The production of 4-adminobutyrate in response to treatments reducing cytosolic pH and the regulation of intracellular pH

GABA (4-aminobutyrate) is synthesized through the decarboxylation of LGlu- (L-Glu-+ H+ ---> GABA + C02), and compared to many free amino acids is present in high concentrations in plant cells. GABA levels rise rapidly and dramatically in response to varied stress conditions including anaerobiosis. Recent papers suggest that GABA production and associated H+ consumption are parts of a metabolic pH-stat mechanism which ameliorates the intracellular pH decline associated with anaerobiosis or other treatments. To test this hypothesis GABA production and efflux have been measured in isolated Asparagus sprengeri cells in response to three treatments which potentially cause intracellular acidification. Acid loads were imposed using 60 min of (i) anaerobiosis, (ii) H+/LGlu- cotransport, and (iii) treatment with permeant weak acids (butyric, acetic and propionic). Both intra- and extracellular GABA concentrations increased more than 100% after anaerobiosis, almost 1000% after H+/L-Glu- cotransport (light or dark) and almost 5000/0 after addition of 5 mM butyric acid at pH 5.0. HPLC analysis of amino acids indicates that as GABA concentrations increased in response to butyric acid addition, glutamate concentrations decreased. Time-course studies demonstrated that added butyric acid stimulates GABA production by 2800/0 within 15 seconds. A fluorescent determination of cytosolic pH indicates that addition of butyric or other weak acids resulted in a rapid reduction in cytosolic pH of 0.6 pH units. The half time for the response to butyric acid addition is 2.1 seconds, indicating that the decline in cytosolic pH is rapid enough to account for the rapid stimulation of GABA production. The acid load in response to butyric acid addition was assayed by measurements of 14C-butyric acid uptake. Calculations indicate that GABA production accounted for 45% of the imposed acid load. The biological significance of GABA efflux is not yet understood. The results support the original hypothesis suggesting a role for GABA production in cellular pH regulation.

Second Welland Canal - Book 1, Survey Map 13 - St. Catharines and Locks 4 and 5

Survey map of the Second Welland Canal created by the Welland Canal Company showing the Town of St. Catharines. Identified structures associated with the Canal include Lock 4, Lock House, Lock 5, Small Lock House, the towing path, and Gasometer for Canal. The surveyors' measurements and notes can be seen in red and black ink and pencil. Local area landmarks are also identified and include streets and roads (ex. Geneva Street, Queenston Street, and Academy Street), C. Phelps Mill and Store House, St. Catharines and Welland Canal Gas Works, William Mahony's Tannery, Cooper Shop, a barrel shed, barn, and gas tanks. Properties and property owners of note are: Concession 6 Lots 14, 14, and 16, Concession 7 Lots 14, 15, and 16, C. Phelps, R. M. Clement, Orson Phelps, R. Collier, D. P. Haynes, W. Chace, and John Soper.

The Gleaner and Niagara Newspaper, January 15, 1818

The Gleaner and Niagara Newspaper was a weekly paper in the town of Niagara-on-the-Lake that began publication December 4, 1817 and ceased in 1830. This bound volume has been divided by publication dates. Other dates included in the volume are: 1817 December 18 1817 December 25 1818 January 1 1818 January 8 1818 January 22 There is also a letter that was found inside the bound volume written by Andrew Heron on October 1 1817.

Baltimore Patriot and Evening Advertiser Vol. 3 No. 89- April 15, 1814

This daily paper, except Sundays, began publication on August 17, 1813 and ceased April 30, 1817. It was also published tri-weekly for a "COUNTRY" edition. The paper later became the Baltimore Patriot. Topics of interest include: Page 1: announcement Capt. M. Simmones Burnbury is seeking men to form a company of Sea Fencibles; Page 2: U.S. Legislature authorizes bill to increase the Marine Corps; U.S. Legislature authorizes bill to purchase vessels captured on Lake Erie; U.S. Senate bill authorizes the appointment of officers for the flotilla service; President of United States authorized to accept service of volunteer corps for defence of United States; Letter from Capt. A. H. Holmes to Lieut. Col. Butler detailing an American victory over the British near Detroit; announcement of British forces building barges for the purpose of attacking Detroit; War sloop Erie returns to Baltimore unable to reach sea due to British forces in the Chesapeake; Page 3: Death notice of Mr. Ward Fairchild of the U.S. Army; Page 4: Commander of the U.S. Flotilla seeks men for the Chesapeake Flotilla.

