Mutations within the Primer Binding Site of the Human Immunodeficiency Virus Type 1 Define Sequence Requirements Essential for Reverse Transcription

AbstractThe primer binding site (PBS) is involved in two stages during the reverse transcription of the retroviral RNA genome. In the early stage, the PBS provides complementary sequences through which tRNALys,3binds the viral RNA genome to initiate minus-strand DNA synthesis; in the later stages, complementarity between the plus- and minus-strand copies of the PBS is required to facilitate the second template transfer needed to complete reverse transcription. We previously constructed a mutant HIV-1 proviral genome, designated as pHXB2PBS(pheC + 5) (now referred to as pheC + 5), which was used to identify regions of the PBS involved in the initiation and second template transfer steps of reverse transcription. To further define the sequence requirements of the PBS for the initiation of reverse transcription, we have made single nucleotide substitutions within the first six nucleotides of the pheC + 5 PBS. Our results demonstrate that mutations within the first five nucleotides of the PBS which disrupt base paring with tRNALys,3-PBS results in an noninfectious virus; a G-U base pair at position six of the tRNALys,3-PBS complex was tolerated. In contrast to the requirements for initiation, we found that complementary binding between only three base pairs of the plus- and minus-strand PBSs was required for the extension of plus-strand DNA during the second template transfer. Furthermore, regions of the minus-strand DNA of up to 24 nucleotides could be looped-out to facilitate the complementarity required for the completion of plus-strand DNA synthesis. Taken together, the results of our studies demonstrate that different features of the PBS with respect to RNA:RNA and DNA:DNA interactions are required for initiation of reverse transcription and the completion of plus-strand DNA synthesis, respectively

Spectroscopy of Rydberg atoms in non-neutral cold plasmas

The electric field in mm-sized one-component non-neutral plasmas is measured using the Stark effect of Rydberg atoms embedded in them. The plasmas are clouds of cold Rb+Rb+-ions, which are produced by UV photoionization of laser-cooled Rb atoms in a magneto-optic trap. The dependence of the electric field on the number of ions and the Coulomb explosion of the ion clouds have been studied. © 2002 American Institute of Physics.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87634/2/89_1.pd

Young children's cognitive achievement: home learning environment, language and ethnic background

For decades, research has shown differences in cognitive assessment scores between White and minority ethnic group(s) learners as well as differences across different minority ethnic groups. More recent data have indicated that the home learning environment and languages spoken can impact cognitive assessment and other corollary outcomes. This study uses the Millennium Cohort Study to jointly assess how minority ethnic group, home learning environment and home languages predict child cognitive assessment scores. Regression analyses were conducted using two assessment measures. The following is hypothesised: (1) cognitive achievement scores vary by minority ethnic group, (2) more home learning environment in early childhood leads to higher cognitive development scores and (3) English only in the home yields the highest cognitive scores while no English in the home yields the lowest. Findings reveal that there are differences in cognitive scores along ethnic group categories although there are also some unexpected findings. Home learning environment does not play as large a role as was predicted in raising the assessment scores overall for learners while speaking English in the home does, irrespective of ethnic background

Ethical and methodological issues in engaging young people living in poverty with participatory research methods

This paper discusses the methodological and ethical issues arising from a project that focused on conducting a qualitative study using participatory techniques with children and young people living in disadvantage. The main aim of the study was to explore the impact of poverty on children and young people's access to public and private services. The paper is based on the author's perspective of the first stage of the fieldwork from the project. It discusses the ethical implications of involving children and young people in the research process, in particular issues relating to access and recruitment, the role of young people's advisory groups, use of visual data and collection of data in young people's homes. The paper also identifies some strategies for addressing the difficulties encountered in relation to each of these aspects and it considers the benefits of adopting participatory methods when conducting research with children and young people

Rydberg-atom trajectories in a ponderomotive optical lattice

Using semiclassical simulations, we investigate the trajectories and the microwave spectra of Rydberg atoms excited in a ponderomotive optical lattice. We relate distinct features found in the microwave spectra to characteristic types of trajectory. Several methods are presented that are designed to greatly improve the trapping efficiency of the lattice and to generalize the trapping from one to three dimensions.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/85435/1/njp10_11_113036.pd

Childhood publics in search of an audience: reflections on the children’s environmental movement

The essay reflects on the children's environmental movement from the perspective of cultural theory, as well as the authors' own and others’ research on children’s encounters, experiences and engagement in public life. The concepts of political knowingness, childhood publics, and listening publics are evoked to think through the surprise that the children's environmental movement generated in the public sphere. The idiom is positioned as an audience ‘hearing aid’ for turning babbling into political messages. In so doing we find that the messages from the children’s environmental movement are not out of place in the current humanities and social sciences literatures on the Anthropocene

Ethanol Activation of Protein Kinase A Regulates GABAA Receptor Subunit Expression in the Cerebral Cortex and Contributes to Ethanol-Induced Hypnosis

Protein kinases are implicated in neuronal cell functions such as modulation of ion channel function, trafficking, and synaptic excitability. Both protein kinase C (PKC) and A (PKA) are involved in regulation of γ-aminobutyric acid type A (GABAA) receptors through phosphorylation. However, the role of PKA in regulating GABAA receptors (GABAA-R) following acute ethanol exposure is not known. The present study investigated the role of PKA in the effects of ethanol on GABAA-R α1 subunit expression in rat cerebral cortical P2 synaptosomal fractions. Additionally, GABA-related behaviors were examined. Rats were administered ethanol (2.0–3.5 g/kg) or saline and PKC, PKA, and GABAA-R α1 subunit levels were measured by western blot analysis. Ethanol (3.5 g/kg) transiently increased GABAA-R α1 subunit expression and PKA RIIβ subunit expression at similar time points whereas PKA RIIα was increased at later time points. In contrast, PKC isoform expression remained unchanged. Notably, lower ethanol doses (2.0 g/kg) had no effect on GABAA-R α1 subunit levels, although PKA type II regulatory subunits RIIα and RIIβ were increased at 10 and 60 min when PKC isozymes are also known to be elevated. To determine if PKA activation was responsible for the ethanol-induced elevation of GABAA-R α1 subunits, the PKA antagonist H89 was administered to rats prior to ethanol exposure. H89 administration prevented ethanol-induced increases in GABAA-R α1 subunit expression. Moreover, increasing PKA activity intracerebroventricularly with Sp-cAMP prior to a hypnotic dose of ethanol increased ethanol-induced loss of righting reflex (LORR) duration. This effect appears to be mediated in part by GABAA-R as increasing PKA activity also increased the duration of muscimol-induced LORR. Overall, these data suggest that PKA mediates ethanol-induced GABAA-R expression and contributes to behavioral effects of ethanol involving GABAA-R