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Waste diversion and reduction continues to be a prominent discussion among Canadian municipalities as we collectively recognize the impact that waste production has on the environment and our future, especially in the context of climate change. Much of the focus in this regard has been on individual waste generation and reduction and the “zero-waste” movement, with less focus on construction, renovation, and demolition (CRD) waste. Research shows that CRD waste contributes between 27% and 40% of total municipal solid waste in Canada and it is estimated that the CRD sector is responsible for 40% of raw material consumption in North America. With an estimated potential of 95% of CRD materials being available for salvage, reuse, repurposing, and recycling, there is a lot of opportunity for growth in responsible CRD waste management. My research shows that deconstruction, rather than demolition of buildings, is an important next step in waste diversion for Canadian municipalities and the waste generated from CRD presents an opportunity to recover a significant amount of resources. This research explores the barriers for deconstruction programs and policies for large, Canadian municipalities, how to overcome those barriers, and establishes a framework for moving forward in a municipal setting, working with the City of Edmonton for a real-world application. The results show that deconstruction has a small foothold in Canada and the US, but there are some leading-edge and developing examples. My framework builds on these and offers a path for actioning residential building deconstruction that can have a significant impact on reducing CRD waste going to landfills.

Questo volume, il secondo della collana, discute l’applicazione del principio di proporzionalità nella disciplina bancaria europea. Modulare le regole in funzione delle caratteristiche di ciascuna banca, in particolare, può essere assai costoso. È questa una delle ragioni per cui la disciplina europea tende spesso a preferire soluzioni regolamentari a “taglia unica” (one-size-fits-all). Anche quando considera le banche di minori dimensioni, il legislatore trascura inoltre le peculiarità che caratterizzano i gruppi bancari di credito cooperativo: pur concepiti come veri e propri “strumenti di vigilanza” e composti principalmente da banche piccole e non complesse, anch’essi finiscono per essere soggetti alle medesime regole dettate per le large institution. Sono queste le premesse da cui muovono gli autori nel formulare possibili soluzioni al problematico arretramento della proporzionalità. Nello stesso tempo, risulta opportuno assicurare che l’applicazione di tale principio non sfoci in un ingiustificato allentamento della disciplina prudenziale, per evitare di minacciare la stabilità dell’intero sistema finanziario. In questo senso, la recente crisi del settore bancario statunitense – culminata con i fallimenti di Silicon Valley Bank, Signature Bank e First Republic Bank – consente di formulare conclusioni articolate, capaci di distinguere tra requisiti prudenziali di capitale, regole di corporate governance e adempimenti di vigilanza.

Waste diversion and reduction continues to be a prominent discussion among Canadian municipalities as we collectively recognize the impact that waste production has on the environment and our future, especially in the context of climate change. Much of the focus in this regard has been on individual waste generation and reduction and the “zero-waste” movement, with less focus on construction, renovation, and demolition (CRD) waste. Research shows that CRD waste contributes between 27% and 40% of total municipal solid waste in Canada and it is estimated that the CRD sector is responsible for 40% of raw material consumption in North America. With an estimated potential of 95% of CRD materials being available for salvage, reuse, repurposing, and recycling, there is a lot of opportunity for growth in responsible CRD waste management. My research shows that deconstruction, rather than demolition of buildings, is an important next step in waste diversion for Canadian municipalities and the waste generated from CRD presents an opportunity to recover a significant amount of resources. This research explores the barriers for deconstruction programs and policies for large, Canadian municipalities, how to overcome those barriers, and establishes a framework for moving forward in a municipal setting, working with the City of Edmonton for a real-world application. The results show that deconstruction has a small foothold in Canada and the US, but there are some leading-edge and developing examples. My framework builds on these and offers a path for actioning residential building deconstruction that can have a significant impact on reducing CRD waste going to landfills.