National Intelligencer Vol. XVI, No. 2193- October 15, 1814

The Intelligencer was an American newspaper that was established, in 1800, in Washington by Samuel Harrison Smith, a young Jeffersonian- Republican from Philadelphia. War of 1812 related content includes: Page 1: Major General Winfield Scott's arrival in Washington escorted by Colonel Laval's cavalry; report from Sackett's Harbour on the American and British navies; a debate on a bill to temporarily remove the seat of government from the City of Washington due to the war; the Military Committee wishing to have the President commission gold medals for distinguished service for Major Generals Brown and Scott and silver medals to Brigadier Generals Ripley, Miller and Porter for their efforts at Chippewa, Erie and Niagara as well as gold medals to Brigadier General Gaines for efforts at Erie and Brigadier General Macomb for efforts at Plattsburg; resolutions of the House on the victory of Commodore Thomas Macdonough at the Battle on Lake Champlain; Page 2: more detail on the proposed removal of the seat of government from Washington; Page 3: a letter from Major General Andrew Jackson to the Secretary of War on repelling the British at Fort Bowyer on the Point of Mobile and naval battle reports on the vessels Hermes, Sophie, and Carron; a letter from Major William Lawrence to Andrew Jackson on the battle at Fort Bowyer; military promotions of Alexander Macomb, Roger Jones, J. Hindman; more on the removal of the seat of government; citizens of Washington offering to volunteer in defense of the city; banks willing to loan the government funds to re-build destroyed government buildings; assignments of Major General Scott, General Winder and Major General Gaines; report of 300 British prisoners from Fort Erie passed through Washington; Montreal newspaper report condemning Provost for his actions at the Battle of Lake Champlain; trouble with stage coaches traveling between New Haven and Bridgeport due to cannon fire; report of a British naval squadron appearing off New Port and by Long Island and New Bedford; report of enemy naval movements from Norfolk; announcement of a book entitled "A Narrative of the Battle of Bladensburg" by and officer of General Smith; Page 4: report on the amassing navy at Kingston under Drummond and the defenses at Sackett's Harbor; report of American and British naval news from Nova Scotia and the east coast; actions of the American sloop of war, The Peacock, in international waters; an enlistment announcement by Captain Perrin Willis of the 2nd Regiment Infantry; listing of American officers and privates released from parole on October 13; Other notices report on stray horses, properties for sale, runaway slaves, imprisonments, missing livestock, medical lectures at the University of Maryland, stage lines, auctions, etc. The paper was a supporter of the Jefferson and Madison administrations until 1810 when it was sold to Joseph Gales Jr. from North Carolina. In 1812 William Seaton joined Gales as a publishing partner. This paper made significant contributions to the nation and wielded considerable influence in political circles during its publication. It has been praised for its "high standard of journalistic excellence and high intellectual level of its contents". (William E. Ames , National Intelligencer: Washington's Leading Political Newspaper) The Intelligencer was, until 1810, named the National Intelligencer, and Washington Advertiser. It was a tri-weekly paper and had a peak circulation of 6, 000. Publication was suspended in 1869.

Chemoenzymatic Total Synthesis of Morphine alkaloids: Synthesis of Dihydrocodeine and Hydrocodone via a Double Claisen Strategy and ent-Hydromorphone via an Oxidative Dearomatization/intramolecular [4+2] Cycloaddition

This thesis describes the chemoenzymatic synthesis of three morphine alkaloids. The total synthesis of dihydrocodeine and hydrocodone was accomplished starting from bromobenzene in 16 and 17 steps, respectively. The key steps included a microbial oxidation of bromobenzene by E. coli JM109 (pDTG601A), a Kazmaier-Claisen rearrangement of glycinate ester to generate C-9 and C-14 stereo centers, a Johnson-Claisen rearrangement to set the C-13 quaternary center, and a C-10/C-11 ring closure via a Friedel-Crafts reaction. In addition, the total synthesis of ent-hydromorphone starting from β-bromoethylbenzene in 12 steps is also described. The key reactions included the enzymatic dihydroxylation of β-bromoethylbenzene to the corresponding cis-cyclohexadienediol, a Mitsunobu reaction, and an oxidative dearomatization followed by an intramolecular [4+2] cycloaddition.

