Latin 14807

Contient : I Questiones super epistolas Pauli ; Questiones theologice (99) ; Objectiones contra eos qui dicunt quod Christus non est aliquid secundum quod est homo (115) ; II Isidori Soliloquia (125) ; Bonaventure itinerarium mentis (155) ; S. Augustini manuale (176v) ; De conflictu virtutum et vitiorum secundum B. (186v)

ALBUM de Caspar Held, étudiant en théologie à Tübingen (1564). In-8 de 106 f., mar. bl. jans. (Hans Asper.)

Cranach (Lucas). Passional, cité

Recueil de lettres et de pièces originales, et de copies de pièces indiquées comme telles dans le dépouillement qui suit.

Contient : 1 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le mareschal de Montmorency,... Escript à Sainct Germain en Laye, le XXVIIe jour de jung » ; 2 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le sire de Montmorency, connestable de France... Escript à Bourg en Bresse, le IIe jour d'octobre, l'an M.V.C.XXI » ; 3 Lettre de « LEONOR [D'AUTRICHE]... à messrs les grand maistre de France et arcevesque de Bourges... De Victorie, le XXIIIIe d'avril » ; 4 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le mareschal de Chabannes,... Escript à Bloys, le XIIIIme jour de may » ; 5 Lettre de « LOYSE [DE SAVOIE]... à monseigneur de Montmorency,... Escript à Paris, le IIIIme jour d'octobre » ; 6 Lettre de « RENEE DE FRANCE [duchesse de Ferrare]... à la royne ma dame et seur » ; 7 Lettre de « MARGUERITE [D'AUTRICHE]... à mon cousin le grant maistre de France... De Malines, ce XXVIIIe de juillet anno [M.D.]XXX » ; 8 Lettre de « DE LALAING,... Sr D'OCTRASTE,... à... monseigneur le grant maistre... De Malines, le penultiesme jour de juillet, l'an [M.D.]XXX » ; 9 Lettre de « CLAUDE [DE LORRAINE, duc] DE GUYSE,... à monseigneur le grant maistre... D'Esclarron, le XXIX jour d'apvril » ; 10 Lettre de « LOYSE [DE SAVOIE]... à monseigneur d'Alby,... Escript à Sainct Germain en Laye, ce XXIXe jour d'aoust » ; 11 Lettre de « l'admiral [PHILIPPES CHABOT, Sr DE] BRYON,... à monseigneur... le grant maistre... Escript à Angoulesme, le XXVe jour de may mil.V.C.XXX » ; 12 Lettre de « GABRIEL DE GRAMONT, evesque de Tarbe... à monseigneur... le grant maistre et mareschal de France... A Boullongne, le XXXe jour de mars, l'an mil cinq cens et trente » ; 13 Lettre, en italien, du « marquis DE MENTOUE », FREDERIC II, à « monseigneur [de Montmore]nsi, marescal [de Fran]cia... Mantue, ultimo maii M.D.XXV » ; 14 Lettre d'«ANTHOYNE DE LORRAINE,... à monseigneur le grant maistre... Escript à Luneville, le XXVIe jour de novembre » ; 15 Lettre, en italien, d'HERCULE D'EST, fils du duc de Ferrare, « duca DE CHARTRES,... à monseigneur lo gran maestro... Datum in Ferrariis, a XV de giugno 1529 » ; 16 « La Capitulation qui a esté faicte avec mon seigneur le conte de Roguendolff, chevalier de l'ordre du roy, pour la levée qu'il va faire presentement en Allemaigne... Du VIIIme jour d'avril 1562, apres Pasques ». Copie ; 17 Lettre du roi « FRANÇOYS [Ier]... au cappitaine Bouzy,... Escript à St Germain en Laye, le IXme jour de juing » ; 18 Lettre du roi « FRANÇOYS [Ier]... à monseigneur Du Fresnoy, cappitaine de Therouanne... Escript à Sainct Germain en Laye, le IXe jour de juing » ; 19 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le mareschal de Montmorancy,... Escript à Sainct Germain en Laye, le XXIIe jour d'avril » ; 20 Lettre du roi « FRANÇOYS [Ier]... à Labarre... Escript à Sainct Germain en Laye, le IXme jour de juing » ; 21 Lettre du roi « FRANÇOYS [Ier]... à monseigneur de