Effects of genotype, nutrient availability, and defoliation on aspen phytochemistry and insect performance

Genetic and environmental variability, and their interactions, influence phytochemical composition and, in turn, herbivore performance. We evaluated the independent and interactive effects of plant genotype, nutrient availability, and defoliation on the foliar chemistry of quaking aspen (Populus tremuloides) and consequences for performance of gypsy moths (Lymantria dispar). Saplings of four genotypes were grown under two conditions of nutrient availability and subjected to three levels of artificial defoliation. Concentrations of all secondary and primary metabolites evaluated responded to at least one or more of the experimental treatments. Of the secondary metabolites, phenolic glycosides were affected strongly by genotype, less so by nutrient availability, and not induced by defoliation. Condensed tannins were strongly dependent upon genotype, soil nutrient availability, and their interaction, and, in contrast to phenolic glycosides, were induced by artificial defoliation. Of the primary metabolites, foliar nitrogen was affected by genotype and soil nutrient availability. Starch concentrations were affected by genotype, nutrient availability, defoliation and interactions among these factors. Foliar water content responded to genotype, nutrient availability, and defoliation, and the effect of nutrient availability depended on genotype. Herbivore performance on these plants was strongly influenced by plant genotype and soil nutrient availability, but much less so by defoliation. Although several of the compound types (condensed tannins, starch, and water) responded to defoliation, quantitative variation in these compounds did not contribute to substantive changes in herbivore performance. Rather, the primary source of variation in insect performance was due to plant genotype (phenolic glycoside levels), while nutrient availability (foliar nitrogen levels) was of secondary importance. These results suggest that genetic variation in aspen plays a major role in determining patterns of insect performance, whereas environmental variation, such as was tested, here is of negligible importance.     

On the syntheses of branched saturated fatty acids

In order to investigate the relation between the structure of mono-branched saturated fatty acids and their physical properties, a three-factor central composite design was constructed. For this purpose sixteen different fatty acids were prepared. The synthetic strategy was to use if possible, a few common starting materials for the preparation of most of the acids. Thus alkylation and hydrolysis of oxazolines were used for the preparation of 2-butylhexanoic acid, 2-methyloctadecanoic acid, 2-hexadecyloctadecanoic acid and 2-pentyloctadecanoic acid. A number of acids were prepared from thiophene derivatives followed by desulfurization with Raney-Nickel alloy under alkaline conditions. Thus, starting from 3-ethylthiophene, 4-ethyl-2-thiophenecarboxylic acid and 4-ethyl-2-methyl-5-thiophenecarboxylic acid were prepared, which upon desulfurization gave the desired 4-methylhexanoic acid. From 3-bromo-2-methylthiophene, 3-ethyl-2-methyl-5-thiophenecarboxylic acid was preparedvia 3-acetyl-2-methylthiophene and 3-ethyl-2-methylthiophene. Desulfurization gave 4-ethylhexanoic acid. Another approach started with 2-acylthiophenes, which were reacted with Grignard reagent to yield the appropriate olefins. By metalation and reaction with carbon dioxide, these were transformed to the corresponding 2-thiophenecarboxylic acids. Upon desulfurization, the desired fatty acids were obtained. In this way 6-propyldecanoic acid, 6-hexyldodecanoic acid, 6-methyldodecanoic acid and 6-pentylpentadecanoic acid were prepared. The remaining four acids were prepared from some of the branched acids described above through Kolbe reactions of dioic acids. Thus 16-methyloctadecanoic acid and 10-methyl-dodecanoic acid were obtained from 4-methylhexanoic acid, 16-ethyloctadecanoic acid from 4-ethylhexanoic acid and 9-pentyloctadecanoic acid from 6-pentylpentadecanoic acid.     

Quantifying the predatory effect of round goby on Saginaw Bay dreissenids

Invasive dreissenid mussels (D. polymorpha and D. r. bugensis) have fundamentally altered Laurentian Great Lake ecosystems, however in many areas their abundances have declined since the mid-1990s. Another invader, the benthic fish round goby (Neogobius melanostomus), is morphologically adapted to feed on dreissenids and likely affects dreissenid populations; however, the degree of this predatory effect is variable. In 2009 and 2010, we examined round goby abundances, size distributions, diet contents, and diet selectivity in Saginaw Bay, Lake Huron; a shallow bay that has been subjected to numerous anthropogenic stressors. We further used a consumption model to estimate dreissenid consumption by three different size classes of round goby. Round gobies were found throughout the bay and most were smaller than 80 mm total length. Round gobies of all sizes consumed dreissenids (including fish as small as 30 mm total length), though dreissenids were rarely preferred. The relative proportion of dreissenids (by biomass) present in diets of round gobies increased with fish size, but also throughout the year for all size classes. Despite this, overall consumptive effects of round gobies on dreissenids in Saginaw Bay were low. Many dreissenids present in the bay were larger than those consumed by round gobies. Bioenergetics-based model estimates suggest that the smallest round gobies are responsible for the majority of dreissenid consumption. While our findings are limited to soft substrates and influenced by sampling restrictions, our study design allowed us to put bounds on our estimates based upon these multiple sources of uncertainty.     

