Cellular and molecular markers of anteroposterior and dorsoventral organisation in the vitellogenic follicles of adult Sarcophaga bullata (Diptera) and dorsoventral orientation of follicles in the ovary

Summary Polar organisation in the follicles of adult Sarcophaga bullata is reflected in the nurse cell-oocyte axis and in the orientation of the two polar cell pairs in the follicular epithelium. The internal organisation of the nurse cell chamber contributes to polarity but not to dorsoventral asymmetry. Dorsoventral asymmetry is correlated with the eccentric position of the germinal vesicle and the orientation of the polar cell pairs; no other follicle cell specialisations are seen. In an ovary, follicles are preferentially orientated with the dorsal side to the centre of the ovary. Cytoskeletal and some haemolymph proteins are molecular markers of polarity. Thus, in pre-vitellogenic stages, tubulin immunoreactivity is higher in the oocyte than in the nurse cells, actin immunoreactivity is the same over the cystocytes and larval serum proteins are restricted to the poles. During vitellogenesis, both actin and tubulin become more concentrated in the nurse cells and larval serum protein 1 accumulated in the polar cells during border cell migration when yolk polypeptides also accumulate in the oocyte. At the end of vitellogenesis a lipophorin is taken up by the oocyte. No molecular marker of dorsoventral asymmetry was identified.     

Staff Nurses’ Perceptions and Experiences about Structural Empowerment: A Qualitative Phenomenological Study

The aim of the study reported in this article was to investigate staff nurses’ perceptions and experiences about structural empowerment and perceptions regarding the extent to which structural empowerment supports safe quality patient care. To address the complex needs of patients, staff nurse involvement in clinical and organizational decision-making processes within interdisciplinary care settings is crucial. A qualitative study was conducted using individual semi-structured interviews of 11 staff nurses assigned to medical or surgical units in a 600-bed university hospital in Belgium. During the study period, the hospital was going through an organizational transformation process to move from a classic hierarchical and departmental organizational structure to one that was flat and interdisciplinary. Staff nurses reported experiencing structural empowerment and they were willing to be involved in decision-making processes primarily about patient care within the context of their practice unit. However, participants were not always fully aware of the challenges and the effect of empowerment on their daily practice, the quality of care and patient safety. Ongoing hospital change initiatives supported staff nurses’ involvement in decision-making processes for certain matters but for some decisions, a classic hierarchical and departmental process still remained. Nurses perceived relatively high work demands and at times viewed empowerment as presenting additional. Staff nurses recognized the opportunities structural empowerment provided within their daily practice. Nurse managers and unit climate were seen as crucial for success while lack of time and perceived work demands were viewed as barriers to empowerment.     

Optimizing culture conditions of a porcine epithelial cell line IPEC-J2 through a histological and physiological characterization

The high similarity between pigs and humans makes pigs a good gastrointestinal (GI) model for humans. Recently an epithelial cell line originating from the jejunum of pig (IPEC-J2) became available. Once validated, this model can be used to investigate the complex interactions occurring in the intestine. The advantages of using IPEC-J2 as in vitro model of the GI tract are the high resemblance between humans and pigs, and the ease of extrapolating in vitro to in vivo characteristics. In this study, the IPEC-J2 cells were functionally characterized by measuring the trans-epithelial electrical resistance (TEER), and by histological and ultrastructural studies. IPEC-J2 cells grown on six different permeable support systems, were investigated. The Transwell^®-COL collagen-coated membrane (1.12 cm²) showed the best results concerning time efficiency and TEER values. The optimum seeding density of 12 × 10⁵ cells/mL ensured that after 9 days of differentiation a confluent monolayer was formed. The decrease in TEER values after a maximum had been reached, coincided with the ultrastructural development of apical microvilli. We conclude that IPEC-J2 cells grown on collagen-coated membranes represent a valuable in vitro model system for the small intestinal epithelium which can be of great interest for intestinal research.     