Torcy,... Escript à Sainct Germain en Laye, le IXme jour de juing » ; 22 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le grant maistre... Escript à St Germain en Laye, ce XXVme jour de mars » ; 23 Lettre du roi « FRANÇOYS [Ier]... à mon cousin le mareschal de Monmorancy,... Escript à Lyon, ce VIe jour de septembre » ; 24 Lettre de « P. DE VILLERS L'YLE ADAM,... maistre de Rhodes... à monseigneur le mareschal de Montmorency, mon nepveu... De Cyvette Vielhe... aoust 1523 » ; 25 Lettre, en italien, du « cardinale CORNELIO,... al' illustrissimo signor... gran mastro... Da Venetia, alli XIIII di giugno M.D.XXVIII° » ; 26 Lettre de FRANÇOIS DE MONTMORENCI, Sr DE « LA ROCHEPOT,... à... monseigneur le grant maistre... De Mello, se Vme jour de mars » ; 27 Lettre d'«ANDREA DORIA,... à monseigneur le grant maistre... De Gennes, ce IIIIe mars » ; 28 Lettre, en italien, de « lo capitaneo GIOVANNI LOISE, Albanese, homo del Sor conte Guido Rangone,... allo illustrissimo... monsegnor grant mestre... Da Paris, alli VI decembre 1536 » ; 29 Lettre de « MONTMORENCY,... à monseigneur de Humyeres, chevalier de l'ordre et lieutenant general du roy en Piemont... De Coucy, le XXIIIe jour de may » ; 30 Lettre de « CLAUDE [DE LORRAINE, duc] DE GUYSE,... à monseigneur le grant maistre... Du camp devant Beaulieu, le IIIIe d'aoust » ; 31 Lettre de « BOUNYVET,... à monseigneur de La Rochepot,... A Lyon, le XXIIe jour de juillet » ; 32 « Double du traicté faict entre messrs les mareschal de Montejehan et marquis del Gouast touchant la tresve... Faict à Montcallier, le XXIme jour de febvrier, l'an mil V.C.XXXVII » ; 33 Lettre, en italien, d'«HERCULES ESTENSIS, [duc] DE CHARTRES,... à monseigneur lo gran maestro... De Alessandria, a ultimo octobre 1528 » ; 34 Lettre, en italien, d'«HANNIBAL GONZAGA,... al' illustrissimo... monsignor gran maestro... Da Chero, lo XVI di feb° M.D.XXXVII » ; 35 Lettre, en italien, de « FR., episcopus laudensis... al mio carissimo fratello il S. Gaspar Sormano,... Murani, 29 7bre 1529 » ; 36 Lettre, en italien, de « GIOVANNE CLEMENTE STANGHA,... al 'illustrissimo gran mastro de Francia et marchial de Memoransi,... Datum in Barletta, a di VIII februarii 1529 » ; 37 Lettre de RENE, « batar de Savoye... à monseigneur le mareschal de Montmorency,... De Lyon, ce XIme de novembre » ; 38 Lettre, en italien, de « JOACHIN [DE VAUX]... al' illustrissimo... Sor gran maestro de Francia... Da Londra, XXII marcio M.D.XXX » ; 39 Lettre d'«HUMYERES,... à monseigneur de Montmorancy, grant maistre et mareschal de France... De Mouchy, ce IIe jour de janvier » ; 40 Lettre de « CLAUDE DE TANDE,... à monseigneur le grant moestre... De Marseille, ce XXme jour d'avril » ; 41 Lettre de « GEORGE D'ARMAGNAC, evesque de Roudez... à monseigneur... de Humieres, lieutenant general du roy en Piedmont... A Venise, le derrenyer de juillet 1537 » ; 42 Lettre de « l'admiral [PHILIPPE CHABOT, Sr DE] BRYON,... à monseigneur... le grant maistre... Escript à Lezay, le XIXe jour d'avril mil V.C.XXX » ; 43 Lettre, en italien, de « J. FREGOSO,... al' illustrissimo... monsignore il gran meistro... In Lione, l'ultimo di juglio M.D.XXVI » ; 44 Lettre, en italien, de « GUIDO RANGONE,... Di Venetia, alli XXIII de martio 1530 » ; 45 Lettre, en italien, d'«ANTONIO DORIA,... allo illustrissimo... monsor il gran mestro... Di galera nel porto de Bachone, a di XIII di agosto de M.D.XXIII » ; 46 Lettre, en italien, de « DE CARPI,... au roy [François Ier]... De Rome, ce XXIXe jour de novembre M.V.C.XXVI » ; 47 Lettre de « MONTMORENCY,... à monseigneur mon cousin, monseigneur le grant maistre... De Montingny, ce VIIIe de janvier XV.C.XXVII » ; 48 Lettre, en italien, de « R. PIO CL. DI FAENZA,... al' illustrissimo... signor... grammastro di Francia... Da Roma, a li XXVII di genaro del M.D.XXX » ; 49 Lettre, en italien, de « GREGORIO CASALE,... allo illustrissimo... mons. lo gran mastro di Francia... In Bologna, alli 12 di marzo 1530 » ; 50 Lettre de « MONTMORENCY,... à monseigneur de La Poumeraye, premier president des comptes en Bretaigne... De Chantilly, le XIIIIe jour de mars 1541 » ; 51 Lettre, en italien, de « JOACHIN [DE VAUX]... al' illustrissimo... Sr gran maestro de Francia... Da Ferrara, el XIII di marzo M.D.XXVIIII » ; 52 Extrait d'un « estat de la maison de monseigneur le duc d'Angoulesme et de mesdames ses seurs... faict le XVme jour de mars, l'an mil cinq cens vingt neuf » ; 53 Lettre, en italien, de « GUIDO RANGONE,... al' illustrissimo... signor... monsignor gran maestro... Di Sorges, l'ultimo di novembre 1536 » ; 54 Lettre, en espagnol, adressée « al muy illustre seññor... gran maestre... De Perpiñan, a XVIIe de dezembre de 1537 » ; 55 Lettre de « PHILIBERTE DE LUXAMBOURG [princesse D'ORANGE]... au roy [François Ier]... De Lons le Saulnier, le XIIIIe jour de decembre 1530 » ; 56 Lettre, en italien, de « CARGUIN DE BOZELO,... all' illustrissimo... Sr de Momoransi,... marscial di Franza... De Pinaruolo, alli due de novembre M.D.XXXVI » ; 57 Lettre, en italien, de « J. FREGOSO,... allo illustrissimo... monsre il gran meistro... In Lione, il di 25 de juglio M.D.XXVI » ; 58 Lettre, en italien, de « MANFREDO DA CORREGIO,... al' illustrissimo... signore Momoransi,... De Corregio, alli III de ginaro 1525 » ; 59 Lettre, en italien, de « J. FREGOSO,... allo illustrissimo... signore il gran meistro... In Marseglia, l'ottavo di augosto M.D.XXVI » ; 60 Lettre, en italien, de « GASPAR SORMANO,... al' illustrissimo... el signor gran maestro di Francia... Da Ferrara, alli 14 di maggio 1529 » ; 61 Lettre, en italien, de « GUIDO RANGONE,... al' illustrissimo... monsignore il gran maestro di Franza... Di Piacenza, il giorno XXV di ottobrio M.D.XXVII » ; 62 Lettre, en latin, des « balivus et consules terre Vallesii... illustrissimo domino magno magistro Francie... Die XXVI mensis octobris M.° V.C.XXVI » ; 63 Lettre, en italien, de « GALEATIO VESCONTE,... allo illustrissimo monsegnor el maestre de Franza... De Lione, a li 20 de julio 1530 » ; 64 Lettre, avec chiffre, de « CLAU [DE] DODIEU DE VELLY,... à monseigneur... le grand maistre... A Fleurence, ce XXVIIme jour de decembre M.V.C.XXVIII » ; 65 Lettre, en italien, d'«HANIBALLE GONZAGA, conte... allo illustrissimo... monseigneur grammaestro di Franza... Da Novelara, alli XIIII di juglio M.D.XXVIII » ; 66 Lettre, en italien, d'«ANTONIO RINCON,... a monsignor de Momoranci, gran maestre di Franza... Da Casxovia, XXII de settembre 1527 » ; 67 Lettre, en italien, d'«ANDREA DORIA,... allo illustrissimo... monseigneur le gran maestro di Fransa... Di Genoa, alli XXI di agosto M.D.XXVII » ; 68 Lettre de « LOYS DE PRAET,... à monseigneur... le grant maistre de France... De Fontarrabye, le XIXe de may » ; 69 Lettre, en latin, des « scultetus et consules urbis Bernensis... domino de Memorence, generali locumtenenti et capitaneo regio... Ex urbe nostra Bernensi, quinta februarii, anno etc. XXIIII° » ; 70 Lettre, en italien, d'«el vescovo di Como... allo illustrissimo... monsignore il gran maestro... Di Angolemo, alli XII di maggio M.D.XXX »