Effects of protein and juglone on gypsy moths: Growth performance and detoxification enzyme activity

The individual and interactive effects of dietary protein and juglone on larval performance and midgut detoxification enxymes were investigated for the gypsy moth,Lymantria dispar. The experimental design was a 2 × 3 factorial, with two levels of protein and three levels of juglone. We monitored survival/development rates from egg hatch to pupation and conducted fourth-instar feeding trials for determination of nutritional indices. Enzyme solutions were prepared from midguts of fifth instars and assayed for polysubstrate monooxygenase, esterase, quinone reductase, and glutathione transferase activities. Results showed that low protein levels prolonged development times, increased consumption rates, and reduced pupal weights. Juglone markedly reduced survival, growth, and consumption rates, increased development times, and reduced pupal weights. The interaction between protein and juglone influenced larval digestion efficiencies and female pupal weights. Polysubstrate monooxygenase activities were unaffected by diet, whereas esterase activities increased in response to both low dietary protein and presence of juglone. Low protein levels increased soluble quinone reductase activities but decreased glutathione transferase activities. Glutathione transferase activities were lowest in larvae fed low-protein, high-juglone diets and may have contributed to the especially poor performance of larvae on those diets. Quinone reductase and glutathione transferase are the systems of importance in detoxification of juglone, and moderate to low activities of these enzymes may explain why gypsy moths perform poorly on members of the Juglandaceae.     

Interaction between a Fab fragment against gp41 of human immunodeficiency virus 1 and its peptide epitope: characterization using a peptide epitope library and molecular modeling

The molecular interaction of the Fab fragment of the human monoclonalantibody 3D6, directed against the transmembrane protein gp41of human immunodeficiency virus (HTV) 1, with its peptide epitopeis characterized by a panel of overlapping peptides, a peptideepitope library and molecular modeling techniques. The sequenceCSGKLICTTAVPW, corresponding to amino acids 605–617 ofgp41, was identified as the best binding peptide (K_D = 1x10^-8mol/1). This peptide served as a starting point to prepare acellulose-bound peptide epitope library in which each residueof the epitope is substituted by all L- and D-amino acids, resultingin 494 epitope peptide variants which were subsequently analyzedfor binding 3D6. The library was synthesized to identify residuescritical for binding and to obtain information about the molecularenvironment of the epitope peptide bound to 3D6. Both cysteineresidues, as well as isoleucine 6, threonine 8 and proline 12,of the epitope were highly sensitive to substitution. Usingthe data obtained from the epitope characterization, as wellas a low-resolution electron density map of a 3D6 Fab-peptidecomplex, a 3-D model of the Fab-peptide complex was generatedby molecular modeling. The modeling experiments predict bindingof the peptide, which is cyclized via the two cysteine residues,to a pocket formed dominantly by the hypervariable loops complementaritydetermining regions CDR3L, CDR2H and CDR3H.     

Naphthalene oxidation over vanadium-modified Pt catalysts supported on γ-Al2O3

Alumina-supported platinum catalysts modified by vanadium were synthesised and tested for the complete oxidation of naphthalene. The catalysts were characterised by BET, pulsed CO chemisorption, powder X-ray diffraction, laser Raman spectroscopy and temperature-programmed reduction. Whilst BET and CO chemisorption results showed that the addition of vanadium modified both the textural properties of the support and the distribution of Pt, XRD and TPR data suggested the presence of V₂O₅ on catalysts with higher V loadings. TPR data showed that the concentration of V₂O₅ and possibly some other vanadium species increased as the V loading increased. Only 0.5%V was found to promote the activity of the 0.5%Pt/γ-Al₂O₃ catalyst. The activity enhancement has been related to the presence of a more easily reducible vanadium species coupled with the enhanced number of surface Pt sites. On the other hand, the reduced activity demonstrated by catalysts with higher vanadium content (1 – 12%) has been attributed to the presence of crystalline V₂O₅.     

Preservation and visualization of molecular structure in detergent-extracted whole mounts of cultured cells.

Today's electron microscopes have a resolution sufficient to resolve supramolecular structures. However, the methods used to prepare biological samples for electron microscopy often limit our ability to achieve the resolution that is theoretically possible. We use whole mounts of detergent-extracted cells grown on Formvar-coated gold grids as a model system to evaluate various steps in the preparation of biological samples for high resolution scanning electron microscopy (SEM). Factors that are important in determining the structure and composition of detergent-extracted cells include the nature of the detergent and the composition of the extraction vehicle. Chelation of calcium is extremely important to stabilize and preserve the cytoskeletal filaments. We have also demonstrated both morphologically and by gel electrophoresis that treatment of cells with bifunctional protein crosslinkers before or during extraction with detergent can significantly enhance the preservation of both proteins and supramolecular structures. The methods used to dry samples are a major determinant of the quality of structural preservation. For cytoskeletons freeze-drying (FD) is superior to critical point-drying (CPD), one reason being that CPD samples have to be dehydrated, thereby causing more shrinkage as compared to FD samples. The high pressures to which samples are exposed during CPD may also cause increased shrinkage, and water contamination during CPD causes severe structural damage. We have obtained the best structural preservation of detergent-extracted and fixed cells by manually plunging them into liquid propane and drying over night in a freeze-dryer. The factor that most limits achievement of high resolution in SEM is the metal coat, which has to be very thin, uniform, and free of grain in order not to hide structures or to create artifactual ones. We have found that sputter-coating with 1-3 nm of tungsten (W) or niobium (Nb) gives extremely fine-grained films as well as satisfactory emission of secondary electrons. These samples can also be examined at high resolution by transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). The best preservation and visualization of supramolecular structures have been obtained using cryosputtering, in which the samples are freeze-dried and then sputter-coated within the freeze-dryer while still frozen.     

