Adaptation and validation of the Portuguese version of the regulation of eating behavior scale (REBSp).

The aim of present study was two-fold: i) to translate and adapt the Regulation of Eating Behavior Scale to Portuguese (REBSp), and ii) to analyze its psychometrics properties (factorial validity with gender invariance analyses, reliability and construct validity). The study sample was composed by 471 Portuguese participants (68.4% females) with a mean age of 30.5 years (SD = 11.2). Structural equation modeling was used to verify the psychometric properties of the scale using SPSS v. 23.0 and AMOS 24.0 software. The analysis showed that the Portuguese 24-item scale presented a good fit, achieving good reliability and convergent validity. Some issues arose with discriminant validity within autonomous and controlled regulations, discussed in light of the simplex pattern expected by self-determination theory literature. Additionally, the scale presented concurrent validity and evidence of gender measurement invariance. Latent mean analysis between genders showed that women presented higher means for intrinsic motivation and integrated regulation when compared to men. In conclusion, analyses suggest that the 24-item Portuguese version of REBS can be used safely to assess the eating regulation in both genders.

Upgrading end-of-line residues of the red seaweed Gelidium sesquipedale to polyhydroxyalkanoates using Halomonas boliviensis.

Agar extraction from Gelidium and Gracilaria red seaweed species produces hundred thousand ton of carbohydrate-rich residues annually. Gelidium sesquipedale waste biomass obtained after agar extraction, still contained 44.2 % w/w total carbohydrates (dry-weight basis). These residues were biologically up-graded to poly-3-hydroxybutyrate (P3HB) after saccharification of their carbohydrate fraction to simple sugars. A combined hydrolysis treatment using sulfamic acid followed by enzymatic hydrolysis with cellulases produced a glucose-rich hydrolysate with a negligible content of inhibitors. With this treatment a sugar yield of circa 30 % (g glucose/g biomass) was attained. The algal hydrolysates were assessed as carbon source for the production of P3HB by the halotolerant bacteria Halomonas boliviensis. A cell concentration of 8.3 g L-1 containing 41 % (w/w) of polymer and a yield (YP/S ) of 0.16 gpolymer/gglucose were attained in shake flask assays. In this work, cellulose-rich seaweed waste was shown to be an upgradable, sustainable source of carbohydrates.

A subpopulation of human Adark spermatogonia behaves as the reserve stem cell.

Human spermatogonial stem cells (SSCs) are an essential source to maintain spermatogenesis as an efficient process for daily sperm production with high self-renewal capacity along adulthood. However, the phenotype and the subpopulation that represent the real reserve SSC for the human testis remain unknown. Moreover, although SSC markers have been described for undifferentiated spermatogonia (Adark and Apale), the existence of a specific subtype that could be identified as the actual/true SSC has not yet been fully determined. Herein we evaluated spermatogonial morphology, kinetics, positioning regarding blood vasculature in relation to protein expression (UTF1, GFRA1, and KIT) as well as proliferative activity (MCM7) and identified a small subpopulation of Adark with nuclear rarefaction zone (AdVac) that behaves as the human reserve SSC. We show that AdVac is the smallest human spermatogonial population (10%), staying quiescent (89%) and positioned close to blood vessels throughout most of the stages of the seminiferous epithelium cycle (SEC) and divides only at stages I and II. Within this AdVac population, we found a smaller pool (2% of A undifferentiated spermatogonia) of entirely quiescent cells exhibiting a high expression of UTF1 and lacking GFRA1. This finding suggests them as the real human reserve SSC (AdVac UTF1+/GFRA1-/MCM7-). Additionally, Adark without nuclear vacuole (AdNoVac) and Apale have similar kinetic and high proliferative capacity throughout the SEC (47%), indicating that they are actively dividing undifferentiated spermatogonia. Identification of human stem cells with evident reserve SSC functionality may help further studies intending to sort SSCs to treat male diseases and infertility.

Osteogenic potential of human dental pulp stem cells cultured onto poly-ε-caprolactone/poly (rotaxane) scaffolds.

