ALBERT

Notes: A polymerase chain reaction (PCR)-based assay to detect nervous necrosis virus (NNV) in fish was developed by using two sets of primers designed on a highly conserved region of the coat protein gene encoded by RNA2 of NNV. The first pair of primers amplified a fragment of 605 bp by one-step reverse-transcription (RT)-PCR, while the second pair amplified an internal segment of 255 bp by nested PCR. Addition of nested PCR increased the assay sensitivity 100-fold when carried out in a separate tube (two-step assay) and 10-fold when performed in the same tube (one-step assay). The sensitivity of the two-step assay was 104 times higher than that of virus cultivation. Nested PCR served also to confirm the specificity of the first amplification, as verified also by Southern hybridization analysis and direct sequencing. In species known to be susceptible to infection, such as European sea bass, Dicentrarchus labrax, and gilthead seabream, Sparus aurata, NNV was often detectable in brain tissue by RT-PCR alone but only by the two-step assay in blood, sperm, ovarian tissue or larvae. The same was true for sperm and ovarian tissue of shi drum, Umbrina cirrosa. NNV was also detected in the brains of Japanese red seabream, Pagrus major and brown meagre, Sciaena umbra, suggesting that these species can also be infected. No NNV was detected in samples of Artemia salina nauplii and rotifers obtained from a fish farm with an NNV outbreak. The inclusion of nested PCR in the assay appears to be necessary to screen out NNV-positive broodfish by blood sampling and testing of their larval progeny.

Notes: The mid-day responses of wheat ear CO2 and water vapour exchange to full-season CO2 enrichment were investigated using a Free-Air CO2 Enrichment (FACE) apparatus. Spring wheat [Triticum aestivum (L). cv. Yecora Rojo] was grown in two experiments under ambient and elevated atmospheric CO2 (Ca) concentrations (approximately 370 μmol mol−1 and 550 μmol mol−1, respectively) combined first with two irrigation (Irr) schemes (Wet: 100% and Dry: 50% replacement of evapotranspiration) and then with two levels of nitrogen (N) fertilization (High: 350, Low: 70 kg ha−1 N). Blowers were used for Ca enrichment. Ambient Ca plots were exposed to blower induced winds as well the Ca× N but not in the Ca× Irr experiment. The net photosynthesis for the ears was increased by 58% and stomatal conductance (gs) was decreased by 26% due to elevated Ca under ample water and N supply when blowers were applied to both Ca treatments. The use of blowers in the Ca-enriched plots only during the Ca× Irr experiment (blower effect) and Low N supply restricted the enhancement of net photosynthesis of the ear due to higher Ca. In the latter case, the increase of net photosynthesis of the ear amounted to 26%. The decrease in gs caused by higher Ca was not affected by the blower effect and N treatment. The mid-day enhancement of net photosynthesis due to elevated Ca was higher for ears than for flag leaves and this effect was most pronounced under ample water and N supply. The contribution of ears to grain filling is therefore likely to increase in higher Ca environments in the future. In the comparison between Wet and Dry, the higher Ca did not alter the response of net photosynthesis of the ear and gs to Irr. However, Ca enrichment increased the drought tolerance of net photosynthesis of the glume and delayed the increase of the awn portion of net photosynthesis of the ear during drought. Therefore, the role of awns for maintaining high net photosynthesis of the ear under drought may decrease when Ca increases.

