Characterization of the light field in laboratory scale enclosures of eutrophic lake water (Lake Loosdrecht,The Netherlands)

Light conditions in laboratory scale enclosures (LSE) of shallow, eutrophic Lake Loosdrecht (The Netherlands), including a method for simulating a natural incident light course, are described. Total PAR (400–700 nm) and spectral irradiance distribution were measured at sestonic chlorophyll a and dry weight concentrations 100 mg m^–3 and 16 g m^–3, respectively. Phytoplankton was dominated by Oscillatoria spp. The euphotic depth (Z _eu) was 0.7–1.0 m. Shortly after filling the LSE with lake water, diffuse attenuation coefficients ranged from 14 m^–1 for blue to 5 m^–1 for red light. Around Z _eu, attenuation in the blue region was markedly lower and irradiance reflectance (R) continued to increase; these anomalies were caused by lateral incident light from the LSE's waterbath. Spectrophotometry indicated that absorption was mainly by particles, but dissolved humic substances (gilvin) were also important. The particles were likely to be dominated by detritus absorbing more blue relative to red light. Subsurface R in lake water in the LSE had a maximum around 705 nm and low values in the blue band, but was lower than that previously reported for measurements in situ. Wash-out of detritus, presumably both dissolved and particulate fractions, by flow-through with synthetic medium, greatly affected the spectral reflectance measured outside the LSE. The maximum value of R decreased from 0.022 to 0.009, and the peak shifted to 550 nm.     

Oxidation of methyl linoleate in micellar solutions induced by the combination of iron(II)/ascorbic acid and iron(II)/H2O2

The oxidation of methyl linoleate (ML) was studied in the presence of Fe(II) alone and its combination with either ascorbic acid (AsAH(2)) or hydrogen peroxide (H(2)O(2)) at different molar ratios. Reactions were carried out in micellar solutions of TTAB (tetradecyltrimethylammonium bromide) and SDS (sodium dodecyl sulfate), respectively, and were monitored by UV spectroscopy and electrospray ionization mass spectrometry (ESI-MS). Fe(II) alone was able to catalyze the oxidation of ML in micellar solutions of TTAB, but not in those of SDS. The combination of H(2)O(2) with Fe(II) showed catalytic effect only in the TTAB medium, leading to different ML and Fe(II) oxidation kinetics compared to the Fe(II)-only catalyzed reactions. The AsAH(2)/Fe(II) combination demonstrated to be a good catalyst for the oxidation of ML in SDS micellar solutions, but not in TTAB micellar solutions; the activity of the catalyst was dependent on the AsAH(2)/Fe(II) molar ratio. The obtained results confirm that, for the ML oxidation to be initiated, the presence of a Fe(II)/Fe(III) couple is essential, which is related to the pH of micellar solutions. The catalytic properties of the AsAH(2)/Fe(II) combination were explained by taking into account the anti-oxidant and pro-oxidant properties of AsAH(2), as well as the possible formation of an iron/ascorbate complex as the initiator of the ML oxidation.     

Cloning and characterization of the bgxA gene from Erwinia chrysanthemi D1 which encodes a β-glucosidase/xylosidase enzyme

A -glucosidase/xylosidase gene from Erwinia chrysanthemi strain D1 was cloned and sequenced. This gene, named bgxA, encodes a ca. 71 kDa protein product which, following removal of the leader peptide, resulted in a ca. 69 kDa mature protein that accumulated in the periplasmic space of E. chrysanthemi strain D1 and Escherichia coli cells expressing the cloned gene. The protein exhibited both -glucosidase and -xylosidase activities but gave no detectable activity on xylan or carboxymethyl cellulose. The enzyme was classified as a type 3 glycosyl hydrolase, but was unusual in having a truncated B region at the carboxyl-terminus. Several E. chrysanthemi strains isolated from corn produced the glucosidase/xylosidase activity but not those isolated from dicot plants. However, bgxA marker exchange mutants of strain D1 were not detectably altered in virulence on corn leaves.     

Energy Efficiency and Structural Change in the Netherlands, 1980–1995

International agreement has been reached to reduce greenhouse gas emissions worldwide. One important way of decoupling CO₂ emissions from economic growth is by introducing technical measures to improve energy efficiency. In this article, we assess the influence of developments in energy efficiency and economic structure on the total primary energy consumption in the Netherlands over the period 1980– 1995. We find a distinct decoupling of the economic growth and energy consumption of 1.5% per year in the 15-year analysis period. We measure (technical) changes in energy efficiency by changes in the energy consumption per physical unit of production or activity. The aggregate rate of (technical) energy-efficiency improvement was 1.4% per year over the period 1980–1995. The use of physical production indicators makes it possible to measure energy-efficiency developments without detailed surveys at a very low level of aggregation. When we look at economic structural changes over this period, we find that (i) no substantial shift took place at the level of the economic sectors that we distinguish; (ii) the most energy intensive subsectors grew much faster than the total economy; and (iii) at the subsector level, on average, a sizable decoupling of physical production and value added occurred. We conclude that structural changes, that is, changes in the composition of the economy, did not lead to a net decrease in the energy intensity of the Netherlands over the period 1980–1995.     

