Design of polyzinc finger peptides with structured linkers

Zinc finger domains are perhaps the most versatile of all known DNA binding domains. By fusing up to six zinc finger modules, which normally recognize up to 18 bp of DNA, designer transcription factors can be produced to target unique sequences within large genomes. However, not all continuous DNA sequences make good zinc finger binding sites. To avoid having to target unfavorable DNA sequences, we designed multizinc finger peptides with linkers capable of spanning long stretches of nonbound DNA. Two three-finger domains were fused by using either transcription factor IIIA for the Xenopus 5S RNA gene (TFIIIA) finger 4 or a non-sequence-specific zinc finger as a “structured” linker. Our gel-shift results demonstrate that these peptides are able to bind with picomolar affinities to target sequences containing 0–10 bp of nonbound DNA. Furthermore, these peptides display greater sequence selectivity and bind with higher affinity than similar six-finger peptides containing long, flexible linkers. These peptides are likely to be of use in understanding the behavior of polydactyl proteins in nature and in the targeting of human, animal, or plant genomes for numerous applications. We also suggest that in certain polydactyl peptides an individual finger can “flip” out of the major groove to allow its neighbors to bind shorter, nontarget DNA sequences.

Patterns of variance in stage-structured populations: Evolutionary predictions and ecological implications

Variability in population growth rate is thought to have negative consequences for organism fitness. Theory for matrix population models predicts that variance in population growth rate should be the sum of the variance in each matrix entry times the squared sensitivity term for that matrix entry. I analyzed the stage-specific demography of 30 field populations from 17 published studies for pattern between the variance of a demographic term and its contribution to population growth. There were no instances in which a matrix entry both was highly variable and had a large effect on population growth rate; instead, correlations between estimates of temporal variance in a term and contribution to population growth (sensitivity or elasticity) were overwhelmingly negative. In addition, survivorship or growth sensitivities or elasticities always exceeded those of fecundity, implying that the former two terms always contributed more to population growth rate. These results suggest that variable life history stages tend to contribute relatively little to population growth rates because natural selection may alter life histories to minimize stages with both high sensitivity and high variation.

Interaction of GroEL with a highly structured folding intermediate: iterative binding cycles do not involve unfolding.

The GroE proteins are molecular chaperones involved in protein folding. The general mechanism by which they facilitate folding is still enigmatic. One of the central open questions is the conformation of the GroEL-bound nonnative protein. Several suggestions have been made concerning the folding stage at which a protein can interact with GroEL. Furthermore, the possibility exists that binding of the nonnative protein to GroEL results in its unfolding. We have addressed these issues that are basic for understanding the GroE-mediated folding cycle by using folding intermediates of an Fab antibody fragment as molecular probes to define the binding properties of GroEL. We show that, in addition to binding to an early folding intermediate, GroEL is able to recognize and interact with a late quaternary-structured folding intermediate (Dc) without measurably unfolding it. Thus, the prerequisite for binding is not a certain folding stage of a nonnative protein. In contrast, general surface properties of nonnative proteins seem to be crucial for binding. Furthermore, unfolding of a highly structured intermediate does not necessarily occur upon binding to GroEL. Folding of Dc in the presence of GroEL and ATP involves cycles of binding and release. Because in this system no off-pathway reactions or kinetic traps are involved, a quantitative analysis of the reactivation kinetics observed is possible. Our results indicate that the association reaction of Dc and GroEL in the presence of ATP is rather slow, whereas in the absence of ATP association is several orders of magnitude more efficient. Therefore, it seems that ATP functions by inhibiting reassociation rather than promoting release of the bound substrate.

Electoral Systems and Ethnic Conciliation: A Structured, Focused Analysis of Vote-Pooling in Northern Ireland Elections 1998-2011

This research project examines the role of electoral system rules in affecting the extent of conciliatory behavior and cross-ethnic coalition making in Northern Ireland. It focuses on the role of the Single Transferable Vote (STV) electoral system in shaping party and voter incentives in a post-conflict divided society. The research uses a structured, focused comparison of the four electoral cycles since the Belfast Agreement of 1998. This enables a systematic examination of each electoral cycle using a common set of criteria focused on conciliation and cross-ethnic coalition making. Whilst preference voting is assumed to benefit moderate candidates, in Northern Ireland centrist and multi-ethnic parties outside of the dominant ethnic communities have received little electoral success. In Northern Ireland the primary effect of STV has not been to encourage inter-communal voting but to facilitate intra-community and intra-party moderation. STV has encouraged the moderation of the historically extreme political parties in each of the ethnic bloc. Patterns across electoral cycles suggest that party elites from the Democratic Unionist Party (DUP) and Sinn Fein have moderated their policy positions due to the electoral system rules. Therefore they have pursued lower-preference votes from within their ethnic bloc but in doing so have marginalized parties of a multi-ethnic or non-ethnic orientation.

A Transdiagnostic Social Skills Intervention; Using a Structured Story to Teach Children How to Ask a Friend to Play

An (independent samples comparison) controlled study was conducted to assess the efficacy of a novel approach to social skills training for children in a local socialization group at Knippenberg, Patterson & Associates (KPA). The treatment condition involved the combination of a Structured Story (i.e., novel bibliotherapy technique for children with social skills deficits), and a behavioral rehearsal (or role-play) segment, where the children practiced the target social skill featured in the Structure Story. The control group did not receive the Structured Story nor the behavioral rehearsal. Children in both groups engaged in ten-minutes of free play that was videorecorded for later observation and scoring by the principal investigator. Two target behaviors were assessed; asking a friend to play, and duration of joint play between two or more peers. The results did not show significant differences for either target variable between the group that received the novel intervention and the control group. Limitations of the current study and implications for further research are discussed.

