Amplification of the DFR1 gene in Saccaromyces cerevisiae

A new system was employed to study amplification of t,he DHF'R gene DFB,1 ) in Sa<,:;charoillYCB§. .Q~~Yi...S!i<;1~. . This system consists of a series of yeast strains containing a casset,te which encodes t he yeast, D..ERl gene ttghtly linked tjO a f usion of the yeast 1EU2. regulat,ory region wi tJ1 the LAQZ str ctural gene from E. cO.1-1 (,) . M. Clement , unpubl i,::;hed) . Th's casset;t e was shown t.o be integrat,ed int o a unj que chromosomal l ocati on in each strain . Yeast cells were se l ected for MTX-resistance and overproduction of ~ galac t osi d se ( B-gal ). Since the inserted DF'Rl and ~ACZ genes are independently regulated, it was thought that cel l s with this phenotype probably contain e d ampl if ications of the cassette. A lar ge variat ion in the f requn y o f MTX-resistance was found between the di ff e r ent str ains. These freqlen c ~ es r anged from about 2 x 10 - 7 fo r a population of cells containing the cassette integrated at, the BI J2.l gene in t,he middle of the long arm of chromosome V, to about 5 x 10-4 for a strain with the cassette i nserted in the r DNA cluster Abo It 85% of the MTX- res i stcmt iso l ates examined showed enhanced B·-gal act i v ity rel a t ive t o the parental strain . For the ma jorit y of strains, the mean B- gal activity in drug-r sistant clones was about 3 times that o f the parent following a single se l ect i on step . I n con t r ast, primary MTX-resistant derivat~ves of cells with the cassette inserted 3 at the rDNA cluster showed inc r eases in B- gal activity ranging from 9 - 14 f old r elative to the parent. Analysis of the latte r s train by Southe rn hybr idization indicated that the cassette was inde e d amplified several fold in MTX-re sistant derivatives. A sing l e strain, in which the cassette was inserted at the !lEA;], loc u.s , was used to examine in more detai 1 , the parameters affecting DFRl gene amplificat~ion in yeast . The mean B- gal activity in drug-resistant derivatives of this strain could be increased from 3 to 6 or 7 fold relative to the parent, by stepwise sel ection using increasing MTX concentrations. B-gal overproduction was found to be un stable in all primary and highly -resistant isolates examined. There was no indication, h owever, of a decrease i n growth r a t e in MTX-res i s tant cells which overproduced B - gal.

Effect of the position of the double-bond in monounsaturated phospholipids on the dynamics of model membranes

Therllloelynalllics of lllodel 11lel1ll)rane systeills containing 1110nollnsaturatecl I)lloSI)holil) ids is strongly infllienced l)y the I)osition of the C==C dOlll)le })ond in tIle acyl chain. The telllI)eratllres of both chain-nlelting (TM) and La -+ HI! (TH) I)hase traIlsitions are lowered by IIp to 20°C when C==C is Inoved froln positions 6 or 11 to I)osition 9 in an 18-carl)on chain. This work is an attellll)t to ellicidate the uIlderlying Illoleclilar Illechanisllls reSI)Onsi])le for tllese draillatic tllerillodynaillic changes. Mixtllres of di-18: 1 l)hoSI)hatidylethanolanline with C==C at l)ositioIlS 6, 9, 11 were llsed, witll a sI1lall aI1lOlint of I)erdellterated tetradecanol, known to })e a gooel rel)Orter of the cllain Illoleclilar order. SI)ectral second 11I0I1lents were llsed to Illonitor tIle La -+ HII I)hase transition, which was fOllnd to ])e ])road (2-6°C), with a slight llysteresis on heatiIlg/cooling. The orientational order I)rofiles were nleasllred 1lSiIlg 2H Illiclear Illagnetic resonance and changes in these order I)rofiles between La aIld HII I)hases silow l)oth a local increase in order in the vicinity of the C==C bonds and an o\Terall decrease ill the average orientational order of the chain as a whole. These Sll])tle changes recluire })oth high-fidelity SI)ectrosCol)y and a careflll data analysis that takes into aCCOllnt the effects due to l)artiall1lagnetically-indllced orientational ordering of the l)ilayers. In tIle COIltext of SOllle recently rel)Orted cross-relaxation 11leaSlirenlents in Silllilar l)llOSI)llolil)iels, 0111' reslilts sllggest that large-anll)litllde conforlllational changes in the interior of tIle I110del 111eI11])ranes I)lay a 1110re significant role than I)reviollsly thOllght.

