The Recombinant Amyloid-β Peptide Aβ1-42 Aggregates Faster and Is More Neurotoxic than Synthetic Aβ1-42

Aggregation of the amyloid-β (Aβ) peptide is considered a central event in the pathogenesis of Alzheimer's disease (AD). In order to bypass methodological bias related to a variety of impurities commonly present in typical preparations of synthetic Aβ, we developed a simple, generally applicable method for recombinant production of human Aβ and Aβ variants in Escherichia coli that provides milligram quantities of Aβ in very high purity and yield. Amyloid fibril formation in vitro by human Aβ1-42, the key amyloidogenic Aβ species in AD, was completed threefold faster with recombinant Aβ1-42 compared to synthetic preparations. In addition, recombinant Aβ1-42 was significantly more toxic to cultured rat primary cortical neurons, and it was more toxic in vivo, as shown by strongly increased induction of abnormal phosphorylation of tau and tau aggregation into neurofibrillary tangles in brains of P301L tau transgenic mice. We conclude that even small amounts of impurities in synthetic Aβ—including a significant fraction of racemized peptides that cannot be avoided due to the technical limitations of peptide synthesis—prevent or slow Aβ incorporation into the regular quaternary structure of growing β-amyloid fibrils. The results validate the use of recombinant Aβ1-42 for both in vitro and in vivo studies addressing the mechanisms underlying Aβ aggregation and its related biological consequences for the pathophysiology, therapy, and prevention of AD.     

Alkali‐based AFEX pretreatment for the conversion of sugarcane bagasse and cane leaf residues to ethanol

Sugarcane is one of the major agricultural crops cultivated in tropical climate regions of the world. Each tonne of raw cane production is associated with the generation of 130 kg dry weight of bagasse after juice extraction and 250 kg dry weight of cane leaf residue postharvest. The annual world production of sugarcane is ～1.6 billion tones, generating 279 MMT tones of biomass residues (bagasse and cane leaf matter) that would be available for cellulosic ethanol production. Here, we investigated the production of cellulosic ethanol from sugar cane bagasse and sugar cane leaf residue using an alkaline pretreatment: ammonia fiber expansion (AFEX). The AFEX pretreatment improved the accessibility of cellulose and hemicelluloses to enzymes during hydrolysis by breaking down the ester linkages and other lignin carbohydrate complex (LCC) bonds and the sugar produced by this process is found to be highly fermentable. The maximum glucan conversion of AFEX pretreated bagasse and cane leaf residue by cellulases was ～85%. Supplementation with hemicellulases during enzymatic hydrolysis improved the xylan conversion up to 95–98%. Xylanase supplementation also contributed to a marginal improvement in the glucan conversion. AFEX‐treated cane leaf residue was found to have a greater enzymatic digestibility compared to AFEX‐treated bagasse. Co‐fermentation of glucose and xylose, produced from high solid loading (6% glucan) hydrolysis of AFEX‐treated bagasse and cane leaf residue, using the recombinant Saccharomyces cerevisiae (424A LNH‐ST) produced 34–36 g/L of ethanol with 92% theoretical yield. These results demonstrate that AFEX pretreatment is a viable process for conversion of bagasse and cane leaf residue into cellulosic ethanol. Biotechnol. Bioeng. 2010;107: 441–450. © 2010 Wiley Periodicals, Inc.     

Cereal rust fungi genomics and the pursuit of virulence and avirulence factors

Rust diseases cause significant reductions annually in yield of cereal crops worldwide. Traditional monoculture cropping systems apply significant selection pressure on the pathogen to cause rapid shifts in pathotypes. Plant breeders strive to stay ahead of the evolving pathogens by releasing new crop genotypes with new rust resistance genes or gene combinations. Owing to the limited number of known resistance genes and the lack of molecular understanding of the plant-pathogen interaction, rusts remain challenging organisms to study, both at organismal and molecular levels. This review discusses recent progress by a number of laboratories towards better understanding the molecular component of rust disease resistance.     

Contribution of leaf surface fungi to the air spora

High concentrations of airborne fungal spores frequently occur from spring through fall in temperate areas of the world. Although it is generally assumed that fungi on leaf surfaces are contributors to the air spora, little data are available comparing the types of fungi found on leaf surfaces with those in the atmosphere. Air sampling was carried out with a Burkard Spore Trap located on the roof of a building on the University of Tulsa campus using standard methods. Leaf samples were aseptically collected from Ulmus americana and Quercus palustris trees on campus, placed in sterile plastic bags, and brought to the lab. For each leaf, 4 cm² areas of both upper and lower leaf surfaces were swabbed and plated on malt extract agar with streptomycin. Cultures were incubated at room temperature for 5–7 days and then examined microscopically. Results were expressed as colony forming units (CFU)/cm². Twenty-one fungal taxa were identified from the air samples. The most abundant taxa were Cladosporium, ascospores, basidiospores, and Alternaria; together these four spore types comprised over 90% of the yearly total. Yeasts were the most abundant fungi isolated from both leaf types. Among the mycelial fungi were Phoma species, followed by Cladosporium and Alternaria. Overall twenty genera of filamentous fungi were identified. Yeasts and Phoma are normally splash dispersed and were not identified in the Burkard air samples. However, 10 taxa isolated from leaf surfaces were registered in air samples. Crude estimates of the leaf surface area of each tree suggest that the total fungal load was approximately 5.04×10⁸ CFU for Ulmus and 2.71×10⁸ CFU for Quercus. Of these levels, 19% were from fungi also detected in air samples. The data suggest that some leaf-surface fungi are major contributors to the air spora.     