BACKGROUND: Bioengineering aims to develop innovative scaffolds to improve cellular activities for tissue regeneration. OBJECTIVES: To evaluate the biological behavior of human dental pulp stem cells (hDPSCs) seeded onto an experimental polymeric-based scaffold comprising poly-ε-caprolactone/poly (rotaxane). MATERIAL AND METHODS: Adhesion, viability, and proliferation as well as alkaline phosphatase (ALP) activity, mineralized nodule formation (alizarin red assay), and expression of genes related to osteogenic differentiation, including ALP, type 1 collagen alpha 1 (COL1A1), Runt-related transcription factor (Runx-2), and osteocalcin (BGLAP/OCN), were evaluated in hDPSCs seeded onto polymeric scaffolds. RESULTS: hDPSCs expressed typical levels of mesenchymal stem cell surface markers. Cell growth increased upon cultivation on polymeric blend scaffold and the cells gained osteoblast-like appearance. Fourteen days after seeding hDPSCs on the scaffolds, irrespective to the culture medium used (clonogenic or mineralization medium), the cells presented ALP activity higher than that of control cells grown in clonogenic medium. The cells cultivated in mineralization medium on the scaffold showed significantly higher expression of all genes than the control cells, except for BGLAP gene expression. At 21 days, the group cultivated on the scaffold and mineralization medium showed maximum level of mineralization. SIGNIFICANCE: Poly-ε-caprolactone/poly (rotaxane) blend is noncytotoxic to hDPSCs and improved genomic and functional osteogenic differentiation. Thus, poly-ε-caprolactone/poly (rotaxane) blend may serve as a promising bioactive biomaterial for bone tissue bioengineering.

Prevalence and predictors of weight loss maintenance: a retrospective population-based survey of European adults with overweight and obesity.

BACKGROUND: The prevalence of weight loss attempts has increased worldwide, although the extent to which sustained weight loss is achieved is unknown. There is insufficient research into weight loss maintenance (WLM) in individuals with overweight or obesity who have recently lost clinically significant amounts of weight (≥5%), particularly in the European general population. The present study aimed to determine the prevalence and retrospective predictors of WLM in population-based samples of European adults with overweight or obesity who had made a recently completed weight loss attempt. METHODS: Participants (N = 2000) in UK, Denmark and Portugal completed an online survey about loss and regain in their most recent completed weight loss attempt, features of their attempt (duration, self-weighing, lapses, strategies), as well as loss of control and binge eating. Multiple regression analysis was used to determine factors retrospectively associated with WLM in those who achieved clinically significant weight loss (n = 1272). RESULTS: Mean (SD) self-reported weight loss was 9% (8%) and mean (SD) regain was 96.3% (9%) of participants' start weight. Twenty-three percent of the total sample had maintained weight loss of ≥5% for at least 1 month. Controlling for weight loss and time since attempt, predictors of better WLM were avoidance of a temporary lapse, infrequent/absent loss of control and binge eating, and use of a greater number of dietary strategies for WLM (r2  = 0.338, P < 0.001). PRINCIPAL CONCLUSIONS: Factors associated with recent successful WLM indicate the importance of the continued use of dietary and other strategies for WLM, particularly in the face of a lapse, as well as the need to manage dysfunctional eating behaviours.

Effects of photobiomodulation therapy on the extracellular matrix of human dental pulp cell sheets.

