Studies in selectivity of lipases in preparation of cyanohydrins, sugar conjugates and secondary alcohols

Biocatalysis can be applied in organic synthetic chemistry to counter challenges posed by increased demands towards chemo-, regio- and stereoselectivity, not forgetting the need for greener chemistry. During the last 30 years, biocatalysis with the use of enzymes as chiral catalysts has become more common in chemistry laboratories and industrial processes. In this thesis, the use of lipases as versatile biocatalysts in the acylation of alcohols is examined both in the light of literature examples and four original publications. In the first part of the work presented in this thesis lipases were utilized in two examples concerning secondary alcohols. First, the kinetic resolution of heterocyclic aromatic secondary alcohols through transesterification was thoroughly examined including the studies of competing hydrolysis and esterification reactions. In another example, lipases were utilized in the formation of a dynamic systemic resolution (DSR) process which in turn was used as a developmental tool in the optimization of the dynamic kinetic resolution (DKR) of five heterocyclic aromatic cyanohydrins in one pot for the preparation of cyanohydrin esters as single enantiomers. In the second part of the work, the regio- and stereoselectivity of lipases was used to form sugar conjugates of glyceric and β-amino acids. The primary hydroxyl groups of methyl α-D-galacto-, -gluco- and -mannopyranosides were now acylated trough lipasecatalyzed transesterification and enantioselective lipase-catalyzed ring-opening of β- lactams, respectively.

Physical properties of structured lipids from lard and soybean oil produced by enzymatic interesterification

The main goal of the present research was to evaluate the physical properties of blends of lard and soybean oil modified by enzymatic interesterification catalyzed by two different commercial (microbial) lipases, viz. from Candida cylindracea (AY30TM) and from Mucor circinelloides (M10TM). Pure lard exhibited a softening point of ca. 31.8 °C before interesterification, and this value shifted towards 29.1 °C after interesterification by AY30 lipase and towards 28.8 °C after interesterification by M10 lipase The interesterified lard exhibited lower consistency after reaction with both lipases, and this decrease was more pronounced for the reaction catalyzed by M10 lipase. This result was most likely due to the sn-1,3-specificity of M10 lipase. Pure lard displayed a lower SFC after interesterification, and M10 lipase proved to be more effective than AY30 lipase. The non-interesterified lard had a SFC of 31.3% at 10 °C, which was reduced to 23.8 and 19.9% after interesterification with AY30 lipase and M10 lipase, respectively. The lard and soybean oil blends were affected by the enzymatic interesterification and dilution with soybean oil.

N-acétyltransférase lysosomale : organisation, fonctionnement et défauts moléculaires chez les patients atteints du syndrome de Sanfilippo type C

L’acétylation des résidus de glucosamine terminaux par la N-acétyltransférase lysosomale (HGSNAT) est une étape essentielle de la dégradation catabolique de l’héparan sulfate. Des défauts dans cette réaction causent une maladie de surcharge lysosomale autosomale récessive rare : le désordre de Sanfilippo type C (SFC). À ce jour, 54 mutations ont été rapportées chez des patients SFC, incluant 13 mutations des sites d’épissage, 11 insertions et délétions, 8 mutations non-sens, 18 mutations faux-sens et 4 polymorphismes, avec différentes manifestations phénotypiques. Nous avons identifié 10 d’entre elles et effectué une étude exhaustive portant sur l’éventail des mutations SFC, leur distribution dans la population de patients, ainsi que leur impact potentiel sur la structure de la HGSNAT. Les erreurs d’épissage, les mutations non-sens, les insertions et les délétions devraient toutes entraîner un ARN non fonctionnel qui est rapidement dégradé par des mécanismes de contrôle qualité cellulaire. Les 4 polymorphismes identifiés sont des changements d'acides aminés qui ne modifient pas l'activité enzymatique, la glycosylation ou la localisation et n'ont donc pas de signification au niveau clinique. Au niveau des enzymes, les polymorphismes sont des changements d’acides aminés qui n’affectent pas la fonction, mais dans un contexte d’acides nucléiques ils peuvent être considérés comme des mutations faux-sens. Les dix-huit mutations faux-sens qui ont été exprimées ont produit des protéines inactives, en raison d'erreurs dans leur repliement. Ceci expliquerait donc la progression sévère de la maladie chez les personnes porteuses de ces mutations. Les protéines mutantes mal repliées sont anormalement glycosylées et conservées dans le réticulum endoplasmique. La thérapie par amélioration de l’activité enzymatique par des chaperonnes est une option thérapeutique potentielle, spécifiquement conçue pour exploiter l'activité enzymatique résiduelle de mutants mal repliés, afin d’éliminer les substrats stockés. Nous avons démontré que le traitement de plusieurs lignées de fibroblastes de patients SFC avec le chlorhydrate de glucosamine, un inhibiteur spécifique de la HGSNAT, a partiellement restauré l’activité de l'enzyme mutante, fournissant une preuve de l’utilité future de la thérapie par des chaperonnes dans le traitement de la maladie de SFC.