Notes: 1. Knowledge of synchrony in trends is important to determining regional responses of lakes to disturbances such as atmospheric deposition and climate change. We explored the temporal coherence of physical and chemical characteristics of two series of mostly alpine lakes in nearby basins of the Colorado Rocky Mountains. Using year-to-year variation over a 10-year period, we asked whether lakes more similar in exposure to the atmosphere be-haved more similarly than those with greater influence of catchment or in-lake processes.2. The Green Lakes Valley and Loch Vale Watershed are steeply incised basins with strong altitudinal gradients. There are glaciers at the heads of each catchment. The eight lakes studied are small, shallow and typically ice-covered for more than half the year. Snowmelt is the dominant hydrological event each year, flushing about 70% of the annual discharge from each lake between April and mid-July. The lakes do not thermally stratify during the period of open water. Data from these lakes included surface water temper-ature, sulphate, nitrate, calcium, silica, bicarbonate alkalinity and conductivity.3. Coherence was estimated by Pearson's correlation coefficient between lake pairs for each of the different variables. Despite close geographical proximity, there was not a strong direct signal from climatic or atmospheric conditions across all lakes in the study. Individual lake characteristics overwhelmed regional responses. Temporal coherence was higher for lakes within each basin than between basins and was highest for nearest neighbours.4. Among the Green Lakes, conductivity, alkalinity and temperature were temporally coherent, suggesting that these lakes were sensitive to climate fluctuations. Water tem-perature is indicative of air temperature, and conductivity and alkalinity concentrations are indicative of dilution from the amount of precipitation flushed through by snowmelt.5. In Loch Vale, calcium, conductivity, nitrate, sulphate and alkalinity were temporally coherent, while silica and temperature were not. This suggests that external influences are attenuated by internal catchment and lake processes in Loch Vale lakes. Calcium and sulphate are primarily weathering products, but sulphate derives both from deposition and from mineral weathering. Different proportions of snowmelt versus groundwater in different years could influence summer lake concentrations. Nitrate is elevated in lake waters from atmospheric deposition, but the internal dynamics of nitrate and silica may be controlled by lake food webs. Temperature is attenuated by inconsistently different climates across altitude and glacial meltwaters.6. It appears that, while the lakes in the two basins are topographically close, geologically and morphologically similar, and often connected by streams, only some attributes are temporally coherent. Catchment and in-lake processes influenced temporal patterns, especially for temperature, alkalinity and silica. Montane lakes with high altitudinal gradients may be particularly prone to local controls compared to systems where coherence is more obvious.

Notes: Saccharomyces cerevisiae strains lacking a functional Pho85 cyclin-dependent kinase (cdk) exhibit a complex phenotype, including deregulation of phosphatase genes controlled by the transcription factor Pho4, slow growth on rich media, failure to grow using galactose, lactate or glycerol as a carbon source and hyperaccumulation of glycogen. The ability of Pho85 to regulate the transcription factor Pho4 is mediated by its association the Pho80 cyclin. Some other regulatory functions of the Pho85 cdk have been shown to be mediated via its interaction with a recently identified family of Pho80-related cyclins (Pcls). Here, we show that the poorly characterized Pho80-like protein Pcl7 forms a functional kinase complex with the Pho85 cdk, and that the activity of this complex is inhibited in response to phosphate starvation. Additionally, we show that Pcl7 interacts with the phosphate-regulated cyclin–cdk inhibitor Pho81, and that the regulation of the Pcl7–Pho85 complex in response to changes in phosphate levels is dependent on Pho81. Thus, we demonstrate for the first time that the Pho81 regulator is not dedicated to regulating Pho80, but may act to co-ordinate the activity of both the Pho80–Pho85 and Pcl7–Pho85 cyclin–cdk complexes in response to phosphate levels. We also demonstrate that expression of Pcl7 is cell cycle regulated, with maximal activity occurring in mid to late S-phase, perhaps suggesting a role for Pcl7 in cell cycle progression. Finally, we describe the phenotype of pcl7Δ and pcl6Δ yeast strains that have defects in carbon source utilization.

Notes: Abstract Two fundamental aspects of frequency analysis shape the functional organization of primary auditory cortex. For one, the decomposition of complex sounds into different frequency components is reflected in the tonotopic organization of auditory cortical fields. Second, recent findings suggest that this decomposition is carried out in parallel for a wide range of frequency resolutions by neurons with frequency receptive fields of different sizes (bandwidths). A systematic representation of the range of frequency resolution and, equivalently, spectral integration shapes the functional organization of the iso-frequency domain. Distinct subregions, or "modules," along the iso-frequency domain can be demonstrated with various measures of spectral integration, including pure-tone tuning curves, noise masking, and electrical cochlear stimulation. This modularity in the representation of spectral integration is expressed by intrinsic cortical connections. This organization has implications for our understanding of psychophysical spectral integration measures such as the critical band and general cortical coding strategies.

