Paracrine effects of oocyte secreted factors and stem cell factor on porcine granulosa and theca cells in vitro

Oocyte control of granulosa and theca cell function may be mediated by several growth factors via a local feedback loop(s) between these cell types. This study examined both the role of oocyte-secreted factors on granulosa and thecal cells, cultured independently and in co-culture, and the effect of stem cell factor (SCF); a granulosa cell derived peptide that appears to have multiple roles in follicle development. Granulosa and theca cells were isolated from 2-6 mm healthy follicles of mature porcine ovaries and cultured under serum-free conditions, supplemented with: 100 ng/ml LR3 IGF-1, 10 ng/ml insulin, 100 ng/ml testosterone, 0-10 ng/ml SCF, 1 ng/ml FSH (granulosa), 0.01 ng/ml LH (theca) or 1 ng/ml FSH and 0.01 ng/ml LH (co-culture) and with/without oocyte conditioned medium (OCM) or 5 oocytes. Cells were cultured in 96 well plates for 144 h, after which viable cell numbers were determined. Medium was replaced every 48 h and spent medium analysed for steroids.Oocyte secreted factors were shown to stimulate both granulosa cell proliferation (P < 0.001) and oestradiol production (P < 0.001) by granulosa cells throughout culture. In contrast, oocyte secreted factors suppressed granulosa cell progesterone production after both 48 and 144 hours (P < 0.001). Thecal cell numbers were increased by oocyte secreted factors (P = 0.02), together with a suppression in progesterone and androstenedione synthesis after 48 hours (P < 0.001) and after 144 hours (P = 0.02), respectively. Oocyte secreted factors also increased viable cell numbers (P < 0.001) in co-cultures together with suppression of progesterone (P < 0.001) and oestradiol (P < 0.001). In granulosa cell only cultures, SCF increased progesterone production in a dose dependent manner (P < 0.001), whereas progesterone synthesis by theca cells was reduced in a dose dependent manner (P = 0.002). Co-cultured cells demonstrated an increase in progesterone production with increasing SCF dose (P < 0.001) and an increase in oestradiol synthesis at the highest dose of SCF (100 ng/ml). In summary, these findings demonstrate the presence of a co-ordinated paracrine interaction between somatic cells and germ cells, whereby oocyte derived signals interact locally to mediate granulosa and theca cell function. SCF has a role in modulating this local interaction. In conclusion, the oocyte is an effective modulator of granulosa-theca interactions, one role being the inhibition of luteinization

Early programming of the oocyte epigenome temporally controls late prophase I transcription and chromatin remodelling

Oocytes are arrested for long periods of time in the prophase of the first meiotic division (prophase I). As chromosome condensation poses significant constraints to gene expression, the mechanisms regulating transcriptional activity in the prophase I-arrested oocyte are still not entirely understood. We hypothesized that gene expression during the prophase I arrest is primarily epigenetically regulated. Here we comprehensively define the Drosophila female germ line epigenome throughout oogenesis and show that the oocyte has a unique, dynamic and remarkably diversified epigenome characterized by the presence of both euchromatic and heterochromatic marks. We observed that the perturbation of the oocyte's epigenome in early oogenesis, through depletion of the dKDM5 histone demethylase, results in the temporal deregulation of meiotic transcription and affects female fertility. Taken together, our results indicate that the early programming of the oocyte epigenome primes meiotic chromatin for subsequent functions in late prophase I.

X-Linked Retinitis Pigmentosa 2 Is a Novel Maternal-Effect Gene Required for Left-Right Asymmetry in Zebrafish

Retinitis pigmentosa 2 (RP2) gene is responsible for up to 20% of X-linked retinitis pigmentosa, a severe heterogeneous genetic disorder resulting in progressive retinal degeneration in humans. In vertebrates, several bodies of evidence have clearly established the role of Rp2 protein in cilia genesis and/or function. Unexpectedly, some observations in zebrafish have suggested the oocyte-predominant expression of the rp2 gene, a typical feature of maternal-effect genes. In the present study, we investigate the maternal inheritance of rp2 gene products in zebrafish eggs in order to address whether rp2 could be a novel maternal-effect gene required for normal development. Although both rp2 mRNA and corresponding protein are expressed during oogenesis, rp2 mRNA is maternally inherited, in contrast to Rp2 protein. A knockdown of the protein transcribed from both rp2 maternal and zygotic mRNA results in delayed epiboly and severe developmental defects, including eye malformations, that were not observed when only the protein from zygotic origin was knocked down. Moreover, the knockdown of maternal and zygotic Rp2 revealed a high incidence of left-right asymmetry establishment defects compared to only zygotic knockdown. Here we show that rp2 is a novel maternal-effect gene exclusively expressed in oocytes within the zebrafish ovary and demonstrate that maternal rp2 mRNA is essential for successful embryonic development and thus contributes to egg developmental competence. Our observations also reveal that Rp2 protein translated from maternal mRNA is important to allow normal heart loop formation, thus providing evidence of a direct maternal contribution to left-right asymmetry establishment.

