The effect of α-mating factor secretion signal mutations on recombinant protein expression in Pichia pastoris

The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins is initially fused to the Saccharomyces cerevisiae α-mating factor secretion signal leader. Extensive site-directed mutagenesis of the prepro-region of the α-mating factor secretion signal sequence was performed in order to determine the effects of various deletions and substitutions on expression. Though some mutations clearly dampened protein expression, deletion of amino acids 57–70, corresponding to the predicted 3rd alpha helix of α-mating factor secretion signal, increased secretion of reporter proteins horseradish peroxidase and lipase at least 50% in small-scale cultures. These findings raise the possibility that the secretory efficiency of the leader can be further enhanced in the future.     

Interference and the ideal free distribution: models and tests

We review the assumptions and predictions of five competitivedistribution models that predict how optimal foragers will bedistributed across resource patches when gains are reduced byinterference. This review revealed a number of previously ignoredpredictions and assumptions: in particular, there should beno change in relative patch use as competitor density changes.A new model is proposed in which interference results from thecosts of encounters with other foragers and where the gainson a patch are independent of the costs of interference. Thismodel predicts that as density increases, there will be increasedproportional use of lower-quality patches. Past empirical studiesof interference distributions are reanalyzed; none of the studiesprovides strong support for any of the existing ideal free-distributionmodels. We suggest that previous results are more consistentwith the predictions of our new model.     

DIFFERENCES IN BIOMASS ALLOCATION TO REPRODUCTIVE AND VEGETATIVE STRUCTURES OF MALE AND FEMALE PLANTS OF A DIOECIOUS,PERENNIAL HERB,SILENE ALBA (MILLER) KRAUSE

The pattern of biomass allocation of males and females and the sex ratio and growth characteristics of plants from three seed-size classes in Silene alba were investigated in a greenhouse study. Seed size significantly affected adult plant size and flower production of both male and female plants, but there was no significant difference in the proportion of males and females emerging in three seed-size categories. Male and female plants differed in the proportion of total biomass allocated to vegetative and reproductive structures and these differences were consistent across all seed-size categories. Males allocated a greater proportion of their biomass to flowers than did females. Female reproductive effort was dependent upon the percentage of flowers producing mature capsules. Only females with greater than 20% fruit set have a higher reproductive expenditure than males. Consequently, female expenditure is potentially greater than males, but is spread out over a longer portion of the growing season. This difference in the timing of reproductive expenditures by males and females allows females to allocate more biomass to growth during the early flowering period and may therefore account for the common pattern in herbaceous perennial dioecious species in which adult females are larger than adult males.     

Evaluation of aid to diagnosis of pigmented skin lesions in general practice: controlled trial randomised by practice

Objectives To determine whether an aid to the diagnosis of pigmented skin lesions reduces the ratio of benign lesions to melanomas excised in general practice.Design Controlled trial randomised by practice.Setting General practices in Perth, Western Australia.Participants 468 general practitioners in 223 practices.Interventions Intervention practices were given an algorithm and instant camera to assist with the diagnosis of pigmented skin lesions. All practices were given national guidelines on managing melanoma.Main outcome measures Ratio of benign pigmented lesions to melanomas excised. Analyses conducted with and without inclusion of seborrhoeic keratoses.Results At baseline the ratios of benign to malignant lesions were lower in the intervention group than in the control group. During the trial period the ratios were higher in the intervention group (19:1 v 17:1 without seborrhoeic keratoses and 29:1 v 26:1 with seborrhoeic keratoses). After adjustment for patients'' age, sex, and socioeconomic status, the ratio was 1.02 times higher (95% confidence interval 0.68 to 1.51, P = 0.94) in the intervention group when seborrhoeic keratoses were not included and 1.03 times higher (0.71 to 1.50, P = 0.88) when seborrhoeic keratoses were included. General practitioners in the intervention group were less likely than those in the control group to excise the most recent pigmented skin lesion they managed (22% v 48%, P < 0.001) and to refer the patient to a specialist (16% v 27%, P = 0.06).Conclusions Provision of the algorithm and camera did not decrease the ratio of benign pigmented skin lesions to melanomas excised by general practitioners.     

The polytene chromosomes in the fat body nuclei ofDrosophila melanogaster

A photo-map of the polytene chromosomes of fat body nuclei from third instar larvae ofDrosophila melanogaster has been constructed and keyed to the revised salivary gland maps of Bridges (1938), Bridges and Bridges (1939) and Bridges (1941a, b, 1942). Apparent variations in banding pattern are discussed in the light of current studies on polytene chromosome structure and function.     

