Primary-secondary Transition Research Articles

Abstract 829: SAS1B protein: Determining whether ASTL or SAS1B has a role in tumor progression

Abstract The matrix metalloproteinase Sperm Acrosomal SLLP1 Binding (SAS1B) is an ideal molecular and cellular candidate for both female contraception and for cancer therapies. SAS1B translation is restricted among adult tissues to the ovary and oocytes, and SAS1B first appears in oocytes of follicles at the primary-secondary transition and also in tumors of varied types and stages. Research conducted using SAS1B antibodies suggested that the protein is localized at the cell membrane of cancer cells and can get internalized in a classical endocytosis assay and can be killed using an immunotoxin linked onto antibodies. The current study aims at deciphering whether the ASTL gene or the SAS1B protein is important for tumor progression, invasion and metastasis through cell biology and molecular biology tools and ensuring tumor cell-killing properties via an ADC-mediated killing using biochemistry and molecular biology tools. PCR, immunohistochemistry, Immunofluorescence, and Western blots revealed that a noncancerous breast cell line, MCF10A5E, is SAS1B null and can be used to study cancer-like properties once stably transfected with the SAS1B construct. Previous work in the lab indicated that an SAS1B-ADC strategy is a viable immunotherapy that could benefit patients who have failed to respond to chemotherapy, if indeed SAS1B plays a role in cancer development and progression. The outcomes of an SAS1B-ADC in a breast cancer model will be presented. Citation Format: Zunair S. Khokhar, Eusebio S. Pires, Alexander J. Kim, Ryan D'Souza. SAS1B protein: Determining whether ASTL or SAS1B has a role in tumor progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 829.

Pupils' Expectations and Experiences of PE across the Primary–secondary Transition in South Wales

This article was published in Cylchgrawn Addysg Cymru/Wales Journal of Education on 1 March 2018, available at https://doi.org/10.16922/wje.20.1.6

“It’s cool but challenging” The Relevance of Basic Need Fulfillment for Students’ School Enjoyment and Emotional Experiences at the Transition from Primary to Secondary Education

Based on the theoretical perspective of self-determination, this study examined the effects of basic need fulfillment (relatedness/competence/autonomy) on students’ school enjoyment before and after the transition from primary to secondary school. Furthermore, the academic and social impact of control beliefs on the emotions triggered by the upcoming transition was tested. The sample consisted of 63 students who filled out questionnaires at different time points before (grade 4; students between the ages of 9 and 10) and after (grade 5) the primary-secondary transition. The results highlight the importance of a positive teacher-student relationship as well as positive relationships amongst the students themselves for students’ school enjoyment before and after the transition. In terms of the emotions evoked by the upcoming transition, the results reveal that students with higher academic self-concept experienced more positive emotions than students with lower academic self-concept. From these findings, it can be concluded that positive relationships and the belief in one’s own academic capabilities can contribute to the likelihood of experiencing the transition to secondary school as an event accompanied by positive emotions such as joy and excitement rather than a stressful or threatening life-event.

Correction: Membrane associated cancer-oocyte neoantigen SAS1B/ovastacin is a candidate immunotherapeutic target for uterine tumors.

The metalloproteinase SAS1B [ovastacin, ASTL, astacin-like] was immunolocalized on the oolemma of ovulated human oocytes and in normal ovaries within the pool of growing oocytes where SAS1B protein was restricted to follicular stages spanning the primary-secondary follicle transition through ovulation. Gene-specific PCR and immunohistochemical studies revealed ASTL messages and SAS1B protein in both endometrioid [74%] and malignant mixed Mullerian tumors (MMMT) [87%] of the uterus. A MMMT-derived cell line, SNU539, expressed cell surface SAS1B that, after binding polyclonal antibodies, internalized into EEA1/LAMP1-positive early and late endosomes. Treatment of SNU539 cells with anti-SAS1B polyclonal antibodies caused growth arrest in the presence of active complement. A saporin-immunotoxin directed to SAS1B induced growth arrest and cell death. The oocyte restricted expression pattern of SAS1B among adult organs, cell-surface accessibility, internalization into the endocytic pathway, and tumor cell growth arrest induced by antibody-toxin conjugates suggest therapeutic approaches that would selectively target tumors while limiting adverse drug effects in healthy cells. The SAS1B metalloproteinase is proposed as a prototype cancer-oocyte tumor surface neoantigen for development of targeted immunotherapeutics with limited on-target/off tumor effects predicted to be restricted to the population of growing oocytes.