Notes: Abstract The prostaglandin endoperoxide H synthases-1 and 2 (PGHS-1 and PGHS-2; also cyclooxygenases-1 and 2, COX-1 and COX-2) catalyze the committed step in prostaglandin synthesis. PGHS-1 and 2 are of particular interest because they are the major targets of nonsteroidal anti-inflammatory drugs (NSAIDs) including aspirin, ibuprofen, and the new COX-2 inhibitors. Inhibition of the PGHSs with NSAIDs acutely reduces inflammation, pain, and fever, and long-term use of these drugs reduces fatal thrombotic events, as well as the development of colon cancer and Alzheimer's disease. In this review, we examine how the structures of these enzymes relate mechanistically to cyclooxygenase and peroxidase catalysis, and how differences in the structure of PGHS-2 confer on this isozyme differential sensitivity to COX-2 inhibitors. We further examine the evidence for independent signaling by PGHS-1 and PGHS-2, and the complex mechanisms for regulation of PGHS-2 gene expression.

Notes: Many abiotic environmental factors elicit the production of stress-ethylene in higher plants. To elucidate the molecular mechanisms underlying the regulation of stress-ethylene production and the physiological roles played by stress-ethylene in stress responses of plants, we studied the gene expression of ACC synthase in tobacco plants that had been subjected to environmental stresses. Four new tobacco ACC synthase cDNA fragments, NT-ACS2, NT-ACS3, NT-ACS4 and NT-ACS5, were identified and sequenced. It was found that NT-ACS2 could be induced by wounding, cold temperature and, especially, sunlight. NT-ACS4 was induced at a faster kinetics by wounding. The multiple environmental stress-responsive (MESR) NT-ACS2 gene was found to contain three introns and four exons and encode a polypeptide of 484 amino acids, 54·6 kDa and pI 6·87. Computer analysis of the 3·4 kb 5′ flanking region upstream of the ACS coding region revealed the existence of a group of putative cis-acting regulatory elements potentially conferring wounding, chilling, and UV light inducibility. Phylogenetic analysis of ACC synthase genes of different plant origins indicated that the chill-inducible NT-ACS2 gene is closely related to a chilling-inducible citrus ACS gene.

Notes: Although exogenous electric fields have been reported to influence the orientation of plant root growth, reports of the ultimate direction of differential growth have been contradictory. Using a high-resolution image analysis approach, the kinetics of electrotropic curvature in Vigna mungo L. roots were investigated. It was found that curvature occurred in the same root toward both the anode and cathode. However, these two responses occurred in two different regions of the root, the central elongation zone (CEZ) and distal elongation zone (DEZ), respectively. These oppositely directed responses could be reproduced individually by a localized electric field application to the region of response. This indicates that both are true responses to the electric field, rather than one being a secondary response to an induced gravitropic stimulation. The individual responses differed in the type of differential growth giving rise to curvature. In the CEZ, curvature was driven by inhibition of elongation, whereas curvature in the DEZ was primarily due to stimulation of elongation. This stimulation of elongation is consistent with the growth response of the DEZ to other environmental stimuli.

Notes: 1. Because people impact lake ecosystems, it is important to consider factors influencing the human use of freshwater resources. We investigated the influence of the landscape position, as well as lake area, recreational facilities, and distance to highways and urban centres, on lake use by boaters in the Northern Highland Lake District of Wisconsin, U.S.A.2. In aerial surveys of ninety-nine randomly selected lakes, we did not see boats on over half of the lakes. Of the lakes with boats, we found a strong correlation between the number of boats and lake area. Recreational boats tended to be found on large, accessible lakes with good boating facilities. Boats were not seen on small, stained lakes with few recreational facilities.3. Regression models showed that lake size and landscape position explained 63% of the variability in the average number of boats per lake and landscape position explained 24% of the variability in boat density on all ninety-nine lakes. Social variables representing the quality of boating facilities and the perception of good fishing explained 70% of the variability in number of boats per lake and 54% of the variation in boat density on all lakes. A combined model using both physical and social variables increased the explanatory power for both number and density. Lake use by boaters was correlated with landscape position, the quality of fishing and the availability of recreational facilities. When the analysis was restricted to the forty-six lakes where boats were found, only the availability of recreational facilities proved a significant predictor of boat density.4. Our results suggest that lake choice by recreational boaters may be best predicted by a combination of the location of a lake in a regional hydrologic landscape, and considerations of available facilities and perceptions regarding fishing.