Country-specific effects of neonicotinoid pesticides on honey bees and wild bees.

Neonicotinoid seed dressings have caused concern world-wide. We use large field experiments to assess the effects of neonicotinoid-treated crops on three bee species across three countries (Hungary, Germany, and the United Kingdom). Winter-sown oilseed rape was grown commercially with either seed coatings containing neonicotinoids (clothianidin or thiamethoxam) or no seed treatment (control). For honey bees, we found both negative (Hungary and United Kingdom) and positive (Germany) effects during crop flowering. In Hungary, negative effects on honey bees (associated with clothianidin) persisted over winter and resulted in smaller colonies in the following spring (24% declines). In wild bees (Bombus terrestris and Osmia bicornis), reproduction was negatively correlated with neonicotinoid residues. These findings point to neonicotinoids causing a reduced capacity of bee species to establish new populations in the year following exposure.

Manganese in the upper Severn mid-Wales.

The concentrations of manganese (Mn) in the Upper River Severn (the Plynlimon catchments) are examined in relation to rainfall, cloud water, throughfall, stemflow and stream water concentrations where there is over 20 years of monitoring data available. Manganese concentrations are particularly low in rainfall and cloud water, with maximum concentrations occurring under low volumes of catch due to atmospheric "washout" of contaminants and dry deposition. There is strong Mn enrichment in throughfall and stemflow and this is probably linked to cycling through the vegetation. Manganese in the streams and groundwaters are primarily supplied from within-catchment sources. The highest concentrations occur within the tree canopy probably due to element cycling and in groundwaters due to mobilisation from the rock. Manganese concentrations in streams are at their lowest during spring and summer following long dry spells, with rapid increases following subsequent rain. There is no clear long-term trend in Mn concentration in the streams although there are increases in Mn concentrations for years when there is extensive felling of spruce plantation forest and in 1995 following a more extensive dry period. New high resolution monitoring picks up the effects of the rising limb of the hydrograph when concentrations rapidly increase, diurnal patterns during summer low-flow periods and contrasting dynamics between moorland and forested catchments.

The biogeochemistry of arsenic in a remote UK upland site: trends in rainfall and runoff, and comparisons with urban rivers.

Ten years of monitoring of rainfall and streams in the remote acidic and acid sensitive moorland and afforested moorland of upland mid-Wales reveals concentrations of arsenic (As) typically <1 µg L(-1). On average, the lowest concentrations occur within rainfall and they have declined over time probably in response to reductions in global emissions. There is a corresponding reduction within the streams except for forested systems where concentrations up to doubled following clear-fell. Within the streams there are both annual cycling and diurnal cycling of As. The annual cycling gives maxima during the summer months and this probably reflects the importance of groundwater inputs and mineralisation/desorption from the surface soil layers. Correspondingly, the diurnal cycling occurs during the summer months at low flow periods with As concentrations highest in the afternoon/evening. For the urban/industrial basins of northern England with historically a much higher As deposition, land contamination and effluent discharges, comparative data indicate As concentrations around three fold higher: strong seasonal patterns are observed for the same reasons as with the uplands. Across the sites, the As concentrations are over an order of magnitude lower than that of environmental concern. Nonetheless, the results clearly show the effects of declining emissions on rainfall deposition and some indication of areas of historic contamination. Arsenic is mainly present in the <0.45 fraction, but cross-flow filtration indicates that approx. 43% is in the colloidal phase at the clean water sites, and 16% in the colloidal phase at the contaminated sites. Part of this colloidal component may well be associated with organic carbon.

Modulation of chaperone gene expression in mutagenized Saccharomyces cerevisiae strains developed for recombinant human albumin production results in increased production of multiple heterologous proteins.

The yeast Saccharomyces cerevisiae has been successfully established as a commercially viable system for the production of recombinant proteins. Manipulation of chaperone gene expression has been utilized extensively to increase recombinant protein production from S. cerevisiae, focusing predominantly on the products of the protein disulfide isomerase gene PDI1 and the hsp70 gene KAR2. Here we show that the expression of the genes SIL1, LHS1, JEM1, and SCJ1, all of which are involved in regulating the ATPase cycle of Kar2p, is increased in a proprietary yeast strain, developed by several rounds of random mutagenesis and screening for increased production of recombinant human albumin (rHA). To establish whether this expression contributes to the enhanced-production phenotype, these genes were overexpressed both individually and in combination. The resultant strains showed significantly increased shake-flask production levels of rHA, granulocyte-macrophage colony-stimulating factor, and recombinant human transferrin.