Expression of Sperm Acrosomal SLLP1 Binding (SAS1B) Protein in a Mammalian System for Crystallization

SAS1B (or Ovastacin) is considered an ideal candidate for both female contraception and for cancer therapies. SAS1B first appears in oocytes of follicles at the primary‐secondary transition in pre‐pubertal rodent ovaries and beyond. Quiescent oocytes within primordial and primary follicles are SAS1B negative. In 7 eutherian adult ovaries, the precise temporal and spatial onset of SAS1B expression was conserved. Crystal studies are a vital tool for drug design and for better understanding the molecular structure of targetable proteins, such as SAS1B. Full length SAS1B and its mature form (lacking the N terminus) were PCR‐cloned using gene specific primers sequence containing a kozak sequence at the N‐terminus and six histidine tags at the C‐terminus, using the TOPO cloning strategy. The PCR products were introduced into a pcDNA 3.4 vector and transformed into chemically competent E. Coli cells. Cells were plated on ampicillin infused Luria agar plates and colonies were allowed to grow overnight. Colonies were inoculated in fresh ampicillin containing Luria broth and DNA was extracted using a miniprep kit. Eluted DNA was sent for DNA sequencing. One of the full length clones, clone #9, showed the correct insert for full length SAS1B while screening of E. Coli colonies for the correct sequence for matured SAS1B is ongoing. Subsequently, once the correct sequences have been confirmed through sequencing, transfection of the correct vector‐insert species will be conducted using Expi293 mammalian cells, followed by protein isolation, detection and purification for the purpose of obtaining soluble SAS1B to generate crystals.

School belongingness and mental health functioning across the primary-secondary transition in a mainstream sample: multi-group cross-lagged analyses.

The relationship between school belongingness and mental health functioning before and after the primary-secondary school transition has not been previously investigated in students with and without disabilities. This study used a prospective longitudinal design to test the bi-directional relationships between these constructs, by surveying 266 students with and without disabilities and their parents, 6-months before and after the transition to secondary school. Cross-lagged multi-group analyses found student perception of belongingness in the final year of primary school to contribute to change in their mental health functioning a year later. The beneficial longitudinal effects of school belongingness on subsequent mental health functioning were evident in all student subgroups; even after accounting for prior mental health scores and the cross-time stability in mental health functioning and school belongingness scores. Findings of the current study substantiate the role of school contextual influences on early adolescent mental health functioning. They highlight the importance for primary and secondary schools to assess students’ school belongingness and mental health functioning and transfer these records as part of the transition process, so that appropriate scaffolds are in place to support those in need. Longer term longitudinal studies are needed to increase the understanding of the temporal sequencing between school belongingness and mental health functioning of all mainstream students.

SAS1B protein [ovastacin] shows temporal and spatial restriction to oocytes in several eutherian orders and initiates translation at the primary to secondary follicle transition

Sperm Acrosomal SLLP1 Binding (SAS1B) protein (ovastacin) is an oolemmal binding partner for the intra-acrosomal sperm protein SLLP1. Immunohistochemical localization revealed that SAS1B translation is restricted among adult tissues to the ovary and oocytes, SAS1B appearing first in follicles at the primary-secondary transition. Quiescent oocytes within primordial follicles and primary follicles did not stain for SAS1B. Examination of neonatal rat ovaries revealed SAS1B expression first as faint signals in postnatal day 3 oocytes, with SAS1B protein staining intensifying with oocyte growth. Irrespective of animal age or estrus stage, SAS1B was seen only in oocytes of follicles that initiated a second granulosa cell layer. The precise temporal and spatial onset of SAS1B expression was conserved in adult ovaries in seven eutherian species, including nonhuman primates. Immunoelectron micrographs localized SAS1B within cortical granules in MII oocytes. A population of SAS1B localized on the oolemma predominantly in the microvillar region anti-podal to the nucleus in ovulated MII rat oocytes and on the oolemma in macaque GV oocytes. The restricted expression of SAS1B protein in growing oocytes, absence in the ovarian reserve, and localization on the oolemma suggest this zinc metalloprotease deserves consideration as a candidate target for reversible female contraceptive strategies.