L’introduzione di una disciplina identica per tutte le banche (one-size-fits-all approach) va spesso a detrimento degli enti più piccoli e rischia altresì di pregiudicare la concorrenza sul mercato. Di qui, il tradizionale utilizzo del principio di proporzionalità quale criterio applicativo di regole altrimenti uniformi, in modo da “calibrare” le norme in base alle particolari caratteristiche di ciascuna banca. Nondimeno, la presenza di notevoli “costi minimi” di compliance e le difficoltà riscontrate dalle autorità di vigilanza nel declinare le proprie strategie di enforcement hanno suggerito di elevare la proporzionalità a vero e proprio metodo per la redazione delle norme. Con il CRD V package, è stata così introdotta la figura degli « enti piccoli e non complessi », cui il legislatore ricollega l’applicazione di una disciplina “più leggera” in tema di SREP e obblighi di reporting. Tuttavia, poiché l’attività di vigilanza deve sempre svolgersi “al più alto livello di consolidamento”, la necessaria parte- cipazione delle BCC a un gruppo bancario cooperativo impedisce l’applicazione delle predette regole e attira tali enti all’interno della più severa disciplina prevista per le large institution. Pur condivisibile per i gruppi bancari tradizionali, espressione della medesima impresa, una simile conclusione non pare invece ragionevole per i gruppi bancari cooperativi, le cui peculiarità impongono all’interprete di adeguare l’applicazione di tali regole alle specificità del caso concreto., The ‘one-size-fits-all’approach to banking regulation may harm smaller institutions and jeopardize competition. For this reason, the proportionality principle requires supervisors to ‘calibrate’ the actual application of prudential regulation according to the size and complexity of each bank. Nonetheless, while economies of scale are still expected to play a fundamental role, even the high costs of enforcement experienced by supervisors may hinder the actual application of the proportionality principle. To address these obstacles, the ‘CRD V package’ adopted a different approach to proportionality and introduced a specific set of tailored rules for “small and non-complex institutions” (for example, providing a lighter reporting requirement or a milder approach to SREP). However, since supervisory activities must always be carried out ‘at the highest level of consolidation’, the mandatory participation of community banks in a ‘cooperative group’ excludes the application of the special regime drawing these institutions within the stricter rules provided for “large banks”. Although reasonable for traditional banking groups — usually the expression of a single economic entity — such a conclusion does not hold for cooperative banking groups, whose peculiarities must be taken into proper consideration by the supervisor.

This Article examines the constitutionality of an Israeli bill that criminalizes the use of PA systems in prayer houses, punishable by a fine of 5000–10,000 NIS (the Muezzin Law). The Bill was presented to the Israeli Parliament (the Knesset) as a religiously-neutral environmental law. This Article asserts that a careful reading of the Bill’s language reveals that it is specifically tailored to apply precisely to Muslim prayer houses, thus criminalizing the Muslim call for prayer (the adhan), especially the call occurring between dawn and sunrise (the Fajer adhan). As such, we perceive the Muezzin law as violating the right to equality and the right to dignity of the Muslim minority in Israel, as well as infringing upon its religious feelings. Additionally, we contend that the Muezzin Law is not truly driven by environmental concern, but rather that it represents a conflict with religious dimension (a CRD)—namely, the perception that the adhan, as a Muslim symbol, poses a threat to the identity of Jews in Israel. Examining the constitutionality of the Muezzin Law introduces a crucial question relating to the interplay between constitutional law and criminal law. Our assertion is that in any constitutional democracy, in order for the legislature to validly classify conduct as a crime, such criminalization must befit the values of constitutional democracy, serve a proper purpose, and be proportionate. The requirement for proportionality consists of three subtests: (a) the rational connection test; (b) the necessity test; and (c) the balancing benefits test. It is our contention that the Muezzin Law comprises an unconstitutional criminalization of the Fajer adhan. It stands in contrast with the basic values of constitutional democracy, primarily that of tolerance towards a religious minority, particularly, the Muslim community. Additionally, we assert that the Muezzin Law’s purpose is improper as it aims at infringing upon the religious feelings of the Muslim