Photobiomodulation therapy (PBMT) and the cell sheet (CS) technology improve processes relevant to tissue regeneration. The aim of this study was to investigate the effects of different PBMT parameters on the architecture (histology), protein composition (Western blotting and immunohistochemistry) and ultrastructure [scanning electron microscopy (SEM) and transmission electron microscopy (TEM)] of the extracellular matrix (ECM) synthesized by CSs composed by human dental pulp stem cells (hDPSCs). METHODS: Thawed cells were recharacterized by the expression profile of the surface molecules of mesenchymal stem cells (MSCs) using flow cytometry. Clonogenic medium supplemented with vitamin C (20 µg/ml) was used for obtaining the CSs. PBMT was performed with continuous-wave diode laser (660 nm, 20 mW, 0.028cm2, 0.71 W/cm2) in punctual and contact mode. The CSs were allocated in 3 experimental groups: Control: no further treatment; PBMT1 [4 s, 3 J/cm2 (lower energy density), 0.08 J/point] and PBMT2 [7 s, 5 J/cm2 (higher energy density), 0.14 J/point]. Statistical comparisons were performed (p ≤ .05). RESULTS: The cells presented the classical immunoprofile of MSCs. Type I and type III collagens and fibronectin were present in the ECM of the CSs. PBMT1 induced higher amount of fibronectin. The overall ultrastructure of the CSs in the PBMT1 was epithelial-like, whereas the PBMT2 leads to CSs with fusiform cells arranged in bundles. TEM identified a more mature ECM and signs of apoptosis and necrosis in the PBMT2 group. CONCLUSION: PBMT influence the composition and ultrastructure of the ECM of CSs of hDPSCs. Thus, PBMT, specifically when applied in the lower energy density, could be of importance in the determination of the mechanical quality of CSs, which may favor cell therapy by improving the CS transplantation approach.

Screening of hydrogel-based scaffolds for dental pulp regeneration-A systematic review.

OBJECTIVE: The aim of this systematic review was to evaluate the most appropriate hydrogel scaffold type (natural, synthetic or hybrid) to be applied with stem cells for dental pulp regeneration. The findings should help clinicians make an informed choice about the appropriate scaffold to be applied for this approach. DESIGN: Three electronic databases were searched (Medline, Web of Science and Scopus). The review was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA). RESULTS: From 4990 potentially relevant studies initially identified, 18 papers fulfilled the eligibility criteria and were considered for this review. Natural scaffolds were applied in most studies. Collagen was the most studied scaffold. In 5 of 10 studies, only growth factors were added to the constructs. Even without growth factors, these scaffolds containing stem cells were able to support the formation of dentin. The synthetic scaffolds were the least studied. Only 4 studies were selected, and in 3 of them, the same scaffold (Puramatrix) was evaluated. Puramatrix by itself was unable to form dental pulp when dental pulp stem cells were not present. Synthetic and hybrid hydrogels were unable to attract stem cells from the host. The presence of growth factors in these constructs seems to be of relevance since dental pulp tissue formation was achieved only when the hybrid scaffold was applied with growth factors. CONCLUSION: All types of hydrogel-based scaffolds, when containing mesenchymal stem cells, are able to form connective tissue with different degrees of similarity to dental pulp. However, current data is too heterogeneous to compare and identify the advantages of any specific scaffold.

The effect of profession on burnout in hospital staff.

Background: A high level of burnout has been described in health professionals. However, literature regarding other hospital employees is scarce. Aims: To assess the prevalence of burnout in different professional groups of hospital staff and how the professional category is associated with levels of burnout. Methods: Employees of a University Hospital in Portugal completed a self-administered online questionnaire in 2014-2015. We used the Portuguese version of the Maslach Burnout Inventory-Human Services Survey and scored three dimensions of burnout (emotional exhaustion, depersonalization, personal accomplishment) as low, average or high. We estimated odds ratios (OR) and 95% confidence intervals (95% CI) by logistic regression, adjusting for age, sex, use of anxiolytic/antidepressant drugs and job satisfaction. Results: There was a 10% response rate; 368 questionnaires were available for analysis. High levels of burnout due to emotional exhaustion were observed in all professional categories. Nurses, administrative staff and technicians more frequently scored higher levels of emotional exhaustion (59%, 50% and 50%, respectively) and lack of personal accomplishment (41%, 52% and 38%, respectively) than physicians and healthcare assistants. Not all professionals scored highly for depersonalization. Emotional exhaustion scores were significantly lower in healthcare assistants than nurses (adjusted OR 0.26, 95% CI 0.10-0.64). Conclusions: Burnout affects all professional categories of hospital staff. Future studies should use larger, more representative samples of hospital staff, perform longitudinal analyses and analyse data on specifics of each professional category and other potential confounders.