Primary–secondary transition: Differences between teachers’ and children’s perceptions

Transition from primary to secondary school is an important but neglected topic. For this review, 88 studies were selected from 325 possible studies, as including substantive data related to transition. The teacher’s perspective and the child’s perspective were very different, the former principally concerned with attainment and the latter principally concerned with socio-emotional issues. Children were concerned with peer relations and bullying, self-esteem and external support networks. Teachers were concerned with the attainment dip on entry to secondary school, curriculum problems, school strategies to ameliorate these, special groups and children with disability. The secondary school was a problem for all children at first, but after a term many children adjusted, though 40 percent still struggled after a year. The difficulty was greater for children from homes in poverty or ethnic minorities, particularly where parental encouragement was lacking. The quality of the evidence was critiqued and evidence-based implications for practice and policy and future research were outlined.

Using creative drama to facilitate primary–secondary transition

Primary–secondary transition can be a period of extreme anxiety for some children, with lowering of self-esteem and confidence. Creative drama has been reported to enhance self-esteem and confidence. This paper explores the possibility of using drama to facilitate successful transition. It focuses on secondary datasets from six primary schools in a local authority in Scotland (357 pupils and 12 teachers) to explore the effectiveness of creative drama. It then reports data collected from four drama professionals to explore whether drama can be effective in facilitating successful transition and the reasons behind that. The data suggest that creative drama can be used effectively in transitions due to its potential to empower pupils, to facilitate an understanding of emotional issues, to provide realistic scenarios with a degree of distance and a safe space to rehearse real life transitions.

Assessing pupil concerns about transition to secondary school

The primary-secondary transition can negatively affect pupils' emotional and psychological adjustment. However, methods for assessing concerns regarding secondary school are limited. A reliable and valid measure of transition-related concerns would be useful both in evaluating and shaping the content and delivery of universal and targeted transition support programmes. To assess the validity of a quantitative self-report measure of school concerns as an assessment tool during the primary-secondary school transition. A UK sample of 147 Year 6 primary school pupils and 263 Year 7 secondary school pupils. Self-reports of school concerns and school liking as well as self-reported and peer assessments on a range of psychological adjustment measures were collected. The School Concerns Questionnaire (SCQ) showed good reliability at primary and secondary school. Secondary concerns reduced significantly post-transition. Three factors emerged from an exploratory factor analysis of the SCQ and the pattern of results was replicated for post-transition concerns. Pupils with higher school concerns at secondary school reported reduced liking of school and reduced trust and respect for teachers. The SCQ was associated with generalized anxiety and female gender both pre-and post-transition. Peer problems and depression were associated with pre- and post-transition concerns, respectively. The SCQ is a simple to complete, reliable, and valid tool for assessing primary-secondary transition concerns. It could serve a valuable role in the evaluation and development of universal and targeted school-based initiatives that aim to promote positive secondary transition.

Structure, expression differentiation and evolution of duplicated fiber developmental genes in Gossypium barbadense and G. hirsutum.