Measuring curriculum integration in the junior grades: the design of an instrument

This study \Alas initiated in response to the Junior Division Review (1985) publ ished by the Ministry of Education for the Province of Ontario. Curriculum integration is an element used within the educational paradigm designed by the Ontario Ministry of Education. It is a term frequent1y verbal ized b>' educators in this province, but because of 1 imi ted resource support regarding this methodology, it was open to broad interpretation resulting in an extreme v ar i at i on i nit simp 1 eme n tat i on • I n de ed, the Min i s try intimated that it was not occurring to any significant degree across the province. The objective of this thes is was· to define integration in the junior classroom and de-:.ign a meas.ur·ement in-:.tr-ument which would in turn high 1 i gh t indicators of curriculum integration. The :.tudy made a prel iminary, field-based survey of educa tiona 1 professionals in order to generate a relevant description of integrated curr-iculum programm i ng as def i ned in the j un i or classroom. The description was a compilation of views expressed by a random selection of teachers, consultants, supervisory officers and principals. The survey revea 1 ed a much more comprehens i ve vi et·<,l of the attributes of integrated programming than tradition would dictate and resulted in a functional definition tha t was broader than past prac t ices. Based on the information generated by this survey, an instrument ou t 1 in i ng program cr iter i a of was devised. an integrated junior cla~·sroom Th i s measuremen t i nstrumen t , designed for all levels of educators, was named uThe Han~.son I nstrumen t for the Measuremen t of Program Integrat ion in the Jun i or Cl assroom". It refl ected five categories intrinsic to the me thodol ogy of integration: Teacher Behaviour, Student Behaviour, Classroom Layout, Cl as~·r oom Environment and Progr amm i ng. Each category and the items therein were successfully tested in val idi ty and rel iabi 1 i ty checKs. Interestingly, the individual class was found to be the major variable programming in in the measuremen t the j un i or d i vis i on • of The integrated instrument demonstrated potential not onl)' a~· an initial measure of the degree of integrated curriculum, but as a guide to strategies to implement such a methodology.

The use of diatom assemblages to determine paleotrophic changes in lakes

The purpose of this study was to develop a classifi cation scheme for l ake trophic status based on the relative abundance of l ake sediment diatom trophic indicator species. A total of 600 diatom frustules were counted from the surface sediments of e a ch of 30 lakes selected to repr e seni~ a continuum from u.lt ra-oligotrophic t,o fairly eutrophic but not hype r-' eutrophic conditions. Published autecological information was used to determine the trophic indicator status of each of the s pecies. A quotieht was derived by dividing the s um of all the e utrophic indicator species by the sum of all oligotrophic indicai.-:.or species. Oligo'- mesotrophic. mesotrophic and meso-eutrophic species were added to both the numerator and denomina tor. Five categories of diatom i.nferred trophic status were recognized : ultra-oligotrophic - 0'-0.2:3, oligotrophic::: 0.24-0 . 70, mesotrophic :: 0.'71 -0.99, meso-elxtrophic :: 1. 00-1. '78 and eutrophic:: 1.. 79-2. 43. But only three of these (oligotrophic:: 0-0.69, mesotrophic ::: 0 . 70'-1.69 j and eutrophic:: 1.70-2.50) proved usef ul. The present study of the relationship between diatom inferred trophic status and the literature-derived trophic status of SO lake s (which were purposely chosen to represent a broad spectrum of lakes in Canada) indicated that: 1) Based on diatom species (assemblages ) it is possible to segregate the lakes from which. th",)se diatoms were taken into three basic categories : o ligotrophic, mesotrophic and eutrophic lake types. ~~) It was not possible t,o separate meso-eutrophic and o l igo-mesotrophic lakes f rom mesotrophic l akes as the the degree of overlap betwee n the diat,om species in these lake types was extremely high. 3) Ha d mo r e ul tra-oligotroph,ic lakes been sampled it might have been possible to more a ccurately s eparate them f rom oligotrophic Jakes. 4 ) Had. more humic lakes been sampled in this s tudy I f eel it would have been possible to identify a unique diatom a ssemblage which would h a ve chara cterized t his lake type . Re gression analyses were performed using the aforementioned diatom inferred trophic index as a f unction of 1) log Sec chi transparency (r = - 0.70) 2) total phosphorus (r = 0. 77 ) and 3) chlorophyll-a (r = 0.74). Once e ach of these rel ationships had been established , it was possible to infer paleotrophic (downcore) changes in an oligotrophic lake (Barbara Lake) and in a eutrophic lake (Chemung Lake) . Barbara Llake was dominated by oligotrophic s pecies and remained oligotrophic throughout the 200-·year history r epresented by i t s 32 em long sediment core. Chemung Lake is currently dominated. by eutrophic species but went through a mesotrophic st,age which was associated with a rise in the water level of the lake followi n g dam construction in its watershed in the early 1.900 J ::;. This was followed by its reversion to it,s present day eutrophic stage (dominated by eutrophic species) possibly as a r esult of shallowing process which can be attributed to " silting' up" of the reservoir and the invasion of the l ake by Myriophyllum spjcatum (Eurasian milfoil) i n the 1970's . In addition, nutrient .:r':l.ch run"'offwhich resulted from increased human a.ctivities associated with cottage development along its shores has contribut ed to its eutrophication. There is some evidence that the rat,e o :f its prog ressive eutrophication has declined during the last decade. This was correlated with legislation enacted in the 60's and 70's in Ontario which was aimed at reducing nutrient loading from cottages.