Role of Phosphorylation of the Oxysterol Binding Protein (OSBP) in (PI(4)P) and Sterol-Containing Membrane Recognition and Binding

Studies have demonstrated that the oxysterol binding protein (OSBP) acts as a phosphatidylinositol phosphate (PIP)-sterol exchanger at membrane contact sites (MCS) of the endoplasmic reticulum (ER) and Golgi. OSBP is known to pick up phosphatidylinositol-4-phosphate (PI(4)P) from the ER, transfer it to the trans-Golgi in exchange for a cholesterol molecule that is then transferred from the trans-Golgi to the ER. Upon further examination of this pathway by Ridgway et al. (1), it appeared that phosphorylation of OSBP played a role in the localization of OSBP. The dephosphorylation state of OSBP was linked to Golgi localization and the depletion of cholesterol at the ER. To mimic the phosphorylated state of OSBP, the mutant OSBP-S5E was designed by Ridgway et al. (1). The lipid and sterol recognition by wt-OSBP and its phosphomimic mutant OSBP-S5E were investigated using immobilized lipid bilayers and dual polarization interferometry (DPI). DPI is a technique in which the protein binding affinity to immobilized lipid bilayers is measured and the binding behavior is examined through real time. Lipid bilayers containing 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and varying concentrations of PI(4)Ps or sterols (cholesterol or 25-hydroxycholesterol) were immobilized on a silicon nitride chip. It was determined that wt-OSBP binds differently to PI(4)P-containing bilayers compared to OSBP-S5E. The binding behavior suggested that wt-OSBP extracts PI(4)P and the change in the binding behavior, in the case of OSBP-S5E, suggested that the phosphorylation of OSBP may prevent the recognition and/or extraction of PI(4)P. In the presence of sterols, the overall binding behavior of OSBP, regardless of phosphorylation state, was fairly similar. The maximum specific bound mass of OSBP to sterols did not differ as the concentration of sterols increased. However, comparing the maximum specific bound mass of OSBP to cholesterol with oxysterol (25-hydroxycholesterol), OSBP displayed nearly a 2-fold increase in bound mass. With the absence of the wt-OSBP-PI(4)P binding behavior, it can be speculated that the sterols were not extracted. In addition, the binding behavior of OSBP was further tested using a fluorescence based binding assay. Using 22-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-23,24-bisnor-5-cholen-3β-ol (22-NBD cholesterol), wt-OSBP a one site binding dissociation constant Kd, of 15 ± 1.4 nM was determined. OSBP-S5E did not bind to 22-NBD cholesterol and Kd value was not obtained.

Deletion of the RNR4 gene causes hyperresistance to the carcinogen 4-NQO in the yeast model

