Regulation and transcript analysis of the ceroperon responsible for quorum sensing in Rhodobacter sphaeroides 2.4.1

Rhodobacter sphaeroides 2.4.1 is a Gram negative facultative photoheterotrophic bacterium that has been shown to have an N-acyl homoserine lactone-based quorum sensing system called cer for c&barbelow;ommunity e&barbelow;scape r&barbelow;esponse. The cer ORFs are cerR, the transcriptional regulator, cerI, the autoinducer synthase and cerA , whose function is unknown. The autoinducer molecule, 7,8- cis-N-(tetradecenoyl) homoserine lactone, has been characterized. The objective of this study was to identify an environmental stimulus that influences the regulation of cerRAI and, to characterize transcription of the cer operon. ^ A cerR::lacZ transcriptional fusion was made and β-Galactosidase assays were performed in R. sphaeroides 2.4.1 strains, wild type, AP3 (CerI−) and AP4 (CerR−). The cerR::lacZ β-Galactosidase assays were used as an initial survey of the mode of regulation of the Cer system. A cerA::lacZ translational fusion was created and was used to show that cerA can be translated. The presence of 7,8-cis-N-(tetradecenoyl) homoserine lactone was detected from R. sphaeroides strains wild type and AP4 (CerR−) using a lasR::lacZ translational fusion autoinducer bioassay. The cerR::lacZ transcriptional fusion in R. sphaeroides 2.4.1 wild type was tested under different environmental stimuli, such as various carbon sources, oxygen tensions, light intensities and culture media to determine if they influence transcription of the cer ORFs. Although lacZ assay data implicated high light intensity at 100 W/m2 to stimulate cer transcription, quantitative Northern RNA data of the cerR transcript showed that low light intensity at 3 W/m2 is at least one environmental stimulus that induces cer transcription. This finding was supported by DNA microarray analysis. Northern analysis of the cerRAI transcript provided evidence that the cer ORFs are co-transcribed, and that the cer operon contains two additional genes. Bioinformatics was used to identify genes that may be regulated by the Cer system by identifying putative lux box homologue sequences in the presumed promoter region of these genes. Genes that were identified were fliQ, celB and calsymin, all implicated in interacting with plants. Primer extension was used to help localize cis-elements in the promoter region. The cerR::lacZ transcriptional fusion was monitored in a subset of different global DNA binding transcriptional regulator mutant strains of R. sphaeroides 2.4.1. Those regulators involved in maintaining an anaerobic photosynthetic lifestyle appeared to have an effect. Collectively, the data imply that R. sphaeroides 2.4.1 activates the Cer system when grown anaerobic photosynthetically at low light intensity, 3 W/m2, and it may be involved in an interaction with plants. ^

Emerging Identities in Colonial Tunisia: "Alliancist" and Zionist Representations in Tunis prior to World War I

By 1900 the Jewish community of Tunisia witnessed the emergence of new competing identities: “assimilationist” of the Alliance Israelite Universelle, termed “Alliancist,” and Zionist. Strikingly, two members of the same family in Tunis, Raymond Valensi, President of the AIU Regional Committee, and Alfred Valensi, President of the Zionist Federation, led the struggle for their separate causes. In his discussion of identity in the modern world, Homi Bhabha asks, "How do strategies of representation or empowerment come to be formulated in the competing claims of communities…where, despite shared histories of …discrimination, the exchange of values, meanings and priorities…may be profoundly antagonistic…?" It is in this context that the claims of the Alliance and Zionism will be examined prior to World War I, based on the Archives of the AIU and on such secondary sources as the indispensable work of Paul Sebag. The tensions between the Alliancists and Zionists continued until the outbreak of World War II, as the French-speaking Jews of Tunisia sought to define their individual and collective identities.

Lost in Translation: Emotion and Expression through Technology

My Thesis study was designed to bring to topic certain issues involved with CMC (computer-mediated communication.) Often we are presented with confusing or misleading situations when it comes to expressing our emotions through technological means. It is important that we are aware of certain issues such as the use of emoticons, expressing sarcasm, and the ongoing trend of Internet slang. These various aspects can create confusion in CMC, leading to a loss in translation. My survey study was designed to probe deeper into these issues by asking general questions and by analyzing sample CMC scenarios.

Applying the Strengths Perspective to Increase Safety and Weil-Being: Views from Families and Providers

Consensus about the value of the strengths perspective is developing among child welfare and family service practitioners. Yet, few first-hand reports are available from the perspectives of family members and interdisciplinary service providers about the principles most important for engaging and supporting family members to achieve needed outcomes. This paper briefly highlights principles most often cited as key to application of the strengths perspective and compares first-hand accounts from family members and service providers. These views were elicited through focus groups facilitated by a community-based family support program.Implications for strengths-based practice with families are discussed.

