Extremely low frequency magnetic fields in residences in Germany. Distribution of measurements, comparison of two methods for assessing exposure, and predictors for the occurrence of magnetic fields above background level

We examined the results of 1,835 magnetic field measurements in German residences conducted between November 1997 and September 1999. The measurements were part of an epidemiological study on the relationship between magnetic fields and childhood leukemia. We performed a fixed-location measurement of the magnetic field at 50 Hz and 16 2/3 Hz (frequency of the German railway system) over 24 h in the child’s bedroom in the residence of each study participant. In addition, we conducted a second 24 h-measurement in the living room at 50 Hz, and spot measurements while walking through all rooms of the respective dwelling. Median 50 Hz magnetic fields above 0.2 μT were found to be infrequent in Germany (only 1.4% of all residences). Fields produced by high-voltage power lines (123–420 kV) were lower than expected: the median magnetic field was above 0.2 μT in only 8 (32.0%) of 25 residences located 50 m or closer to a high-voltage power line indicating that power lines in Germany are usually run well below the maximum power load. We found that magnetic fields were correlated with the type of residence and higher magnetic fields were measured in apartment buildings. There was also some evidence for a positive correlation between magnetic fields and traffic density and an inverse association between magnetic fields and family net income. The 24 h-magnetic field measurements correlated well with the spot measurements (r>0.7). However, when dichotomized with a cut-off point of 0.2 μT, there was only a poor agreement between the two measurement methods. A loss of the strength of the association after categorization was also observed when comparing the arithmetic mean and median of the same 24 h-measurement. In summary, these analyses give a valuable overview of magnetic field distributions in German residences. Received: 31 January 2000 / Accepted: 25 July 2000     

A transgene-centered approach to the biosafety of transgenic phosphinothricin-tolerant plants

The microbial bar and pat genes confer tolerance to the non-selective herbicide phosphinothricin (PPT; sold as Basta or Finale). This tolerance in plants could provide an environmental gain compared to current-day herbicide cocktails, but the safety of such a transgene approach is questioned by many. The biosafety of the presence of these herbicide tolerance genes in plants is evaluated in a transgene-centered approach. Potentially, the introduction of transgenic PPT-tolerant crops could result in acquired PPT tolerance in weedy relatives of these crops. Assuming responsible use of this trait in agronomy, the ecological consequences with respect to weediness or spread of the transgenic PPT tolerance are concluded to be negligible. The key issue for the toxicological evaluation is whether or not the plant has actually been sprayed with PPT. Consumption of the gene and/or gene product from unsprayed transgenic plant material will not have adverse effects. In case of PPT-sprayed material, PPT or its derivatives could be present in food and feed and crop-specific metabolites might be formed. To date, the toxicological impact of such a putative exposure is not sufficiently clear, and further premarket testing is recommended.     

Attenuation of cell cycle regulator p27<Superscript>Kip1</Superscript> expression in vertebrate epithelial cells mediated by extracellular signals in vivo and in vitro

Cell cycle arrest in potentially dividing cells is often mediated by inhibitors of G1/S-phase cyclin-dependent kinases. The cyclin E/CDK2-inhibitor p27^Kip1 has been implicated in this context in epithelial cells. We cloned and sequenced p27^Kip1 of ducklings (Anas platyrhynchos) and used an in vitro assay system to study the mechanism of p27^Kip1 downregulation in the nasal gland which precedes an increase in proliferation rate upon initial exposure of the animals to osmotic stress. Western blot studies revealed that p27^Kip1 is downregulated during 24 h of osmotic stress in ducklings with the steepest decline in protein levels between 5 and 8 h. As indicated by the results of Northern blot and semi-quantitative PCR studies, protein downregulation is not accompanied by similar changes in mRNA levels indicating that Kip1 is regulated mainly at the translational (synthesis) or posttranslational level (degradation). Using recombinant duck Kip1 protein expressed in E. coli, we showed that Kip1 is subject to polyubiquitinylation by cytosolic enzymes from nasal gland cells indicating that loss of Kip1 may be regulated, at least in part, by acceleration of protein degradation. In cultured nasal gland tissue, attenuation of Kip1 expression could be induced by activation of the muscarinic acetylcholine receptor indicating that mAChR-receptor signalling may play a role in the re-entry of quiescent gland cells into the cell cycle.     

