Subdivision of the bacterioferritin comigratory protein family of bacterial peroxiredoxins based on catalytic activity

Peroxiredoxins are ubiquitous proteins that catalyze the reduction of hydroperoxides, thus conferring resistance to oxidative stress. Using high-resolution mass spectrometry, we recently reclassified one such peroxiredoxin, bacterioferritin comigratory protein (BCP) of Escherichia coli, as an atypical 2-Cys peroxiredoxin that functions through the formation of an intramolecular disulfide bond between the active and resolving cysteine. An engineered E. coli BCP, which lacked the resolving cysteine, retained enzyme activity through a novel catalytic pathway. Unlike the active cysteine, the resolving cysteine of BCP peroxiredoxins is not conserved across all members of the family. To clarify the catalytic mechanism of native BCP enzymes that lack the resolving cysteine, we have investigated the BCP homologue of Burkholderia cenocepacia. We demonstrate that the B. cenocepacia BCP (BcBCP) homologue functions through a 1-Cys catalytic pathway. During catalysis, BcBCP can utilize thioredoxin as a reductant for the sulfenic acid intermediate. However, significantly higher peroxidase activity is observed utilizing glutathione as a resolving cysteine and glutaredoxin as a redox partner. Introduction of a resolving cysteine into BcBCP changes the activity from a 1-Cys pathway to an atypical 2-Cys pathway, analogous to the E. coli enzyme. In contrast to the native B. cenocepacia enzyme, thioredoxin is the preferred redox partner for this atypical 2-Cys variant. BCP-deficient B. cenocepacia exhibit a growth-phase-dependent hypersensitivity to oxidative killing. On the basis of sequence alignments, we believe that BcBCP described herein is representative of the major class of bacterial BCP peroxiredoxins. To our knowledge, this is the first detailed characterization of their catalytic activity. These studies support the subdivision of the BCP family of peroxiredoxins into two classes based on their catalytic activity.

Proteomics and phylogenetic analysis of the cathepsin L protease family of the helminth pathogen Fasciola hepatica:Expansion of a repertoire of virulence-associated factors

Cathepsin L proteases secreted by the helminth pathogen Fasciola hepatica have functions in parasite virulence including tissue invasion and suppression of host immune responses. Using proteomics methods alongside phylogenetic studies we characterized the profile of cathepsin L proteases secreted by adult F. hepatica and hence identified those involved in host-pathogen interaction. Phylogenetic analyses showed that the Fasciola cathepsin L gene family expanded by a series of gene duplications followed by divergence that gave rise to three clades associated with mature adult worms (Clades 1, 2, and 5) and two clades specific to infective juvenile stages (Clades 3 and 4). Consistent with these observations our proteomics studies identified representatives from Clades 1, 2, and 5 but not from Clades 3 and 4 in adult F. hepatica secretory products. Clades 1 and 2 account for 67.39 and 27.63% of total secreted cathepsin Ls, respectively, suggesting that their expansion was positively driven and that these proteases are most critical for parasite survival and adaptation. Sequence comparison studies revealed that the expansion of cathepsin Ls by gene duplication was followed by residue changes in the S2 pocket of the active site. Our biochemical studies showed that these changes result in alterations in substrate binding and suggested that the divergence of the cathepsin L family produced a repertoire of enzymes with overlapping and complementary substrate specificities that could cleave host macromolecules more efficiently. Although the cathepsin Ls are produced as zymogens containing a prosegment and mature domain, all secreted enzymes identified by MS were processed to mature active enzymes. The prosegment region was highly conserved between the clades except at the boundary of prosegment and mature enzyme. Despite the lack of conservation at this section, sites for exogenous cleavage by asparaginyl endopeptidases and a Leu-Ser[downward arrow]His motif for autocatalytic cleavage by cathepsin Ls were preserved.

Exploring molecular recognition pathways within a family of gelators with different hydrogen bonding motifs

