Type II aromatic polyketide biosynthetic tailoring enzymes: diversity and adaptation in Streptomyces secondary metabolism.

Members of the bacterial genus Streptomyces are well known for their ability to produce an exceptionally wide selection of diverse secondary metabolites. These include natural bioactive chemical compounds which have potential applications in medicine, agriculture and other fields of commerce. The outstanding biosynthetic capacity derives from the characteristic genetic flexibility of Streptomyces secondary metabolism pathways: i) Clustering of the biosynthetic genes in chromosome regions redundant for vital primary functions, and ii) the presence of numerous genetic elements within these regions which facilitate DNA rearrangement and transfer between non-progeny species. Decades of intensive genetic research on the organization and function of the biosynthetic routes has led to a variety of molecular biology applications, which can be used to expand the diversity of compounds synthesized. These include techniques which, for example, allow modification and artificial construction of novel pathways, and enable gene-level detection of silent secondary metabolite clusters. Over the years the research has expanded to cover molecular-level analysis of the enzymes responsible for the individual catalytic reactions. In vitro studies of the enzymes provide a detailed insight into their catalytic functions, mechanisms, substrate specificities, interactions and stereochemical determinants. These are factors that are essential for the thorough understanding and rational design of novel biosynthetic routes. The current study is a part of a more extensive research project (Antibiotic Biosynthetic Enzymes; www.sci.utu.fi/projects/biokemia/abe), which focuses on the post-PKS tailoring enzymes involved in various type II aromatic polyketide biosynthetic pathways in Streptomyces bacteria. The initiative here was to investigate specific catalytic steps in anthracycline and angucycline biosynthesis through in vitro biochemical enzyme characterization and structural enzymology. The objectives were to elucidate detailed mechanisms and enzyme-level interactions which cannot be resolved by in vivo genetic studies alone. The first part of the experimental work concerns the homologous polyketide cyclases SnoaL and AknH. These catalyze the closure of the last carbon ring of the tetracyclic carbon frame common to all anthracycline-type compounds. The second part of the study primarily deals with tailoring enzymes PgaE (and its homolog CabE) and PgaM, which are responsible for a cascade of sequential modification reactions in angucycline biosynthesis. The results complemented earlier in vivo findings and confirmed the enzyme functions in vitro. Importantly, we were able to identify the amino acid -level determinants that influence AknH and SnoaL stereoselectivity and to determine the complex biosynthetic steps of the angucycline oxygenation cascade of PgaE and PgaM. In addition, the findings revealed interesting cases of enzyme-level adaptation, as some of the catalytic mechanisms did not coincide with those described for characterised homologs or enzymes of known function. Specifically, SnoaL and AknH were shown to employ a novel acid-base mechanism for aldol condenzation, whereas the hydroxylation reaction catalysed by PgaM involved unexpected oxygen chemistry. Owing to a gene-level fusion of two ancestral reading frames, PgaM was also shown to adopt an unusual quaternary sturucture, a non-covalent fusion complex of two alternative forms of the protein. Furthermore, the work highlighted some common themes encountered in polyketide biosynthetic pathways such as enzyme substrate specificity and intermediate reactivity. These are discussed in the final chapters of the work.

A structural bridge between alternant and non-alternant hydrocarbons

A simple set of trimethylene-substituted even, fully-pi-bonded, non-alternant monocycles is shown to have several key features in common with acyclic, even alternant polyenes at the Hückel level. These non-alternant molecules provide a bridge between alternant and non-alternant hydrocarbons. This topic might serve as a useful addition to Hückel theory courses targeted at senior undergraduate students.

Multicointegration, polynomial cointegration and I(2) cointegration with structural breaks. An application to the sustainability of the US external deficit

In this paper we model the multicointegration relation, allowing for one structural break. Since multicointegration is a particular case of polynomial or I(2) cointegration, our proposal can also be applied in these cases. The paper proposes the use of a residualbased Dickey-Fuller class of statistic that accounts for one known or unknown structural break. Finite sample performance of the proposed statistic is investigated by using Monte Carlo simulations, which reveals that the statistic shows good properties in terms of empirical size and power. We complete the study with an empirical application of the sustainability of the US external deficit. Contrary to existing evidence, the consideration of one structural break leads to conclude in favour of the sustainability of the US external deficit.