外源SOD和APX基因在转基因烟草中的表达与遗传

分析转超氧化物歧化酶基因（SOD）或抗坏血酸过氧化物酶基因（APX）烟草及其自交和杂交后代的叶片中超氧化物歧化酶（SOD）和过氧化物酶（POD）活性的结果表明：转基因烟草的SOD和POD活性在终花期最强,不同叶位叶中SOD活性差异不明显,POD活性以下部叶为最高;转基因烟草的SOD或POD活性显著高于近等基因的非转基因品系。杂交后代（F1、F2）的SOD活性能保持稳定,略高于亲本;自交后代（S1～S3）与自交亲本的SOD和POD活性相当。     

我国烟田昆虫群落的划分及其特征分析

根据纬度、海拔、温度、降雨、日照等地理、气象因子指标,对我国71个植烟市和(或)县及生态条件独特的市和(或)县进行了聚类分析,聚类结果与我国烟草的7个一级种植区基本吻合,我国烟田烟草昆虫群落可依此划分为东北、黄淮、长江中上游、长江中下游、西南、南部和北部西部等7个。各群落的重要烟草昆虫种类有所不同,烟草昆虫天敌的种类差别更大。其中西南群落食物网最复杂。     

Excision and transposition of piggyBac transposable element in tobacco budworm embryos

The TTAA-specific lepidopteran transposon piggyBac has already proved useful as a gene-transfer vector for efficient transformation of a wide variety of insects. Transposable element excision and transposition assays are useful indicators of an element's ability to be mobilized in vivo and, thus, potentially serve as a transforming vector. Here, we report that this transposon is capable of excision and transposition in tobacco budworm embryos with relatively low frequency.     

Pheromone Release Behaviour in Females of Phyllonorycter junoniella (Z.) (Lepidoptera, Gracillariidae) Under Constant and Cycling Temperatures

The calling activity of virgin Phyllonorycter junoniella (Z.) females under a cycling thermal regime differed from that under constant temperature in the following ways: the percentage of females calling at the morning activity peak was increased; the morning period of calling activity was prolonged; the total period of calling activity of an individual female was increased; an extra peak of activity occurred at the end of the photophase, and females changed the calling pattern during the active period. We assume that the occurrence of two calling peaks and the extension of the calling period might be adaptive for a Ph. junoniella species with a sex ratio strongly shifted towards females (8:1 females:males), as it could lead to an increased proportion of females mated.     

Determinants of Leaf Litter Nutrient Cycling in a Tropical Rain Forest: Soil Fertility Versus Topography

We investigated the influence of landscape-level variation in soil fertility and topographic position on leaf litter nutrient dynamics in a tropical rain forest in Costa Rica. We sampled across the three main edaphic conditions (ultisol slope, ultisol plateau, and inceptisol) to determine the effect of soil nutrients on leaf litter nutrient concentrations while controlling for topography, and to examine topographic effects while controlling for soil nutrients. Both leaf litter macronutrient [phosphorus (P), nitrogen (N), sulfur (S), calcium (Ca), potassium (K), magnesium (Mg)] and micronutrient concentrations were quantified throughout a 4-year period. Leaf litter [P], [N] and [K] varied significantly among soil types. The variation in [P], [N], and [K] was explained by soil fertility alone. Leaf litter [S], [Ca], and [Mg] did not vary among the three soil types. Macronutrient (P, K, Mg, S, Ca) concentrations in the leaf litter were much less variable than those of Fe and Al. Lower variability in essential plant nutrients suggests a great deal of plant control over the amount of nutrients resorbed before senescense. Leaf litter macronutrient concentrations varied significantly over the 4-year period, but the temporal variation did not differ among the three edaphic types as anticipated. Hence, although the magnitude of nutrient fluxes may be controlled by local factors such as soil fertility, temporal patterns are likely regulated by a common environmental variable such as precipitation or temperature.     

Identifying Differences in Carbon Exchange among Arctic Ecosystem Types

Our objective was to determine how varied is the response of C cycling to temperature and irradiance in tundra vegetation. We used a large chamber to measure C exchange at 23 locations within a small arctic catchment in Alaska during summer 2003 and 2004. At each location, we determined light response curves of C exchange using shade cloths, twice during a growing season. We used data to fit a simple photosynthesis-irradiance, respiration-temperature model, with four parameters. We used a maximum likelihood technique to determine the acceptable parameter space for each light curve, given measurement uncertainty. We then explored which sites and time periods had parameter sets in common—an indication of functional similarity. We found that seven distinct parameter sets were required to explain observed C flux responses to temperature and light variation at all sites and time periods. The variation in estimated maximum photosynthetic rate (P_max) was strongly correlated with measurements of site leaf area index (LAI). The behavior of tussock tundra sites, the dominant vegetation of arctic tundra, could largely be described with a single parameter set, with a P_max of 9.7 μmol m⁻² s⁻¹. Tussock tundra sites had, correspondingly, similar LAI (mean = 0.66). Non-tussock sites (for example, sedge and shrub tundras) had larger spatial and temporal variations in both C dynamic parameters (P_max varying from 9.7–25.7 μmol m⁻² s⁻¹) and LAI (0.6–2.0). There were no clear relationships between dominant non-tussock vegetation types and a particular parameter set. Our results suggest that C dynamics of the acidic tussock tundra slopes and hilltops in northern Alaska are relatively simply described during the peak growing season. However, the foot-slopes and water tracks have more variable patterns of LAI and C exchange, not simply related to the dominant vegetation type.