Bioassay for detecting active site insensitivity in horn fly (Diptera: Muscidae) larvae

A bioassay was used to detect active site insensitivity (knock-down resistance [kdr]) in pyrethroid resistant larvae of the horn fly, Haematobia irritans (L.). The larvae of the resistant population had KD50's 42.0-, 28.1- and 29.2-fold greater to permethrin, fenvalerate and lambda-cyhalothrin, respectively, compared with the susceptible population. In filter paper bioassays, resistant adult horn flies were 17 to 39.1 times less susceptible to the pyrethroids than susceptible adults at LC50. These results further document active site insensitivity as the major mechanism of pyrethroid resistance in the horn fly.     

The charge and structural stability of apolipoprotein A-I in discoidal and spherical recombinant high density lipoprotein particles.

The details of how high density lipoprotein (HDL) microstructure affects the conformation and net charge of apolipoprotein (apo) A-I in various classes of HDL particles have been investigated in homogeneous recombinant HDL (rHDL) particles containing apoA-I, palmitoyl-oleoyl phosphatidylcholine (POPC) and cholesteryl oleate. Isothermal denaturation with guanidine HCl was used to monitor alpha-helix structural stability, whereas electrokinetic analyses and circular dichroism were used to determine particle charge and apoA-I secondary structure, respectively. Electrokinetic analyses show that at pH 8.6 apoA-I has a net negative charge on discoidal (POPC.apoA-I) particles (-5.2 electronic units/mol of apoA-I) which is significantly greater than that of apoA-I either free in solution or on spherical (POPC.cholesteryl oleate.apoA-I) rHDL (approximately -3.5 electronic units). Raising the POPC content (32-128 mol/ml of apoA-I) of discoidal particles 1) increases the particle major diameter from 9.3 to 12.1 nm, 2) increases the alpha-helix content from 62 to 77%, and 3) stabilizes the helical segments by increasing the free energy of unfolding (delta GD degree) from 1.4 to 3.0 kcal/mol of apoA-I. Raising the POPC content (28-58 mol/mol of apoA-I) of spherical particles 1) increases the particle diameter from 7.4 to 12.6 nm, 2) increases the percent alpha-helix from 62 to 69%, and 3) has no significant effect on delta GD degree (2.2 kcal/mol of apoA-I). This study shows that different HDL subspecies maintain particular apoA-I conformations that confer unique charge and structural characteristics on the particles. It is likely that the charge and conformation of apoA-I are critical molecular properties that modulate the metabolism of HDL particles and influence their role in cholesterol transport.     

Quantitative measurement of lipoprotein surface charge by agarose gel electrophoresis.

The electrophoretic mobilities of low density lipoprotein (LDL) and six pure proteins in a 0.5% agarose gel have been compared to literature electrophoretic mobility values determined by the Tiselius moving boundary method. There is a strong correlation (r = 0.99) between the electrophoretic mobilities determined by the two techniques. The electrophoretic behavior of charged particles smaller than very low density lipoproteins (VLDL) is not markedly perturbed by a 0.5% agarose matrix, and variations in mobility primarily reflect differences in particle valence and density of surface charge. Application of electrokinetic theory to derive protein and lipoprotein net charges from the electrophoretic mobilities in agarose yields a quantitative delineation of lipoprotein electrophoretic migration patterns wherein the beta mobility region comprises a surface potential range of -4.5 to -7.0 mV; the pre-beta region a range of -7.0 to -10.5 mV; the alpha mobility region a range of -10.5 to -12.5 mV and the serum albumin region a range of -12.5 to -14.0 mV. Because protein conformation and charge are critical in metabolic regulation, the agarose gel electrophoresis technique provides a valuable analytical tool that should help to elucidate further details of the structure-function relationships of serum lipoprotein particles.     

Construction and characterisation of a yeast artificial chromosome library containing three haploid maize genome equivalents

We have constructed a yeast artificial chromosome (YAC) library using high-molecular-weight DNA prepared from agarose-embedded leaf protoplasts of the maize inbred line UE95. This library contains 79 000 clones with an average insert size of 145 kb and should therefore represent approximately three haploid genome equivalents. The library is organised as an ordered array in duplicate microtitre plates. Forty-one pools of DNA from 1920 individual clones have been prepared for rapid screening of the library by the polymerase chain reaction (PCR). Using this approach, together with conventional colony hybridisation, we have been able to identify between one and eight positive clones for every probe used.     

Patterns of urease synthesis in developing soybeans

An examination of in vivo polysome-bound activity indicates that soybean (Glycine max, cv. Prize) seed urease is synthesized on large polysomes (n ≥ 15). In vitro urease synthesis is directed by a large RNA (3,000-3,300 nucleotides). Urease synthesis occurs throughout the normal protein biosynthetic phase of the developing seed. Surprisingly, the activity/antigen ratios of urease increase throughout development. Urease appears to be in a more highly polymerized state in mature beans versus beans in early development.     

Turnover and trends in butterfly communities on two British tidal islands: stochastic influences and deterministic factors

Aim Community trends were investigated for two small islands and two local mainland butterfly communities within the UK over a period of 20–30 years. Location Hilbre Island off the Wirral Peninsula at 53.33° N, 3.10° W; Lindisfarne, an island off the Northumberland coast at 56.41° N, 1.48° W; Leighton Moss at 54.08° N, 2.26° W; Wyre Forest at 52.23° N, 2.14° W, UK. Methods Butterfly species data were collected on Hilbre and two mainland sites (Leighton Moss and Wyre Forest) from 1983 to 2006, and on Lindisfarne from 1977 to 2006, as part of the National Habitat Survey, the UK Butterfly Monitoring Scheme and ‘Butterflies for the New Millennium Atlas’ recording. Matrices of associations (Sokal and Michener’s matching coefficient S_SM; resemblance coefficient) were computed between years and subject to non‐metric multidimensional scaling (NMDS) and Mantel tests. The pattern of extinctions and colonizations at sites were examined, their heterogeneity tested by applying a Friedman test to fractional incidences for the same years. Regression analysis (multiple regression and logit regression) was used to relate butterfly numbers and incidences to climate variables, time and previous records. Results Significant community trends based on population counts and species’ incidences were found for all four sites. There was a significant climatic signal for Hilbre; although this was not apparent for the remaining sites, significant associations occurred between records for a number of species and climatic variables at all sites. Substantial turnover of species on the islands was inversely related to numbers of records for species but not to their conspicuousness to recorders. Main conclusions We argue that time trends are widespread in butterfly communities, even for relatively short periods; they are largely generated by stochastic influences rather than by more substantive factors such as climate change. Potential biases in surveying and recording history are shown to be unlikely. A clear climate signal was found only for the small Hilbre Island, for which there was also evidence for the significant influence of colonization capability of individual source species. We conclude that for many species, small islands will be sinks or pseudosinks and their ‘populations’ vulnerable to small changes in source–sink dynamics.