Another Look at the Null of Stationary Real Exchange Rates: Panel Data with Structural Breaks and Cross-section Dependence

This paper re-examines the null of stationary of real exchange rate for a panel of seventeen OECD developed countries during the post-Bretton Woods era. Our analysis simultaneously considers both the presence of cross-section dependence and multiple structural breaks that have not received much attention in previous panel methods of long-run PPP. Empirical results indicate that there is little evidence in favor of PPP hypothesis when the analysis does not account for structural breaks. This conclusion is reversed when structural breaks are considered in computation of the panel statistics. We also compute point estimates of half-life separately for idiosyncratic and common factor components and find that it is always below one year.

On the effects of welding parameters on weld quality of plasma arc keyhole welding of structural steels

The possibility and the usefulness of applying plasma keyhole welding to structural steels with different compositions and material thicknesses, and in various welding positions has been examinated. Single pass butt welding with I groove in flat, horizontal vertical and vertical positions and root welding with V , Y and U grooves of thick plate material in flat position have been studied and the welds with high quality has been obtained. The technological conditions for successful welding are presented. The single and interactive effects of welding parameters on weld quality, especially on surface weld defects, geometrical form errors, internal defects and mechanical properties (strength, ductility, impact toughness, hardness and bendability) of weld joint, are presented. Welding parameter combinations providing the best quality welds are also presented.

Predicting overall service quality: a structural equation modelling approach

In this article, the results of a modified SERVQUAL questionnaire (Parasuraman et al., 1991) are reported. The modifications consisted in substituting questionnaire items particularly suited to a specific service (banking) and context (county of Girona, Spain) for the original rather general and abstract items. These modifications led to more interpretable factors which accounted for a higher percentage of item variance. The data were submitted to various structural equation models which made it possible to conclude that the questionnaire contains items with a high measurement quality with respect to five identified dimensions of service quality which differ from those specified by Parasuraman et al. And are specific to the banking service. The two dimensions relating to the behaviour of employees have the greatest predictive power on overall quality and satisfaction ratings, which enables managers to use a low-cost reduced version of the questionnaire to monitor quality on a regular basis. It was also found that satisfaction and overall quality were perfectly correlated thus showing that customers do not perceive these concepts as being distinct

Modelo de comportamiento en sevicio para estructuras reticulares de hormigón. Parte I: Descripción del modelo y ejemplos de aplicación

A general method for instantaneous and time-dependent serviceability analysis of plane concrete frames is presented. The methodology is based in an extension of the classic matrix formulation for bars. The main aspects influencing the behaviour of the structural concrete are considered: cracking, creep, shrinkage or prestress losses. To simulate the effect of cracking a smeared model (developed in Part II) based on the modification of the tensile branch of the concrete stress-strain relationship is adopted. The general approach considered permits the application to different materials and constitutive laws. Sequential construction (sectional and structural), incorporation of reinforcement, consideration of the loads history; placing and removing shores, and restraining or releasing in boundary conditions are considered. Some examples are included to highlight the capabilities of the model

On the application of knoevenagel condensation for the synthesis of benzylidene benzothiazine compounds and structural study

The synthesis and physico-chemical properties of new 6-acetylamino or 6-benzoyl-amino 2-benzylidene-4-methyl-4H-benzo[1,4]thiazin-3-ones and 6-benzoylamino or 6-nitro 2-benzylidene-4H-benzo[1,4]thiazin-3-ones are described. These benzylidene benzothiazine compounds were prepared by the Knoevenagel condensation with benzaldehydes. The configurations and conformations of benzylidene benzothiazine derivatives were optimised using the semi-empirical method AM1.

Structural development of the harvester head saw unit

The main goal of the thesis was to further develop harvester head saw device to the Finnish forest machine manufacturer. The work was done from the basis of the manufacturer´s current production model and the earlier study from this same subject called: “Development of chain saw for harvester” Tero Kaatrasalo, 2004. The work was focused to improving the serviceability and reliability of the saw device, but design also included adding few beforehand determined new features into the saw unit. This was done to give some added value for the end customer. The work includes analysis of the earlier saw devices and ideations of the improvements for the structure.