Obesity and Persistent Organic Pollutants: Possible Obesogenic Effect of Organochlorine Pesticides and Polychlorinated Biphenyls

Persistent organic pollutants (POPs) are endocrine‐disrupting chemicals associated with the development of the metabolic syndrome and type 2 diabetes. In humans, little is known about their role in the potential origin of obesity. This study aims to assess the associations between serum levels of POPs and the prevalence of obesity in a cohort of obese and lean adult men and women. POP serum samples were investigated cross‐sectionally in 98 obese and 47 lean participants, aged ≥18 years. Serum samples were analyzed for the presence of polychlorinated biphenyl (PCB) congeners 153, 138, 180, and 170 and for the organochlorine pesticides, dichloro‐diphenyl‐dichloroethylene (pp‐DDE), and β‐hexachlorocyclohexane (βHCH). We established a significant negative correlation between BMI, waist, fat mass percentage, total and subcutaneous abdominal adipose tissue, and serum levels of PCB 153, 180, 170, and the sumPCBs. For βHCH, we demonstrated a positive correlation with BMI, waist, fat mass percentage, and total and subcutaneous abdominal adipose tissue. PCBs 180, 170, and the sum of PCBs correlated significantly negative with homeostasis model assessment for insulin resistance (HOMA_IR). βHCH correlated significantly positively with HOMA_IR. A strong correlation was established between all POP serum levels and age. We established a positive relationship between high serum levels of βHCH and BMI and HOMA_IR, whereas serum PCB levels were inversely correlated with BMI and HOMA_IR. Combined, these results suggest that the diabetogenic effect of low‐dose exposure to POPs might be more complicated than a simple obesogenic effect.     

Haploid vegetative mycelia of Armillaria gallica show among-cell-line variation for growth and phenotypic plasticity

Vegetative mycelial cells of Armillaria are expected to have diploid nuclei. Cells from a single mycelium therefore would not be expected to differ from one another for ecologically relevant quantitative traits. We isolated two sets of basidiome cell lines (from spores and stipe cells) and one set of vegetative cell lines (from an attached rhizomorph) from a single contiguous Armillaria gallica mycelium. We isolated a second set of vegetative cell lines from the soil 20 cm from the above basidiome-rhizomorph complex. In all four sets of cell lines in situ DAPI-DNA measurements showed cells are haploid and quantitative-trait analyses of cell lines grown at different water potentials revealed high levels of among-cell-line genetic variation for both growth and phenotypic plasticity. Haploidy and the existence of ecologically relevant genetic variation within vegetative individuals are unexpected and mean that a process similar to evolutionary adaptation could take place within the soma of a genetic individual. We believe this is a key to understanding how large A. gallica mycelia survive exposure to variation in ecological conditions during lives that potentially span several tree (host) generations.     

Non-breeding song rate reflects nutritional condition rather than body condition

Numerous studies have focused on song in songbirds as a signal involved in mate choice and intrasexual competition. It is expected that song traits such as song rate reflect individual quality by being dependent on energetic state or condition. While seasonal variation in bird song (i.e., breeding versus non-breeding song) and its neural substrate have received a fair amount of attention, the function and information content of song outside the breeding season is generally much less understood. Furthermore, typically only measures of condition involving body mass are examined with respect to song rate. Studies investigating a potential relationship between song rate and other indicators of condition, such as physiological measures of nutritional condition, are scant. In this study, we examined whether non-breeding song rate in male European starlings (Sturnus vulgaris) reflects plasma metabolite levels (high-density lipoproteins (HDL), albumin, triglycerides and cholesterol) and/or body mass. Song rate was significantly positively related to a principal component representing primarily HDL, albumin and cholesterol (and to a lesser degree plasma triglyceride levels). There was only a trend toward a significant positive correlation between song rate and body mass, and no significant correlation between body mass and the abovementioned principal component. Therefore, our results indicate that nutritional condition and body mass represent different aspects of condition, and that song rate reflects nutritional rather than body condition. Additionally, we also found that intra-individual song rate consistency (though not song rate itself) was significantly positively related to lutein levels, but not to body mass or nutritional condition. Together our results suggest that the relation between physiological measures of nutritional condition and song rate, as well as other signals, may present an interesting line of future research, both inside and outside the breeding season.     

Literature Review and Comparison of Two Statistical Methods to Evaluate the Effect of Botulinum Toxin Treatment on Gait in Children with Cerebral Palsy

Aim

This study aimed at comparing two statistical approaches to analyze the effect of Botulinum Toxin A (BTX-A) treatment on gait in children with a diagnosis of spastic cerebral palsy (CP), based on three-dimensional gait analysis (3DGA) data. Through a literature review, the available expert knowledge on gait changes after BTX-A treatment in children with CP is summarized.