This Article examines the constitutionality of an Israeli bill that criminalizes the use of PA systems in prayer houses, punishable by a fine of 5000–10,000 NIS (the Muezzin Law). The Bill was presented to the Israeli Parliament (the Knesset) as a religiously-neutral environmental law. This Article asserts that a careful reading of the Bill’s language reveals that it is specifically tailored to apply precisely to Muslim prayer houses, thus criminalizing the Muslim call for prayer (the adhan), especially the call occurring between dawn and sunrise (the Fajer adhan). As such, we perceive the Muezzin law as violating the right to equality and the right to dignity of the Muslim minority in Israel, as well as infringing upon its religious feelings. Additionally, we contend that the Muezzin Law is not truly driven by environmental concern, but rather that it represents a conflict with religious dimension (a CRD)—namely, the perception that the adhan, as a Muslim symbol, poses a threat to the identity of Jews in Israel. Examining the constitutionality of the Muezzin Law introduces a crucial question relating to the interplay between constitutional law and criminal law. Our assertion is that in any constitutional democracy, in order for the legislature to validly classify conduct as a crime, such criminalization must befit the values of constitutional democracy, serve a proper purpose, and be proportionate. The requirement for proportionality consists of three subtests: (a) the rational connection test; (b) the necessity test; and (c) the balancing benefits test. It is our contention that the Muezzin Law comprises an unconstitutional criminalization of the Fajer adhan. It stands in contrast with the basic values of constitutional democracy, primarily that of tolerance towards a religious minority, particularly, the Muslim community. Additionally, we assert that the Muezzin Law’s purpose is improper as it aims at infringing upon the religious feelings of the Muslim

L’objectif de cet essai est de proposer des alternatives de gestion durable du bois récupéré postconsommation issu des matériaux de la construction, de la rénovation et de la démolition qui transitent par les centres de tri québécois. Ces derniers gèrent lesdits matériaux, parmi lesquels on retrouve entre 30 et 40 % de bois, dans une perspective de recyclage et de valorisation. Une proportion du bois récupéré a subi un traitement de préservation de types fongicide, algicide et insecticide. La plupart des traitements sont faits par l’application de produits toxiques qui génèrent ensuite une variété importante de contaminants dans l’environnement tels que des métaux lourds, des composés organochlorés, des hydrocarbures aromatiques polycycliques ainsi que des dioxines et des furanes. Se faisant, différentes solutions ont été analysées dans le but d’améliorer la situation actuelle. Une étude des meilleures pratiques en gestion du bois traité a révélé que sa réduction et son tri à la source semblent de bonnes solutions afin de minimiser la quantité totale de bois traité acheminée vers les centres de tri. Une bonne planification et une revue des modes opératoires permettraient ainsi de réduire les excédents, le gaspillage et la surconsommation. Puisque le bois traité est difficile à identifier et à ségréger dans les centres de tri, il serait pertinent de le détourner à la source afin qu’il suive un chemin approprié à sa nature contaminée. Une autre solution serait d’améliorer la performance du tri et de rendre le bois traité plus visible, plus facile à retirer. Aussi, des études comparatives au sujet de différentes solutions de remplacement pour le bois traité pourraient proposer l’utilisation de produits de traitement alternatif ainsi que de matériaux de remplacement comme le béton, l’acier galvanisé ou le bois composite. Au terme de cette analyse, plusieurs recommandations ont été fournies afin de tendre vers une gestion plus durable du bois traité au Québec. Les différ

We report the synthesis of linear and branched (1→4)-D-galactans. Four tetra- and one pentasaccharide were accessed by adopting a procedure of regioselective ring opening of a 4,6-O-naphthylidene protecting group followed by glycosylation using phenyl thioglycoside donros. The binding of the linear pentasaccharide with galectin-3 is also investigated by determination of a co-crystal structure. The binding of the (1→4)-linked galactan to Gal-3 highlights oligosaccharides of pectic galactan, which is abundant in the human diet, as putative Gal-3 ligands.