Biotransformations of water insoluble substrates in aqueous, two-phase and encapsulated systems /

The biotransformation of water insoluble substrates by mammalian and bacterial cells has been problematic, since these whole cell reactions are primarily performed in an aqueous environment The implementation of a twophase or encapsulated system has the advantages of providing a low water system along with the physiological environment the cells require to sustain themselves. Encapsulation of mammalian cells by formation of polyamide capsules via interfacial polymerization illustrated that the cells could not survive this type of encapsulation process. Biotransformation of the steroid spironolactone [3] by human kidney carcinoma cells was performed in a substrate-encapsulated system, yielding canrenone [4] in 70% yield. Encapsulation of nitrile-metabolizing Rhodococcus rhodochrous cells using a polyamide membrane yielded leaky capsules, but biotransformation of 2-(4- chlorophenyl)-3-methylbutyronitrile (CPIN) [6] in a free cell system yielded CPIN amide [7] in 40% yield and 94% ee. A two-phase biotransformation of CPIN consisting of a 5:1 ratio of tris buffer, pH 7.2 to octane respectively, gave CPIN acid [8] in 30% yield and 97% ee. It was concluded that Rhodococcus rhodochrous ATCC 17895 contained a nonselective nitrile hydratase and a highly selective amidase enzyme.

The backwoods of our backyards : Ontario pioneers and the crisis of ecology /

Early in his landmark ecocritical book The Comedy of Survival, Joseph Meeker develops an intriguing hypothesis about human behaviour. He remarks the species Homo sapiens tend to behave like an invasive or pioneering organism, entering a bio-geographical region and aggressively outcompeting all other species for space and resources. Moreover, he suggests, human cultural traditions, at least in the West, have reinforced such behaviour, continually insisting that the impulses he describes are both necessary and right. While Meeker's work goes on to assess a number of literary works in both the tragic and comic modes, his work never fully explores this hypothesis in the context of human pioneers; that is, there is no ~xploration o( how these themes manifest themselves within our culture and what role they might play in the culture of specific pioneering groups. This project is an attempt at just such an analysis, examining the validity of Meeker's hypothesis through a case study of settler literature in Upper Canada/Ontario between the . years 1800-1867. It explores Meeker's work within three main areas: first, Chapter Two situates his book historically within the field of ecocriticism, showing what came before and the explosion of ecocritical inquiry that followed its release. This chapter also delves into the rift between the natural sciences and humanities, arguing that a move towards deeper interdisciplinarity is r:tecessary for the future. Chapter Three examines the biological and ecological ground on which Meeker rests his hypothesis through exploring evolutionary biology as well as invasive and pioneer species behaviour. Lastly, Chapter Four examines how these ecological principles are manifested in the writings of early Canadian settlers, suggesting that Meeker's hypothesis indeed finds itself on stable footing.

The role of water in determining structure and function of macromolecules and macromolecular assemblies /

The interaction of biological molecules with water is an important determinant of structural properties both in molecular assemblies, and in conformation of individual macromolecules. By observing the effects of manipulating the activity of water (which can be accomplished by limiting its concentration or by adding additional solutes, "osmotic stress"), one can learn something about intrinsic physical properties of biological molecules as well as measure an energetic contribution of closely associated water molecules to overall equilibria in biological reactions. Here two such studies are reported. The first of these examines several species of lysolipid which, while present in relatively low concentrations in biomembranes, have been shown to affect many cellular processes involving membrane-protein or membrane-membrane interactions. Monolayer elastic constants were determined by combining X-ray diffraction and the osmotic stress technique. Spontaneous radii of curvature of lysophosphatidylcholines were determined to be positive and in the range +30A to +70A, while lysophosphatidylethanolamines proved to be essentially flat. Neither lysolipid significantly affected the bending modulus of the monolayer in which it was incorporated. The second study examines the role of water in theprocess of polymerization of actin into filaments. Water activity was manipulated by adding osmolytes and the effect on the equilibrium dissociation constant (measured as the criticalmonomer concentration) was determined. As water activity was decreased, the critical concentration was reduced for Ca-actin but not for Mg-actin, suggesting that 10-12 fewer water molecules are associated with Ca-actin in the polymerized state. Thisunexpectedly small amount of water is discussed in the context of the common structural motif of a nucleotide binding cleft.