We report the synthesis of a family of gelators in which alkyl chains are connected to the amino groups of L-lysine methyl ester using a range of different hydrogen bonding linking groups (carbamate, amide, urea, thiourea and diacylhydrazine) using simple synthetic methodology based on isocyanate or acid chloride chemistry. The ability of these compounds to gelate organic solvents such as toluene or cyclohexane can be directly related to the ability of the linking group to form intermolecular hydrogen bonds. In general terms, the ability to structure solvents can be considered as: thiourea <carbamate <amide <urea similar to diacylhydrazine. This process has been confirmed by thermal measurements, scanning electron microscopy (SEM) and infrared and circular dichroism spectroscopies. By deprotecting the methyl ester group, we have demonstrated that a balance between hydrophobic and hydrophilic groups is essential-if the system has too much hydrophilicity (e. g., diacylhydrazine, urea) it will not form gels due to low solubility in the organic media. However, the less effective gelators based on amide and carbamate linkages are enhanced by converting the methyl ester to a carboxylic acid. Furthermore, subsequent mixing of the acid with a second component (diaminododecane) further enhances the ability to form networks, and, in the case of the amide, generates a two-component gel, which can immobilise a wide range of solvents of industrial interest including petrol and diesel (fuel oils), olive oil and sunflower oil (renewable food oils) and ethyl laurate, isopropyl myristate and isopropyl palmitate (oils used in pharmaceutical formulation). The gels are all thermoreversible, and may therefore be useful in controlled release/formulation applications.

Functional and structural characterization of the RNase II-family of enzymes

Dissertation presented to obtain a Doctoral Degree in Biology by Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

Biological and Biochemical Characterization of Mutants of the RNase II family of enzymes

Dissertação para a obtenção de grau de doutor em Biologia pelo Instituto de Tecnologia Química e Biológica. Universidade Nova de Lisboa.

In Search of the Origins of Modernism: Venice and the Crisis of Classical Architecture in the mid-eighteenth century

This paper attempts to prove that in the years 1735 to 1755 Venice was the birthplace and cradle of Modern architectural theory, generating a major crisis in classical architecture traditionally based on the Vitruvian assumption that it imitates early wooden structures in stone or in marble. According to its rationalist critics such as the Venetian Observant Franciscan friar and architectural theorist Carlo Lodoli (1690-1761) and his nineteenth-century followers, classical architecture is singularly deceptive and not true to the nature of materials, in other words, dishonest and fallacious. This questioning did not emanate from practising architects, but from Lodoli himself– a philosopher and educator of the Venetian patriciate – who had not been trained as an architect. The roots of this crisis lay in a new approach to architecture stemming from the new rationalist philosophy of the Enlightenment age with its emphasis on reason and universal criticism.

Schram Family of Wainfleet Township documents, 1839-1901 (not inclusive)

George William Schram (1809-1885), son of Garrett Schram and Leah Van Etten, married Orpha Pearson on Nov.13, 1835. His son Marsena John Schram (farmer) was born in May of 1840, in Canada. He died on Nov. 17, 1926 in Wexford County, Michigan. He was married in 1867 to Sarah (1825-1887).Marsena married again on April 18, 1910 to Ann Clarinda Warner (1861-1924). He was working as a carpenter at this time. They had another son, William who was born about 1838 and he married Sabina Chambers on Jan. 21, 1862. The 1861 census for Wainfleet lists siblings of Marsena John Schram as Sarah J. (age 14), Georgiana (age 5), and William (age 21). The Schrams lived on Concession 5 and owned approximately 144 acres of land. David Thompson was born Feb. 4, 1873 and died Feb. 19, 1951. He married Sally Ann Wilson on Sept. 7, 1825 in Pelham. She died about 1840 in Indiana Ontario (near Cayuga). Lemuel Victor Hogue was born Dec.1, 1854 and died Jan. 12, 1929. He was married to Elizabeth Wills who was born Aug. 2, 1861 and died Mar. 8, 1926. Sources: http://www.findagrave.com/cgi-bin/fg.cgi?page=gr&GRid=99294842 http://wc.rootsweb.ancestry.com/cgi-bin/igm.cgi?op=GET&db=seadragon5&id=I91708

The γ-tocopherol-like family of N-methyltransferases: A taxonomically clustered gene family encoding enzymes responsible for N-methylation of monoterpene indole alkaloids