Dans une ère où la médecine personnalisée prend de plus en plus d’ampleur, il est devenu primordial de mieux comprendre les différents évènements moléculaires qui régulent la progression tumorale. Depuis plusieurs années, les galectines sont au centre d’un intérêt grandissant pour la découverte de biomarqueurs exploitables. De par leur capacité à lier une grande variété de molécules glycosylées, ces lectines animales régulent différents aspects de la progression tumorale, dont la résistance à la chimiothérapie, l’immunoévasion et le potentiel métastatique. Puisque leur expression est souvent dérégulée dans plusieurs types de cancer, il est clair qu’une meilleure caractérisation de leur expression et de leurs fonctions permettra de développer de nouveaux outils pour le diagnostic, le pronostic et le traitement des patients atteints du cancer.L’objectif de ce projet de thèse était, dans un premier temps, de déterminer l’expression et les fonctions de la galectine-7 dans le cancer de la prostate et de l’ovaire. Dans un deuxième temps, nous avons étudié la possibilité d’utiliser une signature de galectines comme biomarqueur et cible thérapeutique dans le cancer ovarien.Pour ce faire, nous avons utilisé des micromatrices tissulaires construites à partir de tissus provenant de patients atteints de cancer. Nous avons d’abord observé que l’expression de la galectine-7 est réprimée dans le cancer de la prostate. Avec l’aide de modèles cellulaires in vitro utilisant les cellules DU-145, nous avons ensuite mis en évidence une activité anti-tumorale de la galectine-7 dans le cancer de la prostate. Nous avons ensuite étudié les fonctions CRD-dépendantes de la galectine-7 à l’aide d’un vecteur d’expression de la galectine-7 dont la séquence codant pour le CRD a été muté. Nous avons établi que la fonction pro-apoptotique de la galectine-7 est indépendante de sa localisation mitochondriale et nucléaire et qu’il s’agit d’une fonction CRD-indépendante. À l’opposé, la réduction du pot

C-type lectins are Ca2+ dependent carbohydrate-recognition proteins that play crucial roles in the invertebrate innate immunity, such as nonself recognition, activation of proPO system, antibacterial activity, promotion of phagocytosis and nodule formation. In this study, a novel C-type lectin of bay scallops Argopecten irradians (Ai Lec) was identified using expressed sequence tag (EST) and RACE techniques. The Ai Lec cDNA encoded a polypeptide of 171 amino acids with a putative signal peptide of 21 amino acid residues and a mature protein of 150 amino acids. The deduced amino acid sequence of Ai Lec was highly similar to those of the C-type lectins from other animals and contained a typical carbohydrate-recognition domain (CRD) of 131 residues, which has four conserved disulfide-bonded cysteine residues that define the CRD and two additional cysteine residues at the amino terminus. The expression of Ai Lec transcript was dominantly detected in the hepatopancreas and slightly detected in the haemocytes of normal scallops. 6 h after Vibrio anguillarum-challenge and 8 h after Micrococcus luteus-challenge, the temporal expression of Ai Lec mRNA in hemocytes was increased by 4.4- and 3.6-folds, respectively. The results suggested that Ai Lec was a constitutive and inducible acute-phase protein and might be involved in immune response to Gram-negative and Gram-positive microbial infection in bay scallop A. irradians.

