Effect of Metalaxyl on the incidence and severity of Pearl millet downy mildew disease in Northern Nigeria

Pearl millet downy mildew (DM) incidence, severity and yield losses of two pearl millet varieties (local and improved) due to the disease were determined in the field. Significant differences in the disease incidence and severity were recorded in the plots sown with metalaxyl-treated seeds and those sown with non-treated seeds, indicating the efficacy of the fungicide on the fungus. Yield losses due to non-treatment of seeds with metalaxyl was 40.88 and 45.39% in a local variety and 43.00 and 18.60% in an improved variety in the 2000 and 2001 cropping seasons respectively. Significant differences between plots sown with metalaxyl-treated and those sown with non-treated seeds were obtained for other yield components such as 1000-grains weight, panicle length and weight.     

Gene expression and the evolution of phenotypic diversity in social wasps

Background  

Organisms are capable of developing different phenotypes by altering the genes they express. This phenotypic plasticity provides a means for species to respond effectively to environmental conditions. One of the most dramatic examples of phenotypic plasticity occurs in the highly social hymenopteran insects (ants, social bees, and social wasps), where distinct castes and sexes all arise from the same genes. To elucidate how variation in patterns of gene expression affects phenotypic variation, we conducted a study to simultaneously address the influence of developmental stage, sex, and caste on patterns of gene expression in Vespula wasps. Furthermore, we compared the patterns found in this species to those found in other taxa in order to investigate how variation in gene expression leads to phenotypic evolution.     

A rapid method for exploring the protein structure universe

We have developed an automatic protein fingerprinting method for the evaluation of protein structural similarities based on secondary structure element compositions, spatial arrangements, lengths, and topologies. This method can rapidly identify proteins sharing structural homologies as we demonstrate with five test cases: the globins, the mammalian trypsinlike serine proteases, the immunoglobulins, the cupredoxins, and the actinlike ATPase domain-containing proteins. Principal components analysis of the similarity distance matrix calculated from an all-by-all comparison of 1,031 unique chains in the Protein Data Bank has produced a distribution of structures within a high-dimensional structural space. Fifty percent of the variance observed for this distribution is bounded by six axes, two of which encode structural variability within two large families, the immunoglobulins and the trypsinlike serine proteases. Many aspects of the spatial distribution remain stable upon reduction of the database to 140 proteins with minimal family overlap. The axes correlated with specific structural families are no longer observed. A clear hierarchy of organization is seen in the arrangement of protein structures in the universe. At the highest level, protein structures populate regions corresponding to the all-alpha, all-beta, and alpha/beta superfamilies. Large protein families are arranged along family-specific axes, forming local densely populated regions within the space. The lowest level of organization is intrafamilial; homologous structures are ordered by variations in peripheral secondary structure elements or by conformational shifts in the tertiary structure.     

Novel cathepsin D inhibitors block the formation of hyperphosphorylated tau fragments in hippocampus

Lysosomal disturbances may be a contributing factor to Alzheimer's disease. We used novel compounds to test if suppression of the lysosomal protease cathepsin D blocks production of known precursors to neurofibrillary tangles. Partial lysosomal dysfunction was induced in cultured hippocampal slices with a selective inhibitor of cathepsins B and L. This led within 48 h to hyperphosphorylated tau protein fragments recognized by antibodies against human tangles. Potent nonpeptidic cathepsin D inhibitors developed using combinatorial chemistry and structure-based design blocked production of the fragments in a dose-dependent fashion. Threshold was in the submicromolar range, with higher concentrations producing complete suppression. The effects were selective and not accompanied by pathophysiology. Comparable results were obtained with three structurally distinct inhibitors. These results support the hypothesis that cathepsin D links lysosomal dysfunction to the etiology of Alzheimer's disease and suggest a new approach to treating the disease.     

The role of exopolysaccharides in dual species biofilm development

A plasmid encoding the green fluorescent protein (GFP) of Aequorea victoria was transformed into a biofilm-forming strain of Enterobacter agglomerans originally isolated from an industrial environment. The transformed strain, EntGFP, could then be identified in dual species biofilms by direct visualization, plate counts and quantitiative fluorescence measurements. A variety of cell constituents and products may be involved in the adhesion and accumulation process and exopolysaccharides (EPS) represent one of these factors. The involvement of EPS in the initial adhesion events and the role in dual species biofilm development was investigated. Cells of EntGFP and Klebsiella pneumoniae Gl interact forming biofilms more successfully in a mixture than in isolation. The co-resistance results in enhanced biofilm formation and increased resistance to disinfection. Microscopic examination showed that the two species were often closely juxtaposed in microcolonies, suggesting the interactions involve surface-associated macromolecules. Fluorescence was used to measure the adhesion of EntGFP cells to Kleb, pneumoniae Gl (Gl) EPS. The results showed EntGFP adhered better to Gl EPS that Ent EPS. Polysaccharde depolymerases isolated from a bacteriophage for Ent. agglomerans were used to degrade Ent EPS specifically. Following polysaccharase treatment, the adhaesion of EntGFP to Gl cells was reduced. This suggests both types of EPS mediate adhesion. The two types of EPS were dissolved in dimethylsulphoxide and when mixed, their viscosity increased, reaching a maximum after ～+40 min. This may partially explain the increased protection of dual species biofilms from disinfectants. The depolymerases were used to treat dual species biofilms and this resulted in the effective removal of both species from the surface. This may suggest Ent contributes more EPS to the biofilm matrix. The EPS play an important role in EntGFP and Gl dual species biofilm formation both as adhesins and as the EPS interact, changing their physical properties.     

