Structure-function analysis of epidermal growth factor: site directed mutagenesis and nuclear magnetic resonance

The role of leucine-47 in determining the structure and activity of human epidermal growth factor was examined using site-directed mutagenesis. Wild type protein and four variants in which Leu47 was replaced by valine, glutamate, aspartate and alanine were produced from yeast. 1H NMR experiments demonstrated that substitution of Leu47 had little effect on the protein structure. The observed reduction in receptor binding affinity caused by the substitutions could thus be attributed to perturbation of a residue directly involved in receptor interactions.     

Dietary variations of predaceous caddisfly larvae (Trichoptera: Rhyacophilidae,Polycentropodidae and Arctopsychidae) from British Columbian streams

The diets of larval Rhyacophilidae (Rhyacophila inculta), Polycentropodidae (Polycentropus variegatus) and Arctopsychidae (Parapsyche almota and P. elsis) from five streams in the University of British Columbia Research Forest, British Columbia (Canada), are recorded and related to feeding mode/constructional activities and prey representation in the habitat. Particular attention was paid to the extent of dietary overlap and the degree of intraspecific dietary variations between streams. An overall similarity of the diets of the study species was notable and all commonly consumed chironomid (Diptera) larvae, Simulium (Diptera) and Zapada (Plecoptera), although there was interspecific variation in the relative importance of these items. Polycentropus variegatus and Parapsyche spp. ate Baetis and Paraleptophlebia, the latter predators also consuming Hydrachnellae (Acarina). Other prey were generally of minor importance and consequently interspecific dietary overlaps were high. Differences in the range of prey consumed by predaceous Trichoptera were apparent. Free-foraging R. inculta which selectively consumed sedentary simuliid larvae had the narrowest niche breadth. Parapsyche spp. and Polycentropus variegatus foreguts generally contained an over-representation of chironomid larvae compared to their proportionate occurrence in the benthos, and these caddisflies exhibited high niche overlap. The relative importance of chironomids as food for Parapsyche spp. and Polycentropus variegatus is attributed to prey behaviour, i.e. drift, and poorly developed escape responses when they are caught on the predator's net.     

The population dynamics of some freshwater carideans (Crustacea: Decapoda) in Hong Kong,with special reference to Neocaridina serrata (Atyidae)

During a 15-month investigation of the population dynamics of three caridean shrimp species in the Lam Tsuen River, New Territories, Hong Kong, the numbers of the commonest species, Neocaridina serrata (Stimpson)(Atyidae), were reduced as water temperatures fell. By contrast, Caridina lanceifrons Yu became more numerous during the winter. The relatively scarce Macrobrachium hainanense (Parisi)(Palaemonidae) tended to be more abundant during the summer, and the abundance of this species was directly correlated with that of N. serrata. The population size of these two species was positively correlated with water temperatures prevailing one and two months prior to the collection of samples, but there was no significant relationship between C. lanceifrons abundance and water temperature.All species exhibited similar growth patterns with an inflection at the attainment of sexual maturity, occurring after the (presumed) 14th moult for N. serrata and in the (presumed) 13th and 18th stadium for C. lanceifrons and M. hainanensis respectively. Ovigerous shrimps of all species were only recorded when water temperatures exceeded 20 °C and the % incidence of ovigerous N. serrata was positively correlated with prevailing temperatures. It is suggested that restriction of freshwater caridean breeding periods by low temperatures may be common in the subtropics while perennial breeding is more likely to be typical of tropical regions.The mean carapace length of N. serrata populations was negatively correlated with prevailing temperatures, but more strongly correlated with temperatures recorded one and two months prior to sampling when gametogenesis may have taken place. Smallest mean sizes were recorded in late summer upon the cessation of juvenile recruitment. N. serrata has a maximum longevity of approximately 12 months and reproduction occurs in the 7th month of life or later. The hatchlings are well developed and clutch size is not related to the size of the brooding female. An annual life cycle, as seen in N. serrata, may be typical of many small freshwater carideans.Department of Zoology, The University of Hong Kong     

The life cycle, population dynamics and productivity of Melanoides tuberculata (Muller, 1774) (Gastropoda: Prosobranchia: Thiaridae) in Hong Kong

An investigation of the life cycle, population dynamics and secondary productivity of the viviparous snail Melanoides tuberculuta inhabiting flooded furrows and irrigation ditches at Ping Long, New Territories, Hong Kong, was undertaken between March 1982 and March 1983. There was a single peak in juvenile recruitment coinciding with the warmer months; hatchlings grew quickly and were sexually mature before the next breeding season. Large animals experienced high mortality and few individuals survived until a second breeding season. Secondary productivity and the production: biomass (P:B) ratio of the population (13.43 g shell–free dry weight m⁻² yr⁻¹ and 4.81, respectively) were high and exceeded values of these parameters recorded for freshwater prosobranch populations in temperate regions. Ping Long M. tuberculata were exclusively female, as is typical of most populations of this species. Animals > 3.0 mm shell width (90–120 days old) had eggs and developing larvae in the brood pouch, and fully–developed larvae were recorded from brood pouches throughout the study although release of hatchlings was distinctly seasonal.
A comparison of life cycle parameters of Malaysian M. turberculata with those of the Ping Long population revealed a marked similarity. The significance of such similarity and the importance of parthenogenicity in the life cycle of this widely distributed snail is discussed. Melanoides tuberculata exhibits the characteristics of an opportunist or 'fugitive' species and, by virtue of its reproductive strategies and high productivity, is able to colonize rapidly and temporarily dominate vacant habitats.     

