Two recombination-dependent DNA replication pathways of bacteriophage T4, and their roles in mutagenesis and horizontal gene transfer

Two major pathways of recombination-dependent DNA replication, “join-copy” and “join-cut-copy,” can be distinguished in phage T4: join-copy requires only early and middle genes, but two late proteins, endonuclease VII and terminase, are uniquely important in the join-cut-copy pathway. In wild-type T4, timing of these pathways is integrated with the developmental program and related to transcription and packaging of DNA. In primase mutants, which are defective in origin-dependent lagging-strand DNA synthesis, the late pathway can bypass the lack of primers for lagging-strand DNA synthesis. The exquisitely regulated synthesis of endo VII, and of two proteins from its gene, explains the delay of recombination-dependent DNA replication in primase (as well as topoisomerase) mutants, and the temperature-dependence of the delay. Other proteins (e.g., the single-stranded DNA binding protein and the products of genes 46 and 47) are important in all recombination pathways, but they interact differently with other proteins in different pathways. These homologous recombination pathways contribute to evolution because they facilitate acquisition of any foreign DNA with limited sequence homology during horizontal gene transfer, without requiring transposition or site-specific recombination functions. Partial heteroduplex repair can generate what appears to be multiple mutations from a single recombinational intermediate. The resulting sequence divergence generates barriers to formation of viable recombinants. The multiple sequence changes can also lead to erroneous estimates in phylogenetic analyses.

Organ-specific and Agamous-regulated expression and glycosylation of a pollen tube growth-promoting protein.

Transmitting tissue-specific (TTS) protein is a pollen tube growth-promoting and attracting glycoprotein located in the stylar transmitting tissue extracellular matrix of the pistil of tobacco. The TTS protein backbones have a deduced molecular mass of about 28 kDa, whereas the glycosylated stylar TTS proteins have apparent molecular masses ranging between 50 and 100 kDa. TTS mRNAs and proteins are ectopically produced in transgenic tobacco plants that express either a cauliflower mosaic virus (CaMV) 35S promoter-TTS2 transgene or a CaMV 35S-promoter-NAG1 (NAG1 = Nicotiana tabacum Agamous gene) transgene. However, the patterns of TTS mRNA and protein accumulation and the quality of the TTS proteins produced are different in these two types of transgenic plants. In 35S-TTS transgenic plants, TTS mRNAs and proteins accumulate constitutively in vegetative and floral tissues. However, the ectopically expressed TTS proteins in these transgenic plants accumulate as underglycosylated protein species with apparent molecular masses between 30 and 50 kDa. This indicates that the capacity to produce highly glycosylated TTS proteins is restricted to the stylar transmitting tissue. In 35S-NAG transgenic plants, NAG1 mRNAs accumulate constitutively in vegetative and floral tissues, and TTS mRNAs are induced in the sepals of these plants. Moreover, highly glycosylated TTS proteins in the 50- to 100-kDa molecular mass range accumulate in the sepals of these transgenic, 35S-NAG plants. These results show that the tobacco NAGI gene, together with other yet unidentified regulatory factors, control the expression of TTS genes and the cellular capacity to glycosylate TTS proteins, which are normally expressed very late in the pistil developmental pathway and function in the final stage of floral development. The sepals in the transgenic 35S-NAG plants also support efficient pollen germination and tube growth, similar to what normally occurs in the pistil, and this ability correlates with the accumulation of the highest levels of the 50- to 100-kDa glycosylated TTS proteins.

Trinucleotide repeats and long homopeptides in genes and proteins associated with nervous system disease and development.

