Fusion protein-based epitope mapping of phytochrome. Precise identification of an evolutionarily conserved domain

Fusion proteins are used to define with precision an evolutionarily conserved domain on the carboxyl-terminal portion of the chromoprotein phytochrome. Simultaneously, assignments of two other epitopes are made with significantly greater precision, while the location of a fourth is confirmed. The epitope-mapping method that is described here is systematic, using complementary, overlapping nested sets of fusion proteins of predefined sequence rather than randomly generated peptides. Moreover, in contrast to previous methods, this approach yields rigorous and unambiguous assignments because it relies solely upon the ability of an antibody to detect a given polypeptide. A cDNA fragment encoding phytochrome amino acids 464-1129, which is its carboxyl terminus, was identified in lambda gt11 and subcloned in frame into the lacZ alpha sequence of pUC18. Four nested sets of subclones in pUC18 were created by digestion with selected restriction endonucleases and with the exonuclease Bal31. Fusion proteins were analyzed by immunoblotting following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The epitope for monoclonal antibody Oat-13 was confirmed to be between residues 551 and 617, while the epitopes for Oat-8 and Oat-28 were narrowed to 624-686 and 624-747, respectively. The epitope recognized by Pea-25, Pea-2, and Oat-15 was resolved unequivocally to a sequence of only seven residues (residues 765-771): N-Pro-Ile-Phe-Gly-Ala-Asp-Glu-C.     

Effects of Schistosoma mansoni on the nutrition of its intermediate host, Biomphalaria glabrata

Biomphalaria glabrata infected with Schistosoma mansoni and maintained on an artificial diet of Spirulina alga displayed reduced growth during the 5 wk following patency. Food consumption per unit snail weight was unaffected. Infected snails also failed to lay eggs. No difference in percentage of assimilation was observed between control and infected individuals, but infected snails had significantly decreased gross conversion efficiencies. The effects of infection on nutrition of B. glabrata were similar to those observed in nutrient-deprived snails fed diets containing low Spirulina levels. Nutrient deprivation, however, did not alter conversion efficiency.     

Aerosolization of recombinant SLPI to augment antineutrophil elastase protection of pulmonary epithelium

In a variety of lung diseases the respiratory epithelial surface must contend with an increased burden of neutrophil elastase (NE). One candidate for augmenting epithelial anti-NE protection is the secretory leukoprotease inhibitor (SLPI). In vitro evaluation demonstrated that 96 +/- 1% of the recombinant SLPI (rSLPI) molecules were capable of inhibiting NE, with an association rate constant of 7.1 +/- 0.1 X 10(6) M-1.s-1. Evaluation of rSLPI after in vitro and in vivo aerosolization showed that aerosolization did not alter rSLPI. Aerosolization of a single dose of 50 mg rSLPI to sheep resulted in a fourfold increase of the anti-NE capacity in epithelial lining fluid (ELF) at 3 h, with a half-life in ELF of 12 h. After aerosolization some rSLPI appeared in lung lymph. Simultaneous aerosolization of rSLPI and recombinant alpha 1-antitrypsin (rAAT) demonstrated a molar ratio of the concentration in lymph to the concentration in ELF 3 h after the aerosol eightfold higher for rAAT than for rSLPI. Overall, these observations demonstrate that it is feasible to use aerosolized rSLPI to directly augment the anti-NE capacity of the lung, particularly on the pulmonary epithelial surface.     

Meteorological factors and the time of onset of chest pain in acute myocardial infarction

Analysis of the time of onset of chest pain in 2254 patients with a myocardial infarction admitted to a coronary care unit in Leicester during a 10-year period shows an association with temperature and humidity. During both the most cold and humid times of the year, the relationship is a strong one. A generalized linear model with a log link was used to fit the data and the backward elimination selection procedure suggested a humid, cold day might help to trigger the occurrence of myocardial infarction. In addition, cold weather was found to have a stronger effect on the male population while those men aged between 50 and 70 years were more sensitive to the effect of high humidity.     