The influence of riparian vegetation on macroinvertebrate community structure and functional organization in six new Guinea streams

Information on the ecology of New Guinea streams is meagre, and data are needed on the trophic basis of aquatic production in rivers such as the Sepik in Papua New Guinea which have low fish yields. This study investigates the relationship between riparian shading (from savanna grassland to primary rainforest), algal and detrital food, and macroinvertebrate abundance and community structure in 6 Sepik River tributary streams. A particular aim was to elucidate macroinvertebrate community responses to changes in riparian conditions. All streams supported diverse benthic communities, but morphospecies richness (overall total 64) was less than in streams on the tropical Asian mainland; population densities of benthic invertebrates, by contrast, were similar to those recorded elsewhere. Low diversity could reflect limited taxonomic penetration, but may result from the absence of major groups (Plecoptera, Heptageniidae, Ephemerellidae, Psephenidae, Megaloptera, etc.) which occur on the Asian mainland. Population densities of all 19 of the most abundant macroinvertebrate taxa varied significantly among the 6 study streams, but community composition in each was broadly similar with dominance by Baetidae and (in order of decreasing importance), Leptophlebiidae, Orthocladiinae, Elmidae and Hydropsychidae. Principal components analysis (PCA) undertaken on counts of abundant macroinvertebrate taxa clearly separated samples taken in two streams from the rest. Both streams contained high detrital standing stocks and one was completely shaded by rainforest. Stepwise multiple-regression analysis indicated that population densities of the majority of abundant taxa (11 out of 19) across streams (10 samples per stream; n = 60) were influenced by algae and/or detritus, although standing stocks of these variables were not clearly related to riparian conditions. When regression analysis was repeated on mean counts of taxa per stream (dependent variables) versus features of each stream as a whole (thus n = 6), % shading and detritus were the independent variables yielding significant regression models most frequently, but pH, total-nitrogen loads and algae were also significant predictors of faunal abundance. Further regression analysis, undertaken separately on samples (n = 10) from each stream, confirmed the ability of algae and detritus to account for significant portions of the variance in macroinvertebrate abundance, but the significance of these variables varied among streams with the consequence that responses of individual taxa to algae or detritus was site-specific.Community functional organization — revealed by investigation of macroinvertebrate functional feeding groups (FFGs) — was rather conservative, and streams were codominated by collector-gatherers (mean across 6 streams = 43%) and grazers (36%), followed by filter-feeders (15%) and predators (7%). The shredder FFG was species-poor and comprised only 0.4% of total macroinvertebrate populations; shredders did not exceed 2% of benthic populations in any stream. PCA of FFG abundance data was characterized by poor separation among streams, although there was some evidence of clustering of samples from unshaded sites. The first 2 PCA axes accounted for 84% of the variation in the data suggesting that the poor separation resulted from the general similarity of FFG representation among streams. Although stepwise multiple-regression analysis indicated that algae and detritus accounted for significant proportions of the variations in population density and relative abundance of some FFGs, the response of community functional organization to changes in riparian conditions and algal and detrital food base was weak — unlike the deterministic responses that may be typical of north-temperate streams.     

Effects of decreased atmospheric deposition on the sulfur budgets of two Dutch moorland pools

The chemical composition of surface waters of two Dutch moorland pools and of incident precipitation, was monitored from 1982 to 1990. For this period, sulfur and water budgets were calculated using a hydrochemical model developed for well-mixed non-stratifying lakes. Total atmospheric deposition of S decreased significantly after 1986 at both locations. A model describing the sulfur budget in terms of input, output and reduction/oxidation processes predicted a fast decrease of pool water SO₄ ²⁻ concentrations after a decrease of atmospheric input. However, SO₄ ²⁻ concentrations in the surface water was lowered only slightly or remained constant. Apparently a source within the lake caused the unexpectedly high SO₄ ²⁻ concentrations. The possible supply of SO₄ ²⁻ from the sediment through regulation by (K-)Al-SO₄ containing minerals or desorption of SO₄ ²⁻ from positively charged surfaces in the sediment was evaluated. Solubility calculations of pore water with respect to alunite, basaluminite and jurbanite indicated that SO₄ ²⁻ concentration was not regulated by these minerals. It is suggested here (1) that desorption of SO₄ ²⁻ from peaty sediments may account for the estimated SO₄ ²⁻ supply provided that the adsorption complex is periodically recharged by partial oxidation of the upper bottom sediments and (2) that because of exposure of a part of the pool bottom to the atmosphere during dry summers and subsequent oxidation of reduced S, the amount of SO₄ ²⁻ may be provided which complements the decreasing depositional SO₄ ²⁻ input. In future research these two mechanisms need to be investigated.     

