Slow conduction in mixed cultured strands of primary ventricular cells and stem cell-derived cardiomyocytes

Modern concepts for the treatment of myocardial diseases focus on novel cell therapeutic strategies involving stem cell-derived cardiomyocytes (SCMs). However, functional integration of SCMs requires similar electrophysiological properties as primary cardiomyocytes (PCMs) and the ability to establish intercellular connections with host myocytes in order to contribute to the electrical and mechanical activity of the heart. The aim of this project was to investigate the properties of cardiac conduction in a co-culture approach using SCMs and PCMs in cultured cell strands. Murine embryonic SCMs were pooled with fetal ventricular cells and seeded in predefined proportions on microelectrode arrays to form patterned strands of mixed cells. Conduction velocity (CV) was measured during steady state pacing. SCM excitability was estimated from action potentials measured in single cells using the patch clamp technique. Experiments were complemented with computer simulations of conduction using a detailed model of cellular architecture in mixed cell strands. CV was significantly lower in strands composed purely of SCMs (5.5 ± 1.5 cm/s, n = 11) as compared to PCMs (34.9 ± 2.9 cm/s, n = 21) at similar refractoriness (100% SCMs: 122 ± 25 ms, n = 9; 100% PCMs: 139 ± 67 ms, n = 14). In mixed strands combining both cell types, CV was higher than in pure SCMs strands, but always lower than in 100% PCM strands. Computer simulations demonstrated that both intercellular coupling and electrical excitability limit CV. These data provide evidence that in cultures of murine ventricular cardiomyocytes, SCMs cannot restore CV to control levels resulting in slow conduction, which may lead to reentry circuits and arrhythmias.

Learning -dependent plasticity of movement representations in the primate primary motor cortex

Primary motor cortex (M1) is involved in the production of voluntary movement and contains a complete functional representation, or map, of the skeletal musculature. This functional map can be altered by pathological experiences, such as peripheral nerve injury or stroke, by pharmacological manipulation, and by behavioral experience. The process by which experience-dependent alterations of cortical function occur is termed plasticity. In this thesis, plasticity of M1 functional organization as a consequence of behavioral experience was examined in adult primates (squirrel monkeys). Maps of movement representations were derived under anesthesia using intracortical microstimulation, whereby a microelectrode was inserted into the cortex to electrically stimulate corticospinal neurons at low current levels and evoke movements of the forelimb, principally of the hand. Movement representations were examined before and at several times after training on behavioral tasks that emphasized use of the fingers. Two behavioral tasks were utilized that dissociated the repetition of motor activity from the acquisition of motor skills. One task was easy to perform, and as such promoted repetitive motor activity without learning. The other task was more difficult, requiring the acquisition of motor skills for successful performance. Kinematic analysis indicated that monkeys used a consistent set of forelimb movements during pellet extractions. Functional mapping revealed that repetitive motor activity during the easier task did not produce plastic changes in movement representations. Instead, map plasticity, in the form of selective expansions of task-related movement representations, was only produced following skill acquisition on the difficult task. Additional studies revealed that, in general, map plasticity persisted without further training for up to three months, in parallel with the retention of task-related motor skills. Also, extensive additional training on the small well task produced further improvements in performance, and further changes in movement maps. In sum, these experiments support the following three conclusions regarding the role of M1 in motor learning. First, behaviorally-driven plasticity is learning-dependent, not activity-dependent. Second, plastic changes in M1 functional representations represent a neural correlate of acquired motor skills. Third, the persistence of map plasticity suggests that M1 is part of the neural substrate for the memory of motor skills. ^

Chemistry of sediment profile GeoB4906

Mineralization of organic matter and the subsequent dissolution of calcite were simulated for surface sediments of the upper continental slope off Gabon by using microsensors to measure O2, pH, pCO2 and Ca2+ (in situ), pore-water concentration profiles of NO3-, NH4+, Fe2+, and Mn2+ and SO42- (ex situ), as well as sulfate reduction rates derived from incubation experiments. The transport and reaction model CoTReM was used to simulate the degradation of organic matter by O2, [NO3]-, Fe(OH)3 and [SO4]2-, reoxidation reactions involving Fe2+ and Mn2+, and precipitation of FeS. Model application revealed an overall rate of organic matter mineralization amounting to 50 µmol C cm**-2 yr**-1, of which 77% were due to O2, 17% to [NO3]- and 3% to Fe(OH)3 and 3% to [SO4]2-. The best fit for the pH profile was achieved by adapting three different dissolution rate constants of calcite ranging between 0.01 and 0.5% d-1 and accounting for different calcite phases in the sediment. A reaction order of 4.5 was assumed in the kinetic rate law. A CaCO3 flux to the sediment was estimated to occur at a rate of 42 g m**-2 yr**-1 in the area of equatorial upwelling. The model predicts a redissolution flux of calcite amounting to 36 g m**-2 yr**-1, thus indicating that ~90% of the calcite flux to the sediment is redissolved.