Methods

Part 1—Intervention studies on BTX-A treatment in children with CP between 4–18 years that used 3DGA data as an outcome measure and were written in English, were identified through a broad systematic literature search. Reported kinematic and kinetic gait features were extracted from the identified studies. Part 2—A retrospective sample of 53 children with CP (6.1 ± 2.3years, GMFCS I-III) received 3DGA before and after multilevel BTX-A injections. The effect of BTX-A on gait was interpreted by comparing the results of paired samples t-tests on the kinematic gait features that were identified from literature to the results of statistical parametric mapping analysis on the kinematic waveforms of the lower limb joints.

Results

Part 1–53 kinematic and 33 kinetic features were described in literature. Overall, there is no consensus on which features should be evaluated after BTX-A treatment as 49 features were reported only once or twice. Part 2—Post-BTX-A, both statistical approaches found increased ankle dorsiflexion throughout the gait cycle. Statistical parametric mapping analyses additionally found increased knee extension during terminal stance. In turn, feature analyses found increased outtoeing during stance after BTX-A.

Conclusion

This study confirms that BTX-A injections are a valuable treatment option to improve gait function in children with CP. However, different statistical approaches may lead to different interpretations of treatment outcome. We suggest that a clear, definite hypothesis should be stated a priori and a commensurate statistical approach should accompany this hypothesis.     

BMP4 plays a role in apoptosis during human preimplantation development

Comprehensive understanding of lineage differentiation and apoptosis processes is important to increase our knowledge of human preimplantation development in vitro. We know that BMP signaling is important for different processes during mammalian development. In mouse preimplantation embryos, BMP signaling has been shown to play a role in the differentiation into extra‐embryonic trophectoderm (TE) and primitive endoderm (PE). In this study, we aimed to investigate the effect of bone morphogenetic protein 4 (BMP4) supplementation on human preimplantation embryos cultured in vitro. The BMP4 treatment impaired human blastocyst formation. No differences in the expression of the early lineage markers NANOG, CDX2, GATA3, and GATA6 were found between BMP4‐treated embryos and controls. Instead, BMP4 supplementation triggered apoptosis in the human blastocyst. We focused on P53, which is known to play a major role in the apoptosis. In BMP4‐treated embryos, the P53 responsive gene expression was not altered; however, the P53 deacetylase SIRT1 was downregulated and acetylated P53 was increased in mitochondria. Altogether, our findings suggest that BMP4 plays a role in the apoptosis during human preimplantation development.     

Differences in dichogamy and herkogamy contribute to higher selfing in contrasting environments in the annual Blackstonia perfoliata (Gentianaceae)

Background and Aims

The establishment of plant populations in novel environments may generate pronounced shifts in floral traits and plant mating systems, particularly when pollinators are scarce. In this study, floral morphology and mating system functioning are compared between recently established and older populations of the annual plant Blackstonia perfoliata that occur in different pollinator environments.

Methods

Hand-pollination and emasculation experiments were conducted to assess the extent of pollinator-mediated pollen deposition and pollen limitation, and the contribution of autonomous selfing to total seed production. Detailed measurements of key floral traits were performed to compare the flower morphology and mating system functioning between plants from both pollination environments.

Key Results

Pollinator-mediated pollen deposition was about twice as low in the recently colonized and pollinator-poor environment compared with the old and pollinator-rich sites, but total seed set was little affected by any type of pollen limitation. The contribution of autonomous selfing to total seed production was higher in the pollinator-poor sites than in the pollinator-rich sites (index of reproductive assurance = 0·56 and 0·17, respectively), and seed production was only poorly affected by selfing, whereas in the pollinator-rich populations selfing reduced total reproductive output by about 40 % compared with outcross pollination. Plants originating from pollinator-poor environments produced smaller flowers that showed significantly lower levels of dichogamy (i.e. protogyny) and herkogamy. These reductions resulted in a 2-fold higher capacity for autonomous selfing under pollinator-free conditions (index of autonomous selfing = 0·81 and 0·41 in plants originating from the pollinator-poor and pollinator-rich environment, respectively).

Conclusions

The results illustrate that plant populations colonizing novel environments can differ markedly in floral morphology and mating system functioning. Due to a temporal shift in the male phase, the breeding system of B. perfoliata shifted from delayed selfing under pollinator-rich conditions towards competing selfing in recently established populations, providing greater reproductive assurance when pollinators and/or reproductive partners are limited.