Using delta15N values to characterise the nitrogen nutrient pathways from intensive animal units.

Previous studies on foliar delta15N values, in certain bryophytes, have indicated signature similarities to source pollutants. The object of this study was to investigate the effect further, by examining the mechanisms whereby isotopic fractionation occurs in systems such as atmospheric ammonia (NH3), throughfall, vegetation and soil. Measurements taken in and around point emission sources will then be used to characterise the various fractionation effects associated with these N transformations, as well as to demonstrate some of the issues associated with using delta15N values as pollution indicators. The atmospheric dispersion model UK-ADMS has also been used to model atmospheric delta15NH3 emissions, with signatures exhibiting marked negative shifts immediately downwind of an agricultural NH3 source. Similar dispersion patterns were mapped for NH3 concentration data illustrating the link between these two forms of measurement.

Export of dissolved organic carbon from peatlands under elevated carbon dioxide levels.

Peatlands represent a vast store of global carbon. Observations of rapidly rising dissolved organic carbon concentrations in rivers draining peatlands have created concerns that those stores are beginning to destabilize. Three main factors have been put forward as potential causal mechanisms, but it appears that two alternatives--warming and increased river discharge--cannot offer satisfactory explanations. Here we show that the third proposed mechanism, namely shifting trends in the proportion of annual rainfall arriving in summer, is similarly unable to account for the trend. Instead we infer that a previously unrecognized mechanism--carbon dioxide mediated stimulation of primary productivity--is responsible. Under elevated carbon dioxide levels, the proportion of dissolved organic carbon derived from recently assimilated carbon dioxide was ten times higher than that of the control cases. Concentrations of dissolved organic carbon appear far more sensitive to environmental drivers that affect net primary productivity than those affecting decomposition alone.

Sequential administration enhances the effect of apomorphine SL in men with erectile dysfunction.

The response to Uprima (apomorphine sublingual, (apo SL)) has been well documented in conventional clinical trials. Apo SL produces a predictable, consistent and durable response across a wide variety of patients. The positive reinforcement of a successful outcome should further support clinical benefit. Apo SL with its rapid onset affords a greater opportunity for spontaneity, which can be an important factor in influencing patient choice. It is recognised that patient counselling and the setting of realistic expectations are vital to a successful outcome. The impact of persisting with sequential treatment on outcome has been calculated from the clinical data. While apo SL is effective de novo in 50% of single doses, additional benefit is observed with repeat dosing. Full benefit may not be achieved until four or more treatments have been taken in an optimal setting. The data also confirm that 3 mg has superior activity. Patients should therefore be encouraged to try a minimum of 4 doses at 3 mg.

Yeast 2 microm plasmid copy number is elevated by a mutation in the nuclear gene UBC4.

The copy number of the Saccharomyces cerevisiae endogenous 2 microm plasmid is under strict control to ensure efficient propagation to the daughter cell without significantly reducing the growth rate of the mother or the daughter cell. A recessive mutation has been identified that resulted in an elevated but stable 2 microm plasmid copy number, which could be complemented by a genomic DNA clone containing the UBC4 gene, encoding an E2 ubiquitin-conjugating enzyme. A ubc4::URA3 deletion resulted in the same elevated 2 microm plasmid copy number. An analysis of the endogenous 2 microm transcripts revealed that the steady-state abundance of REP1, REP2, FLP and RAF were all increased 4-5-fold in the mutant. Analysis of the mutant ubc4 allele identified a single base pair mutation within the UBC4 coding region, which would generate a glutamic acid to lysine amino acid substitution within a region of conserved tertiary structure located within the first alpha-helix of Ubc4p. These investigations represent the first molecular characterization of a mutation within a Saccharomyces cerevisiae nuclear gene shown to affect 2 microm steady-state plasmid copy number and implicate the ubiquitin-dependent proteolytic pathway in host control of 2 microm plasmid copy number.

Carbon assimilation and turnover in grassland vegetation using an in situ (13)CO(2) pulse labelling system.