Le syndrome de fatigue chronique : méconnu mais pas sans solution

Travail d'intégration réalisé dans le cadre du cours PHT-6113.

"La socialización de género en docentes y su posible interacción con adolescentes víctimas de abuso sexual"

La presente investigación aborda el tema de la socialización de género en docentes y su posible interacción con adolescentes víctimas de abuso sexual. La socialización es un proceso de construcción social de la realidad que se desarrolla en diferentes ámbitos y con diferentes agentes; de esta manera, pueden existir procesos de socialización en torno a distintas temáticas que se insertan en la vida social, como por ejemplo, el género, la familia, la escuela o el trabajo. En este trabajo se estudia el proceso de socialización del género de los/las docentes del colegio La Victoria, y de orientadoras-docentes del mismo colegio y del Colegio San Cristóbal. Además de estudiar dichos procesos de socialización (socialización primaria y secundaria), se estudia cómo afecta el proceso de socialización de género en los/las docentes al momento de interactuar con adolescentes víctimas de abuso sexual. Esta investigación se contextualiza dentro del marco legal sobre la protección de los derechos de los/las niños/as y adolescentes. Se parte desde la perspectiva de derechos humanos ya que dentro de esos derechos se encuentran los derechos sexuales y reproductivos (DSR). Estos son derechos humanos, ya que su vulneración es una forma de violentar la dignidad humana y el abuso sexual a menores es una forma de vulnerar los DSR, con el agravante de que la gran mayoría de las víctimas son niños, niñas y adolescentes.

Perdurabilidad y competitividad en bancos comerciales segmento masivo, tamaño intermedio: caso Colpatria

Teniendo en cuenta la complejidad del sistema financiero colombiano y su alto impacto en la economía nacional, surge el interés de realizar un estudio y análisis cualitativo y cuantitativo de diversos agentes de este sector. A partir de allí, se genera un diagnóstico de morbilidad, hacinamiento e imitación en el sector, así como de perdurabilidad del Banco Colpatria Durante el documento se hará énfasis en la estrategia de redes y alianzas del sector estudiado, el cual, según revisión, promueve la bancarización y acceso al sistema financiero en todo el país. A su vez, se analizarán indicadores financieros del sector, se utilizará análisis retrospectivo y se realizarán entrevistas y encuestas a los colaboradores del Banco Colpatria y a los usuarios de sus redes, con el propósito de entender el estado actual del sector, de la organización estudiada, así como de identificar características o elementos del pasado, presente y futuro de la banca y de sus agentes. Por lo tanto, se emite un diagnóstico del sector estratégico: bancos comerciales segmento masivo, tamaño intermedio y finalmente se generan recomendaciones para evitar el hacinamiento del sector y generar perdurabilidad en la entidad estudiada.