BackgroundBoth Gossypium hirsutum and G. barbadense probably originated from a common ancestor, but they have very different agronomic and fiber quality characters. Here we selected 17 fiber development-related genes to study their structures, tree topologies, chromosomal location and expression patterns to better understand the interspecific divergence of fiber development genes in the two cultivated tetraploid species.ResultsThe sequence and structure of 70.59% genes were conserved with the same exon length and numbers in different species, while 29.41% genes showed diversity. There were 15 genes showing independent evolution between the A- and D-subgenomes after polyploid formation, while two evolved via different degrees of colonization. Chromosomal location showed that 22 duplicate genes were located in which at least one fiber quality QTL was detected. The molecular evolutionary rates suggested that the D-subgenome of the allotetraploid underwent rapid evolutionary differentiation, and selection had acted at the tetraploid level. Expression profiles at fiber initiation and early elongation showed that the transcripts levels of most genes were higher in Hai7124 than in TM-1. During the primary-secondary transition period, expression of most genes peaked earlier in TM-1 than in Hai7124. Homeolog expression profile showed that A-subgenome, or the combination of A- and D-subgenomes, played critical roles in fiber quality divergence of G. hirsutum and G. barbadense. However, the expression of D-subgenome alone also played an important role.ConclusionIntegrating analysis of the structure and expression to fiber development genes, suggests selective breeding for certain desirable fiber qualities played an important role in divergence of G. hirsutum and G. barbadense.

A Challenge of Living? Understanding the Psycho-social Processes of the Child During Primary-secondary Transition Through Resilience and Self-esteem Theories

The transition from primary to secondary school can be a period of anxiety for many children. Although most schools have developed systems to ease this process, it has been argued that the emphasis is often on administrative and organisational procedures. In contrast, children and parents are typically more concerned with personal and social issues. It may be that such concerns have received insufficient attention in the past. This paper employs theoretical perspectives from the literature on resilience and self-esteem to examine key aspects of the process of transition from the perspective of the individual child. It is argued that for some children at least, transition can be legitimately considered ‘a challenge of living’ because of the social and personal messages which are received from a range of experiences within a concentrated period of time. The nature and source of these messages are discussed, and some implications for practitioners suggested. It is argued that in order to help vulnerable individuals cope with, and even benefit from, the period of transition, we need to focus more on the way social and personal experiences are interpreted at this time.

A holistic approach to primary—secondary transitions

A longitudinal study was conducted to follow children, in a city in Scotland, to understand their experiences of primary—secondary transition. The children were purposively selected as known to be having/potentially having problems with transition. Initial interviews were conducted with children, their parents and professionals when five were in the first year of secondary school, two in the second year of secondary school and two in the last year of primary school. Follow-up interviews were undertaken with six children again after six months when four were in the second year and two in the first year of the secondary school. Three out of the nine children were finding it difficult to deal with transition even at the follow-up. The issue of transition was complex, with considerable impact of a child's internal attributes, family, peers, school systems, professionals and community on his/her primary—secondary transition being successful or not.

Teachers’ views of the primary–secondary transition in music education in England

This article investigates the transition and transfer process from primary to secondary school in music from the teacher perspective. Thirteen secondary music teachers from 13 secondary schools throughout England were interviewed along with seven primary-school teachers representing five primary schools. This paper reports interview data from both groups of teachers, which together provide accounts of the transfer and transition process from both sides of what was referred to as ‘the humpback bridge’ in our previous report on the pupil data (Marshall and Hargreaves 2007). The research investigated teachers attitudes and current practice in the process of transfer and transition and explored whether or not the claims made in advance of the National Curriculum have been met through the creation of a common language and a progressive curriculum covering all ages and stages of education. Furthermore, we looked for any evidence of the continuing existence of the five approaches commonly used by secondary teachers to the introduction of music in the secondary school suggested 10 years ago by Mills (1996). Our results suggest that while some types of liaison activity between schools have increased, the majority of liaison carried out between primary- and secondary-school teachers in music specifically, tends to focus on administration of instrumental lessons. The results also suggest that, in common with findings from Mills (1996), most secondary teachers in music still feel compelled to start music in secondary school by ‘starting from scratch’. However, we further suggest that far from being viewed in a negative way, this approach can be developed into a beneficial experience enabling pupils to become familiar with and confident within their new secondary environment.