A mobile laboratory was developed to administer a controlled flow of (13)C labelled CO(2) at ambient concentrations ( approximately 350 ppm) in the field. The stable isotope delivery (SID) system consists of an isotope-mixing unit with flow control to a series of 12 independent labelling chambers. In-line CPU controlled infrared gas analysers allow automated measurement of chamber CO(2) concentrations and gas flow management. A preliminary experiment was established on an upland pasture located at the NERC Soil Biodiversity experimental site, Sourhope, UK, in August 1999. The objective of this investigation was to determine the magnitude of pulse-derived C incorporation into a typical upland plant community. To achieve this, the SID system was deployed to pulse-label vegetation with CO(2) enriched with (13)C (50 atom %) at ambient concentrations ( approximately 350 ppm) on two consecutive days in August 1999. Samples of headspace CO(2), shoot and root were taken on four occasions over a period of 28 days after (13)C labelling. These materials were then prepared for (13)C/(12)C ratio determination by continuous-flow/combustion/isotope ratio mass spectrometry (CF-C-IRMS). Results showed that pulse derived CO(2)-C was assimilated at a rate of 128 +/- 32 microg g shoot-C hour(-1). Dynamic samplings showed that pulse-derived (13)C concentrations in the labelled plant tissues declined by 77.4 +/- 6% after 48 hours. The rapid decline in (13)C concentrations in plant matter was the result of C loss from the plant in the form of respired CO(2) and root exudates, and dilution by subsequent unlabelled C assimilates. This novel system offers considerable potential for in situ tracer investigations.

Combination of symptom score, flow rate and prostate volume for predicting bladder outflow obstruction in men with lower urinary tract symptoms.

PURPOSE: The severity of lower urinary tract symptoms associated with benign prostatic enlargement correlates poorly with bladder outlet obstruction. Since urodynamic studies are presumed to be relatively complex, invasive and not cost-effective, they are not routinely performed by physicians treating men with lower urinary tract symptoms. As a result, a large number of patients are treated for bladder outlet obstruction when in fact obstruction may not be present. Since other noninvasive methods have not been effective for predicting bladder outlet obstruction, we investigated whether a combination of prostate volume, uroflowmetry and the American Urological Association (AUA) symptom index would be reliable for predicting this condition. MATERIALS AND METHODS: We prospectively evaluated 204 men with a mean age plus or minus standard deviation of 66.7 +/- 7.5 years who presented with lower urinary tract symptoms. Each patient completed an AUA symptom index questionnaire and underwent uroflowmetry, post-void residual urine volume measurement, pressure flow study and transrectal ultrasound of the prostate to estimate prostatic volume. We constructed receiver operating characteristics curves using various threshold values for maximum urine flow and prostate volume. Threshold values for maximum urine flow and prostate volume were used alone and combined with the AUA symptom index for predicting bladder outlet obstruction. We selected a cutoff value for maximum urine flow of 10 or less ml. per second and prostate volume of 40 gm. or greater, and used these values with an AUA symptom index of greater than 20 to predict bladder outlet obstruction in the group overall. RESULTS: Differences in the mean symptom index score in men with and without bladder outlet obstruction were not statistically significant. There was no obstruction in 19%, 28.9% and 35% of those with severe, moderate and mild symptoms, respectively. The selected cutoff values of maximum urine flow, prostate volume and symptom score combined correctly predicted obstruction in all 39 patients. Therefore, our combination of cutoff values proved to be highly accurate for predicting bladder outlet obstruction. Sensitivity, specificity, and positive and negative predictive values were 26%, 100%, 100% and 32%, respectively. CONCLUSIONS: Our study showed that combining the AUA symptom index, maximum urine flow and prostate volume reliably predicted bladder outlet obstruction in a small subset of patients only. Although bladder outlet obstruction was correctly predicted by our threshold values of AUA symptom index, maximum urine flow and prostate volume in only 39 men (26%) with obstruction, these patients represent a substantial group in any large urological practice treating male lower urinary tract symptoms.

Stable isotope analysis of the origins of zooplankton carbon in lakes of differing trophic state.