State-transitions facilitate robust quantum yields and cause an over-estimation of electron transport in Dunaliella tertiolecta cells held at the CO2 compensation point and re-supplied with DIC

Photosynthetic energy consumption and non-photosynthetic energy quenching processes are inherently linked. Both processes must be controlled by the cell to allow cell maintenance and growth, but also to avoid photodamage. We used the chlorophyte algae Dunaliella tertiolecta to investigate how the interactive regulation of photosynthetic and non-photosynthetic pathways varies along dissolved inorganic carbon (DIC) and photon flux gradients. Specifically, cells were transferred to DIC-deplete media to reach a CO₂ compensation before being re-supplied with DIC at various concentrations and different photon flux levels. Throughout these experiments we monitored and characterized the photophysiological responses using pulse amplitude modulated fluorescence, oxygen evolution, 77 K fluorescence emission spectra, and fast-repetition rate fluorometry. O₂ uptake was not significantly stimulated at DIC depletion, which suggests that O₂ production rates correspond to assimilatory photosynthesis. Fluorescence-based measures of relative electron transport rates (rETRs) over-estimated oxygen-based photosynthetic measures due to a strong state-transitional response that facilitated high effective quantum yields. Adoption of an alternative fluorescence-based rETR calculation that accounts for state-transitions resulted in improved linear oxygen versus rETR correlation. This study shows the extraordinary capacity of D. tertiolecta to maintain stable effective quantum yields by flexible regulation of state-transitions. Uncertainties about the control mechanisms of state-transitions are presented.     

Patient Referral Patterns and the Spread of Hospital-Acquired Infections through National Health Care Networks

Rates of hospital-acquired infections, such as methicillin-resistant Staphylococcus aureus (MRSA), are increasingly used as quality indicators for hospital hygiene. Alternatively, these rates may vary between hospitals, because hospitals differ in admission and referral of potentially colonized patients. We assessed if different referral patterns between hospitals in health care networks can influence rates of hospital-acquired infections like MRSA. We used the Dutch medical registration of 2004 to measure the connectedness between hospitals. This allowed us to reconstruct the network of hospitals in the Netherlands. We used mathematical models to assess the effect of different patient referral patterns on the potential spread of hospital-acquired infections between hospitals, and between categories of hospitals (University medical centers, top clinical hospitals and general hospitals). University hospitals have a higher number of shared patients than teaching or general hospitals, and are therefore more likely to be among the first to receive colonized patients. Moreover, as the network is directional towards university hospitals, they have a higher prevalence, even when infection control measures are equally effective in all hospitals. Patient referral patterns have a profound effect on the spread of health care-associated infections like hospital-acquired MRSA. The MRSA prevalence therefore differs between hospitals with the position of each hospital within the health care network. Any comparison of MRSA rates between hospitals, as a benchmark for hospital hygiene, should therefore take the position of a hospital within the network into account.     

Fixation and distribution of bacterial activity in sand to induce carbonate precipitation for ground reinforcement

The mechanical properties of soil (cohesion, friction, stiffness and permeability) are important parameters for engineering constructions and ecosystems in sedimentary environments. BioGrout is an in situ soil strengthening technique involving microbial-induced carbonate precipitation (MICP). This process involves hydrolysis of urea by bacteria containing the enzyme urease in the presence of dissolved calcium ions, resulting in calcium carbonate precipitation. In order to control the BioGrout process for engineering applications, it is necessary to improve understanding of the relevant phenomena and develop efficiencies to enable up-scaling of the technology to suit commercial applications. Control of a homogeneous distribution of bacterial activity in a sand bed is considered crucial in order to prevent clogging during injection and provide homogeneous reinforcement results. This paper describes a methodology to distribute and fix bacteria (with their enzyme activity) relatively homogeneously in a sand bed, before supplying cementation reagents. The methodology is based on a two-phase injection procedure: a bacterial suspension is injected into the sand body, immediately followed by a fixation fluid (i.e. a solution with high salt content). It is proposed that bacteria are retarded by adsorption and filtration processes and are permanently adsorbed to the sand grains when overtaken by the fixation fluid. The presented experimental approach for optimizing bacterial fixation in porous media can be used as a tool to design the treatment protocol for engineering applications in practice.     

