Photosynthesis and calcification in the calcifying algae Halimeda discoidea studied with microsensors

With microsensors, we measured the steady‐state microprofiles of O2, pH and Ca2+ on the topside of young segments of Halimeda discoidea, as well as the surface dynamics upon light–dark shifts. The effect of several inhibitors was studied. The steady‐state measurements showed that under high light intensity, calcium and protons were taken up, while O2 was produced. In the dark, O2 was consumed, the pH decreased to below seawater level and Ca2+ uptake was reduced to 50%. At low light intensity (12 mmol photons m‐2 s‐1), Ca2+ efflux was observed. Upon light–dark shifts, a complicated pattern of both the pH and calcium surface dynamics was observed. Illumination caused an initial pH decrease, followed by a gradual pH increase: this indicated that the surface pH of H. discoidea is determined by more than one light‐induced process. When photosynthesis was inhibited by dichlorophenyl dimethyl urea (DCMU), a strong acidification was observed upon illumination. The nature and physiological function of this putative pump is not known. The calcium dynamics followed all pH dynamics closely, both in the presence and absence of DCMU. The Ca‐channel blockers verapamil and nifedipine had no effect on the Ca2+ dynamics and steady‐state profiles. Thus, in H. discoidea, calcification is not regulated by the alga, but is a consequence of pH increase during photosynthesis. Acetazolamide had no effect on photosynthesis, whereas ethoxyzolamide inhibited photosynthesis at higher light intensities. Therefore, all carbonic anhydrase activity is intracellular. Carbonic anhydrase is required to alleviate the CO2 limitation. Calcification cannot supply sufficient protons and CO2 to sustain photosynthesis

Technical report on chemical sensors performance

The report will present an overview of sensors that are useful for the study of seeps, and present a detailed comparison of commercial and prototype CO2 and pH sensors. CO2 sensors: 1) Severinghaus CO2 sensors (Microelectrodes ltd), 2) the HydroC-­‐CO2 logger sensor (CONTROS GmbH), 3) the “GasPro-­‐pCO2” (developed by the team coordinated by Prof. S. Lombardi; Sapienza -­‐ University of Rome-­‐CERI), 4) the MuFo (Multifiber Optode logger, developed in this project in a collaboration between Prof. I. Klimant University Graz and the MPI-­‐MM). pH sensors: 1) glass electrodes Microelectrodes ltd), 2) optodes (developed in this project in a collaboration between Prof. I. Klimant University Graz and the MPI-­‐MM), 3) SeaFET (ion-­‐selective field-­‐effect transistor, Satlantic)

Structure and function of natural sulphide-oxidizing microbial mats under dynamic input of light and chemical energy

We studied the interaction between phototrophic and chemolithoautotrophic sulphide-oxidizing microorganisms in natural microbial mats forming in sulphidic streams. The structure of these mats varied between two end-members: one characterized by a layer dominated by large sulphur-oxidizing bacteria (SOB; mostly Beggiatoa-like) on top of a cyanobacterial layer (B/C mats) and the other with an inverted structure (C/B mats). C/B mats formed where the availability of oxygen from the water column was limited (<5 mu M). Aerobic chemolithotrophic activity of the SOB depended entirely on oxygen produced locally by cyanobacteria during high light conditions. In contrast, B/C mats formed at locations where oxygen in the water column was comparatively abundant (445 mu M) and continuously present. Here SOB were independent of the photosynthetic activity of cyanobacteria and outcompeted the cyanobacteria in the uppermost layer of the mat where energy sources for both functional groups were concentrated. Outcompetition of photosynthetic microbes in the presence of light was facilitated by the decoupling of aerobic chemolithotrophy and oxygenic phototrophy. Remarkably, the B/C mats conserved much less energy than the C/B mats, although similar amounts of light and chemical energy were available. Thus ecosystems do not necessarily develop towards optimal energy usage. Our data suggest that, when two independent sources of energy are available, the structure and activity of microbial communities is primarily determined by the continuous rather than the intermittent energy source, even if the time-integrated energy flux of the intermittent energy source is greater