(A) Photoregulation of DNA Functions by Cyclic Azobenzene-tethered Oligonucleotides (B) Site-specific Fluorescent Labeling of DNA using Inverse Electron Demand Diels-Alder Reaction between trans-Cyclooctene Derivatives and BODIPY-Tetrazine Adducts

(A) Most azobenzene-based photoswitches require UV light for photoisomerization, which limit their applications in biological systems due to possible photodamage. Cyclic azobenzene derivatives, on the other hand, can undergo cis-trans isomerization when exposed to visible light. A shortened synthetic scheme was developed for the preparation of a building block containing cyclic azobenzene and D-threoninol (cAB-Thr). trans-Cyclic azobenzene was found to thermally isomerize back to the cis-form in a temperature-dependent manner. cAB-Thr was transformed into the corresponding phosphoramidite and subsequently incorporated into oligonucleotides by solid phase synthesis. Melting temperature measurement suggested that incorporation of cis-cAB into oligonucleotides destabilizes DNA duplexes, these findings corroborate with circular dichroism measurement. Finally, Fluorescent Energy Resonance Transfer experiments indicated that trans-cAB can be accommodated in DNA duplexes. (B) Inverse Electron Demand Diels-Alder reactions (IEDDA) between trans-olefins and tetrazines provide a powerful alternative to existing ligation chemistries due to its fast reaction rate, bioorthogonality and mutual orthogonality with other click reactions. In this project, an attempt was pursued to synthesize trans-cyclooctene building blocks for oligonucleotide labeling by reacting with BODIPY-tetrazine. Rel-(1R-4E-pR)-cyclooct-4-enol and rel-(1R,8S,9S,4E)-Bicyclo[6.1.0]non-4-ene-9-ylmethanol were synthesized and then transformed into the corresponding propargyl ether. Subsequent Sonogashira reactions between these propargylated compounds with DMT-protected 5-iododeoxyuridine failed to give the desired products. Finally a methodology was pursued for the synthesis of BODIPY-tetrazine conjugates that will be used in future IEDDA reactions with trans-cyclooctene modified oligonucleotides.

Les voix électorales, la collection Baby comme témoin de notre histoire politique (1792-1870).

Les nombreux scrutins qui ont eu lieu dernièrement ont suscité plusieurs réflexions tant en ce qui concerne les taux de participation que l’impact réel du vote individuel sur la sphère politique. Récemment, une commission spéciale de l’Assemblée nationale du Québec a parcouru seize villes pour consulter la population sur le mode de scrutin et la loi électorale. De fait, cette dernière a changé radicalement depuis 1792, année où a eu lieu la première élection parlementaire. À ce sujet, la collection Baby de l’Université de Montréal est une source d’information inestimable sur les mœurs électorales du 19e siècle québécois. Les pressions et les sollicitations dont les électeurs font l’objet, les stratégies adoptées par les candidats et leurs partisans, l’ambiance enfiévrée autour des bureaux de scrutin, tous ces éléments trouvent des exemples dans les documents disponibles. En plus de témoigner des vicissitudes du scrutin ouvert et public, ces derniers révèlent des aspects insoupçonnés de la culture politique de l’époque. Cette présentation se veut donc une exploration de la sphère politique de notre passé à partir des traces multiples offertes par la collection Baby.