Distribution and breeding biology of offshore cichlids in Lake Malawi/ Niassa

Synopsis Lake Malawi/Niassa is the second largest rift valley lake in Africa, with an area of 28 800 km², and an average and maximum depth of 292 m and>700 m, respectively. The lake is well known for the great diversity of fish occurring in the inshore zone. However, the offshore fish community is poorly documented. To rectify this, regular sampling was undertaken over two years, using trawl and gillnets at six offshore locations. This paper reports on the species composition, spatial distribution and breeding biology of the dominant cichlids species from the offshore pelagic zone. Cichlids formed approximately 88% of the offshore fish biomass. Most abundant were two species of zooplanktivores in the genus Diplotaxodon that made up 71% of the offshore fish biomass. An undescribed species, given the cheironym D. bigeye, was mainly found at a depth of 220 m during the day, but moved into near surface waters at night when the moon was full. This species was absent from the shallow regions of the lake. The most abundant offshore species was D. limnothrissa, which was distributed evenly throughout the lake to depths of 220 m. A less common offshore zooplanktivore was Copadichromis quadrimaculatus that formed 5% of the biomass and was confined to the upper 100 m of the water column. The main piscivores were in the genus Rhamphochromis and formed approximately 10% of the offshore fish biomass. The two dominant taxa were R. longiceps and the large Rhamphochromis group, and both were more common in the southern half of the lake. The former occurred mainly in the upper 100 m of the water column and the latter mainly at depths of 100–150 m. The length at maturity and fecundity for the dominant offshore species were estimated and seasonal breeding cycles determined from gonad activity and gonado-somatic indices.     

Evolutionary ecology and the conservation of biodiversity

Studies on evolving interactions among species and the coevolutionary process have suggested that the conservation of biodiversity requires a broad geographic perspective, if the `interaction biodiversity' of the earth is to be conserved with its species diversity. Continued maintenance of the geographic mosaic of specialization, defense and population structure appears to be crucial to the coevolutionary process and the long-term persistence of some interspecific interactions.     

The analysis of the NSW wheat variety database. I. Modelling trial error variance

The retrospective analysis of a large database on wheat variety testing in New South Wales (NSW) is considered. This analysis involved three key steps. Initially error variance heterogeneity is modelled, indicating significant differences in error variance due to trial location, year of trialling, sowing date and trial mean yield. The implication of this modelling for the estimaion of variance components is discussed.     

The Probability Distribution of the Amount of an Individual''s Genome Surviving to the following Generation

The probability that at least p% of an individual's genome is passed on collectively to his children is calculated. With data availability the consideration of the chromosome as a whole rather than discrete loci becomes of increasing practical importance. Assuming the genomic continuum model, which allows for recombination, the crossover process in a chromosome pedigree is viewed as a continuous-time Markov random walk on the vertices of a hypercube with time parameter map distance along the chromosome. The desired probability corresponds to the probability of sojourn times of the process in a small set of vertices, which are well approximated via the Poisson clumping heuristic. Results are given for the human genome. It is very likely that an individual with at least four children passes on at least 90% of his genome. There exists no ``equivalent' number of independently segregating loci for this distribution.     

Fluoranthene as a model toxicant in sediment studies with Chironomus riparius

The bioavailability of fluoranthene (as a model toxicant) in freshwater sediment tests with Chironomus riparius was investigated in relation to the partitioning of the chemical between water and sediment. Effects on survival and successful development of newly hatched larvae through to adult emergence were measured over a range of sediment concentrations. Radiolabelled fluoranthene was employed to enable sediment, aqueous and tissue concentrations to be determined. A significant effect on emergence was obtained at a measured sediment concentration of 80 mg kg^-1 (dry weight). The measured concentration in the overlying water was 88 g l^-1. Although effects at 170 mg kg^-1 (dry weight) were more severe, the measured concentration in the overlying water was similar (89 g l^-1), probably reflecting the approach towards the solubility limit. Bioaccumulation of fluoranthene, determined by analysis of emerging adults, was approximately proportional to the sediment concentration. Although the aqueous phase concentration appeared to be the principal factor controlling bioavailability, the increasing toxicity, when water concentrations had reached a plateau, suggested that other mechanisms may also be involved. Different sediment spiking procedures resulted in broadly similar partitioning of the chemical between the sediment and the overlying water. There were only minor differences in toxicity between the procedures, two of which allowed the carrier solvent used for spiking to be eliminated before addition of the test organisms. It was concluded that, for most purposes, all four procedures were equally acceptable, and by retaining a flexible approach, even for standardised regulatory tests, spiking procedures may be tailored to the physico-chemical properties of the test material.     

The effects of tail autotomy on survivorship and body growth of Uta stansburiana under conditions of high mortality

We examined the effects of tail autotomy on survivorship and body growth of both adult and juvenile Uta stansburiana by directly manipulating tail condition. Tail loss decreased neither survivorship nor rate of body growth for individuals in two natural populations. Lack of an influence of tail loss on survivorship in these two populations may be the result of high mortality. Under high mortality any differential effects of tail loss will be lower than in populations facing lower mortality. Growth experiments in the laboratory demonstrated that, under conditions of minimal environmental variation and social interactions, there is no tradeoff between body growth and tail regeneration as has been suggested for other species of lizards. One possible reason for this difference is that U. stansburiana does not use the tail as a storage organ for lipids. The original and regenerated tails are composed mainly of protein. In general, any differential body growth between tailed and tailless individuals may be due to social interactions and not a diversion of limited energy into tail regeneration.