Variables associated with nest survival of Golden-winged Warblers (Vermivora chrysoptera) among vegetation communities commonly used for nesting

Among shrubland- and young forest-nesting bird species in North America, Golden-winged Warblers (Vermivora chrysoptera) are one of the most rapidly declining partly because of limited nesting habitat. Creation and management of high quality vegetation communities used for nesting are needed to reduce declines. Thus, we examined whether common characteristics could be managed across much of the Golden-winged Warbler’s breeding range to increase daily survival rate (DSR) of nests. We monitored 388 nests on 62 sites throughout Minnesota, Wisconsin, New York, North Carolina, Pennsylvania, Tennessee, and West Virginia. We evaluated competing DSR models in spatial-temporal (dominant vegetation type, population segment, state, and year), intraseasonal (nest stage and time-within-season), and vegetation model suites. The best-supported DSR models among the three model suites suggested potential associations between daily survival rate of nests and state, time-within-season, percent grass and Rubus cover within 1 m of the nest, and distance to later successional forest edge. Overall, grass cover (negative association with DSR above 50%) and Rubus cover (DSR lowest at about 30%) within 1 m of the nest and distance to later successional forest edge (negative association with DSR) may represent common management targets across our states for increasing Golden-winged Warbler DSR, particularly in the Appalachian Mountains population segment. Context-specific adjustments to management strategies, such as in wetlands or areas of overlap with Blue-winged Warblers (Vermivora cyanoptera), may be necessary to increase DSR for Golden-winged Warblers.

The influence of bottom topography on the decay of modeled Agulhas rings

The influence of a large meridional submarine ridge on the decay of Agulhas rings is investigated with a 1 and 2-layer setup of the isopycnic primitive-equation ocean model MICOM. In the single-layer case we show that the SSH decay of the ring is primarily governed by bottom friction and secondly by the radiation of Rossby waves. When a topographic ridge is present, the effect of the ridge on SSH decay and loss of tracer from the ring is negligible. However, the barotropic ring cannot pass the ridge due to energy and vorticity constraints. In the case of a two-layer ring the initial SSH decay is governed by a mixed barotropic–baroclinic instability of the ring. Again, radiation of barotropic Rossby waves is present. When the ring passes the topographic ridge, it shows a small but significant stagnation of SSH decay, agreeing with satellite altimetry observations. This is found to be due to a reduction of the growth rate of the m = 2 instability, to conversions of kinetic energy to the upper layer, and to a decrease in Rossby-wave radiation. The energy transfer is related to the fact that coherent structures in the lower layer cannot pass the steep ridge due to energy constraints. Furthermore, the loss of tracer from the ring through filamentation is less than for a ring moving over a flat bottom, related to a decrease in propagation speed of the ring. We conclude that ridges like the Walvis Ridge tend to stabilize a multi-layer ring and reduce its decay.

The origin of brines underlying Lake Kinneret

Lake Kinneret (LK) is a relatively fresh water take situated in the Dead Sea Rift (DSR) Valley. The pore water (PW) in the sediments underlying LK pelagic zone have significantly higher salinity than that of the lake. The concentrations of major ion solutes (Cl, Br, Na, K, Mg) in PW from six 2.4 m to 5.1 m long sediment cores increase linearly with depth, indicating the occurrence of saline, deep seated brines. The upper part of the PW column is affected by the much fresher boundary with LK water and in most cores is characterized by gradually increasing Br/Cl and decreasing Na, Mg, K/Cl molar ratios, which tend to stabilize at about 2.0 m below the sediment surface. The 'stable' molar ratios in the deeper PW vary spatially and are supposed to represent the ratios in the deep underlying brines at each site. When plotted as Na/Cl vs. Br/Cl, the stable ratios of the northern and central part of the lake fall close to a straight line which characterizes many of the brines in the DSR Valley. However, the respective ratios in the southern part of the lake fall markedly off the DSR line. Moreover, Na/Cl and K/Cl molar ratios in the south are significantly higher than in the central and northern parts. delta Cl-37 measured in present LK water is ca. 0.0 parts per thousand. Along the PW column at the lake center, delta Cl-37 is becoming more positive with depth, reaching values of about +0.5 parts per thousand to +0.6 parts per thousand at 3 m depth. Even more positive values (+0.7 parts per thousand to +0.8 parts per thousand) are detected further north, in PW from deeper sediment layers. In contrast, in PW from the southeastern part of the lake, delta Cl-37 is becoming more negative with depth (-1.0 parts per thousand at similar to 2.6 m). It is suggested that these isotopic differences are also indicative of spatial variability in the PW brine sources. O-18 and D values in the PW of all 3 m long cores are similar and resemble the respective levels in LK. The source of H2O in 3 m deep, bed sediments is claimed to be the overlying lake water, and therefore water isotopes do not provide a clue regarding the original water isotopic composition in the underlying brines. PW from the southeast with higher K/Cl and Na/Cl but lower concentrations of these solutes, suggest leaching by meteoric water of sub-surface halite and post-halite salt formations, while the more saline PW from the northern and central parts, that have lower K/Cl and Na/Cl, and higher Br/Cl, are similar to DSR brines and represent underlying residual brines. (C) 2009 Elsevier B.V. All rights reserved.