Validity and reliability of a pictorial instrument for assessing perceived motor competence in Portuguese children

It is important to assess young children's perceived Fundamental Movement Skill (FMS) competence in order to examine the role of perceived FMS competence in motivation toward physical activity. Children's perceptions of motor competence may vary according to the culture/country of origin; therefore, it is also important to measure perceptions in different cultural contexts. The purpose was to assess the face validity, internal consistency, test–retest reliability and construct validity of the 12 FMS items in the Pictorial Scale for Perceived Movement Skill Competence for Young Children (PMSC) in a Portuguese sample. Methods Two hundred one Portuguese children (girls, n = 112), 5 to 10 years of age (7.6 ± 1.4), participated. All children completed the PMSC once. Ordinal alpha assessed internal consistency. A random subsamples (n = 47) were reassessed one week later to determine test–retest reliability with Bland–Altman method. Children were asked questions after the second administration to determine face validity. Construct validity was assessed on the whole sample with a Bayesian Structural Equation Modelling (BSEM) approach. The hypothesized theoretical model used the 12 items and two hypothesized factors: object control and locomotor skills. Results The majority of children correctly identified the skills and could understand most of the pictures. Test–retest reliability analysis was good, with an agreement ration between 0.99 and 1.02. Ordinal alpha values ranged from acceptable (object control 0.73, locomotor 0.68) to good (all FMS 0.81). The hypothesized BSEM model had an adequate fit. Conclusions The PMSC can be used to investigate perceptions of children's FMS competence. This instrument can also be satisfactorily used among Portuguese children.

A competence model for environmental education

The goal of environmental education is ultimately to enable a person to strive for and to attain a more ecological way of life. In this article, we begin by distinguishing three forms of environmental knowledge and go on to predict that people's attitude toward nature represents the force that drives their ecological behavioral engagement. Based on data from 1,907 students, we calibrated previously established instruments to measure ecological behavior, environmental knowledge, and attitude toward nature with Rasch-type models. Using path modeling, we corroborated our theoretically anticipated competence structure. While environmental knowledge revealed a modest behavioral effect, attitude toward nature turned out to be, as expected, the stronger determinant of behavior. Overall, we propose a competence model that has the potential to guide us into more evidence-based ways of promoting the overall ecological engagement of individuals. (DIPF/Orig.)

Quality of implementation in an evidence-based family prevention program: the Family Competence Program

Family prevention programs need to be evidence-based in order to guarantee the success of their implementation. The Family Competence Program (FCP), a Spanish cultural adaptation of the Strengthening Families Program (SFP), has developed different measures and processes to gauge the quality of the implementation. This article is dedicated specifically to two of these measures: the evaluation of the facilitators and the assessment of the family engagement techniques. For evaluating the facilitators, a Delphi technique with experts and professionals is undertaken. For assessing the family techniques, both self-evaluation of trainers and evaluation by families are used. Finding underpin that, in the case of facilitators, is important that, after to skills and experience, they need to understand the theory of change of the program. In the case of family engagement techniques, more detailed, comprehensive talks, discussions and group activities lead to better family engagement outcomes.

Effects of two teaching interventions on nursing students’ acquisition of competence in ECG interpretation