Several human neurological disorders are associated with proteins containing abnormally long runs of glutamine residues. Strikingly, most of these proteins contain two or more additional long runs of amino acids other than glutamine. We screened the current human, mouse, Drosophila, yeast, and Escherichia coli protein sequence data bases and identified all proteins containing multiple long homopeptides. This search found multiple long homopeptides in about 12% of Drosophila proteins but in only about 1.7% of human, mouse, and yeast proteins and none among E. coli proteins. Most of these sequences show other unusual sequence features, including multiple charge clusters and excessive counts of homopeptides of length > or = two amino acid residues. Intriguingly, a large majority of the identified Drosophila proteins are essential developmental proteins and, in particular, most play a role in central nervous system development. Almost half of the human and mouse proteins identified are homeotic homologs. The role of long homopeptides in fine-tuning protein conformation for multiple functional activities is discussed. The relative contributions of strand slippage and of dynamic mutation are also addressed. Several new experiments are proposed.

Neurotrophin 3 rescues neuronal precursors from apoptosis and promotes neuronal differentiation in the embryonic metanephric kidney.

We analyzed the developmental regulation and role of the neurotrophins during metanephric kidney morphogenesis. RNase protection assay revealed the presence of nerve growth factor, neurotrophin 3 (NT-3), and brain-derived neurotrophic factor mRNAs and the regulation of their expression during embryonic development of rat metanephros. NT-3 induced differentiation (neurite outgrowth) and survival (inhibition of apoptosis) of the neuronal precursors in cultured nephrogenic mesenchymes and neuronal differentiation in cultured whole kidneys, whereas NT-4/5, brain-derived neurotrophic factor, and nerve growth factor were without effect. The neurotrophins did not trigger tubular differentiation of isolated nephrogenic cells, which underwent apoptosis when cultured with or without the neurotrophins. NT-3 is thus an inducer of differentiation and a survival factor for renal neuronal cells, but none of the neurotrophins is a morphogen in kidney tubule induction.

Transcription factor GATA-1 permits survival and maturation of erythroid precursors by preventing apoptosis.

The transcription factor GATA-1 recognizes a consensus motif present in regulatory regions of numerous erythroid-expressed genes. Mouse embryonic stem cells lacking GATA-1 cannot form mature red blood cells in vivo. In vitro differentiation of GATA-1- embryonic stem cells gives rise to a population of committed erythroid precursors that exhibit developmental arrest and death. We show here that the demise of GATA-1- erythroid cells is accompanied by several features characteristics of apoptosis. This process occurs despite normal expression of all known GATA target genes examined, including the erythropoietin receptor, and independent of detectable accumulation of the tumor suppressor protein p53. Thus, in addition to its established role in regulating genes that define the erythroid phenotype, GATA-1 also supports the viability of red cell precursors by suppressing apoptosis. These results illustrate the multifunctional nature of GATA-1 and suggest a mechanism by which other hematopoietic transcription factors may ensure the development of specific lineages.

The product of the ataxia-telangiectasia group D complementing gene, ATDC, interacts with a protein kinase C substrate and inhibitor.

Ataxia-telangiectasia (AT) is an autosomal recessive human genetic disease characterized by immunological, neurological, and developmental defects and an increased risk of cancer. Cells from individuals with AT show sensitivity to ionizing radiation, elevated recombination, cell cycle abnormalities, and aberrant cytoskeletal organization. The molecular basis of the defect is unknown. A candidate AT gene (ATDC) was isolated on the basis of its ability to complement the ionizing radiation sensitivity of AT group D fibroblasts. Whether ATDC is mutated in any AT patients is not known. We have found that the ATDC protein physically interacts with the intermediate-filament protein vimentin, which is a protein kinase C substrate and colocalizing protein, and with an inhibitor of protein kinase C, hPKCI-1. Indirect immunofluorescence analysis of cultured cells transfected with a plasmid encoding an epitope-tagged ATDC protein localizes the protein to vimentin filaments. We suggest that the ATDC and hPKCI-1 proteins may be components of a signal transduction pathway that is induced by ionizing radiation and mediated by protein kinase C.