Evaluation of the toxicity and hypoglycemic effect of the aqueous extracts of Cnidoscolus quercifolius Pohl.

Diabetes mellitus is one of the most common chronic degenerative diseases, and it is estimated to increase worldwide to around 415 million and to impact 642 million in 2040. Research shows that some plants are sources of bioactive compounds against diabetes. Thus, the objective of this work was to evaluate the oral toxicity and the hypoglycemic effect of the aqueous extract of the leaves of Cnidoscolus quercifolius Pohl. Diabetes was induced in Swiss mice with streptozotocin and the mice were treated with an aqueous extract of C. quercifolius leaves for a period of 30 days. Phytochemical analysis showed that the extract was rich in flavonoids, catechins and triterpenoid, which did not show any mortality and behavioral alterations in mice treated with 200, 1000, and 2000 mg/kg body weight of the extract for 14 days. Histopathological analysis of organs (kidney, pancreas, liver) from mice treated with the 2000 mg/kg extract revealed no architectural change. In the present study, we found a 29% reduction in glucose levels in animals receiving 200 mg/kg body weight. These results are very promising because they showed that C. quercifolius had a hypoglycemic effect and did not present oral toxicity, thus being a new source of compounds for the control of diabetes.

Efficacy of a randomized controlled self-regulation based physical activity intervention for chronic fatigue: Mediation effects of physical activity progress and self-regulation skills.

OBJECTIVE: Examine the medium-term effects of a brief physical activity (PA) self-regulation (SR) based intervention (4-STEPS program) for chronic fatigue, and explore the mediating effects of PA related variables and SR skills. METHODS: A two-arm randomized controlled trial (Usual Care vs 4-STEPS) was carried out. The 4-STEPS program consisted of Motivational Interviewing and SR-skills training. Fatigue severity (primary outcome) and impact, PA, health-related quality of life (HrQoL), and somatic and psychological distress were assessed at baseline, post-treatment (12weeks) and 12months follow-up. RESULTS: Ninety-one patients (45 intervention and 46 controls) were included. At follow-up, there were significant treatment effects on fatigue severity (g=0.72) and fatigue impact, leisure-time PA, and physical and psychological HrQoL. No significant effects were found for number of daily steps and somatic and psychological distress. Fatigue severity at follow-up was partially mediated by post-treatment progress on a personal PA goal (effect ratio=18%). CONCLUSION: Results suggest that a brief intervention, focusing on the formulation and pursuit of personal PA goals and the use of SR skills, produces sustained benefits for fatigue severity. Despite these promising results, dropout was high and the intervention was not beneficial for all secondary outcomes.

Prevalence of personal weight control attempts in adults: a systematic review and meta-analysis.

The purpose of this systematic review and meta-analysis was to estimate the prevalence of personal weight control attempts (weight loss and/or maintenance) worldwide and to identify correlates, personal strategies used and the underlying motives. We included epidemiological/observational studies of adults (≥18 years) reporting prevalence of weight control attempts in the past-year. Seventy-two studies (n = 1,184,942) met eligibility criteria. Results from high quality studies showed that 42% of adults from general populations and 44% of adults from ethnic-minority populations reported trying to lose weight, and 23% of adults from general populations reported trying to maintain weight annually. In general population studies, higher prevalence of weight loss attempts was observed in the decade of 2000-2009 (48.2%), in Europe/Central Asia (61.3%) and in overweight/obese individuals and in women (p < 0.01). Of the 37 strategies (grouped in 10 domains of the Oxford Food and Activity Behaviours Taxonomy) and 12 motives reported for trying to control weight, exercising and dieting (within the energy compensation and restraint domains, respectively) and wellbeing and long-term health were the most prevalent. To our knowledge, this is the first systematic review to investigate weight control attempts worldwide. Key strategies and motives were identified which have implications for future public health initiatives on weight control.