Flux comparison of Eulerian and Lagrangian estimates of Agulhas leakage: A case study using a numerical model

Estimating the magnitude of Agulhas leakage, the volume flux of water from the Indian to the Atlantic Ocean, is difficult because of the presence of other circulation systems in the Agulhas region. Indian Ocean water in the Atlantic Ocean is vigorously mixed and diluted in the Cape Basin. Eulerian integration methods, where the velocity field perpendicular to a section is integrated to yield a flux, have to be calibrated so that only the flux by Agulhas leakage is sampled. Two Eulerian methods for estimating the magnitude of Agulhas leakage are tested within a high-resolution two-way nested model with the goal to devise a mooring-based measurement strategy. At the GoodHope line, a section halfway through the Cape Basin, the integrated velocity perpendicular to that line is compared to the magnitude of Agulhas leakage as determined from the transport carried by numerical Lagrangian floats. In the first method, integration is limited to the flux of water warmer and more saline than specific threshold values. These threshold values are determined by maximizing the correlation with the float-determined time series. By using the threshold values, approximately half of the leakage can directly be measured. The total amount of Agulhas leakage can be estimated using a linear regression, within a 90% confidence band of 12 Sv. In the second method, a subregion of the GoodHope line is sought so that integration over that subregion yields an Eulerian flux as close to the float-determined leakage as possible. It appears that when integration is limited within the model to the upper 300 m of the water column within 900 km of the African coast the time series have the smallest root-mean-square difference. This method yields a root-mean-square error of only 5.2 Sv but the 90% confidence band of the estimate is 20 Sv. It is concluded that the optimum thermohaline threshold method leads to more accurate estimates even though the directly measured transport is a factor of two lower than the actual magnitude of Agulhas leakage in this model.

Detection and enumeration of sulphate-reducing bacteria in estuarine sediments by competitive PCR

The distribution of sulphate-reducing bacteria (SRB) in the sediments of the Colne River estuary, Essex, UK covering different saline concentrations of sediment porewater was investigated by the use of quantitative competitive PCR. Here, we show that a new PCR primer set and a new quantitative method using PCR are useful tools for the detection and the enumeration of SRB in natural environments. A PCR primer set selective for the dissimilatory sulphite reductase gene (dsr) of SRB was designed. PCR amplification using the single set of dsr-specific primers resulted in PCR products of the expected size from all 27 SRB strains tested, including Gram-negative and positive species. Sixty clones derived from sediment DNA using the primers were sequenced and all were closely related with the predicted dsr of SRB. These results indicate that PCR using the newly designed primer set are useful for the selective detection of SRB from a natural sample. This primer set was used to estimate cell numbers by dsr selective competitive PCR using a competitor, which was about 20% shorter than the targeted region of dsr. This procedure was applied to sediment samples from the River Colne estuary, Essex, UK together with simultaneous measurement of in situ rates of sulphate reduction. High densities of SRB ranging from 0.2 - 5.7 × 108 cells ml-1 wet sediment were estimated by the competitive PCR assuming that all SRB have a single copy of dsr. Using these estimates cell specific sulphate reduction rates of 10-17 to 10-15 mol of SO42- cell-1 day-1 were calculated, which is within the range of, or lower than, those previously reported for pure cultures of SRB. Our results show that the newly developed competitive PCR technique targeted to dsr is a powerful tool for rapid and reproducible estimation of SRB numbers in situ and is superior to the use of culture-dependent techniques.