On the Edge of the World: Trauma and Twinship Selfobject Needs in Mrs. Dalloway

Alienation and aloneness appear as common themes in the experience of those impacted by trauma. Self psychology theorists, including contemporary proponents of intersubjectivity theory, have also discussed the ways in which alienation and disconnection from others permeate the experience of post-traumatic stress disorder. This discussion has highlighted the importance and centrality of twinship selfobject needs in providing a relational home for the emotional pain associated with trauma. These phenomena are especially apparent when one encounters the experiences of those combat veterans who have attempted to readjust to society upon returning home from military service. Using self psychology and intersubjectivity theory, this paper explores the ways that fiction, specifically Virginia Woolf's Mrs. Dalloway, can illuminate the ways that trauma negatively impacts twinship selfobject needs in combat veterans. In examining the character of Septimus Smith, this paper illustrates the estrangement, singularity, and alienation associated with post-traumatic stress, and how this state of being can collude with societal misunderstanding and repression to shatter the self's sense of belongingness with and connection to others.

Cultural conflict: Conveyance of traditional Iranian values and the effects on sense of self of Iranian-American women

The literature on Iranians who immigrated to countries with modern cultures is sparse. This paper presents a theoretical understanding of the research with a particular appreciation on issues related to gender roles, sexuality, and acculturation within this population. In addition, treatment suggestions with this Iranian immigrant population will be presented through the lens of self psychology and a new perspective about the effects on the sense of self of young immigrant Iranian females will be discussed.

A Student's Perspective on Graduate Training in Sport Psychology

This article aims to add to the limited literature related to graduate training in applied sport psychology – specifically from the perspective of a current graduate student. The paper begins with a discussion about the evolving definitions of sport psychology and applied sport psychology as well as a brief history of sport psychology. Next, reasons why graduate students pursue the fields of sport psychology and applied sport psychology along with how this impacts their decisions related to graduate training are examined. Then, literature as well as personal experiences from the author related to education, supervision, practica and internships, mentorship, and certification are explored. Finally, recommendations are provided for students considering graduate training in applied sport psychology.

Evaluation of hybrid and distance education learning environments in Spain

This article describes the adaptation and validation of the Distance Education Learning Environments Survey (DELES) for use in investigating the qualities found in distance and hybrid education psycho-social learning environments in Spain. As Europe moves toward post-secondary student mobility, equanimity in access to higher education, and more standardised degree programs across the European Higher Education Area (EHEA) the need for a high quality method for continually assessing the excellence of distance and hybrid learning environments has arisen. This study outlines how the English language DELES was adapted into the new Spanish-Distance Education Learning Environments Survey (S-DELES) for use with a Bachelor of Psychology and Criminology degree program offering both distance and hybrid education classes. We present the relationships between psycho-social learning environment perceptions and those of student affect. We also present the asynchronous aspects of the environment, scale means, and a comparison between the perceptions of distance education students and their hybrid education counterparts that inform the university about the baseline health of the information and communication technologies (ICT) environment within which the study was conducted.

Culturable aerobic and facultative bacteria from the gut of the polyphagic dung beetle Thorectes lusitanicus Jeckel

Unlike other dung beetles, the Iberian geotrupid Thorectes lusitanicus exhibits polyphagous behavior; for example, it is able to eat acorns, fungi, fruits, and carrion in addition to the dung of different mammals. This adaptation to digest a wider diet has physiological and developmental advantages and requires key changes in the composition and diversity of the beetle's gut microbiota. In this study, we isolated aerobic, facultative anaerobic, and aerotolerant microbiota amenable to grow in culture from the gut contents of T. lusitanicus and resolved isolate identity to the species level by sequencing 16S rRNA gene fragments. Using BLAST similarity searches and maximum likelihood phylogenetic analyses, we were able to reveal that the analyzed fraction (culturable, aerobic, facultative anaerobic, and aerotolerant) of beetle gut microbiota is dominated by the phyla Proteobacteria, Firmicutes and Actinobacteria. Among Proteobacteria, members of the order Enterobacteriales (Gammaproteobacteria) were the most abundant. The main functions associated with the bacteria found in the gut of T. lusitanicus would likely include nitrogen fixation, denitrification, detoxification, and diverse defensive roles against pathogens.