MicroRNA profiling in human breast cancer cell lines exposed to the anti-neoplastic drug cediranib.

Cediranib, a pan-tyrosine kinase inhibitor is showing promising results for the treatment of several solid tumours. In breast cancer, its effects remain unclear, and there are no predictive biomarkers. Several studies have examined the expression profiles of microRNAs (miRNAs) in response to different chemotherapy treatments and found that the expression patterns may be associated with the treatment response. Therefore, our aim was to evaluate the cellular behaviour and differential expression profiles of miRNAs in breast cancer cell lines exposed to cediranib. The biological effect of this drug was measured by viability, migration, invasion and cell death in in vitro assays. Signaling pathways were assessed using a human phospho-receptor tyrosine kinase array. Furthermore, using a miRNA array and quantitative real-time PCR (qRT­PCR), we assessed the relative expression of miRNAs following cediranib treatment. The breast cancer cell lines exhibited a distinct cytotoxic response to cediranib treatment. Cediranib exposure resulted in a decrease in the cell migration and invasion of all the breast cancer cell lines. Treatment with cediranib appeared to be able to modulate the activation of several RTKs that are targets of cediranib such as EGFR and a new potential target ROR2. Furthermore, this drug was able to modulate the expression profile of different microRNAs such as miR-494, miR-923, miR-449a, miR-449b and miR-886-3 in breast cancer cell lines. These miRNAs are reported to regulate genes involved in important molecular processes, according to bioinformatics prediction tools.

Cytotoxicity, genotoxicity and antibacterial effectiveness of a bioceramic endodontic sealer.

AIM: To compare the characteristics of bioceramic endodontic sealer Endosequence BC sealer with those of AH Plus sealer. METHODOLOGY: Cytotoxicity and genotoxicity were analysed on human gingival fibroblasts submitted to cell culture medium conditioned by sealers using the MTT reduction assay and micronucleus formation test (MNT), respectively. Cells grown on fresh medium served as controls. Cell viabilities were measured at 1, 3, 5 and 7 days. The antibacterial activity was analysed on an Enterococcus faecalis strain (ATCC 29212) using both on agar diffusion test (ADT) and a direct contact test (DCT). The inhibition zones in ADT were measured after 48 h and the colony-forming units counting in the DCT after 1, 24, 72 and 168 h. Data were compared by anova and Tukey's test and MNT by Fisher's exact test (P < 0.05). RESULTS: Cultures submitted to Endosequence BC sealer had a significantly higher number of viable cells (P < 0.01) and less micronucleus formation (P < 0.05) than AH Plus sealer. Endosequence BC sealer exhibited significantly smaller inhibition zones (6.00 ± 0.03 mm) than AH Plus sealer (10.31 ± 0.21 mm) (P < 0.05). Moreover, Endosequence BC sealer had significantly smaller antibacterial activity than AH Plus sealer up to 1 h of direct contact (P < 0.05). On other exposure times, both materials had similar antibacterial effectiveness (P > 0.05). CONCLUSIONS: Bioceramic-based sealer had less cytotoxicity and genotoxicity and similar antibacterial effect against E. faecalis in comparison with AH Plus sealer.

p73 is required for ependymal cell maturation and neurogenic SVZ cytoarchitecture.

The adult subventricular zone (SVZ) is a highly organized microenvironment established during the first postnatal days when radial glia cells begin to transform into type B-cells and ependymal cells, all of which will form regenerative units, pinwheels, along the lateral wall of the lateral ventricle. Here, we identify p73, a p53 homologue, as a critical factor controlling both cell-type specification and structural organization of the developing mouse SVZ. We describe that p73 deficiency halts the transition of the radial glia into ependymal cells, leading to the emergence of immature cells with abnormal identities in the ventricle and resulting in loss of the ventricular integrity. p73-deficient ependymal cells have noticeably impaired ciliogenesis and they fail to organize into pinwheels, disrupting SVZ niche structure and function. Therefore, p73 is essential for appropriate ependymal cell maturation and the establishment of the neurogenic niche architecture. Accordingly, lack of p73 results in impaired neurogenesis. Moreover, p73 is required for translational planar cell polarity establishment, since p73 deficiency results in profound defects in cilia organization in individual cells and in intercellular patch orientation. Thus, our data reveal a completely new function of p73, independent of p53, in the neurogenic architecture of the SVZ of rodent brain and in the establishment of ependymal planar cell polarity with important implications in neurogenesis. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 730-747, 2016.