The influence of pore-water advection, benthic photosynthesis, and respiration on calcium carbonate dynamics in reef sands

To investigate diel calcium carbonate (CaCO₃) dynamics in permeable coral reef sands, we measured pore-water profiles and fluxes of oxygen (O₂), nutrients, pH, calcium (Ca²⁺), and alkalinity (TA) across the sediment-water interface in sands of different permeability at Heron Reef, Australia. Background flushing rates were high, most likely as a result of infaunal burrow irrigation, but flux chamber stirring enhanced pore-water exchange. Light and pore-water advection fueled high rates of benthic primary production and calcification in sunlit surface sediments. In the light, benthic photosynthesis and calcification induced surface minima in Ca²⁺ and TA and peaks in pH and O₂. Oxygen penetration depth in coarse sands decreased from ~ 1.2 cm during the day to ~ 0.6 cm at night. Total oxygen uptake (TOU) in dark chambers was three to fourteen times greater than diffusive uptake and showed a direct effect of pore-water advection. Greater sediment oxygen consumption rates were observed in higher permeability sands. In the dark, TA release was not stimulated by increasing TOU because of a damping effect of pore-water advection on metabolic CaCO₃ dissolution efficiency. On a daily basis, CaCO₃ undergoes net dissolution in Heron Reef sands. However, pore-water advection can reverse the CaCO₃ budget and promote CaCO₃ preservation under the most energetic conditions

Fine‐scale measurement of diffusivity in a microbial mat with nuclear magnetic resonance imaging

Noninvasive 1H‐nuclear magnetic resonance (NMR) imaging was used to investigate the diffusive properties of microbial mats in two dimensions. Pulsed field gradient NMR was used to acquire images of the H2O diffusion coefficient, Ds, and multiecho imaging NMR was used to obtain images of the water density in two structurally different microbial mats sampled from Solar Lake (Egypt). We found a pronounced lateral and vertical variability of both water density and water diffusion coefficient, correlated with the laminated and heterogeneous distribution of microbial cells and exopolymers within the mats. The average water density varied from 0.5 to 0.9, whereas the average water diffusion coefficient ranged from 0.4 to 0.9 relative to the values obtained in the stagnant water above the mat samples. The apparent water diffusivities estimated from NMR imaging compared well to apparent O2 diffusivities measured with a diffusivity microsensor. Analysis of measured O2 concentration profiles with a diffusion‐reaction model showed that both the magnitude of calculated rates and the depth distribution of calculated O2 consumption/production zones changed when the observed variations of diffusivity were taken into account. With NMR imaging, diffusivity can be determined at high spatial resolution, which can resolve inherent lateral and vertical heterogeneities found in most natural benthic systems

Postmortem tissue distribution of morphine and its metabolites in a series of heroin related deaths

The abuse of heroin (diamorphine) and heroin deaths are growing around the world. The interpretation of the toxicological results from suspected heroin deaths is notoriously difficult especially in cases where there may be limited samples. In order to help forensic practitioners with heroin interpretation we determined the concentration of morphine (M), morphine‐3‐glucuronide (M3G) and morphine‐6‐glucuronide (M6G) in blood (femoral and cardiac), brain (thalamus), liver (deep right lobe), bone marrow (sternum), skeletal muscle (psoas) and vitreous humor in 44 heroin related deaths. The presence of 6‐monoacetylmorphine (6‐MAM) in any of the postmortem samples was used as confirmation of heroin use. Quantitation was carried out using a validated LC‐MS/MS method with solid phase extraction. We also determined the presence of papaverine, noscapine and codeine in the samples, substances often found in illicit heroin and that may help determine illicit heroin use. The results of this study show that vitreous is the best sample to detect 6‐MAM (100% of cases), and thus heroin use. The results of the M, M3G and M6G quantitation in this study allow a degree of interpretation when samples are limited. However in some cases it may not be possible to determine heroin/morphine use as in 4 cases in muscle (3 cases in bone marrow) no morphine, morphine‐3‐glucuronide or morphine‐6‐glucuronide was detected, even though they were detected in other case samples. As always postmortem cases of suspected morphine/heroin intoxication should be interpreted with care and with as much case knowledge as possible