Predicting the Electron Requirement for Carbon Fixation in Seas and Oceans

Marine phytoplankton account for about 50% of all global net primary productivity (NPP). Active fluorometry, mainly Fast Repetition Rate fluorometry (FRRf), has been advocated as means of providing high resolution estimates of NPP. However, not measuring CO₂-fixation directly, FRRf instead provides photosynthetic quantum efficiency estimates from which electron transfer rates (ETR) and ultimately CO₂-fixation rates can be derived. Consequently, conversions of ETRs to CO₂-fixation requires knowledge of the electron requirement for carbon fixation (Φ_e,C, ETR/CO₂ uptake rate) and its dependence on environmental gradients. Such knowledge is critical for large scale implementation of active fluorescence to better characterise CO₂-uptake. Here we examine the variability of experimentally determined Φ_e,C values in relation to key environmental variables with the aim of developing new working algorithms for the calculation of Φ_e,C from environmental variables. Coincident FRRf and ¹⁴C-uptake and environmental data from 14 studies covering 12 marine regions were analysed via a meta-analytical, non-parametric, multivariate approach. Combining all studies, Φ_e,C varied between 1.15 and 54.2 mol e⁻ (mol C)⁻¹ with a mean of 10.9±6.91 mol e⁻ mol C)⁻¹. Although variability of Φ_e,C was related to environmental gradients at global scales, region-specific analyses provided far improved predictive capability. However, use of regional Φ _e,C algorithms requires objective means of defining regions of interest, which remains challenging. Considering individual studies and specific small-scale regions, temperature, nutrient and light availability were correlated with Φ _e,C albeit to varying degrees and depending on the study/region and the composition of the extant phytoplankton community. At the level of large biogeographic regions and distinct water masses, Φ _e,C was related to nutrient availability, chlorophyll, as well as temperature and/or salinity in most regions, while light availability was also important in Baltic Sea and shelf waters. The novel Φ _e,C algorithms provide a major step forward for widespread fluorometry-based NPP estimates and highlight the need for further studying the natural variability of Φ_e,C to verify and develop algorithms with improved accuracy.     

Unbalancing the Phosphatidylinositol-4,5-bisphosphate–Cofilin Interaction Impairs Cell Steering

Cofilin is a key player in actin dynamics during cell migration. Its activity is regulated by (de)phosphorylation, pH, and binding to phosphatidylinositol-4,5-bisphosphate [PI(4,5)P₂]. Here, we here use a human cofilin-1 (D122K) mutant with increased binding affinity for PI(4,5)P₂ and slower release from the plasma membrane to study the role of the PI(4,5)P₂–cofilin interaction in migrating cells. In fibroblasts in a background of endogenous cofilin, D122K cofilin expression negatively affects cell turning frequency. In carcinoma cells with down-regulated endogenous cofilin, D122K cofilin neither rescues the drastic morphological defects nor restores the effects in cell turning capacity, unlike what has been reported for wild-type cofilin. In cofilin knockdown cells, D122K cofilin expression promotes outgrowth of an existing lamellipod in response to epidermal growth factor (EGF) but does not result in initiation of new lamellipodia. This indicates that, next to phospho- and pH regulation, the normal release kinetics of cofilin from PI(4,5)P₂ is crucial as a local activation switch for lamellipodia initiation and as a signal for migrating cells to change direction in response to external stimuli. Our results demonstrate that the PI(4,5)P₂ regulatory mechanism, that is governed by EGF-dependent phospholipase C activation, is a determinant for the spatial and temporal control of cofilin activation required for lamellipodia initiation.     

The death receptor CD95 activates the cofilin pathway to stimulate tumour cell invasion

The death receptor CD95 promotes apoptosis through well-defined signalling pathways. In colorectal cancer cells, CD95 primarily stimulates migration and invasion through pathways that are incompletely understood. Here, we identify a new CD95-activated tyrosine kinase pathway that is essential for CD95-stimulated tumour cell invasion. We show that CD95 promotes Tyr 783 phosphorylation of phospholipase C-γ1 through the platelet-derived growth factor receptor-β, resulting in ligand-stimulated phosphatidylinositol (4,5)-bisphosphate (PIP(2)) hydrolysis. PIP(2) hydrolysis liberates the actin-severing protein cofilin from the plasma membrane to initiate cortical actin remodelling. Cofilin activation is required for CD95-stimulated formation of membrane protrusions and increased tumour cell invasion.