Genetic patterning of the developing mouse tail at the time of posterior neuropore closure

Posterior neuropore (PNP) closure coincides with the end of gastrulation, marking the end of primary neurulation and primary body axis formation. Secondary neurulation and axis formation involve differentiation of the tail bud mesenchyme. Genetic control of the primary-secondary transition is not understood. We report a detailed analysis of gene expression in the caudal region of day 10 mouse embryos during primary neuropore closure. Embryos were collected at the 27-32 somite stage, fixed, processed for whole mount in situ hybridisation, and subsequently sectioned for a more detailed analysis. Genes selected for study include those involved in the key events of gastrulation and neurulation at earlier stages and more cranial levels. Patterns of expression within the tail bud, neural plate, recently closed neural tube, notochord, hindgut, mesoderm, and surface ectoderm are illustrated and described. Specifically, we report continuity of expression of the genes Wnt5a, Wnt5b, Evx1, Fgf8, RARgamma, Brachyury, and Hoxb1 from primitive streak and node into subpopulations of the tail bud and caudal axial structures. Within the caudal notochord, developing floorplate, and hindgut, HNF3alpha, HNF3beta, Shh, and Brachyury expression domains correlate directly with known genetic roles and predicted tissue interdependence during induction and differentiation of these structures. The patterns of expression of Wnt5a, Hoxb1, Brachyury, RARgamma, and Evx1, together with observations on proliferation, reveal that the caudal mesoderm is organised at a molecular level into distinct domains delineated by longitudinal and transverse borders before histological differentiation. Expression of Wnt5a in the ventral ectodermal ridge supports previous evidence that this structure is involved in epithelial-mesenchymal interaction. These results provide a foundation for understanding the mechanisms facilitating transition from primary to secondary body axis formation, as well as the factors involved in defective spinal neurulation.

Xylary Union Between the New Shoot and Old Stem During Terminal Bud Break in Populus deltoides

Bursting terminal buds and subtending stems of Populus deltoides Bartr. ex Marsh. plants were examined at several stages to determine the pattern of xylary union between the 1st- and 2nd-yr growth increments. Metaxylem vessels differentiated first in traces serving the basalmost leaf in the bud. As successively younger leaves began growth, metaxylem vessels differentiated in their traces. The site of metaxylem reactivation was in the second or third internode beneath the bud (i.e., in the 1st-yr stem), and subsequent differentiation progressed bidirectionally in each set of traces as the leaves they served expanded. Traces leading either to abscised leaf positions on the 1st-yr stem or to bud-scale leaf positions were reactivated by the tangential “spread” of activity from adjacent traces serving expanding leaves. The new elements were all secondary xylem vessels, as were those of the basipetal trace components, although they were functionally continuous with the metaxylem vessels that differentiated acropetally. Xylem fibers were initiated in the same position and differentiated in the same sequence as vessels. However, fiber differentiation lagged behind that of vessels. Whereas vessel differentiation was associated with leaf expansion, fiber differentiation was associated with leaf maturation. As each leaf matured in sequence, the primary-secondary transition zone advanced acropetally to the bud base and then in the new shoot until it attained a positional relation with leaf maturation comparable to that of 1st-yr plants.

Expectation of Gifted Children in the Primary-Secondary Transition.

Despite the comprehensiveness of the creativity literature, there are few studies that deal directly with the perceptions of creative children in their everyday concerns. Increasingly, however, educators are turning to ethnomethodology as providing wider and more appropriate knowledge for their purposes.

Anatomy of the primary-secondary transition zone in stems of Populus deltoides

The transition from primary to secondary stem tissues occurs as a continuum, and a precise anatomical definition of the transition does not exist. A definition was derived for Populus deltoides based on the birefringent properties of the fiber wall. This definition was quantitatively reproducible in the 9 plants tested, and the secondary transition was found to occur in the internode associated with the first mature leaf from the apex. The primary-secondary transition did not occur uniformly around the periphery. It was first observed in the vascular bundles opposite the incoming trace, and from there it progressed in a counter-clockwise direction. Within the transition internode, each vascular bundle and each tissue comprising the bundle differentiated in accord with the physiological age and the phyllotactic disposition of the developing leaf to which it led. Within any one vascular bundle, differentiation occurred first in the metaxylem vessels and associated fibers, followed closely by extension of fibers into the interfascicular regions and centripetal differentiation of the phloem fibers. The ontogenetic sequence of differentiation for each of the principal tissues of the secondary transition zone is described.