Purpose: Nurse ability to recognise patient arrhythmias could contribute to preventing in-hospital cardiac arrest. Research suggests that nurses and nursing students lack competence in electrocardiogram (ECG) interpretation. The aim of this study was to compare the effects of two training strategies on nursing students’ acquisition of competence in ECG interpretation. Materials and methods: A controlled randomised trial with 98 nursing students. Divided in groups of 12–16, participants were randomly allocated to one of the following 3-h teaching intervention groups: 1) traditional instructor-led (TILG), and 2) flipped classroom (FCG). Participants’ competence in ECG interpretation was measured in terms of knowledge (%), skills (%) and self-efficacy (%) using a specifically designed and previously validated toolkit at pre-test and post-test. Two-way MANOVA explored the interaction effect between ‘teaching group’ and ‘time of assessment’ and its impact on participants’ competence. Within-group differences at pre-test and post-test were explored by carrying out paired t-tests. Between-group differences at pre- and post-test were examined by performing independent t-test analysis. Results: There was a statistically significant interaction effect between ‘teaching group’ and ‘time of assessment’ on participants’ competence in ECG interpretation (F(3,190) = 86.541, p = 0.001; Wilks’ Λ = 0.423). At pre-test, differences in knowledge (TILG = 35.12 ± 12.07; FCG = 35.66 ± 10.66), skills (TILG = 14.05 ± 10.37; FCG = 14.82 ± 14.14), self-efficacy (TILG = 46.22 ± 23.78; FCG = 40.01 ± 21.77) and all other variables were non-significant (p > 0.05). At post-test, knowledge (TILG = 55.12 ± 14.16; FCG = 94.2 ± 7.31), skills (TILG = 36.90 ± 16.45; FCG = 86.43 ± 14.32) and self-efficacy (TILG = 70.78 ± 14.55; FCG = 79.98 ± 10.35) had significantly improved, regardless of the training received (p < 0.05). Nonetheless, participants in the FCG scored significantly higher than participants in the TILG in knowledge, skills and self-efficacy (p < 0.05). Conclusion: Flipping the classroom for teaching ECG interpretation to nursing students may be more effective than using a traditional instructor-led approach in terms of immediate acquisition of competence in terms of knowledge, skills and self-efficacy. Further research on the effects of both teaching strategies on the retention of the competence will be undertaken.

Neustonic distribution of decapod planktonic stages and competence of brachyuran megalopae in coastal waters

Settlement rate may not reflect larval supply to coastal waters in different marine invertebrates and demersal fishes. The importance of near-shore oceanography and behaviour of late larval stages may be underestimated. The present study conducted neustonic sampling over station grids and along full-length transects at two embayments in south-eastern Brazil to (1) compare diurnal and nocturnal occurrence of most frequent decapod stages to assess their vertical movements, (2) describe the formation of larval patches and (3) measure competence of crab megalopae according to their distance to recruitment grounds. Several shrimp species apparently undergo a diel vertical migration, swimming crab megalopae showed no vertical movements and megalopae of the intertidal crab Pachygrapsus transversus revealed a reversed vertical migration. During the day, crab megalopae aggregated in convergence zones just below surface slicks. These larvae consisted of advanced, pre-moult stages, at both mid-bay and near-shore patches. Competence, measured as the time to metamorphosis in captivity, was similar between larval patches within each taxon. Yet, subtidal portunids moulted faster to juveniles than intertidal grapsids, possibly because they were closer to settlement grounds. Megalopae of Pachygrapsus from benthic collectors moulted faster than those from bay areas. These results suggest that alternative vertical migration patterns of late megalopae favour onshore transport, and actual competence takes place very close to suitable substrates, where larvae may remain for days before settlement. Lack of correlation between larval supply and settlement for Pachygrapsus suggests that biological processes, besides onshore transport, may play an important role in determining settlement success of coastal crabs.

STORYTELLING, EMOTION UNDERSTANDING, AND KINDERGARTNERS’ SOCIAL COMPETENCE

Interpreting others’ emotions is theoretically foundational for children’s social competence, yet little research contrasts Emotion Understanding (EU) types against their theoretical correlates. This study investigated kindergartners’ situationistic EU (attributing emotions based on external events) and mentalistic EU (attributing emotions from others’ mental states) in relation to Theory of Mind (ToM) and social skills, as rated by parents and teachers. The EU measures were expected to have low associations with one another and to relate differently to ToM and select social skills. Mentalistic EU was expected to be an important predictor of teacher-rated social skills. Results supported the hypothesis that mentalistic EU and situationistic EU are distinct constructs. However, both relate to ToM. Furthermore, while ToM and situationistic EU variables were included in the regression model, only vocabulary and mentalistic EU were significant predictors for teacher-rated social skills. Results indicate the importance of mentalistic EU in aspects of kindergartners’ social competence.

Development of motor competence in childhood and adolescence

This dissertation aimed to investigate the development of motor competence in childhood and adolescence. As a starting point, a systematic review was conducted to investigate the instruments used for the evaluation of motor competence. This review identified a gap in the literature regarding the existence of an instrument to assess motor competence based on the three theoretical constructs (stability, locomotor, manipulative). In an attempt to fill this gap, a valid quantitative instrument was proposed. To meet this purpose, 584 children, between 6 and 14 years of age, were evaluated in nine motor tasks, three for each construct. The final instrument comprised two motor tasks for each construct (stability, locomotor, manipulative) and presented very good fit indexes. This instrument was used to analyse the motor competence behaviour by gender in different age groups, indicating that generally boys outperformed girls and both genders increased their performance across age groups. However, different motor competence growth rates appear in both genders across age groups. In addition, children with high and low motor competence were compared regarding some of their health related fitness variables. We found that, regardless of age and gender, the group with better motor proficiency always showed better results in health related fitness. We found positive moderate to high correlations between motor competence and the variables of cardiovascular fitness and muscular fitness, and an inverse correlation with body composition across the four age groups. We also found that motor competence explained 75% of the variance of the health related fitness for the total sample, with locomotion as the primary predictor. However, when analysing the four age groups, stability skills seem to play an important role in health related fitness in the transition from childhood to adolescence. In conclusion, educational and health policies should consider the development of motor competence as an essential strategy to promote healthy development throughout life.