3D plane-based egomotion for SLAM on semi-structured environment

Several works deal with 3D data in SLAM problem. Data come from a 3D laser sweeping unit or a stereo camera, both providing a huge amount of data. In this paper, we detail an efficient method to extract planar patches from 3D raw data. Then, we use these patches in an ICP-like method in order to address the SLAM problem. Using ICP with planes is not a trivial task. It needs some adaptation from the original ICP. Some promising results are shown for outdoor environment.

Self-heating function of carbon nanofiber cement pastes

The viability of carbon nanofiber (CNF) composites in cement matrices as a self-heating material is reported in this paper. This functional application would allow the use of CNF cement composites as a heating element in buildings, or for deicing pavements of civil engineering transport infrastructures, such as highways or airport runways. Cement pastes with the addition of different CNF dosages (from 0 to 5% by cement mass) have been prepared. Afterwards, tests were run at different fixed voltages (50, 100 and 150V), and the temperature of the specimens was registered. Also the possibility of using a casting method like shotcrete, instead of just pouring the fresh mix into the mild (with no system’s efficiency loss expected) was studied. Temperatures up to 138 °C were registered during shotcrete-5% CNF cement paste tests (showing initial 10 °C/min heating rates). However a minimum voltage was required in order to achieve a proper system functioning.

Structured carbons as supports for hydrogenation hybrid catalysts prepared by the immobilization of a Rh diamine complex

A CNF-monolith sample (carbon nanofibres grown on a ceramic monolith), and a granular carbon xerogel have been used as supports for hybrid catalysts where the active species is an Rh diamine complex. The advantages of these supports are their open porous structure and their morphology, which make catalyst handling easier and avoid difficult separation processes. The obtained catalysts are noticeably more active than the homogeneous Rh complex and are stable against leaching. At first use, partial reduction of the Rh complex takes place and nanometer-sized Rh particles develop, which increases the catalyst activity. Despite the open porous structure, mass transport limitations are present, especially in the case of the carbon xerogel based catalyst. Differences in internal mass transfer limitations are essentially due to the different diffusional path lengths.

The Developmental Experiences of Adolescent Females in Structured Basketball Programs

The purpose of the present study was to determine whether adolescent females had unique developmental experiences in different types of basketball programs. The Youth Experiences Survey 2.0 [YES] (Hansen & Larson, 2005) was used to measure the learning experiences of 14 and 15 year old females (n = 212) who were enrolled in a school, recreational, or competitive basketball program. Interviews with organization representatives were conducted to determine the structure of each basketball program (n= 16) from which participants were drawn. One-way ANOVAs and Bonferroni comparisons were used to compare YES 2.0 positive experience scale scores of participants in school, recreational and competitive basketball programs. Results revealed that females in recreational programs had significantly lower scores than those in competitive and school programs on numerous positive experiences scales. Mann-Whitney U tests found that those in school and competitive programs reported higher stress levels. Interview results indicate that four characteristics of competitive and school programs may contribute to participants in these programs reporting more growth experiences: 1) time commitment, 2) coaches’ training and background, 3) competition, and 4) volunteer opportunities.

The Contribution of Structured Activity and Deliberate Play to the Development of Expert Perceptual and Decision-Making Skill

The developmental histories of 32 players in the Australian Football League (AFL), independently classified as either expert or less skilled in their perceptual and decision- making skills, were collected through a structured interview process and their year-on-year involvement in structured and deliberate play activities retrospectively determined. Despite being drawn from the same elite level of competition, the expert decision-makers differed from the less skilled in having accrued, during their developing years, more hours of experience in structured activities of all types, in structured activities in invasion-type sports, in invasion-type deliberate play, and in invasion activities from sports other than Australian football. Accumulated hours invested in invasion-type activities differentiated between the groups, suggesting that it is the amount of invasion-type activity that is experienced and not necessarily intent (skill development or fun) or specificity that facilitates the development of perceptual and decision-making expertise in this team sport.

Structured Sports and Physical Activities: Their Critical Role

Even though one recognizes the physical and psychological kindness of h more and more involvement of the children to the physical and sporty activities, these programs seem more and more inaccessible to some. This article underlines the importance school déprogrammes centered on structured physical and sporty activities. In the setting of a recent survey, one asked youngsters to 5th year to take in note all their activities (as diary calendar) during two days of consecutive week. L·s children that participated in structured physical and sporty activities before and after the hours class sembkient to pull a lot more phisir of their day that those that didn't participate in these activities. Besides, seuk the young that took to structured physical and sporty activities reached the rates recommended of daily physical activity. The article proposes to the school administrators, to the parents, to the children and to the collectivities various measures encouraging the adoption of additional programs centered on the physical and sporty activities in school environment.

Military Capabilities: From Pooling & Sharing to a Permanent and Structured Approach. Security Policy Brief No. 37, September 2012

In 2013 the European Council for the first time since long will deal with European defence. An excellent opportunity: to move key Pooling & Sharing projects to the implementation stage in the short term, and to launch a permanent and structured approach to the development of European military capabilities for the long term.