Some Implications of the Zero Lower Bound on Interest Rates for the Term Structure and Monetary Policy

In an economy where cash can be stored costlessly (in nominal terms), the nominal interest rate is bounded below by zero. This paper derives the implications of this nonnegativity constraint for the term structure and shows that it induces a nonlinear and convex relation between short- and long-term interest rates. As a result, the long-term rate responds asymmetrically to changes in the short-term rate, and by less than predicted by a benchmark linear model. In particular, a decrease in the short-term rate leads to a decrease in the long-term rate that is smaller in magnitude than the increase in the long-term rate associated with an increase in the short-term rate of the same size. Up to the extent that monetary policy acts by affecting long-term rates through the term structure, its power is considerably reduced at low interest rates. The empirical predictions of the model are examined using data from Japan.

Habit Formation and the Persistence of Monetary Shocks

This paper studies the persistent effects of monetary shocks on output. Previous empirical literature documents this persistence, but standard general equilibrium models with sticky prices fail to generate output responses beyond the duration of nominal contracts. This paper constructs and estimates a general equilibrium model with price rigidities, habit formation, and costly capital adjustment. The model is estimated via Maximum Likelihood using US data on output, the real money stock, and the nominal interest rate. Econometric results suggest that habit formation and adjustment costs to capital play an important role in explaining the output effects of monetary policy. In particular, impulse response analysis indicates that the model generates persistent, hump-shaped output responses to monetary shocks.

Les fils RSS en bibliothèques : pourquoi et comment?

Présentation donnée le 10 novembre 2006 dans le cadre du Programme de formation continue de la Corporation des bibliothécaires professionnels du Québec (CBPQ).

Les fils RSS en bibliothèque de droit

Présentation donnée le 1er mars 2007 à l'Association des bibliothèques de droit de Montréal (ABDM) / Montreal Association of Law Libraries (MALL).

De l'économie informelle à la morphologie sociale de l'économie. L'étude de cas des Mafy ady à Tananarive (Madagascar).

Ce travail porte sur l'organisation sociale de l'économie dans deux quartiers de la ville basse de Tananarive, capitale de Madagascar. Il s'agit de secteurs de très grande pauvreté caractérisés par des pratiques socioéconomiques généralement définies à travers la notion d'économie informelle. Dans cette thèse, la problématique de l'économie informelle est recadrée dans les termes plus généraux d'une sociologie de l'économie et est abordée sous l'angle d'une reconstruction sociographique des propriétés de cette économie. Il s'agit d'étudier les activités de production, les circulations des richesses et la redistribution de ces richesses sous l'angle de la reproduction sociale, celle des groupes qui opèrent ces pratiques et qui se reproduisent dans ces pratiques. Ce travail de reconstruction s'appuie sur une approche de la morphologie sociale, c'est-à-dire sur l'exploration des relations sociales empiriques, leurs enracinements dans le monde sensible et les espaces sociaux construits par ces relations. Les résultats de cette recherche sociographique mettent en évidence la présence de six espaces sociaux différenciés de pratiques socioéconomiques qui composent l'économie générale de ces quartiers. Ils mettent également en évidence la présence de trois grands cadres sociaux d'intelligibilité du réel qui sont coprésents dans la régulation des activités localisées dans ces espaces. L'hypothèse générale qui est dégagée de la thèse est que cette coprésence est au fondement des discontinuités systématiquement observées dans les activités, empêchant les personnes et groupes sociaux vivants dans ces quartiers de reproduire des conditions d'existences qui seraient tant soit peu conformes aux idéalités sous-tendues par ces cadres sociaux d'intelligibilité du réel.

La protection des données personnelles en France

Un résumé en anglais est également disponible.

La teneur du standard de fiabilité des moyens électroniques de signature

Un résumé en anglais est également disponible. Le présent document a été présenté pour l'obtention du diplôme de Maîtrise en droit. Le ménoire a été accepté et classé parmi les 5% de la discipline.