Carbon stable isotope analysis was carried out on zooplankton from 24 United Kingdom lakes to examine the hypothesis that zooplankton dependence on allochthonous sources of organic carbon declines with increasing lake trophy. Stable isotope analysis was also carried out on particulate and dissolved organic matter (POM and DOM) and, in 11 of the lakes, of phytoplankton isolates. In 21 of the 24 lakes, the zooplankton were depleted in 13C relative to bulk POM, consistent with previous reports. δ13C for POM showed relatively little variation between lakes compared to high variation in values for DOM and phytoplankton. δ13C values for phytoplankton and POM converged with increasing lake trophy, consistent with the expected greater contribution of autochthonous production to the total organic matter pool in eutrophic lakes. The difference between δ13C for zooplankton and that for POM was also greatest in oligotrophic lakes and reduced in mesotrophic lakes, in accordance with the hypothesis that increasing lake trophic state leads to greater dependence of zooplankton on phytoplankton production. However, the difference increased again in hypertrophic lakes, where higher δ13C values for POM may have been due to greater inputs of 13C-enriched organic matter from the littoral zone. The very wide variation in phytoplankton δ13C between lakes of all trophic categories made it difficult to detect robust patterns in the variation in δ13C for zooplankton.

A dynamic study of earthworm feeding ecology -using stable isotopes.

Changes in the specific diet of earthworms with time in relation to landuse changes and two different climates were studied by analysing (13)C and (15)N natural abundance in soils and animals. Soil samples from three depths (0-10, 10-20 and 20-30 cm) and earthworms were collected from two sites: Santiago (Northwest Spain) and North Wyke (Southwest England) both consisting of replicated long-term grasslands and recently converted to maize plots. Earthworms were hand-sorted in the field at the peak of the maize growth and after harvesting at both sites. In the Spanish plots, nine and eight earthworm species, all belonging to the three ecological categories (epigeic, anecic and endogeic), were found under maize and permanent pasture, whereas at the English site five and seven different species were, respectively, identified. At both sites (13)C isotopic values of the earthworm tissues reflected changes in diet from C(3) to C(4) with epigeic and epi/anecic worms in the maize plots showing one delta unit difference in relation to the ones found in the grassland plots. Anecic worms seemed to be less responsive to landuse changes. The higher (13)C values of the Spanish soils were also reflected in the earthworm tissues when compared with the English samples. (15)N values showed no clear relationship with the cropping treatments but were clearly related to the ecological grouping, with endogeic worms reaching the highest values whereas for the epigeic and epi/anecic species the lowest values were obtained. This finding was also previously recorded by other authors1 and suggests that, in the future, stable isotope techniques could also be a useful tool in taxonomic studies. Copyright 1999 John Wiley & Sons, Ltd.

Transurethral needle ablation of the prostate: a urodynamic based study with 2-year followup.

PURPOSE: We evaluated the efficacy of transurethral needle ablation of the prostate for the treatment of lower urinary tract symptoms related to benign prostatic hyperplasia (BPH). This study was urodynamic based with 2-year followup to determine whether transurethral needle ablation of the prostate could reduce bladder outlet obstruction and, if so, whether the effect was durable. MATERIALS AND METHODS: A total of 47 patients with symptomatic BPH underwent transurethral needle ablation of the prostate under local anesthesia and intravenous sedation. All patients were evaluated subjectively using the American Urological Association symptom index and the quality of life score. Patients were evaluated objectively with uroflowmetry, post-void residual volume and pressure-flow studies. All patients underwent subjective and objective evaluation before treatment. Followup was conducted at 1, 3, 6, 12 and 24 months after treatment. Short and long-term complications were assessed. RESULTS: At 6-month followup there was 71% improvement in mean cases (22.4 to 6.6, 42 patients symptom index, p < 0.05), and 66% improvement in mean quality of life score (4.6 to 1.56, 42 patients, p < 0.05). Maximum flow rate, post-void residual volume and detrusor pressure at maximum flow rate also showed statistically significant improvements throughout the study. At 12-month followup there was a 55% increase in maximum flow rate (6.6 to 10.23 ml. per second, 29 patients, p < 0.05). A 37% reduction in mean detrusor pressure at maximum flow rate (92.4 cm. to 58 cm. water, 31 patients, p < 0.05) was recorded at 24-month followup, thus indicating that transurethral needle ablation of the prostate can lower bladder pressure-significantly. Post-void residual volume decreased from a pretreatment mean of 76.1 ml. to a mean of 36.9 ml. (31 patients, p < 0.05) at 24 months. Short-term complications (3 months) included transient posttreatment urinary retention in 8 patients (17%), duration 1 to 9 days, mild to moderate transient frequency dysuria all patients which resolved in more than 90% by 5 weeks and epididymitis in 1. A patient questionnaire was used to evaluate changes in sexual function and there were no reports of disturbances in erectile function or retrograde ejaculation. There were no long-term complications. However, 6 patients (12.7%) had persistent bothersome symptoms during the followup period and underwent transurethral prostate resection. Further analysis of this subset of patients with respect to pretreatment evaluation and transurethral needle ablation procedure did not reveal significant differences between them and patients with successful outcomes. CONCLUSIONS: Transurethral prostate resection is a safe and effective technique for treating lower urinary tract symptoms related to benign prostatic hyperplasia. The technique can be performed in the office as an outpatient, or as a same day surgical procedure, using topical anesthesia with intravenous sedation, if necessary. In the majority of patients subjective and objective improvements were sustained for the duration of this study, which included 2-year followup with pressure-flow studies.