Full Text, Thesis (PhD Doctorate), Doctor of Philosophy (PhD), Institute for Glycomics, Dengue Fever virus (DENV) is a very old mosquito-borne flavivirus that has made a modern worldwide re-emergence as a result of population movement and growth, urbanisation and lapse of vector control. The World Health Organisation estimates that 2.5 billion people, or two-fifths of the world’s population are at risk from DENV, which can cause serious illness and in its severe forms, death. Despite the humanitarian and economic burden that DENV and its flaviviral relatives create, there are no chemotherapeutic drugs available and vaccine development is challenging. Mammalian host cell infection by DENV is mediated by the Envelope glycoprotein (EGP), which covers the entire exposed surface of the mature virus particle and is comprised of three exposed protein domains (DI, DII and DIII) and a transmembrane anchor. While significant effort has been invested to better understand how DIII of EGP participates in receptor mediated endocytosis of DENV into host cells, including the site and structure of the receptor binding site, or carbohydrate recognition domain (CRD), and the structure of ligands involved remain undefined. A recent study of mammalian cell surface glycans involved in DENV infection by Dr Kazuya Hidari and co-workers identified DENV inhibition by the glycolipid Paragloboside, which includes the tetrasaccharide Lacto-N-neotetraose (nLc4)1. This thesis reports an investigation of DENV-2 EGP DIII ligand specificity and characterisation of the DIII CRD involved in mammalian cell infection. To achieve this, soluble and high level expression of DENV-2 ThNH-7/93 EGP DIII was established from Pichia pastoris (P. pastoris) yeast and the recombinant DIII was successfully purified to near homogeneity by single step affinity chromatography. The biological activity of DIII was assessed by DENV permissible cell based assays and the recombinant protein was shown to have retained its wildtype host cell receptor binding activity. Recombinant DIII protein was utilised to suc

C-type lectins are Ca2+ dependent carbohydrate-recognition proteins that play crucial roles in the invertebrate innate immunity, such as nonself recognition, activation of proPO system, antibacterial activity, promotion of phagocytosis and nodule formation. In this study, a novel C-type lectin of bay scallops Argopecten irradians (Ai Lec) was identified using expressed sequence tag (EST) and RACE techniques. The Ai Lec cDNA encoded a polypeptide of 171 amino acids with a putative signal peptide of 21 amino acid residues and a mature protein of 150 amino acids. The deduced amino acid sequence of Ai Lec was highly similar to those of the C-type lectins from other animals and contained a typical carbohydrate-recognition domain (CRD) of 131 residues, which has four conserved disulfide-bonded cysteine residues that define the CRD and two additional cysteine residues at the amino terminus. The expression of Ai Lec transcript was dominantly detected in the hepatopancreas and slightly detected in the haemocytes of normal scallops. 6 h after Vibrio anguillarum-challenge and 8 h after Micrococcus luteus-challenge, the temporal expression of Ai Lec mRNA in hemocytes was increased by 4.4- and 3.6-folds, respectively. The results suggested that Ai Lec was a constitutive and inducible acute-phase protein and might be involved in immune response to Gram-negative and Gram-positive microbial infection in bay scallop A. irradians.

Dengue Fever virus (DENV) is a very old mosquito-borne flavivirus that has made a modern worldwide re-emergence as a result of population movement and growth, urbanisation and lapse of vector control. The World Health Organisation estimates that 2.5 billion people, or two-fifths of the world’s population are at risk from DENV, which can cause serious illness and in its severe forms, death. Despite the humanitarian and economic burden that DENV and its flaviviral relatives create, there are no chemotherapeutic drugs available and vaccine development is challenging. Mammalian host cell infection by DENV is mediated by the Envelope glycoprotein (EGP), which covers the entire exposed surface of the mature virus particle and is comprised of three exposed protein domains (DI, DII and DIII) and a transmembrane anchor. While significant effort has been invested to better understand how DIII of EGP participates in receptor mediated endocytosis of DENV into host cells, including the site and structure of the receptor binding site, or carbohydrate recognition domain (CRD), and the structure of ligands involved remain undefined. A recent study of mammalian cell surface glycans involved in DENV infection by Dr Kazuya Hidari and co-workers identified DENV inhibition by the glycolipid Paragloboside, which includes the tetrasaccharide Lacto-N-neotetraose (nLc4)1. This thesis reports an investigation of DENV-2 EGP DIII ligand specificity and characterisation of the DIII CRD involved in mammalian cell infection. To achieve this, soluble and high level expression of DENV-2 ThNH-7/93 EGP DIII was established from Pichia pastoris (P. pastoris) yeast and the recombinant DIII was successfully purified to near homogeneity by single step affinity chromatography. The biological activity of DIII was assessed by DENV permissible cell based assays and the recombinant protein was shown to have retained its wildtype host cell receptor binding activity. Recombinant DIII protein was utilised to suc, Thesis (PhD Doctorate), Doctor of Philosophy (PhD), Institute for Glycomics, Full Text