The population dynamics and sexual strategy of Anodonta woodiana (Bivalvia: Unionacea) in Plover Cove Reservoir, Hong Kong

A 12-month investigation of the population dynamics, growth, ageing, reproduction and sexuality of Anodonta woodiana (Lea, 1834) in Plover Cove Reservoir, Hong Kong, has been undertaken. Recruitment occurs in the summer, May through July being the period of peak incidence of glochidia on captive fish hosts. Juveniles grow rapidly to a shell length of 30 mm and are sexually mature by the following spring. A study of the population structure of A. woodiana indicated that most animals were less than 8 years old. Results of a Walford plot for this species indicated a maximum theoretical shell length (in Plover Cove) of 162·5 mm and a life span of approximately 12 years; the latter is in good agreement with age estimates for European unionids.
Anodonta woodiana is dioecious with females predominating in a ratio of 60:40. A very small number of individuals (0·3%) are hermaphrodite. Males possess mature gonads throughout the year whereas females come into reproductive condition during the spring. There was no evidence of shell form sexual dimorphism, as has been recorded for other unionaceans.
The reproductive strategy and time of gonad maturation of A. woodiana is compared with that of unionids from elsewhere and with other fresh water bivalves from southern China.     

Sulfate reduction and S-oxidation in a moorland pool sediment

In an oligotrophic moorland pool in The Netherlands, S cycling near the sediment/water boundary was investigated by measuring (1) SO₄ ^2– reduction rates in the sediment, (2) depletion of SO₄ ^2– in the overlying water column and (3) release of³⁵S from the sediment into the water column. Two locations differing in sediment type (highly organic and sandy) were compared, with respect to reduction rates and depletion of SO₄ ^2– in the overlying water.Sulfate reduction rates in sediments of an oligotrophic moorland pool were estimated by diagenetic modelling and whole core³⁵SO₄ ^2– injection. Rates of SO₄ ^2– consumption in the overlying water were estimated by changes in SO₄ ^2– concentration over time in in situ enclosures. Reduction rates ranged from 0.27–11.2 mmol m^–2 d^–1. Rates of SO₄ ^2– uptake from the enclosed water column varied from –0.5, –0.3 mmol m^–2 d^–1 (November) to 0.43–1.81 mmol m^–2 d^–1 (July, August and April). Maximum rates of oxidation to SO₄ ^2– in July 1990 estimated by combination of SO₄ ^2– reduction rates and rates of in situ SO₄ ^2– uptake in the enclosed water column were 10.3 and 10.5 mmol m^–2 d^–1 at an organic rich and at a sandy site respectively.Experiments with³⁵S^2– and³⁵SO₄ ^2– tracer suggested (1) a rapid formation of organically bound S from dissimilatory reduced SO₄ ^2– and (2) the presence of mainly non SO₄ ^2–-S derived from reduced S transported from the sediment into the overlying water. A³⁵S^2– tracer experiment showed that about 7% of³⁵S^2– injected at 1 cm depth in a sediment core was recovered in the overlying water column.Sulfate reduction rates in sediments with higher volumetric mass fraction of organic matter did not significantly differ from those in sediments with a lower mass fraction of organic matter.Corresponding author     

The Active and the Inactive Form of the hAT1 Receptor Have an Identical Ligand-Binding Environment: An MPA Study on a Constitutively Active Angiotensin II Receptor Mutant

Several models of activation mechanisms were proposed for G protein-coupled receptors (GPCRs), yet no direct methods exist for their elucidation. The availability of constitutively active mutants has given an opportunity to study active receptor conformations within acceptable limits using models such as the angiotensin II type 1 (AT₁)¹ receptor mutant N111G-hAT₁ which displays an important constitutive activity. Recently, by using methionine proximity assay, we showed for the hAT₁ receptor that TMD III, VI, and VII form the ligand-binding pocket of the C-terminal amino acid of an antagonistic AngII analogue. In the present contribution, we investigated whether the same residues would also constitute the ligand-binding contacts in constitutively activated mutant (CAM) receptors. For this purpose, the same Met mutagenesis strategy was carried out on the N111G double mutants. Analysis of 43 receptors mutants in the N111G-hAT₁ series, photolabeled and CNBr digested, showed that there were only subtle structural changes between the wt-receptor and its constitutively active form.