Analysis of carbon substrates used by Listeria monocytogenes during growth in J774A.1 macrophages suggests a bipartite intracellular metabolism

Intracellular bacterial pathogens (IBPs) are dependent on various nutrients provided by the host cells. Different strategies may therefore be necessary to adapt the intracellular metabolism of IBPs to the host cells. The specific carbon sources, the catabolic pathways participating in their degradation, and the biosynthetic performances of IBPs are still poorly understood. In this report, we have exploited the technique of C-13-isotopologue profiling to further study the carbon metabolism of Listeria monocytogenes by using the EGDe wild-type strain and mutants (defective in the uptake and/or catabolism of various carbon compounds) replicating in J774A.1 macrophages. For this goal, the infected macrophages were cultivated in the presence of [1, 2-C-13(2)]glucose, [U-C-13(3)]glycerol, [U-C-13(3)]pyruvate, [U-C-13(3)]lactate, or a mix of [U-C-13]amino acids. GC/MS-based isotopologue profiling showed efficient utilization of amino acids, glucose 6-phosphate, glycerol, and (at a low extent) also of lactate but not of pyruvate by the IBPs. Most amino acids imported from the host cells were directly used for bacterial protein biosynthesis and hardly catabolized. However, Asp was de novo synthesized by the IBPs and not imported from the host cell. As expected, glycerol was catabolized via the ATP-generating lower part of the glycolytic pathway, but apparently not used for gluconeogenesis. The intermediates generated from glucose 6-phosphate in the upper part of the glycolytic pathway and the pentose phosphate shunt likely serve primarily for anabolic purposes (probably for the biosynthesis of cell wall components and nucleotides). This bipartite bacterial metabolism which involves at least two major carbon substrates-glycerol mainly for energy supply and glucose 6-phosphate mainly for indispensible anabolic performances-may put less nutritional stress on the infected host cells, thereby extending the lifespan of the host cells to the benefit of the IBPs

Reakcija 133Cs(d, 3He)132Xe

The 133Cs(d, 3He)132Xe reaction was measured at the Munich Q3D magnetic spectrograph with high resolution. The spectra were obtained at beam energies of Ed = 27 MeV and Ed = 28 MeV. The corresponding angles of the spectrograph were 30◦ and 50◦ , respectively. The achieved FWHM was between 7 keV and 10 keV and the energies of the levels could be determined with a precision between 0.5 keV and 1 keV.Pomoću Q3D magnetskog spektrografa istraživali smo reakciju 133Cs(d, 3He)132Xe pri visokom razlučivanju. Izmjereni su spektri za energiju upadnih deuterona od 27 i 28 MeV, na kutovima raspršenja 30◦ odnosno 50◦ . Postignuto je razlučivanje između 7 i 10 keV, a energije stanja u 132Xe su određene na točnost između 0.5 i 1 keV

Feedback control of arm movements using Neuro-Muscular Electrical Stimulation (NMES) combined with a lockable, passive exoskeleton for gravity compensation

Within the European project MUNDUS, an assistive framework was developed for the support of arm and hand functions during daily life activities in severely impaired people. This contribution aims at designing a feedback control system for Neuro-Muscular Electrical Stimulation (NMES) to enable reaching functions in people with no residual voluntary control of the arm and shoulder due to high level spinal cord injury. NMES is applied to the deltoids and the biceps muscles and integrated with a three degrees of freedom (DoFs) passive exoskeleton, which partially compensates gravitational forces and allows to lock each DOF. The user is able to choose the target hand position and to trigger actions using an eyetracker system. The target position is selected by using the eyetracker and determined by a marker-based tracking system using Microsoft Kinect. A central controller, i.e., a finite state machine, issues a sequence of basic movement commands to the real-time arm controller. The NMES control algorithm sequentially controls each joint angle while locking the other DoFs. Daily activities, such as drinking, brushing hair, pushing an alarm button, etc., can be supported by the system. The robust and easily tunable control approach was evaluated with five healthy subjects during a drinking task. Subjects were asked to remain passive and to allow NMES to induce the movements. In all of them, the controller was able to perform the task, and a mean hand positioning error of less than five centimeters was achieved. The average total time duration for moving the hand from a rest position to a drinking cup, for moving the cup to the mouth and back, and for finally returning the arm to the rest position was 71 s.EC/FP7/248326/EU/MUltimodal Neuroprostesis for Daily Upper limb Support/MUNDU