Synthesis of resorcinolic lipids bearing structural similarities to cytosporone A

Inspired by the structure and biological activities of resorcinolic lipids and, particularly cytosporone A- a potent inhibitor of plantule germination and growth, we have performed the synthesis of the analogs 3-heptyl-3-hydroxy-5,7-dimethoxy-2-benzofuran-1(3H)-one (1) and 3-heptyl-3-hydroxy-4,6-dimethoxy-2-benzofuran-1(3H)-one (2). The intermediates and products were submitted to allelopathic test using Lactuca sativa L. seeds. Target compound 1 showed an inhibitory effect on germination and growth of hypocotyl and radicle in milimolar range.

Structural and reactivity analyses of 2-benzylamino-1,4-naphthoquinone by X-ray characterization, electrochemical measurements, and dft single-molecule calculations

This study represents an integrated approach towards understanding the electronic and structural aspects of 2-benzylamino-1,4-naphthalenedione, a representative 2-amino-napfthoquinone. To this end, theoretical calculations performed at the B3PW91/6-31+G(d) level of density functional theory, electrochemical and X-ray structural investigation were employed. Two intramolecular H-bonds and other two intermolecular H-bonds were observed, including non-classical interactions. Cyclic voltammogram (CV) and differential pulse voltammetry (DPV) show two pairs of peaks, being each one a monoelectronic process.

Thermal and structural study of guava (Psidium guajava L) powders obtained by two dehydration methods

Two food products (powders) were obtained by hot-air drying or lyophilisation methods on the whole guava fruits. The powders were characterised by sensory and thermal analyses (TGA-DSC), infrared spectroscopy (IR), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Thermal, morphological and structural characterisations showed a similar behaviour for the two solids. TGA-DSC and IR showed the presence of pectin as the main constituent of solids. A semi-crystalline profile was evidenced by XRD, and lamellar/spherical morphologies were observed by SEM. Sensory analyses revealed an aroma highly related to guava. These value-added food products are an alternative to process guava and avoid loss during postharvest handling.

Encapsulation of naproxen in nanostructured system: structural characterization and in vitro release studies

Nanoparticles were produced by solvent emulsification evaporation method with the following characteristics: nanometric size (238 ± 3 nm), narrow polydispersity index (0.11), negative zeta potential (-15.1 mV), good yield of the process (73 ± 1.5%), excellent encapsulation efficiency (81.3 ± 4.2%) and spherical shape. X-rays diffraction demonstrated the loss of drug crystallinity after encapsulation; however, the profile of the diffractograms of the poly-ε-caprolactone (PCL) nanoparticles was kept. Differential scanning calorimetry thermograms, correspondingly, exhibited the loss of drug melting peak and the increasing of the melting point of the PCL nanoparticles, evidencing an interaction drug-polymer. Naproxen release was low and sustained obeying the Higuchi´s kinetic. The results show that nanoparticles are promising sustained release system to the naproxen.

Preparation of glasses and glass ceramics of heavy metal oxides containing silver: optical, structural and electrochemical properties

Silver containing heavy metal oxide glasses and glass ceramics of the system WO3-SbPO4-PbO-AgCl with different AgCl contents have been prepared and their thermal, structural and optical properties characterized. Glass ceramics containing metallic silver nanoparticles have been prepared by annealing glass samples at temperatures above the glass transition and analyzed by transmission electron microscopy and energy dispersive X-ray microanalysis. The presence of the metallic clusters has been also confirmed by the observation of a surface plasmon resonance band in the visible range. Cyclic voltammetric measurements indicated the presence of metallic silver into the glasses, even before to perform the thermal treatment.

Complete ¹H and 13C NMR structural assignments for a group of four goyazensolide-type furanoheliangolides

Four goyazensolide-type sesquiterpene lactones - lychnofolide, centratherin, goyazensolide and goyazensolide acetate - were thoroughly studied by NMR experimental techniques. ¹H NMR, 13C NMR {¹H}, COSY, HMQC, HMBC, J-res. and NOE experiments were performed to provide the needed structural information. Complete and unequivocal assignment, including the determination of all multiplicities, was obtained for each structure and the data collections are presented in tables.