G -rich oligodeoxyribonucleotides: Triplexes and transcription

Formation of a triple helix resulting from oligonucleotide binding to the DNA double helix offers new possibilities to control gene expression at the transcriptional level. Purine-motif triplexes can be formed under physiological pH. Nevertheless, this formation was inhibited by certain monovalent cations during the association but not during dissociation. Since triplexes are very stable, it was possible to assemble them in the absence of KCl and have them survive throughout the course of an in vitro transcription reaction. As for the design of a better triplex-forming oligonucleotide, 12 nucleotides in length afforded the highest binding affinity. G/T-rich oligonucleotides can be very polymorphic in solution. The conditions for forming purine-motif triplexes, duplexes or G-quartets were determined. Understanding these parameters will be important for the practical use of G-rich oligonucleotides in the development of DNA aptamers where the structure of the oligonucleotide is paramount in dictating its function. Finally, purine-motif triplexes were demonstrated to significantly inhibit gene transcription in vitro. The optimal effect on this process was dependent on the location of triplexes within the promoter, i.e., whether upstream or proximally downstream of the transcription start site. The mechanism for the inhibition of transcription appeared to be interference with initiation through preventing engagement by RNA polymerase. This finding is revolutionary when compared to the conventional model where triplexes inhibit transcription only by occluding binding by trans-acting proteins. Our findings broaden the utility of triplexes and support a strategy for antigene therapy by triplexes. ^

Preliminary ichthyolith biostratigraphy and age profiles at DSDP Sites 91-595 and 91-596

Seventy meters of Cenozoic and Mesozoic pelagic clay cored at DSDP Sites 595 and 596 provide the basis for a preliminary analysis of ichthyolith biostratigraphy in the southwest Pacific. A most likely order of the more reliable ichthyolith events is compared with a synthesis of ichthyolith biostratigraphy in the North Pacific and with dated composite ranges. The resultant preliminary ichthyolith stratigraphy suggests that the Cenozoic is represented by the upper 20 m at Site 596 and 16 to 22 m at Site 595. Mixing of taxa precludes a clear recognition of the Cretaceous/Tertiary boundary at Site 595. The occurrence of 13 newly described subtypes is recorded in Mesozoic sediments at Sites 595 and 596. These new subtypes and previously described Mesozoic forms may be useful for recognizing Mesozoic subdivisions when their occurrences in sequences dated by other microfossils are investigated.

Nitration of γ-tocopherol and oxidation of α-tocopherol by copper-zinc superoxide dismutase/H2O2/NO2−: Role of nitrogen dioxide free radical

Copper-zinc superoxide dismutase (Cu,ZnSOD) is the antioxidant enzyme that catalyzes the dismutation of superoxide (O2•−) to O2 and H2O2. In addition, Cu,ZnSOD also exhibits peroxidase activity in the presence of H2O2, leading to self-inactivation and formation of a potent enzyme-bound oxidant. We report in this study that lipid peroxidation of l-α-lecithin liposomes was enhanced greatly during the SOD/H2O2 reaction in the presence of nitrite anion (NO2−) with or without the metal ion chelator, diethylenetriaminepentacetic acid. The presence of NO2− also greatly enhanced α-tocopherol (α-TH) oxidation by SOD/H2O2 in saturated 1,2-dilauroyl-sn-glycero-3-phosphatidylcholine liposomes. The major product identified by HPLC and UV-studies was α-tocopheryl quinone. When 1,2-diauroyl-sn-glycero-3-phosphatidylcholine liposomes containing γ-tocopherol (γ-TH) were incubated with SOD/H2O2/NO2−, the major product identified was 5-NO2-γ-TH. Nitrone spin traps significantly inhibited the formation of α-tocopheryl quinone and 5-NO2-γ-TH. NO2− inhibited H2O2-dependent inactivation of SOD. A proposed mechanism of this protection involves the oxidation of NO2− by an SOD-bound oxidant to the nitrogen dioxide radical (•NO2). In this study, we have shown a new mechanism of nitration catalyzed by the peroxidase activity of SOD. We conclude that NO2− is a suitable probe for investigating the peroxidase activity of familial Amyotrophic Lateral Sclerosis-linked SOD mutants.

Structure, Properties, and Tissue Localization of Apoplastic α-Glucosidase in Crucifers1

Apoplastic α-glucosidases occur widely in plants but their function is unknown because appropriate substrates in the apoplast have not been identified. Arabidopsis contains at least three α-glucosidase genes; Aglu-1 and Aglu-3 are sequenced and Aglu-2 is known from six expressed sequence tags. Antibodies raised to a portion of Aglu-1 expressed in Escherichia coli recognize two proteins of 96 and 81 kD, respectively, in vegetative tissues of Arabidopsis, broccoli (Brassica oleracea L.), and mustard (Brassica napus L.). The acidic α-glucosidase activity from broccoli flower buds was purified using concanavalin A and ion-exchange chromatography. Two active fractions were resolved and both contained a 96-kD immunoreactive polypeptide. The N-terminal sequence from the 96-kD broccoli α-glucosidase indicated that it corresponds to the Arabidopsis Aglu-2 gene and that approximately 15 kD of the predicted N terminus was cleaved. The 81-kD protein was more abundant than the 96-kD protein, but it was not active with 4-methylumbelliferyl-α-d-glucopyranoside as the substrate and it did not bind to concanavalin A. In situ activity staining using 5-bromo-4-chloro-3-indolyl-α-d-glucopyranoside revealed that the acidic α-glucosidase activity is predominantly located in the outer cortex of broccoli stems and in vascular tissue, especially in leaf traces.

Marketing and compromising for sustainability : Competing orders of worth in the North Atlantic

Funding The author(s) disclosed receipt of the following financial support for the research, authorship, and/ or publication of this article: The research upon which this article reports was funded by the Leverhulme Trust, grant F/00 273/N.

Interpreting prophecy and the appearing of Christ /

"Psalm of joy": p. [1] at end.

The souls of black folk : essays and sketches /

Of our spiritual strivings -- Of the dawn of freedom -- Of Mr. Booker T. Washington and others -- Of the meaning of progress -- Of the wings of Atalanta -- Of the training of black men -- Of the black belt -- Of the quest of the golden fleece -- Of the sons of master and man -- Of the faith of the fathers -- Of the passing of the first-born -- Of Alexander Crummell -- Of the coming of John -- The sorrow songs.