Spectroscopija 125Te (n,γ), (d,p) i (3He,α) reakcijama

Single γ-ray spectra and γγ-coincidences, following thermal neutron capture in 124Te, were measured with semiconductor detectors at the light-water reactor LWR - 15 at Řež. Intensities of g transitions in 125Te were normalized using the absolute intensity of 7.8% of the 6620 keV line in 36Cl. The high resolution (d,p) measurements were performed with 17 MeV deuterons, using the Q3D spectrograph at two scattering angles of 15° and 30°. Spectra were recorded in the range up to 3.3 MeV and calibrated using the proton peaks with l =1, and the corresponding level energies were determined in the thermal neutron capture reaction. The (3He,α) experiment was carried out with a 32 MeV He beam at the angle of 10°. The spectrum was recorded in one run by means of a large detector in a range up to approximately 4.7 MeV. The absolute intensities were determined by measuring the total beam current.Mjerili su se izravni γ- i sudesni γ-γ-spektri nakon uhvata termičkih neutrona u 124Te. Mjerenja (d,p) reakcija visokog razlučivanja na toj jezgri izvela su se s deuteronima 17 MeV na kutovima od 15° i 30° , pomoću Q3D spektrografa. Također se mjerio spektar u reakciji (3He,α) na 10° . Na osnovi tih mjerenja utvrđena su mnoga nova stanja 125Te na energijama uzbude do oko 4.5 MeV. Razlika grananja 403/443 keV iz izravnih i sudesnih γ-spektara ukazuje na jaku primjesu E2 u primarnom prijelazu od 6125 keV

Spectroscopija 125Te (n,γ), (d,p) i (3He,α) reakcijama

Single γ-ray spectra and γγ-coincidences, following thermal neutron capture in 124Te, were measured with semiconductor detectors at the light-water reactor LWR - 15 at Řež. Intensities of g transitions in 125Te were normalized using the absolute intensity of 7.8% of the 6620 keV line in 36Cl. The high resolution (d,p) measurements were performed with 17 MeV deuterons, using the Q3D spectrograph at two scattering angles of 15° and 30°. Spectra were recorded in the range up to 3.3 MeV and calibrated using the proton peaks with l =1, and the corresponding level energies were determined in the thermal neutron capture reaction. The (3He,α) experiment was carried out with a 32 MeV He beam at the angle of 10°. The spectrum was recorded in one run by means of a large detector in a range up to approximately 4.7 MeV. The absolute intensities were determined by measuring the total beam current.Mjerili su se izravni γ- i sudesni γ-γ-spektri nakon uhvata termičkih neutrona u 124Te. Mjerenja (d,p) reakcija visokog razlučivanja na toj jezgri izvela su se s deuteronima 17 MeV na kutovima od 15° i 30° , pomoću Q3D spektrografa. Također se mjerio spektar u reakciji (3He,α) na 10° . Na osnovi tih mjerenja utvrđena su mnoga nova stanja 125Te na energijama uzbude do oko 4.5 MeV. Razlika grananja 403/443 keV iz izravnih i sudesnih γ-spektara ukazuje na jaku primjesu E2 u primarnom prijelazu od 6125 keV

Widely Targeted Metabolomics Based on Large-Scale MS/MS Data for Elucidating Metabolite Accumulation Patterns in Plants

Metabolomics is an ‘omics’ approach that aims to analyze all metabolites in a biological sample comprehensively. The detailed metabolite profiling of thousands of plant samples has great potential for directly elucidating plant metabolic processes. However, both a comprehensive analysis and a high throughput are difficult to achieve at the same time due to the wide diversity of metabolites in plants. Here, we have established a novel and practical metabolomics methodology for quantifying hundreds of targeted metabolites in a high-throughput manner. Multiple reaction monitoring (MRM) using tandem quadrupole mass spectrometry (TQMS), which monitors both the specific precursor ions and product ions of each metabolite, is a standard technique in targeted metabolomics, as it enables high sensitivity, reproducibility and a broad dynamic range. In this study, we optimized the MRM conditions for specific compounds by performing automated flow injection analyses with TQMS. Based on a total of 61,920 spectra for 860 authentic compounds, the MRM conditions of 497 compounds were successfully optimized. These were applied to high-throughput automated analysis of biological samples using TQMS coupled with ultra performance liquid chromatography (UPLC). By this analysis, approximately 100 metabolites were quantified in each of 14 plant accessions from Brassicaceae, Gramineae and Fabaceae. A hierarchical cluster analysis based on the metabolite accumulation patterns clearly showed differences among the plant families, and family-specific metabolites could be predicted using a batch-learning self-organizing map analysis. Thus, the automated widely targeted metabolomics approach established here should pave the way for large-scale metabolite profiling and comparative metabolomics

Identification of metabolic and biomass QTL in Arabidopsis thaliana in a parallel analysis of RIL and IL populations