C-type lectins are Ca2+ dependent carbohydrate-recognition proteins that play crucial roles in the invertebrate innate immunity, such as nonself recognition, activation of proPO system, antibacterial activity, promotion of phagocytosis and nodule formation. In this study, a novel C-type lectin of bay scallops Argopecten irradians (Ai Lec) was identified using expressed sequence tag (EST) and RACE techniques. The Ai Lec cDNA encoded a polypeptide of 171 amino acids with a putative signal peptide of 21 amino acid residues and a mature protein of 150 amino acids. The deduced amino acid sequence of Ai Lec was highly similar to those of the C-type lectins from other animals and contained a typical carbohydrate-recognition domain (CRD) of 131 residues, which has four conserved disulfide-bonded cysteine residues that define the CRD and two additional cysteine residues at the amino terminus. The expression of Ai Lec transcript was dominantly detected in the hepatopancreas and slightly detected in the haemocytes of normal scallops. 6 h after Vibrio anguillarum-challenge and 8 h after Micrococcus luteus-challenge, the temporal expression of Ai Lec mRNA in hemocytes was increased by 4.4- and 3.6-folds, respectively. The results suggested that Ai Lec was a constitutive and inducible acute-phase protein and might be involved in immune response to Gram-negative and Gram-positive microbial infection in bay scallop A. irradians.

La gestion des résidus de construction, rénovation et démolition dans l’ensemble des MRC du Québec est fortement influencée par la mise en place de la Politique québécoise de la gestion des matières résiduelles du Québec 1998-2008. Celle-ci prévoyait la mise en valeur de 60 % des résidus de CRD de la province. Cette politique, jumelée à celle de la disparition progressive des Dépôts de matériaux secs prévue dans le Règlement sur l’élimination et l’incinération des matières résiduelles, aura eu des répercussions sur l’offre de service de la récupération. Sans oublier l’impact indéniable qu’elles ont eu sur les performances des infrastructures d’élimination, de récupération et de recyclage présentes dans les 17 régions administratives du Québec. Ces modifications du cadre législatif poussent les acteurs de ce milieu à trouver des solutions de rechange afin de diversifier leurs débouchés mais surtout d’assurer leur survie économique.

C-type lectins are Ca2+-dependent carbohydrate-recognition proteins that play crucial roles in innate immunity. The cDNA of C-type lectin (AiCTL1) in the bay scallop Argopecten irradians was cloned by expressed sequence tag (EST) and RACE techniques. The full-length cDNA of AiCTL1 was 660 bp, consisting of a T-terminal. untranslated region (UTR) of 30 bp and a 3' UTR of 132 bp with a polyadenylation signal sequence AATAAA and a poly(A) tail. The AiCTL1 cDNA encoded a polypeptide of 166 amino acids with a putative signal peptide of 20 amino acid residues and a mature protein of 146 amino acids. The deduced amino acid sequence of AiCTL1 was highly similar to those of the C-type lectins from other animals and contained a typical carbohydrate-recognition domain (CRD) of 121 residues, which has four conserved disulfide-bonded cysteine residues that define the CRD and two additional cysteine residues at the amino terminus. AiCTL1 mRNA was dominantly expressed in the hemocytes of the bay scallop. The temporal expression of AiCTL1 mRNA in hemocytes was increased by 5.7-and 4.9-fold at 6 h after injury and 8 h after injection of bacteria, respectively. The structural features, high similarity and expression pattern of AiCTL1 indicate that the gene may be involved in injury heating and the immune response in A. irradians. (c) 2008 Elsevier Ltd. All rights reserved.