Flexible lock-in detection system based on synchronized computer plug-in boards applied in sensitive gas spectroscopy

We present a flexible and compact, digital, lock-in detection system and its use in high-resolution tunable diode laser spectroscopy. The system involves coherent sampling, and is based on the synchronization of two data acquisition cards running on a single standard computer. A software-controlled arbitrary waveform generator is used for laser modulation, and a four-channel analog/digital board records detector signals. Gas spectroscopy is performed in the wavelength modulation regime. The coherently detected signal is averaged a selected number of times before it is stored or analyzed by software-based, lock-in techniques. Multiple harmonics of the modulation signal (1f, 2f, 3f, 4f, etc.) are available in each single data set. The sensitivity is of the order of 10(-5), being limited by interference fringes in the measurement setup. The capabilities of the system are demonstrated by measurements of molecular oxygen in ambient air, as well as dispersed gas in scattering materials, such as plants and human tissue.     

Equilibrium approaches to natural water systems—6. Acid-base properties of a concentrated bog-water and its complexation reactions with aluminium(III)

A filtered bog-water, concentrated by means of a freezing technique, has been studied with respect to acid-base properties and aluminium(III) complexation reactions. Sampling was performed during autumn and winter periods with a resulting acidity or alkalinity due to oxic (autumn) or anoxic (winter) conditions. The measurements were performed as potentiometric titrations in constant ionic media (0.02, 0.1 and 0.6 M NaCl) with the use of a glass electrode. The samples show buffer ranges at 3 pH 5 and pH 7.5. The first is ascribed to the presence of carboxylate groups and is characterized by fast equilibria. The second is due to phenolic OH⁻ groups and precipitation reactions with resulting sluggish equilibria. For one sampling period comprehensive measurements were undertaken to study the possible polyelectrolytic character of the organic acids. Due to the small increase in apparent carboxylate pKa-values with the degree of dissociation at low (0.02 M) as well as at high (0.6 M) ionic strength, the possible polyelectrolytic feature of the acids was neglected. Instead, a good fit to data was obtained by introducing a di-protic acid (H₂L) as a model compound. Furthermore the medium dependence of the two acidity constants could satisfactorily be fitted to the expression:

Full-size image (1K)

, where K is the constant at infinite dilution, a and b parameters of which b has been adjusted to present data. The following K values were obtained: pK₁ = 3.65 and pK₂ = 4.30. The complexation with Al(III) could be described by the formation of AlL⁺, AlL₂⁻ and the ternary species AlLH₋₁. The stability constants (log k₁ = 4.4 (winter), 4.2 (autumn); log k₂ = 4.2, 4.7; pKa (AlL⁺) = 4.2, 4.2) show no significant trend with sampling period but indicate a stability of the complexes greater than for phtalic acid but lower than for oxalic acid. Finally, the theoretical solubility of the clay mineral kaolinite in the presence of bog-water was computer modelled. The calculations show up on a 10-fold increase with respect to soluble aluminium at pH = 5.     

Building a new supply chain position: an exploratory study of companies in the timber housing industry

Innovation is an important source of growth for many companies. It is also challenging, as it may require that the companies build a new position in the supply chain. The literature is devoid of analysis of different ways to build such positions in the construction industry. The purpose here is to explore the challenges involved in implementing innovations that require companies to establish new positions in a construction supply chain. Three in‐depth case studies in the Swedish timber housing industry illustrate different challenges that arise with the different modes of repositioning of organic growth, collaboration and acquisition, and with moving either backward or forward in the supply chain. Organic growth is a slow process that prevents companies from taking a first‐mover advantage. Acquisitions can involve difficulties with exploiting acquired competences. Collaboration brings the risks of diversity of interests and transient solutions. Furthermore, and as shown in the cases studied, challenges are associated with pre‐existing competencies and relationships, with technology‐oriented companies facing customer‐related difficulties and market‐oriented companies mainly facing ones involving technological solutions. The construction industry’s idiosyncrasies amplify these challenges, as its fragmentation makes it difficult to connect with external companies at new positions. Stronger ties with business partners are likely to enable construction companies to exploit more easily innovations that require repositioning in the supply chain.