Metabolic changes in the lungs after ischaemia

CITATION: Engelbrecht, F. M., Edwards, I. J. & De Beer, D. P. 1980. Metabolic changes in the lungs after ischaemia. South African Medical Journal, 58:409-413.The original publication is available at http://www.samj.org.zaThe effects of variable periods of ischaemia on the isolated lungs of rats and rabbits, stored for up to 6 hours at 4°C, 21°C and 37°C under standardized conditions, were investigated in vitro in terms of oxygen consumption, the rate of 1-14C-leucine incorporation into soluble proteins, and 1-14C-palmitate incorporation into total phospholipids and lipid fractions. The endogenous oxygen uptake of rat lung slices in an air phase, from tissues stored at 4°C and 21°C under ischaemic conditions for 6 hours and at 37°C for 4 hours, was significantly different from the control values. The oxygen uptake of lungs from animals anaesthetized with pentobarbitone prior to exsanguination and stored for only 2 hours at 37°C differed significantly from control values. Judged by the rate of incorporation of radiolabelled leucine into soluble proteins and that of palmitate into total lipids and phospholipids of lungs after storage for increasing periods at 4°C and 37°C, significant differences were already found after 1 1/2 hours. From this observation it would appear that these parameters are very sensitive indicators for assessing irreversible lung damage due to ischaemia.Publisher’s versio

Rotavirus genetic diversity, disease association, and temporal change in hospitalized rural Kenyan children

Background. The effectiveness of rotavirus vaccines will be dependent on the immunity conferred against prevalent and emergent variants causing severe diarrheal disease. Longitudinal surveillance of disease-causing strains is a prerequisite to intervention. Methods. Molecular characterization was conducted on rotavirus-positive stool samples from children admitted with diarrhea to a rural district hospital during 2002-2004. Extracted viral RNA was separated by polyacrylamide gel electrophoresis, and rotavirus VP4 (P types) and VP7 (G types) specificities were determined. Results. Among 558 investigated cases, the predominant genotype was P[8]G1 (42%), followed by P[8]G9 (15%), P[4]G8 (7%), P[6]G8 (6%), and P[8]G8 (4%), with 10% mixed strains. Overall, there were 6 different P types and 7 G types. No association was identified between genotype and child age, sex, or severity of diarrhea. The P and G genotypes and polyacrylamide gel electropherotypes showed significant temporal variation in frequency: P[8]G1 decreased from 51% (95% confidence interval [CI], 43%-58%) in 2002 to 30% (95% CI, 24%-37%) in 2004, and P[4]G8 increased from 2% (95% CI, 0%-5%) in 2002 to 13% (95% CI, 9%-19%). Quarterly data revealed seasonally endemic and emergence and/or decay patterns. Conclusions. Our study of rotavirus strains causing severe diarrhea in rural Kenyan children showed a predominance of P[8]G1 and confirms the importance of G8 and G9 strains in sub-Saharan Africa. Considerable genetic diversity of rotavirus strains was observed, including substantial mixed and unusual types, coupled with significant temporal strain variation and emergence. These results warn of variable vaccine efficacy and the need for long-term surveillance of circulating rotavirus genotypes

Beyond persons: extending the personal / subpersonal distinction to non-rational animals and artificial agents

The distinction between personal level explanations and subpersonal ones has been subject to much debate in philosophy. We understand it as one between explanations that focus on an agent’s interaction with its environment, and explanations that focus on the physical or computational enabling conditions of such an interaction. The distinction, understood this way, is necessary for a complete account of any agent, rational or not, biological or artificial. In particular, we review some recent research in Artificial Life that pretends to do completely without the distinction, while using agent-centered concepts all the way. It is argued that the rejection of agent level explanations in favour of mechanistic ones is due to an unmotivated need to choose among representationalism and eliminativism. The dilemma is a false one if the possibility of a radical form of externalism is considered