An investigation into the effects of homopteran honeydew sugars versus floral nectar sugars on black fly longevity, flight performance and digestion /

Floral nectar is thought to be the primary carbohydrate source for most dipteran species. However, it has been shown that black flies (Burgin & Hunter 1997 a,b,c), mosquitoes (Foster 1995; Burkett et al. 1999; Russell & Hunter 2002), deer flies (Magnarelli & Burger 1984; Janzen & Hunter 1998; Ossowski & Hunter 2000), horse flies (Schutz & Gaugler 1989; Hunter & Ossowski 1999) and sand flies (MacVicker et al. 1990; Wallbanks et al. 1990; Cameron et al. 1992, 1995; Schlein & Jacobson 1994, 1999; Hamilton & EI Naiem 2000) feed on homopteran honeydew as well as floral nectar. Prior to 1997 floral nectar was thought to be the main source of carbohydrates for black flies. However, Burgin & Hunter (1 997a) demonstrated that up to 35% of black flies had recently consumed meals of homo pte ran honeydew. This information has necessitated a re-assessment of many life history aspects of black flies. Attempts are being made to examine the effects of nectar versus honeydew on black fly fecundity and parasite transmission (Hazzard 2003). Recently, Stanfield and Hunter (unpublished data) have shown that in female black flies, honeydew sugars produce flights of longer distance and duration than do nectar sugars. This thesis examines two aspects of black fly biology as it relates to sugar meal consumption. First, the effects of honeydew and nectar on black fly longevity are examined. Second, the proximate causation behind longer flight performances in honeydew-fed flies will be examined. The comparison between these two sources is important because nectar is composed of mainly simple sugars (monosaccharides and disaccharides) whereas honeydew is composed of both simple and complex sugars (including trisaccharides and tetrasaccharides ).

Spin labile conducting metallopolymers : a new architecture for hybrid multifunctional materials

The synthesis of 3-ethynylthienyl- (2.07), 3-ethynylterthienyl- (2.19) substituted qsal [qsalH = N-(8-quinolyl)salicylaldimine] and 3,3' -diethynyl-2,2' -bithienyl bridging bisqsal (5.06) ligands are described along with the preparation and characterization of eight cationic iron(III) complexes containing these ligands with a selection of counteranions [(2.07) with: SCN- (2.08), PF6- (2.09), and CI04- (2.10); (2.19) with PF6 - (2.20); (5.06) with: cr (5.07), SeN- (5.08), PF6- (5.09), and CI04- (5.10)]. Spin-crossover is observed in the solid state for (2.08) - (2.10) and (5.07) - (5.10), including a ve ry rare S = 5/2 to 3/2 spin-crossover in complex (2.09). The unusal reduction of complex (2.10) produces a high-spin iron(I1) complex (2.12). Six iron(II) complexes that are derived from thienyl analogues of bispicen [bispicen = bis(2-pyridylmethyl)-diamine] [2,5-thienyl substituents = H- (3.11), Phenyl- (3.12), 2- thienyl (3.13) or N-phenyl-2-pyridinalimine ligands [2,5-phenyl substituents = diphenyl (3.23), di(2-thienyl) (3.24), 4-phenyl substituent = 3-thienyl (3.25)] are reported Complexes (3.11), (3.23) and (3.25) display thermal spin-crossover in the solid state and (3.12) remains high-spin at all temperatures. Complex (3.13) rearranges to form an iron(II) complex (3.14) with temperature dependent magnetic properties be s t described as a one-dimensional ferromagnetic chain, with interchain antiferromagnetic interactions and/or ZFS dominant at low temperatures. Magnetic succeptibility and Mossbauer data for complex (3.24) display a temperature dependent mixture of spin isomers. The preparation and characterization of two cobalt(II) complexes containing 3- ethynylthienyl- (4.04) and 3-ethynylterhienyl- (4.06) substituted bipyridine ligands [(4.05): [Co(dbsqh(4.04)]; (4.07): [Co(dbsq)2(4.06)]] [dbsq = 3,5-dbsq=3,5-di-tert-butylI ,2-semiquinonate] are reported. Complexes (4.05) and (4.07) exhibit thermal valence tautomerism in the solid state and in solution. Self assembly of complex (2.10) into polymeric spheres (6.11) afforded the first spincrossover, polydisperse, micro- to nanoscale material of its kind. . Complexes (2.20), (3.24) and (4.07) also form polymers through electrochemical synthesis to produce hybrid metaUopolymer films (6.12), (6.15) and (6.16), respectively. The films have been characterized by EDX, FT-IR and UV-Vis spectroscopy. Variable-temperature magnetic susceptibility measurements demonstrate that spin lability is operative in the polymers and conductivity measurements confirm the electron transport properties. Polymer (6.15) has a persistent oxidized state that shows a significant decrease in electrical resistance.