El humor verbal en niños de Educación Primaria: desarrollo de la conciencia metapragmática

Este trabajo constituye un estudio sobre el humor verbal en niños y niñas de 4.º curso de Educación Primaria (9-10 años). Tres grandes vías articulan el análisis. En primer lugar, el debate sobre las presuntas diferencias de género en la interpretación y el uso del humor verbal obtiene un interés especial en el caso de los niños, dado su menor nivel de «contaminación» por las convenciones y prejuicios sociales. En segundo lugar, se detecta la práctica ausencia de estudios sobre el humor infantil desde el punto de vista lingüístico, ya que el grueso de las investigaciones sobre el tema proviene de la psicología evolutiva. En tercer lugar, la etapa de edad objeto de este estudio responde a un periodo crucial en el desarrollo lingüístico y, especialmente, metalingüístico del niño.

An Integrative Definition of Coaching Effectiveness and Expertise

The purpose of the current paper is to present an integrative definition of coaching effectiveness and expertise that is both specific and conceptually grounded in the coaching, teaching, positive psychology, and athletes' development literature. The article is organized into six sections. The first section is used to situate the proposed definition in the predominant conceptual models of coaching. The second, third, and fourth sections provide detailed discussion about each of the three components of the proposed definition of coaching effectiveness: (a) coaches' knowledge, (b) athletes' outcomes, and (c) coaching contexts. The proposed definition is presented in the fifth section along with a clarification of common terminology and guiding postulates. The final section includes implications for practice and research.

Season-Long Examination of the Intervention Tone of Coach-Athlete Interactions and Athlete Development in Youth Sport

Objectives: Coaches are a primary influence on athletes' development in youth sport (Horn, 2008). However, the intervention tone of coaches' behaviour has not been directly observed. The purpose of this study was to examine associations between the intervention tone exhibited by youth sport coaches and athletes' individual developmental trajectories over the course of a season. Design: Short-term longitudinal study with behavioural observation. Method: Fifty-five athletes and their coaches from five youth volleyball teams were observed at three time points, and the intervention tone of interactive behaviour was systematically coded and organized by coach-athlete dyad. Athletes completed measures of the 4C's of athlete development (competence, confidence, connection, character) at each time point, which were used to create individualized developmental trajectories. Person-centred analyses were used to examine associations between athletes' developmental trajectories and their unique interactive experiences with their coach. Results: Cluster analysis revealed the presence of three distinct clusters based on athletes' developmental trajectories: 1) high and increasing, 2) low and decreasing, and 3) moderate and maintaining, with athletes from each team distributed across clusters. Analysis of dyadic interaction profiles revealed significant differences in interactive behaviour between clusters. Conclusions: Results suggest that differences in coach-athlete interactive experiences are associated with different developmental trajectories over the course of a season, even for athletes working with the same coach, highlighting the individualized nature of coaches' influence on young athletes. Practical implications for coaches include a critical awareness of their unique interactive relationship with each athlete independently, as well as the importance of fostering these relationships with regard to young people as more than just athletes.

Experience in Competitive Youth Sport and Needs Satisfaction: The Singapore Story

The purpose of this study was to examine the relationship between sport experiences and psychological needs satisfaction of Singapore high school athletes who were involved in inter-school competition. A total of 1250 school athletes from 22 sports participated in the study. The athletes were between 13 and 18 years old and had an average of 3 years of experience in school sport (SD=.18). Cluster analysis was employed to identify homogenous groups based on the seven developmental experiences domains of the Youth Experience Survey (YES 2.0; Hansen & Larson, 2005). A one-way analysis of variance (ANOVA) was conducted to determine whether differences existed among the clusters in terms of psychological needs satisfaction (i.e., sense of autonomy, perceived competence and relatedness). The results of the cluster analysis showed that there were different subgroups of athletes with distinct developmental experiences, and they varied in the degree to which their psychological needs were satisfied. Generally, subgroups that had high levels of positive experiences and low levels of negative experiences in sport had better fulfillment of psychological needs. It is important to ensure that policies and programmes are formulated, delivered and monitored effectively to promote positive experiences for youth who are involved in competitive sports.