Spawning failure in Brycon amazonicus may be associated with ovulation and not with final oocyte maturation

Avaliaram-se os possíveis mecanismos envolvidos com a falha na desova de matrinxãs (Brycon amazonicus), submetidas à indução hormonal por extrato bruto de hipófise de carpa. Para tal, após a extrusão, os ovários foram coletados e analisados histomorfometricamente. Nas fêmeas que não desovaram (FNDs), a maioria dos ovócitos vitelogênicos remanescentes nos ovários atingiu a maturação final, apresentando quebra de vesícula germinativa, mas não foram ovulados (NOs). Consequentemente, estas fêmeas apresentaram frequências mais baixas de folículos pós ovulatórios (5%) quando comparadas com a que desovou (FD) (23%). Com relação aos NOs, os valores se inverteram e a frequência destes nas FNDs (21%) foi maior do que na FD (3%). Estes dados indicam que as falhas na desova desta espécie estão provavelmente relacionadas com a ovulação, uma vez que a maturação final dos ovócitos ocorre de forma similar tanto nas FNDs como na FD. Os dados sugerem que as substâncias que promovem a ovulação, como as prostaglandinas, podem aumentar o sucesso de desova em peixes reofílicos.

Artificial fertilization of oocytes and sperm activation in pacu: effects of the spermatozoa:oocyte ratio, water volume, and in natura semen preservation

Objetivou-se foi avaliar a fertilização artificial e a duração da motilidade espermática em pacus com diferentes doses inseminantes, volumes de água e preservação do sêmen in natura. Foram realizados quatro experimentos para avaliação do efeito de doses inseminantes (7x10³, 7x10(4), 7x10(5), 7x10(6) e 7x10(7) espermatozoides ovócito-1) sobre a fertilização artificial dos ovócitos; do efeito do volume de água (0,5; 15,0; 30,0; 45,0 e 60,0 mL de água mL-1 de ovócitos) com doses inseminantes de 105.481 e 210.963 espermatozoides ovócito-1; do efeito de diluição do sêmen (0,005; 0,05; 0,5 e 5,0 µL de sêmen mL-1 de água) sobre a duração da motilidade espermática; e do efeito do armazenamento a 15 ºC por 9 h sobre a duração da motilidade espermática e o índice de sobrevivência espermática. Os maiores resultados obtidos foram: doses inseminantes entre 7x10³ e 7x10(7) espermatozoides ovócito-1; 15 a 60 mL de água mL-1 de ovócitos; diluição de 0.005 µL sêmen mL-1 de água e 98,65% de sobrevivência espermática até o tempo de preservação de 2h45min36s. A preservação a 15ºC por 9 horas não influencia a duração da motilidade espermática. As maiores taxas de fertilização podem ser observadas no emprego de 0,27 a 270 µL de sêmen mL-1 de ovócitos, com 15 a 60 mL de água para ativação.

Effects of the spermatozoa: oocyte ratio, water volume and water temperature on artificial fertilization and sperm activation of cascudo-preto

The objective of this study was to evaluate the effects of water volume and water temperature on the sperm motility duration and the number of spermatozoa, and the water volume on the fertilization rates of oocytes of Rhinelepis aspera. Experiments were carried out to evaluate the effect of semen dilutions (1.74×10-5, 1.74×10-4, 1.74×10-3, 1.74×10-2, 1.74×10-1 and 1.00 mL of sperm.mL-1 of water) and water temperature (5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 ºC) on spermatozoa motility duration. In addition, the effects of insemination dose (7×10³, 7×10(4), 7×10(5), 7×10(6) and 7×10(7) spermatozoa.oocyte-1) and water volume (1.0, 30.0, 60.0, 90.0 and 120.0 mL water.2.0 mL-1 oocytes) on the artificial fertilization rates of oocytes were evaluated. The longest sperm motility duration were observed for the semen dilution of 1.74×10-5 mL semen.mL-1 water and in water at 5 ºC. The highest fertilization rates were obtained for insemination doses between 7.00×10³ and 1.23×10(7) spermatozoa. oocyte-1 and water volume of 28.11 mL water.2.0 mL-1 oocytes.