(A) Optimization of solid body phase synthesis of RNA using the Cpep chemistry. (B) Synthesis of BODIPY labeled oligonucleotides

(A) Solid phase synthesis of oligonucleotides are well documented and are extensively studied as the demands continue to rise with the development of antisense, anti-gene, RNA interference, and aptamers. Although synthesis of RNA sequences faces many challenges, most notably the choice of the 2' -hydroxy protecting group, modified 2' -O-Cpep protected ribonucleotides were synthesized as alternitive building blocks. Altering phosphitylation procedures to incorporate 3' -N,N-diethyl phosphoramidites enhanced the overall reactivity, thus, increased the coupling efficiency without loss of integrety. Furthermore, technical optimizations of solid phase synthesis cycles were carried out to allow for successful synthesis of a homo UIO sequences with a stepwise coupling efficiency reaching 99% and a final yield of 91 %. (B) Over the past few decades, dipyrrometheneboron difluoride (BODIPY) has gained recognition as one of the most versatile fluorophores. Currently, BODIPY labeling of oligonucleotides are carried out post-synthetically and to date, there lacks a method that allows for direct incorporation of BODIPY into oligonucleotides during solid phase synthesis. Therefore, synthesis of BODIPY derived phosphoramidites will provide an alternative method in obtaining fluorescently labelled oligonucleotides. A method for the synthesis and incorporation of the BODIPY analogues into oligonucleotides by phosphoramidite chemistry-based solid phase DNA synthesis is reported here. Using this approach, BODIPY-labeled TlO homopolymer and ISIS 5132 were successfully synthesized.

Alternative splicing of DNA polymerase beta in vertebrates

Alternative splicing (AS) is the predominant mechanism responsible for increasing eukaryotic transcriptome and proteome complexity. In this phenomenon, numerous mRNA transcripts are produced from a single pre-mRNA sequence. AS is reported to occur in 95% of human multi-exon genes; one specific gene that undergoes AS is DNA polymerase beta (POLB). POLB is the main DNA repair gene which performs short patch base excision repair (BER). In primate untransformed primary fibroblast cell lines, it was determined that the splice variant (SV) frequency of POLB correlates positively with species lifespan. To date, AS patterns of POLB have only been examined in mammals primarily through the use of cell lines. However, little attention has been devoted to investigating if such a relationship exists in non-mammals and whether cell lines reflect what is observed in vertebrate tissues. This idea was explored through cloning and characterization of 1,214 POLB transcripts from four non-mammalian species (Gallus gallus domesticus, Larus glaucescens, Xenopus laevis, and Pogona vitticeps) and two mammalian species (Sylvilagus floridanus and Homo sapiens) in two tissue types, liver and brain. POLB SV frequency occurred at low frequencies, < 3.2%, in non-mammalian tissues relative to mammalian (>20%). The highest POLB SV frequency was found in H. sapiens liver and brain tissues, occurring at 65.4% and 91.7%, respectively. Tissue specific AS of POLB was observed in L. glaucescens, P. vitticeps, and H. sapiens, but not G. gallus domesticus, X. laevis and S. floridanus.The AS patterns of a second gene, transient receptor potential cation channel subfamily V member 1 (TRPV1), were compared to those of POLB in liver and brain tissues of G. gallus domesticus, X. laevis and H. sapiens. This comparison was performed to investigate if any changes (either increase or decrease) observed in the AS of POLB were gene specific or if they were tissue specific, in which case similar changes in AS would be seen in POLB and TRPV1. Analysis did not reveal an increase or decrease in both the AS of POLB and TRPV1 in either the liver or brain tissues of G. gallus domesticus and H. sapiens. This result suggested that the AS patterns of POLB were not influenced by tissue specific rates of AS. Interestingly, an increase in the AS of both genes was only observed in X. laevis brain tissue. This result suggests that AS in general may be increased in the X. laevis brain as compared to liver tissue. No positive correlation between POLB SV frequency and species lifespan was found in non-mammalian tissues. The AS patterns of POLB in human primary untransformed fibroblast cell lines were representative of those seen in human liver tissue but not in brain tissue. Altogether, the AS patterns of POLB from vertebrate tissues and primate cell lines revealed a positive correlation between POLB SV frequency and lifespan in mammals, but not in non-mammals. It appears that this positive correlation does not exist in vertebrate species as a whole.