Crystallization and X-ray diffraction study of recombinant platelet-derived endothelial cell growth factor.

Crystals of recombinant platelet-derived endothelial cell growth factor (PD-ECGF) were obtained by the hanging drop vapour diffusion technique. The crystals belong to the space group P2(1)2(1)2(1) with unit cell dimensions a = 63.7 A, b = 70.4 A, c = 219.6, alpha = beta = gamma = 90 degrees, and probably contain a single dimer in the asymmetric unit. Diffraction to a minimum Bragg spacing of 3.5 A has been obtained using a synchrotron X-ray source.

Thymidine phosphorylase activity of platelet-derived endothelial cell growth factor is responsible for endothelial cell mitogenicity.

Recombinant human platelet-derived endothelial cell growth factor, expressed in the yeast Saccharomyces cerevisiae was purified to greater than 98% purity by anion-exchange and hydroxyapatite chromatography. It was shown to possess thymidine phosphorolytic activity in vitro (pH optimum, pH 5.3; Km, 0.11 mM; Vmax, 12.5 mmol min-1 mg-1; turnover number, 9.4 s-1). Covalent modification simultaneously inhibited the enzymatic and mitogenic properties of the protein, while interaction with a cell-surface receptor was not required to stimulate mitogenesis. Purified Escherichia coli thymidine phosphorylase was also mitogenic toward endothelial cells. It is proposed that platelet-derived endothelial cell growth factor is human thymidine phosphorylase which promotes endothelial cell proliferation by reducing thymidine levels that would otherwise be inhibitory to endothelial cell growth.

Expression of recombinant platelet-derived endothelial cell growth factor in the yeast Saccharomyces cerevisiae.

A platelet-derived endothelial cell growth factor cDNA has been cloned, sequenced and expressed using the Saccharomyces cerevisiae PRB1 promoter. Soluble recombinant platelet-derived endothelial cell growth factor constituted 0.5-1.0% of total soluble protein. Yeast soluble protein extracts containing recombinant platelet-derived endothelial cell growth factor stimulate the growth of calf pulmonary artery endothelial cells in vitro.

Cloning and characterisation of the Saccharomyces cerevisiae glycerol-3-phosphate dehydrogenase (GUT2) promoter.

The Saccharomyces cerevisiae glycerol-3-phosphate dehydrogenase (GUT2) promoter and part of the protein-coding region have been isolated on a 6.3-kb genomic DNA fragment. Nucleotide sequence analysis shows that the promoter has many structural features in common with yeast glycolytic enzyme promoters. Chromosomal mapping indicates that this genomic fragment is located on chromosome XII. The GUT2 promoter has been used to construct a recombinant human albumin (reHA) secretion vector; yeast transformed with this vector secrete reHA into the culture supernatant.

Saccharomyces cerevisiae strains that overexpress heterologous proteins.

We describe a system that facilitates the selection of host mutants that overproduce a range of secreted and internally produced heterologous proteins in Saccharomyces cerevisiae. These mutants were initially selected for their ability to oversecrete recombinant human albumin (rHA), as detected by a direct visual assay that relies upon antibody precipitation in solid media. Yeast strains that were able to synthesize and secrete increased levels of rHA also produced elevated levels of internally expressed proteins including alpha 1-antitrypsin Pittsburgh variant and plasminogen activator inhibitor type 2.