Local coexpression domains of two to four genes in the genome of Arabidopsis

Expression of genes in eukaryotic genomes is known to cluster, but cluster size is generally loosely defined and highly variable. We have here taken a very strict definition of cluster as sets of physically adjacent genes that are highly coexpressed and form so-called local coexpression domains. The Arabidopsis (Arabidopsis thaliana) genome was analyzed for the presence of such local coexpression domains to elucidate its functional characteristics. We used expression data sets that cover different experimental conditions, organs, tissues, and cells from the Massively Parallel Signature Sequencing repository and microarray data (Affymetrix) from a detailed root analysis. With these expression data, we identified 689 and 1,481 local coexpression domains, respectively, consisting of two to four genes with a pairwise Pearson's correlation coefficient larger than 0.7. This number is approximately 1- to 5-fold higher than the numbers expected by chance. A small (5%-10%) yet significant fraction of genes in the Arabidopsis genome is therefore organized into local coexpression domains. These local coexpression domains were distributed over the genome. Genes in such local domains were for the major part not categorized in the same functional category (GOslim). Neither tandemly duplicated genes nor shared promoter sequence nor gene distance explained the occurrence of coexpression of genes in such chromosomal domains. This indicates that other parameters in genes or gene positions are important to establish coexpression in local domains of Arabidopsis chromosomes.     

The release of genetically modified crops into the environment. Part II. Overview of ecological risk assessment

Despite numerous future promises, there is a multitude of concerns about the impact of GM crops on the environment. Key issues in the environmental assessment of GM crops are putative invasiveness, vertical or horizontal gene flow, other ecological impacts, effects on biodiversity and the impact of presence of GM material in other products. These are all highly interdisciplinary and complex issues. A crucial component for a proper assessment is defining the appropriate baseline for comparison and decision. For GM crops, the best and most appropriately defined reference point is the impact of plants developed by traditional breeding. The latter is an integral and accepted part of agriculture. In many instances, the putative impacts identified for GM crops are very similar to the impacts of new cultivars derived from traditional breeding. When assessing GM crops relative to existing cultivars, the increased knowledge base underpinning the development of GM crops will provide greater confidence in the assurances plant science can give on the risks of releasing such crops.     

Ovarian aging in two species of long-lived rockfish, Sebastes aleutianus and S. alutus

Little is known about the ovary during aging in long-lived fish with respect to follicular stages and de novo oogenesis. We examined two species of rockfish, Sebastes aleutianus (rougheye rockfish) and Sebastes alutus (Pacific ocean perch). Fish were sampled offshore of British Columbia, age was estimated by otolith annuli, and the ovaries were examined histologically. In S. aleutianus, age up to 80 yr did not markedly alter the frequency distribution of oocytes, follicles, or their total numbers. Similarly, in a larger sample of S. alutus, the abundance of oocytes and follicles showed little age trend up through 77 yr. However, fish older than 50 yr lacked the largest and smallest oocyte size classes (40-60, >80 microm) and the smallest follicle size class (200-350 microm), which results from the later seasonal developmental state of these older fish. These data provide evidence that oogenesis continues at advanced ages in these two species, in contrast with long-held assumptions about mammals. These species represent an iteroparous extreme in the spectrum of life history strategies and merit investigation to determine the mechanisms for such an extended reproductive life span.     

The Q Exactive HF,a Benchtop Mass Spectrometer with a Pre-filter,High-performance Quadrupole and an Ultra-high-field Orbitrap Analyzer