The identification and characterization of inter- and intra-species genetic diversity derived from retrotransposons in humans

Retrotransposons, which used to be considered as “junk DNA”, have begun to reveal their immense value to genome evolution and human biology due to recent studies. They consist of at least ~45% of the human genome and are more or less the same in other mammalian genomes. Retrotransposon elements (REs) are known to affect the human genome through many different mechanisms, such as generating insertion mutations, genomic instability, and alteration in gene expression. Previous studies have suggested several RE subfamilies, such as Alu, L1, SVA and LTR, are currently active in the human genome, and they are an important source of genetic diversity between human and other primates, as well as among humans. Although several groups had used Retrotransposon Insertion Polymorphisms (RIPs) as markers in studying primate evolutionary history, no study specifically focused on identifying Human-Specific Retrotransposon Element (HS-RE) and their roles in human genome evolution. In this study, by computationally comparing the human genome to 4 primate genomes, we identified a total of 18,860 HS-REs, among which are 11,664 Alus, 4,887 L1s, 1,526 SVAs and 783 LTRs (222 full length entries), representing the largest and most comprehensive list of HS-REs generated to date. Together, these HS-REs contributed a total of 14.2Mb sequence increase from the inserted REs and Target Site Duplications (TSDs), 71.6Kb increase from transductions, and 268.2 Kb sequence deletion of from insertion-mediated deletion, leading to a net increase of ~14 Mb sequences to the human genome. Furthermore, we observed for the first time that Y chromosome might be a hot target for new retrotransposon insertions in general and particularly for LTRs. The data also allowed for the first time the survey of frequency of TE insertions inside other TEs in comparison with TE insertion into none-TE regions. In summary, our data suggest that retrotransposon elements have played a significant role in the evolution of Homo sapiens.

ORGANOSILICON BIOTECHNOLOGY: A BIO-INSPIRED APPROACH TO THE HYDROLYSIS OF ALKOXYSILANES and THE LIPASE-CATALYZED SYNTHESIS OF SILOXANE-CONTAINING POLYESTERS AND POLYAMIDES

The first part of this thesis studied the capacity of amino acids and enzymes to catalyze the hydrolysis and condensation of tetraethoxysilane and phenyltrimethoxysilane. Selected amino acids were shown to accelerate the hydrolysis and condensation of tetraethoxysilane under ambient temperature, pressure and at neutral pH (pH 7±0.02). The nature of the side chain of the amino acid was important in promoting hydrolysis and condensation. Several proteases were shown to have a capacity to hydrolyze tri- and tet-ra- alkoxysilanes under the same mild reaction conditions. The second part of this thesis employed an immobilized Candida antarctica lipase B (Novozym-435, N435) to produce siloxane-containing polyesters, polyamides, and polyester amides under solvent-free conditions. Enzymatic activity was shown to be temperature dependent, increasing until enzyme denaturation became the dominant pro-cess, which typically occurred between 120-130ᵒC. The residual activity of N435 was, on average, greater than 90%, when used in the synthesis of disiloxane-containing polyesters, regardless of the polymerization temperature except at the very highest temperatures, 140-150ᵒC. A study of the thermal tolerance of N435 determined that, over ten reaction cycles, there was a decrease in the initial rate of polymerization with each consecutive use of the catalyst. No change in the degree of monomer conversion after a 24 hour reaction cycle was found.

Cyclotetrasiloxane Frameworks for the Chemoenzymatic Synthesis of Oligoesters

Immobilized lipase B from Candida antarctica (Novozym® 435, N435) was utilized as part of a chemoenzymatic strategy for the synthesis of branched polyesters based on a cyclotetrasiloxane core in the absence of solvent. Nuclear magnetic resonance spectroscopy and matrix-assisted laser desorption ionization time-of-flight mass spectrometry were utilized to monitor the reactions between tetraester cyclotetrasiloxanes and aliphatic diols. The enzyme-mediated esterification reactions can achieve 65– 80% consumption of starting materials in 24–48 h. Longer reaction times, 72–96 h, resulted in the formation of cross-linked gel-like networks. Gel permeation chromatography of the polymers indicated that the masses were Mw ¼ 11 400, 13 100, and 19 400 g mol 1 for the substrate pairs of C7D4 ester/ octane-1,8-diol, C10D4 ester/pentane-1,5-diol and C10D4 ester/octane-1,8-diol respectively, after 48 h. Extending the polymerization for an additional 24 h with the C10D4 ester/octane-1,8-diol pair gave Mw ¼ 86 800 g mol 1. To the best of our knowledge this represents the first report using lipase catalysis to produce branched polymers that are built from a cyclotetrasiloxane core.