C-type lectins are Ca2+-dependent carbohydrate-recognition proteins that play crucial roles in innate immunity. The cDNA of C-type lectin (AiCTL1) in the bay scallop Argopecten irradians was cloned by expressed sequence tag (EST) and RACE techniques. The full-length cDNA of AiCTL1 was 660 bp, consisting of a T-terminal. untranslated region (UTR) of 30 bp and a 3' UTR of 132 bp with a polyadenylation signal sequence AATAAA and a poly(A) tail. The AiCTL1 cDNA encoded a polypeptide of 166 amino acids with a putative signal peptide of 20 amino acid residues and a mature protein of 146 amino acids. The deduced amino acid sequence of AiCTL1 was highly similar to those of the C-type lectins from other animals and contained a typical carbohydrate-recognition domain (CRD) of 121 residues, which has four conserved disulfide-bonded cysteine residues that define the CRD and two additional cysteine residues at the amino terminus. AiCTL1 mRNA was dominantly expressed in the hemocytes of the bay scallop. The temporal expression of AiCTL1 mRNA in hemocytes was increased by 5.7-and 4.9-fold at 6 h after injury and 8 h after injection of bacteria, respectively. The structural features, high similarity and expression pattern of AiCTL1 indicate that the gene may be involved in injury heating and the immune response in A. irradians. (c) 2008 Elsevier Ltd. All rights reserved.

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant

Immunoglobulin E (IgE)-mediated allergy occurs when our immune system causes a reaction to otherwise harmless substances (allergens). Allergens are predominantly proteins present in biological materials such as pollens, mites, animal epithelia, moulds and foods. In vitro tests for specific IgE antibodies usually employ an allergen source extract as an antibody capturing reagent. The proportion of allergenic molecules in these biochemically complex extracts may vary. Recombinant allergens may be obtained in large quantities with biotechnological techniques. These proteins can be characterized biochemically and immunologically, resulting in tests with minimal batch-to-batch variation. This thesis describes different uses of recombinant allergens in component-resolved diagnostics (CRD). In CRD, single allergenic proteins are used to establish a sensitization profile of the patient. Two timothy grass (Phleum pratense) pollen allergens, Phl p 11 and Phl p 4, were cloned and expressed as recombinant proteins. They were subsequently characterized and can, for example, be used in a panel for grass pollen CRD. Single allergens may be useful as diagnostic markers for allergic sensitization. This phenomenon was studied using tropomyosin, a major allergen from the shrimp Penaeus aztecus (Pen a 1). The characteristics of the recombinant and natural proteins were compared. The recombinant tropomyosin was then extensively tested using specific competition for IgE binding against extracts of other crustacean species, house dust mite and cockroach. In cases when an important allergen is missing or underrepresented in a natural extract, the corresponding recombinant allergen may be added to the extract as a spiking reagent. Previous studies have shown that latex extracts for diagnostic testing may lack the allergen Hev b 5. Recombinant Hev b 5 was expressed from a synthetic gene construct, incorporating several adaptations to enable efficient large scale production of the recombinant

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The ABCA4 gene has been involved in several forms of inherited macular dystrophy. In order to further characterize the complex genotype–phenotype relationships involving this gene, we have performed a mutation analysis of ABCA4 in 14 Spanish patients comprising eight STGD (Stargardt), four FFM (fundus flavimaculatus), and two CRD (Cone-rod dystrophy) patients. SSCP (single-strand conformation polymorphism) analysis and DNA sequencing of the coding and 5′ upstream regions of this gene allowed the identification of 16 putatively pathogenic alterations, nine of which are novel. Most of these were missense changes, and no patient was found to carry two null alleles. Overall, the new data agree with a working model relating the different pathogenic phenotypes to the severity of the mutations. When considering the information presented here together with that of previous reports, a picture of the geographic distribution of three particular mutations emerges. The R212C change has been found in French, Italian, Dutch, German, and Spanish but not in British patients. In the Spanish collection, R212C was found in a CRD patient, indicating that it may be a rather severe change. In contrast, c.2588G>C, a very common mild allele in the Dutch population, is rarely found in Southern Europe. Interestingly, the c.2588G>C mutation has been found in a double mutant allele together with the missense R1055W. Finally, the newly described L1940P was found in two unrelated Spanish patients, and may be a moderate to severe allele.