The plant family Apocynaceae accumulates thousands of monoterpene indole alkaloids (MIAs) which originate, biosynthetically, from the common secoiridoid intermediate, strictosidine, that is formed from the condensation of tryptophan and secologanin molecules. MIAs demonstrate remarkable structural diversity and have pharmaceutically valuable biological activities. For example; a subunit of the potent anti-neoplastic molecules vincristine and vinblastine is the aspidosperma alkaloid, vindoline. Vindoline accumulates to trace levels under natural conditions. Research programs have determined that there is significant developmental and light regulation involved in the biosynthesis of this MIA. Furthermore, the biosynthetic pathway leading to vindoline is split among at least five independent cell types. Little is known of how intermediates are shuttled between these cell types. The late stage events in vindoline biosynthesis involve six enzymatic steps from tabersonine. The fourth biochemical step, in this pathway, is an indole N-methylation performed by a recently identified N-methyltransfearse (NMT). For almost twenty years the gene encoding this NMT had eluded discovery; however, in 2010 Liscombe et al. reported the identification of a γ-tocopherol C-methyltransferase homologue capable of indole N-methylating 2,3-dihydrotabersonine and Virus Induced Gene Silencing (VIGS) suppression of the messenger has since proven its involvement in vindoline biosynthesis. Recent large scale sequencing initiatives, performed on non-model medicinal plant transcriptomes, has permitted identification of candidate genes, presumably involved, in MIA biosynthesis never seen before in plant specialized metabolism research. Probing the transcriptome assemblies of Catharanthus roseus (L.)G.Don, Vinca minor L., Rauwolfia serpentine (L.)Benth ex Kurz, Tabernaemontana elegans, and Amsonia hubrichtii, with the nucleotide sequence of the N-methyltransferase involved in vindoline biosynthesis, revealed eight new homologous methyltransferases. This thesis describes the identification, molecular cloning, recombinant expression and biochemical characterization of two picrinine NMTs, one from V. minor and one from R. serpentina, a perivine NMT from C. roseus, and an ajmaline NMT from R. serpentina. While these TLMTs were expressed and functional in planta, they were active at relatively low levels and their N-methylated alkaloid products were not apparent our from alkaloid isolates of the plants. It appears that, for the most part, these TLMTs, participate in apparently silent biochemical pathways, awaiting the appropriate developmental and environmental cues for activity.

Composite Photographs - Woodruff Family of St. Davids, Upper Canada

Composite of the Woodruff Family of St. Davids, Upper Canada. This composite contains 6 4 ½ cm x 4 cm black and white photographs which include: Samuel DeVeaux Woodruff (1819-1904) [the birth and death dates listed on this composite are 1827-1912, but these are actually his wife’s dates] son of William Woodruff. Joseph Woodruff (1820-1882) [1820-1886] son of William Woodruff. Helena Woodruff (1828-1892) daughter of William Woodruff. Julia Woodruff (1825-1870) daughter of William Woodruff. Dr. William Woodruff (1830-1908) son of William Woodruff. Margaret Clements [Clement] Woodruff (1794-1882) wife of William Woodruff

Genealogical and Historical Sketch of the Name and Family of Woodruff

Genealogical and Historical Sketch of the name and family of Woodruff compiled by the Media Research Bureau of Washington, D.C. This document traces the Woodruff name to its Anglo-Saxon origins and lists members of the family who have distinguished themselves in America in more recent times, n.d.

Structural bioinformatics analysis of the family of human ubiquitin-specific proteases

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

Systematics and biology of Fishes of family of the hemiramphidae of Cochin coast

This overall focus of the thesis involves the systematics and biology of fishes of the family hemiramphidae of cochin coast.India is one of the leading fish producing nations in the world with an average annual production of 6.1 million tonnes of fish and shell fish from capture and culture fisheries in 2001 (Ayyappan and Biradar, 2002).Fisheries play a very significant role in the Indian economy by providing employment to nearly 7 million people directly or indirectly, supplying rich protein food and earning valued foreign exchange.Fishes of the family Hemiramphidae are commonly called ‘half beaks‘.In India, studies on hemiramphids commenced with the work of Day (1878,1889) who recorded thirteen species of hemiramphids from the Indian waters.The study area, which is part of Cochin coast is located between Lat.9°28’ and 10° N and Long.76° 13’ and 76° 31 E. Lying parallel to it is an estuary which is commonly called the Cochin backwaters which has a total area of about 200 sq.miles.The study area is subjected to wide variations in salinity from place to place, season and surface to bottom.The Cochin coast and the adjacent back water system of Kerala has a rich and diversified fish fauna. The hemiramphid fishes constitute a minor fishery of this area.The study on the distribution and availability of hemiramphid fishes present in the Cochin coast shows that they evince different patterns of distribution.In the present study it is noticed that fecundity has high correlation with weight than length of the fish.Histological studies revealed that the spermatogenesis in both H. (H) limbatus and H.(H) xanthopterus, consists of spermatogonia, primary spermatocytes secondary, spermatocytes, spermatids and spermatozoa whereas in the oogenesis of both the species.biological study will be useful in implementation of proper measures of conservation and management so that further devastation of the hemiramphids of Cochin Coast can be controlled.

Spin-Peierls vs Peierls distortions in a family of conjugated polymers

Distortions in a family of conjugated polymers are studied using two complementary approaches: within a many-body valence bond approach using a transfer-matrix technique to treat the Heisenberg model of the systems, and also in terms of the tight-binding band-theoretic model with interactions limited to nearest neighbors. The computations indicate that both methods predict the presence or absence of the same distortions in most of the polymers studied.