La stabilité génomique, qui est essentielle à la vie, est possible grâce à la réplication et la réparation de l’ADN. Une des enzymes responsables de la réplication et de la réparation de l’ADN est la ribonucleotide reductase (RNR), qui est retrouvée chez la levure et chez l’humain. Cette enzyme catalyse la formation de déoxyribonucléotides et maintien le pool de dNTP requis pour la réparation et la réplication de l’ADN. L’enzyme RNR est un tétramère α2β2 constitué d’une grande (R1, α2) et d’une petite (R2, β2) sous-unité. Chez S. cerevisiae, les gènes RNR1 et RNR3 encodent la sous-unité α2 (R1). L’activité catalytique de RNR dépend d’une interaction avec le fer et de la formation d’un complexe entre R1 et R2. L’expression de toutes les sous-unités est inductible par les dommages causés à l’ADN. Dans cette étude, nous démontrons que des cellules qui n’expriment pas une des sous-unités, Rnr4, du complexe RNR sont sensibles à divers agents endommageant l’ADN, tels que le méthyl méthane sulfonate, la bléomycine, le péroxyde d’hydrogène et les rayons ultraviolets (UVC 254 nm). Au contraire, le mutant est résistant au 4-nitroquinoline-1- oxide (4-NQO), un composé qui engendre des lésions encombrantes. Par conséquent, le mutant rnr4Δ démontre une réduction marquée en mutations induites par le 4-NQO comparativement à la souche parentale. Nous voulions identifier la voie de réparation de l’ADN qui conférait cette résistance au 4-NQO ainsi que les protéines impliquées. Les voies BER, NER et MMR n’ont pas aboli la résistance au 4-NQO de la souche rnr4Δ. La protéine recombinante Rad51 ne joue pas un rôle critique dans la réparation de l’ADN et dans la résistance au 4-NQO. La délétion du gène REV3, qui encode une polymérase de contournement, impliquée dans la réparation post-réplication, a partiellement aboli la résistance au 4-NQO dans rnr4Δ. Ces résultats suggèrent que la polymérase Rev3 et possiblement d’autres polymérases translésion (Rev1, Rev7, Rad30) pourraient être impliquées dans la réparation de lésions encombrantes dans l’ADN dans des conditions de carence en dNTP. La réparation de l’ADN, un mécanisme complexe chez la levure, implique une vaste gamme de protéines, dont certaines encore inconnues. Nos résultats indiquent qu’il y aurait plus qu’une protéine impliquée dans la résistance au 4-NQO. Des investigations plus approfondies seront nécessaires afin de comprendre la recombinaison et la réparation post-réplication.

L’orientation mentale maternelle : relation avec le développement du langage et de la théorie de l’esprit chez les enfants de 2 à 4 ans

L’étude de l’orientation mentale (OM), soit l’habileté des parents à concevoir et à traiter leur enfant comme un individu disposant d’une vie mentale autonome et active (Meins, 1997), a gagné en intérêt grâce à la démonstration de ses associations avec plusieurs sphères du développement de l’enfant, entre autres avec le style d’acquisition linguistique chez les enfants de 20 mois ainsi que le développement de la théorie de l’esprit chez les enfants d’âge préscolaire (p. ex., Meins & Fernyhough, 1999). En dépit de ces résultats, l’étude du développement du vocabulaire et des stades initiaux de la théorie de l’esprit chez les enfants a été négligée dans cette littérature. La présente thèse est composée de trois articles empiriques et vise à combler certaines lacunes de la littérature actuelle. Le premier article vise à examiner les liens longitudinaux entre l’OM maternelle et le vocabulaire expressif chez les enfants de 2 ans. Les deuxième et troisième articles ont pour but d’évaluer les relations longitudinales entre l’OM maternelle, la sécurité d’attachement des enfants et les toutes premières articulations de la théorie de l’esprit chez les enfants de 2 ans, puis 2 ans plus tard, avec la théorie de l’esprit chez les enfants de 4 ans. Les données des trois articles proviennent de cinq visites effectuées au domicile de 84 dyades mère-enfant. À 12 mois, l’OM maternelle a été mesurée lors de jeux libres mère-enfant à l’aide du système de codification de Meins et al. (2001). À 15 mois, la sécurité d’attachement de l’enfant a été mesurée par un observateur avec le Q-Sort d’attachement (Waters & Deane, 1985). À 2 ans, les mères ont évalué le langage de leur enfant à l’aide des inventaires MacArthur du développement de la communication (Dionne, Tremblay, Boivin, Laplante, & Pérusse, 2003). Finalement, la théorie de l’esprit de l’enfant a été mesurée à 2 et 4 ans à l’aide de diverses tâches expérimentales (Carlson, Mandell, & Williams, 2004). Les résultats du premier article indiquent que l’OM maternelle est associée au développement du vocabulaire expressif des enfants de 2 ans. Les résultats des deuxième et troisième articles indiquent que la compréhension des enfants de 2 et 4 ans aux tâches de théorie de l’esprit est associée à l’OM maternelle. De plus, ces deux articles ont démontré que les garçons, mais pas les filles, présentant des comportements d’attachement davantage sécures ont obtenu de meilleures performances à une tâche demandant la compréhension des perspectives visuelles de leur mère à 2 ans et de celles d’une étrangère à 4 ans. En conclusion, les résultats de la thèse suggèrent que l’utilisation que les mères font de commentaires mentaux à 12 mois semble favoriser l’acquisition de mots dans le vocabulaire expressif de leur enfant à 2 ans ainsi que le développement d’une meilleure compréhension aux tâches de théorie de l’esprit à 2 et 4 ans.