Interactions protéiques et relation dynamique entre phosphorylation / sumoylation / ubiquitination des protéines TIF1α, β et PML: détection in vivo par BRET

Trois protéines de la famille TRIM (Motif TRIpartite), TIF1α, β (Transcriptional Intermediary Factor 1) et PML (ProMyelocytic Leukaemia¬), font l’objet de cette étude. TIF1α est connu comme un coactivateur des récepteurs nucléaires et TIF1β comme le corépresseur universel des protéines KRAB-multidoigt de zinc dont le prototype étudié ici est ZNF74. PML possède divers rôles dont le plus caractérisé est celui d’être l’organisateur principal et essentiel des PML-NBs (PML-Nuclear Bodies), des macrostructures nucléaires très dynamiques regroupant et coordonnant plus de 40 protéines. Il est à noter que la fonction de TIF1α, β et PML est régulée par une modification post-traductionnelle, la sumoylation, qui implique le couplage covalent de la petite protéine SUMO (Small Ubiquitin like MOdifier) à des lysines de ces trois protéines cibles. Cette thèse propose de développer des méthodes utilisant le BRET (Bioluminescence Resonance Energy Transfert) afin de détecter dans des cellules vivantes et en temps réel des interactions non-covalentes de protéines nucléaires mais aussi leur couplage covalent à SUMO. En effet, le BRET n’a jamais été exploré jusqu’alors pour étudier les interactions non-covalentes et covalentes de protéines nucléaires. L’étude de l’interaction de protéines transcriptionnellement actives est parfois difficile par des méthodes classiques du fait de leur grande propension à agréger (famille TRIM) ou de leur association à la matrice nucléaire (ZNF74). L’homo et l’hétérodimérisation de TIF1α, β ainsi que leur interaction avec ZNF74 sont ici testées sur des protéines entières dans des cellules vivantes de mammifères répondant aux résultats conflictuels de la littérature et démontrant que le BRET peut être avantageusement utilisé comme alternative aux essais plus classiques basés sur la transcription. Du fait de l’hétérodimérisation confirmée de TIF1α et β, le premier article présenté ouvre la possibilité d’une relation étroite entre les récepteurs nucléaires et les protéines KRAB- multidoigt de zinc. Des études précédentes ont démontré que la sumoylation de PML est impliquée dans sa dégradation induite par l’As2O3 et dépendante de RNF4, une E3 ubiquitine ligase ayant pour substrat des chaînes de SUMO (polySUMO). Dans le second article, grâce au développement d’une nouvelle application du BRET pour la détection d’interactions covalentes et non-covalentes avec SUMO (BRETSUMO), nous établissons un nouveau lien entre la sumoylation de PML et sa dégradation. Nous confirmons que le recrutement de RNF4 dépend de SUMO mais démontrons également l’implication du SBD (Sumo Binding Domain) de PML dans sa dégradation induite par l’As2O3 et/ou RNF4. De plus, nous démontrons que des sérines, au sein du SBD de PML, qui sont connues comme des cibles de phosphorylation par la voie de la kinase CK2, régulent les interactions non-covalentes de ce SBD mettant en évidence, pour la première fois, que les interactions avec un SBD peuvent dépendre d’un évènement de phosphorylation (“SBD phospho-switch”). Nos résultats nous amènent à proposer l’hypothèse que le recrutement de PML sumoylé au niveau des PML-NBs via son SBD, favorise le recrutement d’une autre activité E3 ubiquitine ligase, outre celle de RNF4, PML étant lui-même un potentiel candidat. Ceci suggère l’existence d’une nouvelle relation dynamique entre phosphorylation, sumoylation et ubiquitination de PML. Finalement, il est suggéré que PML est dégradé par deux voies différentes dépendantes de l’ubiquitine et du protéasome; la voie de CK2 et la voie de RNF4. Enfin une étude sur la sumoylation de TIF1β est également présentée en annexe. Cette étude caractérise les 6 lysines cibles de SUMO sur TIF1β et démontre que la sumoylation est nécessaire à l’activité répressive de TIF1β mais n’est pas impliquée dans son homodimérisation ou son interaction avec la boîte KRAB. La sumoylation est cependant nécessaire au recrutement d’histones déacétylases, dépendante de son homodimérisation et de l’intégrité du domaine PHD. Alors que l’on ne connaît pas de régulateur physiologique de la sumoylation outre les enzymes directement impliquées dans la machinerie de sumoylation, nous mettons en évidence que la sumoylation de TIF1β est positivement régulée par son interaction avec le domaine KRAB et suggérons que ces facteurs transcriptionnels recrutent TIF1β à l’ADN au niveau de promoteur et augmentent son activité répressive en favorisant sa sumoylation.