Plant growth and development are tightly linked to primary metabolism and are subject to natural variation. In order to obtain an insight into the genetic factors controlling biomass and primary metabolism and to determine their relationships, two Arabidopsis thaliana populations [429 recombinant inbred lines (RIL) and 97 introgression lines (IL), derived from accessions Col-0 and C24] were analyzed with respect to biomass and metabolic composition using a mass spectrometry-based metabolic profiling approach. Six and 157 quantitative trait loci (QTL) were identified for biomass and metabolic content, respectively. Two biomass QTL coincide with significantly more metabolic QTL (mQTL) than statistically expected, supporting the notion that the metabolic profile and biomass accumulation of a plant are linked. On the same basis, three out the six biomass QTL can be simulated purely on the basis of metabolic composition. QTL based on analysis of the introgression lines were in substantial agreement with the RIL-based results: five of six biomass QTL and 55% of the mQTL found in the RIL population were also found in the IL population at a significance level of P ≤ 0.05, with >80% agreement on the allele effects. Some of the differences could be attributed to epistatic interactions. Depending on the search conditions, metabolic pathway-derived candidate genes were found for 24–67% of all tested mQTL in the database AraCyc 3.5. This dataset thus provides a comprehensive basis for the detection of functionally relevant variation in known genes with metabolic function and for identification of genes with hitherto unknown roles in the control of metabolism

MUNDUS project : MUltimodal neuroprosthesis for daily upper limb support

Background: MUNDUS is an assistive framework for recovering direct interaction capability of severely motor impaired people based on arm reaching and hand functions. It aims at achieving personalization, modularity and maximization of the user’s direct involvement in assistive systems. To this, MUNDUS exploits any residual control of the end-user and can be adapted to the level of severity or to the progression of the disease allowing the user to voluntarily interact with the environment. MUNDUS target pathologies are high-level spinal cord injury (SCI) and neurodegenerative and genetic neuromuscular diseases, such as amyotrophic lateral sclerosis, Friedreich ataxia, and multiple sclerosis (MS). The system can be alternatively driven by residual voluntary muscular activation, head/eye motion, and brain signals. MUNDUS modularly combines an antigravity lightweight and non-cumbersome exoskeleton, closed-loop controlled Neuromuscular Electrical Stimulation for arm and hand motion, and potentially a motorized hand orthosis, for grasping interactive objects. Methods: The definition of the requirements and of the interaction tasks were designed by a focus group with experts and a questionnaire with 36 potential end-users. Five end-users (3 SCI and 2 MS) tested the system in the configuration suitable to their specific level of impairment. They performed two exemplary tasks: reaching different points in the working volume and drinking. Three experts evaluated over a 3-level score (from 0, unsuccessful, to 2, completely functional) the execution of each assisted sub-action. Results: The functionality of all modules has been successfully demonstrated. User’s intention was detected with a 100% success. Averaging all subjects and tasks, the minimum evaluation score obtained was 1.13 ± 0.99 for the release of the handle during the drinking task, whilst all the other sub-actions achieved a mean value above 1.6. All users, but one, subjectively perceived the usefulness of the assistance and could easily control the system. Donning time ranged from 6 to 65 minutes, scaled on the configuration complexity. Conclusions: The MUNDUS platform provides functional assistance to daily life activities; the modules integration depends on the user’s need, the functionality of the system have been demonstrated for all the possible configurations, and preliminary assessment of usability and acceptance is promising

Atlantic Water advection versus sea-ice advances in the eastern Fram Strait during the last 9 ka - multiproxy evidence for a two-phase Holocene

A sediment core from the West Spitsbergen continental margin was studied to reconstruct climate and paleoceanographic variability during the last ~9 ka in the eastern Fram Strait. Our multiproxy evidence suggests that the establishment of the modern oceanographic configuration in the eastern Fram Strait occurred stepwise, in response to the postglacial sea-level rise and the related onset of modern sea-ice production on the shallow Siberian shelves. The late Early and Mid Holocene interval (9 to 5 ka) was generally characterized by relatively unstable conditions. High abundance of the subpolar planktic foraminifer species Turborotalita quinqueloba implies strong intensity of Atlantic Water (AW) inflow with high productivity and/or high AW temperatures, resulting in a strong heat flux to the Arctic. A series of short-lived cooling events (8.2, 6.9. and 6.1 ka) occurred superimposed on the warm late Early and Mid Holocene conditions. Our proxy data imply that simultaneous to the complete postglacial flooding of Arctic shallow shelves and the initiation of modern sea-ice production, strong advance of polar waters initiated modern oceanographic conditions in the eastern Fram Strait at ~5.2 ka. The Late Holocene was marked by the dominance of the polar planktic foraminifer species Neogloboquadrina pachyderma, a significant expansion of sea ice/icebergs, and strong stratification of the water column. Although planktic foraminiferal assemblages as well as sea surface and subsurface temperatures suggest a return of slightly strengthened advection of subsurface Atlantic Water after 3 ka, a relatively stable cold-water layer prevailed at the sea surface and the study site was probably located within the seasonally fluctuating marginal ice zone during the Neoglacial period