Compact Wideband Antenna for 2.4 GHz WLAN Applications

A novel compact wideband antenna for wireless local area network (WLAN) applications in the 2.4 GHz band are presented. The proposed low profile antenna of dimensions 15 x 14.5 x 1.6 mm offers 18.6% bandwidth and an average gain of~5 dBi. The antenna can be excited directly using a 50 coaxial probe

Compact Wideband Antenna for 2.4 GHz WLAN Applications

A novel compact wideband antenna for wireless local area network (WLAN) applications in the 2.4 GHz band is presented. The proposed low profile antenna of dimensions 15 x 14.5 x 1.6 mm offers 18.6% bandwidth and an average gain of -5 dBi. The antenna can be excited directly using a 50 coaxial probe

An Efficient Multi-component Synthesis of 6-Amino-3-methyl-4-Aryl-2,4- dihydropyrano[2,3-c]Pyrazole-5-carbonitriles

Multi-component reactions are effective in building complex molecules in a single step in a minimum amount of time and with facile isolation procedures; they have high economy1–7 and thus have become a powerful synthetic strategy in recent years.8–10 The multicomponent protocols are even more attractive when carried out in aqueous medium. Water offers several benefits, including control over exothermicity, and the isolation of products can be carried out by single phase separation technique. Pyranopyrazoles are a biologically important class of heterocyclic compounds and in particular dihydropyrano[2,3-c]pyrazoles play an essential role in promoting biological activity and represent an interesting template in medicinal chemistry. Heterocyclic compounds bearing the 4-H pyran unit have received much attention in recent years as they constitute important precursors for promising drugs.11–13 Pyrano[2,3-c]pyrazoles exhibit analgesic,14 anti-cancer,15 anti-microbial and anti-inflammatory16 activity. Furthermore dihydropyrano[2,3-c]pyrazoles show molluscidal activity17,18 and are used in a screening kit for Chk 1 kinase inhibitor activity.19,20 They also find applications as pharmaceutical ingredients and bio-degradable agrochemicals.21–29 Junek and Aigner30 first reported the synthesis of pyrano[2,3-c]pyrazole derivatives from 3-methyl-1-phenylpyrazolin-5-one and tetracyanoethylene in the presence of triethylamine. Subsequently, a number of synthetic approaches such as the use of triethylamine,31 piperazine,32 piperidine,33 N-methylmorpholine in ethanol,34 microwave irradiation,35,36 solvent-free conditions,37–39 cyclodextrins (CDs),40 different bases in water,41 γ -alumina,42 and l-proline43 have been reported for the synthesis of 6-amino-4-alkyl/aryl-3-methyl- 2,4-dihydropyrano[2,3-c]pyrazole-5-carbonitriles. Recently, tetraethylammonium bromide (TEABr) has emerged as mild, water-tolerant, eco-friendly and inexpensive catalyst. To the best of our knowledge, quaternary ammonium salts, more specifically TEABr, have notbeen used as catalysts for the synthesis of pyrano[2,3-c]pyrazoles, and we decided to investigate the application of TEABr as a catalyst for the synthesis of a series of pyrazole-fused pyran derivatives via multi-component reactions

Inclusive probability calculations for the K-vacancy transfer in collisions of S{^15+} on Ar

Using the single-particle amplitudes from a 20-level coupled-channel calculation with ab initio relativistic self consistent LCAO-MO Dirac-Fock-Slater energy eigenvalues and matrix elements we calculate within the frame of the inclusive probability formalism impact-parameter-dependent K-hole transfer probabilities. As an example we show results for the heavy asymmetric collision system S{^15+} on Ar for impact energies from 4.7 to 16 MeV. The inclusive probability formalism which reinstates the many-particle aspect of the collision system permits a qualitative and quantitative agreement with the experiment which is not achieved by the single-particle picture.