The Ups and Downs of Life on the Edge: The Influence of Water Level Fluctuations on Biomass Allocation in two Contrasting Aquatic Plants

Plant communities at wetland edges typically exhibit strong zonation along water depth gradients, although community structure is not as simple as this common observation may suggest. Water levels fluctuate at many temporal scales and in varied patterns. Understanding of how water regime controls plant community structure requires information on how individual species are affected by water regime. The allocation of biomass to roots, tillers, stolons and inflorescences was measured over 16 weeks in two species of aquatic plants with contrasting life histories. Juncus articulatus L. produces large numbers of small seeds while Glyceria australis C. E. Hubb produces small numbers of large seeds. Both species also reproduce vegetatively. Changes in biomass allocation were measured in response to duration, frequency and amplitude of water level fluctuations. The fluctuations in water level were achieved by suspending plants in pots from chains and raising and lowering the pots in and out of water in replicate tanks. There were complex responses to water regime components. As is common to many emergents, both species increased allocation to above ground vegetative structures in response to increasing water depth. Differences between the species in overall patterns of allocation reflected differences in life history. Juncus articulatus showed greater overall biomass production and greater proportional resource allocation to inflorescences than Glyceria australis. There were other differences in response to components of water regime that distinguished the species, mainly in allocation to reproduction in response to the range of depths (amplitude) and extended dry periods. These different functional responses to aspects of water level fluctuations indicate it may be possible to manipulate species composition if water levels are controllable. The most important characteristics of water regime were threshold events such as extremes of depth and dry periods. These may not be captured in measures of water regime such as average water levels.     

The discovery of <Emphasis Type="Italic">Caligus</Emphasis><Emphasis Type="Italic">temnodontis</Emphasis> Brian, 1924 (Copepoda: Caligidae) from the bluefish <Emphasis Type="Italic">Pomatomus saltatrix</Emphasis> (Linnaeus) in the eastern Mediterranean Sea

The temnodontis variety of Caligus mauritanicus Brian, 1924 described by Brian (1924) is a valid species known only from a single host species, Pomatomus saltatrix (Linnaeus). New material of this species has been examined from the same host fish caught from Abuqir Bay, Alexandria (Egypt), from Iskenderun Bay (Turkey) and from off the coast of South Africa. Using this material, C. temnodontis Brian, 1924 is redescribed and compared with related species. It is most closely related to the Indo-Pacific species C. pagrosomi Yamaguti, 1939.     

Sequential gene activation by ecdysteroids in polytene chromosomes ofDrosophila melanogaster

Summary Changes in polytene chromosome 3 L puffing patterns in the fat body ofDrosophila melanogaster larvae and prepupae are compared to those in the salivary gland. While some general features are common to the two tissues, there are differences which reflect their different developmental roles. In vitro experiments with fat body chromosomes show that they have a distinct response to ecdysteroids which is different from that of salivary gland chromosomes, and which does not,in this culture system, reproduce the changes observed in normal development. In short term culture experiments, the fat body chromosomes appear more sensitive to ecdysteroids than the salivary gland chromosomes and, although 20-OH ecdysone is more active than ecdysone in these assays, the possibility is not excluded that ecdysone has a role in normal development as it appears to alter gene activity at physiological levels in these cells.     

Climate drivers alter nitrogen availability in surface peat and decouple N2 fixation from CH4 oxidation in the Sphagnum moss microbiome

Peat mosses (Sphagnum spp.) are keystone species in boreal peatlands, where they dominate net primary productivity and facilitate the accumulation of carbon in thick peat deposits. Sphagnum mosses harbor a diverse assemblage of microbial partners, including N₂-fixing (diazotrophic) and CH₄-oxidizing (methanotrophic) taxa that support ecosystem function by regulating transformations of carbon and nitrogen. Here, we investigate the response of the Sphagnum phytobiome (plant + constituent microbiome + environment) to a gradient of experimental warming (+0°C to +9°C) and elevated CO₂ (+500 ppm) in an ombrotrophic peatland in northern Minnesota (USA). By tracking changes in carbon (CH₄, CO₂) and nitrogen (NH₄-N) cycling from the belowground environment up to Sphagnum and its associated microbiome, we identified a series of cascading impacts to the Sphagnum phytobiome triggered by warming and elevated CO₂. Under ambient CO₂, warming increased plant-available NH₄-N in surface peat, excess N accumulated in Sphagnum tissue, and N₂ fixation activity decreased. Elevated CO₂ offset the effects of warming, disrupting the accumulation of N in peat and Sphagnum tissue. Methane concentrations in porewater increased with warming irrespective of CO₂ treatment, resulting in a ~10× rise in methanotrophic activity within Sphagnum from the +9°C enclosures. Warming's divergent impacts on diazotrophy and methanotrophy caused these processes to become decoupled at warmer temperatures, as evidenced by declining rates of methane-induced N₂ fixation and significant losses of keystone microbial taxa. In addition to changes in the Sphagnum microbiome, we observed ~94% mortality of Sphagnum between the +0°C and +9°C treatments, possibly due to the interactive effects of warming on N-availability and competition from vascular plant species. Collectively, these results highlight the vulnerability of the Sphagnum phytobiome to rising temperatures and atmospheric CO₂ concentrations, with significant implications for carbon and nitrogen cycling in boreal peatlands.