-phase susceptibility non-linearly increases with decreasing phase angle. This may result in imprecise determination of the opAMS in specimens with very low phase angle. The degree of opAMS is higher than that of ipAMS, which may in contrast result in slightly increasing precision n the opAMS determination. It is highly recommended to inspect the results of the statistical tests of each specimen and to exclude the specimens whose opAMS is determined with insufficient precision from further processing. In rocks, whose magnetism is dominated by the mineral with non-zero out-of-phase susceptibility, the principal directions of the opAMS and ipAMS are virtually coaxial, while the degree of opAMS is higher than that of ipAMS. In some specific cases, the opAMS provides us with similar data to those provided by anisotropies of low-field dependent susceptibility and frequency-dependent susceptibility. The advantage of the opAMS compared to the other two anisotropies is its simultaneous measurement with the ipAMS during one measuring process, while the other two anisotropies require the AMS measurements in several fields or at least at two operating frequencies.

Description: The spectral complex conductivity of a water-bearing sand during interaction with carbon dioxide (CO 2 ) is influenced by multiple, simultaneous processes. These processes include partial saturation due to the replacement of conductive pore water with CO 2 and chemical interaction of the reactive CO 2 with the bulk fluid and the grain-water interface. We present a laboratory study on the spectral induced polarization of water-bearing sands during exposure to and flow-through by CO 2 . Conductivity spectra were measured successfully at pressures up to 30 MPa and 80 °C during active flow and at steady-state conditions concentrating on the frequency range between 0.0014 and 100 Hz. The frequency range between 0.1 and 100 Hz turned out to be most indicative for potential monitoring applications. The presented data show that the impact of CO 2 on the electrolytic conductivity may be covered by a model for pore-water conductivity, which depends on salinity, pressure and temperature and has been derived from earlier investigations of the pore-water phase. The new data covering the three-phase system CO 2 -brine-sand further show that chemical interaction causes a reduction of surface conductivity by almost 20 per cent, which could be related to the low pH-value in the acidic environment due to CO 2 dissolution and the dissociation of carbonic acid. The quantification of the total CO 2 effect may be used as a correction during monitoring of a sequestration in terms of saturation. We show that this leads to a correct reconstruction of fluid saturation from electrical measurements. In addition, an indicator for changes of the inner surface area, which is related to mineral dissolution or precipitation processes, can be computed from the imaginary part of conductivity. The low frequency range between 0.0014 and 0.1 Hz shows additional characteristics, which deviate from the behaviour at higher frequencies. A Debye decomposition approach is applied to isolate the feature dominating the data at low frequencies. We conclude from our study that electrical conductivity is not only a highly sensitive indicator for CO 2 saturation in pore space. When it is measured in its full spectral and complex form it contains additional information on the chemical state of the system, which holds the potential of getting access to both saturation and interface properties with one monitoring method.

Description: The 69 Ma Prospector Mountain stock is located in southwestern Yukon in the northern Canadian Cordillera. This massive monzonite-syenogranite stock is thought to be the intrusive volcanic centre for the surrounding coeval Carmacks Group volcanics. Anomalous palaeomagnetic data from these volcanics have provided the only evidence for the commonly posited hypothesis that the Yukon-Tanana terrane (YTT) was part of the far-travelled (1950 ± 600 km northward) ‘Baja BC’ terrane from 70 to 50 Ma. All other geologic evidence and averaged palaeomagnetic data support a northward displacement of ~415 ± 15 km. This study provides a direct test of the Carmacks volcanics’ estimate and examines the possible causes of its anomalous results. Both the stock and volcanics are unmetamorphosed and rest unconformably on metamorphosed basement rocks of the YTT. Palaeomagnetic and mineral magnetic results from 17 of 19 tested sites (218 specimens) in the Prospector stock and its peripheral skarn isolated a stable thermoremanent magnetization (TRM) in magnetite or low-Ti titanomagnetite that was mostly determined on demagnetization between temperatures of 500 and 580 °C. The TRM has a direction of Decl. = 8.3°, Incl. = 82.4° ( N = 17, k = 71.9, α 95 = 4.2°), providing a non-significant northwards translation estimate of 70 ± 880 km for the YTT. The normal-polarity TRM direction at Prospector Mountain provides a highly significant palaeomagnetic reversals test with the reversed-polarity TRM of the 70 Ma Swede Dome stock, another volcanic centre of the Carmacks Group about 190 km to the north. The test affirms to a high probability that both stocks carry primary TRMs and have not been tectonically tilted significantly since emplacement. Combining the palaeopoles for these two stocks with that for the 75 Ma Mount Lorne volcanic centre stock about 210 km south of Prospector Mountain yields a combined northward translation estimate of 330 ± 400 km for the YTT since ~71 Ma. This estimate agrees closely with the 415 ± 15 km estimate from geological constraints on strike-slip motion along the inboard Tintina fault zone since ~70 Ma, and with the palaeomagnetic estimate of 650 ± 450 km northward since ~108 Ma from four mid-Cretaceous batholiths. In contrast, the palaeomagnetic directions from four areas of the ~70 Ma Carmacks volcanics are poorly clustered. Nonetheless their estimates of ~1900 km northward have been used for a quarter of a century to support a far-travelled ‘Baja BC’ tectonic model for the YTT. This paper discusses and concludes that the anomalous far-travelled estimates from the Carmacks volcanics are attributable to the unfortunate additive effects of inadequate averaging of secular variation, dipole offset error, unrecognized primary dip and other possible causes.