Mats of psychrophilic thiotrophic bacteria associated with cold seeps of the Barents Sea

This study focused on the bacterial diversity associated with microbial mats of deep-sea cold seeps at the Norwegian continental margin. Study sites included the Storegga and Nyegga areas as well as the Håkon Mosby mud volcano, where the mats occurred at temperatures permanently close to the freezing point of seawater. Two visually different mat types, i.e. small gray mats and extensive white mats, were studied with the aim to determine the identity of the mat-forming sulfide oxidizers, and to investigate which environmental factors (e.g. sulfate reduction and methane oxidation rates) shown here could explain the observed diversity. Sequence data have been submitted to the EMBL database under accession No. FR847864-FR847887 (giant sulfur bacteria), No. FR827864 (Menez Gwen filament; see Supplementary Material) and No. FR875365-FR877509 (except FR875905; remaining partial sequences).

Geochemistry and soil characteristics of sediment core GeoB4234

Stable isotopes of sedimentary nitrogen and organic carbon are widely used as proxy variables for biogeochemical parameters and processes in the water column. In order to investigate alterations of the primary isotopic signal by sedimentary diagenetic processes, we determined concentrations and isotopic compositions of inorganic nitrogen (IN), organic nitrogen (ON), total nitrogen (TN), and total organic carbon (TOC) on one short core recovered from sediments of the eastern subtropical Atlantic, between the Canary Islands and the Moroccan coast. Changes with depth in concentration and isotopic composition of the different fractions were related to early diagenetic conditions indicated by pore water concentrations of oxygen, nitrate, and ammonium. Additionally, the nature of the organic matter was investigated by Rock-Eval pyrolysis and microscopic analysis. A decrease in ON during aerobic organic matter degradation is accompanied by an increase of the 15N/14N ratio. Changes in the isotopic composition of ON can be described by Rayleigh fractionation kinetics which are probably related to microbial metabolism. The influence of IN depleted in 15N on the bulk sedimentary (TN) isotope signal increases due to organic matter degradation, compensating partly the isotopic changes in ON. In anoxic sediments, fixation of ammonium between clay lattices results in a decrease of stable nitrogen isotope ratio of IN and TN. Changes in the carbon isotopic composition of TOC have to be explained by Rayleigh fractionation in combination with different remineralization kinetics of organic compounds with different isotopic composition. We have found no evidence for preferential preservation of terrestrial organic carbon. Instead, both TOC and refractory organic carbon are dominated by marine organic matter. Refractory organic carbon is depleted in 13C compared to TOC.

Benthic oxygen flixes on the Washington shelf and slope

Benthic oxygen fluxes calculated from in situ microelectrode profiles arc compared with benthic flux chamber O2 uptake measurements on a transect of eight stations across the continental shelf and three stations on the slope of Washington State. Station depths ranged from 40 to 630 m and bottom-water oxygen concentrations were 127-38 µM. The fluxes measured by the two methods were similar on the slope, but on the shelf, the chamber flux exceeded the microelectrode flux by as much as a factor of 3-4. We attribute this difference to pore-water irrigation, a process which apparently accounts for the oxidation of a significant amount of organic C in the continental shelf sediments. Combining our diffusive flux data with other data demonstrates clearly that the bottomwater oxygen concentration must play some significant role in determining the sedimentary oxygen consumption rate. Numerical simulation of the microelectrode 0, profiles suggests that roughly half the diffusive 0, flux must be consumed within - 1 mm of the sediment surface. If this conclusion is correct, then the magnitude of the diffusive flux depends both on the bottom-water oxygen concentration and on the supply rate of labile C to the sediment surf'ace.

Pore-water chemistry of abyssal Arabian Sea surface sediments

Benthic fluxes and pore-water compositions of silicic acid, nitrate and phosphate were investigated for surface sediments of the abyssal Arabian Sea during four cruises (1995-1998). Five sites located in the northern (NAST), western (WAST), central (CAST), eastern (EAST), and southern (SAST) Arabian Sea were revisited during intermonsoonal periods after the NE- and SW-Monsoon. At these sites, benthic fluxes of remineralized nutrients from the sediment to the bottom water of 36-106, 102-350 and 4-16 mmol/m**2/yr were measured for nitrate, silicic acid and phosphate, respectively. The benthic fluxes and pore-water compositions showed a distinct regional pattern. Highest fluxes were observed in the western and northern region of the Arabian Sea, whereas decreasing fluxes were derived towards the southeast. At WAST, the general temporal pattern of primary production, related to the NE- and SW-Monsoon, is reflected by benthic fluxes. In contrast, at sites NAST, SAST, CAST, and EAST a temporal pattern of fluxes in response to the monsoon is not obvious. Our results reveal a clear coupling between the general regional pattern of production in surface waters and the response of the benthic environment, as indicated by the flux of remineralized nutrients, though a spatially differing degree of decoupling during transport and remineralization of particulate organic matter and biogenic opal was observed. This has to be taken into account regarding budget calculations and paleoceanographic topics.

Hydrochemistry measured on water bottle samples during the METEOR International Indian Ocean Expedition 1964/65

The present volume gives the observed physical and chemical data obtained by R.V. "Meteor" in the Indian Ocean during cruise 1964/65. The tables are based on the computations made by the National Oceanographic Data Center (NODC) in Washington. In addition to the normally communicated data, the tables contain four chemical parameters: alkalinity, ammonia, fluoride, and calcium.