Quantitative oral dosing of water soluble and lipophilic contaminants in the Japanese medaka (Oryzias latipes)

Quantitative oral dosing in fish can be challenging, particularly with water soluble contaminants, which can leach into the aquarium water prior to ingestion. We applied a method of bioencapsulation using newly hatched brine shrimp (Artemia franciscana) nauplii to study the toxicokinetics of five chlorinated and brominated halogenated acetic acids (HAAs), which are drinking water disinfection by-products. These results are compared to those obtained in a previous study using a polybrominated diphenyl ether (PBDE-47), a highly lipophilic chemical. The HAAs and PBDE-47 were bioencapsulated using freshly hatched A. franciscana nauplii after incubation in concentrated solutions of the study chemicals for 18 h. Aliquots of the brine shrimp were quantitatively removed for chemical analysis and fed to individual fish that were able to consume 400-500 nauplii in less than 5 min. At select times after feeding, fish were euthanized and the HAA or PBDE-47 content determined. The absorption of HAAs was quantitatively similar to previous studies in rodents: rapid absorption with peak body levels occurring within 1-2 h, then rapidly declining with elimination half-life of 0.3-3 h depending on HAA. PBDE-47 was more slowly absorbed with peak levels occurring by 18 h and very slowly eliminated with an elimination half-life of 281 h.     

Development and Validation of a Real-Time PCR Method To Quantify Rumen Protozoa and Examination of Variability between Entodinium Populations in Sheep Offered a Hay-Based Diet

PCR and real-time PCR primers for the 18S rRNA gene of rumen protozoa (Entodinium and Dasytricha spp.) were designed, and their specificities were tested against a range of rumen microbes and protozoal groups. External standards were prepared from DNA extracts of a rumen matrix containing known numbers and species of protozoa. The efficiency of PCR () was calculated following amplification of serial dilutions of each standard and was used to calculate the numbers of protozoa in each sample collected; serial dilutions of DNA were used similarly to calculate PCR efficiency. Species of Entodinium, the most prevalent of the rumen protozoa, were enumerated in rumen samples collected from 100 1-year-old merino wethers by microscopy and real-time PCR. Both the counts developed by the real-time PCR method and microscopic counts were accurate and repeatable, with a strong correlation between them (R² = 0.8), particularly when the PCR efficiency was close to optimal (i.e., two copies per cycle). The advantages and disadvantages of each procedure are discussed. Entodinium represented on average 98% of the total protozoa, and populations within the same sheep were relatively stable, but greater variation occurred between different sheep (10⁰ and 10⁶ entodinia per gram of rumen contents). With this inherent variability, it was estimated that, to detect a statistically significant (P = 0.05) 20% change in Entodinium populations, 52 sheep per treatment group would be required.     

Toxoplasma gondii profilin acts primarily to sequester G-actin while formins efficiently nucleate actin filament formation in vitro

Apicomplexan parasites employ gliding motility that depends on the polymerization of parasite actin filaments for host cell entry. Despite this requirement, parasite actin remains almost entirely unpolymerized at steady state; formation of filaments required for motility relies on a small repertoire of actin-binding proteins. Previous studies have shown that apicomplexan formins and profilin exhibit canonical functions on heterologous actins from higher eukaryotes; however, their biochemical properties on parasite actins are unknown. We therefore analyzed the impact of T. gondii profilin (TgPRF) and FH1-FH2 domains of two formin isoforms in T. gondii (TgFRM1 and TgFRM2) on the polymerization of T. gondii actin (TgACTI). Our findings based on in vitro assays demonstrate that TgFRM1-FH1-FH2 and TgFRM2-FH1-FH2 dramatically enhanced TgACTI polymerization in the absence of profilin, making them the sole protein factors known to initiate polymerization of this normally unstable actin. In addition, T. gondii formin domains were shown to both initiate polymerization and induce bundling of TgACTI filaments; however, they did not rely on TgPRF for these activities. In contrast, TgPRF sequestered TgACTI monomers, thus inhibiting polymerization even in the presence of formins. Collectively, these findings provide insight into the unusual control mechanisms of actin dynamics within the parasite.     

High Photosynthetic Capacity in a Shade-Tolerant Crassulacean Acid Metabolism Plant (Implications for Sunfleck Use,Nonphotochemical Energy Dissipation,and Susceptibility to Photoinhibition)

Aechmea magdalenae Andre ex Baker, a constitutive Crassulacean acid metabolism (CAM) plant from the shaded Panamanian rain forest understory, has a maximum photosynthesis rate 2 to 3 times that of co-occurring C3 species and a limited potential for photosynthetic acclimation to high light. Chlorophyll fluorescence measurements indicated that (a) compared with co-occurring C3 species, photosynthetic electron transport in A. magdalenae responded more rapidly to light flecks of moderate intensity, attained a higher steady-state rate, and maintained a lower reduction state of plastoquinone during light flecks; (b) these characteristics were associated with phase III CO2 fixation of CAM; (c) when grown in full sun, A. magdalenae was chronically photoinhibited despite a remarkably high nonphotochemical quenching capacity, indicating a large potential for photoprotection; and (d) the degree of photoinhibition was inversely proportional to the length of phase III. Results from the light fleck studies suggest that understory A. magdalenae plants can make more efficient use of sun flecks for leaf carbon gain over most of the day than co-occurring C3 species. The association between the duration of phase III and the degree of photoinhibition for A. magdalenae in high light is discussed in relation to the limited photosynthetic plasticity in this species.