Micro-computed tomography and histomorphometric analysis of human alveolar bone repair induced by laser phototherapy: a pilot study.

Immediate dental implant placement in the molar region is critical, because of the high amount of bone loss and the discrepancy between alveolar crest thickness and the implant platform. Laser phototherapy (LPT) improves bone repair. The aim of this study was to evaluate the human alveolar bone repair 40 days after molar extraction in patients submitted to LPT. Twenty patients were selected for this randomized controlled clinical trial; 10 underwent LPT (laser group) with a GaAlAs diode laser (808 nm, 100 mW, 0.04 cm(2), 75 J/cm(2), 30s per point, 3 J per point, at five points). The control group patients (n=10) were not irradiated. Forty days later, the tissue formed inside the sockets was analyzed by micro-computed tomography and histomorphometry. Data from the two groups were compared with Student's t-test and Pearson's correlation test. The relative bone volume was significantly higher in the laser group (P<0.0001). The control group showed negative correlations (P<0.01) between number and thickness, and between number and separation of trabeculae, and a positive correlation between thickness and separation of trabeculae. The laser group showed a significant negative correlation between the number and thickness of trabeculae (P<0.01). The results suggest that LPT is able to accelerate alveolar bone repair after molar extraction, leading to a more homogeneous trabecular configuration represented by thin and close trabeculae.

Differential effects of behavioral interventions with a graded physical activity component in patients suffering from Chronic Fatigue (Syndrome): An updated systematic review and meta-analysis.

An updated systematic review and meta-analysis was conducted to (1) evaluate the effects of behavioral and psychological interventions containing a graded physical activity component upon fatigue severity, physical functioning, physical activity and psychological distress, and to (2) examine potential moderator effects of trial characteristics (type of control, setting, provider, length of treatment, psychological component, flexibility in physical activity, and minimal face to face patient-provider contact). Pertinent content of selected studies was extracted and rated on a scale of methodological quality. Sixteen randomized controlled trials (N=2004) were included in the meta-analyses. Significant small to medium effect sizes (Hedge's g=0.25 to g=0.66) were found for all outcomes at post-treatment (M=5.2months) and follow-up (M=11.7months), with the exception of physical activity at post-treatment (g=0.11). The largest effects were found for fatigue severity (g=0.61 to g=0.66). Subgroup analyses revealed that minimal contact interventions had additional beneficial effects upon fatigue (g=0.96) and depression (g=0.85). Interventions provided by psychologists-psychotherapists and interventions conducted in secondary-tertiary settings also resulted in more beneficial effects on fatigue. We found some indication of publication bias. The small number of studies and variability between them are limitations of this study. Future research should explore additional moderating effects in order to improve the effectiveness of interventions.

Sex differences in hepatic and intestinal contributions to nevirapine biotransformation in rats.