The quadrupole Orbitrap mass spectrometer (Q Exactive) made a powerful proteomics instrument available in a benchtop format. It significantly boosted the number of proteins analyzable per hour and has now evolved into a proteomics analysis workhorse for many laboratories. Here we describe the Q Exactive Plus and Q Exactive HF mass spectrometers, which feature several innovations in comparison to the original Q Exactive instrument. A low-resolution pre-filter has been implemented within the injection flatapole, preventing unwanted ions from entering deep into the system, and thereby increasing its robustness. A new segmented quadrupole, with higher fidelity of isolation efficiency over a wide range of isolation windows, provides an almost 2-fold improvement of transmission at narrow isolation widths. Additionally, the Q Exactive HF has a compact Orbitrap analyzer, leading to higher field strength and almost doubling the resolution at the same transient times. With its very fast isolation and fragmentation capabilities, the instrument achieves overall cycle times of 1 s for a top 15 to 20 higher energy collisional dissociation method. We demonstrate the identification of 5000 proteins in standard 90-min gradients of tryptic digests of mammalian cell lysate, an increase of over 40% for detected peptides and over 20% for detected proteins. Additionally, we tested the instrument on peptide phosphorylation enriched samples, for which an improvement of up to 60% class I sites was observed.Mass spectrometry (MS)-based¹ proteomics aims at the comprehensive analysis of proteins present in a biological sample (1), and the field has expanded in many surprising directions (2). Application of the developed techniques has revealed novel insights into fundamental biology, as well as produced analysis techniques with implications for clinical applications. A major hurdle, however, is the complexity of the systems under scrutiny, as it has been shown that human cell lines, for instance, express at least 10,000 genes that are detectable as proteins (3–5). If we further consider all the peptides produced in bottom-up proteomics experiments, this hurdle is compounded, as ideally many hundreds of thousands of analytes should be characterized in order for the proteins giving rise to them to be fully reconstructed (6). In principle, issues of sample complexity and dynamic range could be addressed by a very high degree of up-front fractionation. However, this strategy faces diminishing returns and leads to unacceptably long analysis times for most purposes. Given the fact that even with optimal chromatographic resolution many peptides with abundance differences of many orders of magnitude elute within the same time frame, there remains a need to improve the mass spectrometric detection in terms of speed, resolution, and sensitivity.Nanoscale liquid chromatography coupled online to mass spectrometry is the current technique of choice for the analysis of complex peptide mixtures. In a top-N shotgun strategy, a full scan, providing a complete overview of isotope patterns resulting from ionized peptides, is followed by N fragmentation scans performed on the most abundant not-yet-sequenced isotope patterns currently visible in the full scan. During fragmentation, the goal is to cleanly isolate the intended precursor peptide ion, which today is generally done either by a linear ion trap or by a quadrupole mass filter. Fragment ions are then mass measured by an Orbitrap mass analyzer, a time-of-flight analyzer, or, less often, ion cyclotron resonance–Fourier transform or linear or three-dimensional ion traps.Apart from the MS instrumentation, recent developments in the proteomics workflow include a move toward automated online quality control systems (7, 8) and single-run analyses (9), which require very high-performance peptide chromatography (10, 11).The Orbitrap mass analyzer was introduced commercially almost 10 years ago, and hybrid instruments based on this tool have become very popular in proteomics (12). They consist of an upfront mass spectrometer coupled to a so-called C-trap, which stores and compresses the ion population (generally up to one million charges) prior to injection into the Orbitrap analyzer. Up to the Orbitrap Velos and Elite members of this family of instruments, the precursor selection (and usually the fragmentation) occurred in the linear ion trap (13), but a few years ago an instrument based on a quadrupole front end—the Q Exactive mass spectrometer—was developed (14). Compared with the linear ion trap, quadrupole mass filters have the advantage of being capable of nearly instantaneously selecting a small mass region by modulating the RF field, allowing only a select set of ions to have stable trajectories when passing through the rod assembly. As we described previously, this near-instantaneous mass selection capability and the C-trap''s capability of storing ions enable multiplexing of different ion populations (e.g. fragment ions of two or more distinct precursor ions) prior to analysis in the Orbitrap mass analyzer (14). However, that instrument did not use the highest efficiency quadrupole technology (15), and it did not use the compact high-field Orbitrap analyzer that had been introduced in the Orbitrap Elite instrument (16).Here we describe the advances incorporated into the Q Exactive Plus and the Q Exactive HF instruments. These include improved robustness effected by a low-resolution filter upstream of the quadrupole, a segmented quadrupole, and, in the case of the Q Exactive HF instrument, an ultra-high-field mass Orbitrap analyzer, doubling resolution or acquisition speed. We describe these capabilities in the context of single-shot complex mixture analysis of peptides and phosphopeptides.     

Biodiversity and vegetation of small-sized inselbergs in a West African rain forest (Taı, Ivory Coast)

The vegetation of small granitic rock outcrops (geomorphologically small-sized inselbergs) which do not reach the canopy was studied in the Taı rain forest (southwestern Ivory Coast) under aspects of species diversity and phytogeographical affinities. Rock outcrops form edaphically arid (due to absent or very sparse soil cover) and microclimatologically xeric (i.e. low air humidity, temperature regularly exceeding 50°C) islands with cryptogamic crusts, succulents and poikilohydric vascular plants as characteristic elements of their vegetation which differs totally from the surrounding forest. Altogether sixty-six species of vascular plants out of twenty-nine families occur, the number of species correlates positively with inselberg size. Compared with large inselbergs the microclimatic attributes of small-sized rock outcrops are less pronounced. This is accompanied by a decrease of typical inselberg taxa (i.e. species mainly occurring on inselbergs). Low beta diversity between inselbergs indicates deterministic influences as important regulators of species composition. Annual Poaceae and Cyperaceae are richly represented. It can be hypothesized that inselbergs may represent natural growing sites of widely distributed tropical weeds today. Inselbergs might provide habitat resources for savanna elements in rain forest zones.