Effect of dextransucrase cellobiose acceptor products on the growth of human gut bacteria

The selective fermentation by human gut bacteria of gluco-oligosaccharides obtained from the reaction between the glucosyl group of sucrose and cellobiose, catalyzed by dextransucrases (DSR) from Leuconostoc mesenteroides, has been evaluated. Oligosaccharides were fractionated according to their molecular weight, and their effect on the growth of different bacterial groups was studied. To determine the structure (position and configuration of glycosidic linkages)�function relationship, their properties were compared to those of DSR maltose acceptor products (DSRMal) and of recognized prebiotic carbohydrates (fructo-oligosaccharides, FOS). Cellobiose acceptor products (DSRCel) showed bifidogenic properties similar to those of FOS. However, no significant differences related to molecular weight or isomeric configurations were found for DSRCel and DSRMal products.

Demand side response: patterns in Europe and future policy perspectives under capacity mechanisms

Demand Side Response (DSR) has been slow to emerge in European electricity markets. This paper aims to both examine the reasons for low levels of DSR in Europe and reflect on factors that might affect the participation of DSR in capacity mechanisms. It relies on available evidence from the literature, secondary data on existing DSR programmes and energy aggregator's data from industries participating in DSR. Findings show that changes to the duration of contracted loads under existing or new programmes might increase the penetration of DSR. The introduction of capacity mechanisms may increase DSR from demand turn down if longer response times were available.

Financial implications for companies running standby generators in support of a smart grid

Integrating renewable energy into built environments requires additional attention to the balancing of supply and demand due to their intermittent nature. Demand Side Response (DSR) has the potential to make money for organisations as well as support the System Operator as the generation mix changes. There is an opportunity to increase the use of existing technologies in order to manage demand. Company-owned standby generators are a rarely used resource; their maintenance schedule often accounts for a majority of their running hours. DSR encompasses a range of technologies and organisations; Sustainability First (2012) suggest that the System Operator (SO), energy supply companies, Distribution Network Operators (DNOs), Aggregators and Customers all stand to benefit from DSR. It is therefore important to consider impact of DSR measures to each of these stakeholders. This paper assesses the financial implications of organisations using existing standby generation equipment for DSR in order to avoid peak electricity charges. It concludes that under the current GB electricity pricing structure, there are several regions where running diesel generators at peak times is financially beneficial to organisations. Issues such as fuel costs, Carbon Reduction Commitment (CRC) charges, maintenance costs and electricity prices are discussed.

Hydrogen peroxide is a novel mediator of inflammatory hyperalgesia, acting via transient receptor potential vanilloid 1-dependent and independent mechanisms

Inflammatory diseases associated with pain are often difficult to treat in the clinic due to insufficient understanding of the nociceptive pathways involved. Recently, there has been considerable interest in the role of reactive oxygen species (ROS) in inflammatory disease, but little is known of the role of hydrogen peroxide (H(2)O(2)) in hyperalgesia. In the present study, intraplantar injection of H(2)O(2)-induced a significant dose- and time-dependent mechanical and thermal hyperalgesia in the mouse hind paw, with increased c-fos activity observed in the dorsal horn of the spinal cord. H(2)O(2) also induced significant nociceptive behavior Such as increased paw licking and decreased body liftings. H(2)O(2) levels were significantly raised in the carrageenan-induced hind paw inflammation model, showing that this ROS is produced endogenously in a model of inflammation. Moreover, superoxide dismutase and catalase significantly reduced carrageenan-induced mechanical and thermal hyperalgesia, providing evidence of a functionally significant endogenous role. Thermal, but not mechanical, hyperalgesia in response to H(2)O(2) (i.pl.) Was longer lasting in TRPV1 wild type mice compared to TRPV1 knockouts. It is unlikely that downstream lipid peroxidation was increased by H(2)O(2). In conclusion, we demonstrate a notable effect of H(2)O(2) in mediating inflammatory hyperalgesia, thus highlighting H(2)O(2) removal as a novel therapeutic target for anti-hyperalgesic drugs in the clinic. (C) 2008 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.