The understanding of the intestine contribution to drug biotransformation improved significantly in recent years. However, the sources of inter-individual variability in intestinal drug biotransformation, namely sex-differences, are still elusive. Nevirapine (NVP) is an orally taken anti-HIV drug associated with severe idiosyncratic reactions elicited by toxic metabolites, with women at increased risk. As such, NVP is a good model to assess sex-dimorphic metabolism. The aim of this study was to perform a comparative profiling of NVP biotransformation in rat intestine and liver and evaluate whether or not it is organ- and sex-dependent. Therefore, nevirapine-containing solutions were perfused through the intestine, in a specially designed chamber, or incubated with liver slices, from male and female Wistar rats. The levels of NVP and its Phase I metabolites were quantified by HPLC-UV. Liver incubation experiments yielded the metabolites 2-, 3-, 8-, and 12-OH-NVP, being 12-OH-NVP and 2-OH-NVP the major metabolites in males and females, respectively. Inter-sex differences in the metabolic profile were also detected in the intestine perfusion experiments. Herein, the metabolites 3- and 12-OH-NVP were only found in male rats, whereas 2-OH-NVP levels were higher in females, both in extraluminal (p<0.01) and intraluminal media. The metabolite 8-OH-NVP was not detected in the intraluminal media from either males or females. In this study, important inter-sex differences were detected in both organs, providing further clues to the sex-dimorphic profile of NVP toxicity. Moreover, an extra-hepatic contribution to NVP biotransformation was observed, strengthening the relevance of the intestinal contribution in the biotransformation of orally taken-drugs.

p73 is required for endothelial cell differentiation, migration and the formation of vascular networks regulating VEGF and TGFß signaling.

Vasculogenesis, the establishment of the vascular plexus and angiogenesis, branching of new vessels from the preexisting vasculature, involves coordinated endothelial differentiation, proliferation and migration. Disturbances in these coordinated processes may accompany diseases such as cancer. We hypothesized that the p53 family member p73, which regulates cell differentiation in several contexts, may be important in vascular development. We demonstrate that p73 deficiency perturbed vascular development in the mouse retina, decreasing vascular branching, density and stability. Furthermore, p73 deficiency could affect non endothelial cells (ECs) resulting in reduced in vivo proangiogenic milieu. Moreover, p73 functional inhibition, as well as p73 deficiency, hindered vessel sprouting, tubulogenesis and the assembly of vascular structures in mouse embryonic stem cell and induced pluripotent stem cell cultures. Therefore, p73 is necessary for EC biology and vasculogenesis and, in particular, that DNp73 regulates EC migration and tube formation capacity by regulation of expression of pro-angiogenic factors such as transforming growth factor-ß and vascular endothelial growth factors. DNp73 expression is upregulated in the tumor environment, resulting in enhanced angiogenic potential of B16-F10 melanoma cells. Our results demonstrate, by the first time, that differential p73-isoform regulation is necessary for physiological vasculogenesis and angiogenesis and DNp73 overexpression becomes a positive advantage for tumor progression due to its pro-angiogenic capacity.

Novel in vitro systems for prediction of veterinary drug residues in ovine milk and dairy products.

A new in vitro tool was developed for the identification of veterinary substrates of the main drug transporter in the mammary gland. These drugs have a much higher chance of being concentrated into ovine milk and thus should be detectable in dairy products. Complementarily, a cell model for the identification of compounds that can inhibit the secretion of drugs into ovine milk, and thus reduce milk residues, was also generated. The ATP-binding cassette transporter G2 (ABCG2) is responsible for the concentration of its substrates into milk. The need to predict potential drug residues in ruminant milk has prompted the development of in vitro cell models over-expressing ABCG2 for these species to detect veterinary drugs that interact with this transporter. Using these models, several substrates for bovine and caprine ABCG2 have been found, and differences in activity between species have been reported. However, despite being of great toxicological relevance, no suitable in vitro model to predict substrates of ovine ABCG2 was available. New MDCKII and MEF3.8 cell models over-expressing ovine ABCG2 were generated for the identification of substrates and inhibitors of ovine ABCG2. Five widely used veterinary antibiotics (marbofloxacin, orbifloxacin, sarafloxacin, danofloxacin and difloxacin) were discovered as new substrates of ovine ABCG2. These results were confirmed for the bovine transporter and its Y581S variant using previously generated cell models. In addition, the avermectin doramectin was described as a new inhibitor of ruminant ABCG2. This new rapid assay to identify veterinary drugs that can be concentrated into ovine milk will potentially improve detection and monitoring of veterinary drug residues in ovine milk and dairy products.