In Vitro Development and Cell Allocation After Aggregation of Syngeneic Wild Type and Fluorescence-Expressing Bovine Cloned Embryos

Background: The in vitro production (IVP) of embryos by in vitro fertilization or cloning procedures has been known to cause epigenetic changes in the conceptus that in turn are associated with abnormalities in pre- and postnatal development. Handmade cloning (HMC) procedures and the culture of zona-free embryos in individual microwells provide excellent tools for studies in developmental biology, since embryo development and cell allocation patterns can be evaluated under a wide range of embryo reconstruction arrangements and in in vitro embryo culture conditions. As disturbances in embryonic cell allocation after in vitro embryo manipulations and unusual in vivo conditions during the first third of pregnancy appear to be associated with large offspring, embryo aggregation procedures may allow a compensation for epigenetic defects between aggregated embryos or even may influence more favorable cell allocation in embryonic lineages, favoring subsequent development. Thus, the aim of this study was to evaluate in vitro embryo developmental potential and the pattern of cell allocation in blastocysts developed after the aggregation of handmade cloned embryos produced using syngeneic wild type and/or transgenic somatic cells. Materials, Methods & Results: In vitro-matured bovine cumulus-oocyte complexes (COC) were manually bisected after cumulus and zona pellucida removal; then, two enucleated hemi-oocytes were paired and fused with either a wild type (WT) or a GFP-expressing (GFP) fetal skin cell at the 11th and 19th passages, respectively. Following chemical activation, reconstructed cloned embryos and zona-free parthenote embryos were in vitro-cultured in microwells, for 7 days, either individually (1 x 100%) or after the aggregation of two structures (2 x 100%) per microwell, as follows: (G1) one WT cloned embryo; (G2) two aggregated WT embryos; (G3) one GFP cloned embryo; (G4) two aggregated GFP embryos; (G5) aggregation of a WT embryo and a GFP embryo; (G6) one parthenote embryo; or (G7) two aggregated parthenote embryos. Fusion (clones), cleavage (Day 2), and blastocyst (Day 7) rates, and embryonic cell allocation were compared by the. 2 or Fisher tests. Total cell number (TCN) in blastocysts was analyzed by the Student's test (P < 0.05). Fusion and cleavage rates, and cell allocation were similar between groups. On a per WOW basis, development to the blastocyst stage was similar between groups, except for lower rates of development seen in G3. However, when based on number of embryos per group (one or two), blastocyst development was higher in G1 than all other groups, which were similar between one another. Cloned GFP embryos had lower in vitro development to the blastocyst stage than WT embryos, which had more TCN than parthenote or aggregated chimeric WT/GFP embryos. Aggregated GFP embryos had fewer cells than the other embryo groups. Discussion: The in vitro development of GFP cloned embryos was lower than WT embryos, with no effects on cell allocation in resulting blastocysts. Differences in blastocyst rate between groups were likely due to lower GFP-expressing cell viability, as GFP donor cells were at high population cell doublings when used for cloning. On a per embryo basis, embryo aggregation on Day 1 resulted in blastocyst development similar to non-aggregated embryos on Day 7, with no differences in cell proportion between groups. The use of GFP-expressing cells was proven a promising strategy for the study of cell allocation during embryo development, which may assist in the elucidation of mechanisms of abnormalities after in vitro embryo manipulations, leading to the development of improved protocols for the in vitro production (IVP) of bovine embryos.

The impact of dental caries and trauma in children on family quality of life

Objectives: To assess the impact of childrens dental caries (DC) and traumatic dental injuries (TDI) on parents quality of life (QoL), adjusted by family income. Methods: Parents of 219 children aged 5 and 6 years answered the Family Impact Scale (FIS) on their perception of QoL and data about income. Three calibrated dentists examined the severity of DC according to decayed, missing and filled permanent teeth index, and children were categorized into: 0 = caries free; 15 = low severity; and =6 = high severity. TDI were classified into uncomplicated and complicated injuries. QoL was measured through FIS items and total score, and Poisson regression was used to associate the variables with the outcome. Results: Severity of DC showed a negative impact on the total score and subscales on parental/family activities, parental emotions and financial burden (P < 0.001). TDI showed a negative impact on total score and in some FIS items. The multivariate-adjusted model showed that only the increase in the severity of childrens DC (RR = 3.19; 95% CI = 2.36, 4.31; P < 0.001) was associated with a greater negative impact on parents QoL, while high family income was a protective factor (RR = 0.68; 95% CI = 0.48, 0.95; P < 0.001). Conclusions: The severity of childrens DC has a negative impact on parents QoL, whereas TDI do not. A lower family income might have a negative impact on parents QoL.

Measurement of prompt J/psi and beauty hadron production cross sections at mid-rapidity in pp collisions at root s=7 TeV

The ALICE experiment at the LHC has studied J/psi production at mid-rapidity in pp collisions at root s = 7 TeV through its electron pair decay on a data sample corresponding to an integrated luminosity L-int = 5.6 nb(-1). The fraction of J/psi from the decay of long-lived beauty hadrons was determined for J/psi candidates with transverse momentum p(t) > 1,3 GeV/c and rapidity vertical bar y vertical bar < 0.9. The cross section for prompt J/psi mesons, i.e. directly produced J/psi and prompt decays of heavier charmonium states such as the psi(2S) and chi(c) resonances, is sigma(prompt J/psi) (p(t) > 1.3 GeV/c, vertical bar y vertical bar < 0.9) = 8.3 +/- 0.8(stat.) +/- 1.1 (syst.)(-1.4)(+1.5) (syst. pol.) mu b. The cross section for the production of b-hadrons decaying to J/psi with p(t) > 1.3 GeV/c and vertical bar y vertical bar < 0.9 is a sigma(J/psi <- hB) (p(t) > 1.3 GeV/c, vertical bar y vertical bar < 0.9) = 1.46 +/- 0.38 (stat.)(-0.32)(+0.26) (syst.) mu b. The results are compared to QCD model predictions. The shape of the p(t) and y distributions of b-quarks predicted by perturbative QCD model calculations are used to extrapolate the measured cross section to derive the b (b) over bar pair total cross section and d sigma/dy at mid-rapidity.

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.

Plant invasions research in Latin America: fast track to a more focused agenda

While many developed countries have invested heavily in research on plant invasions over the last 50 years, the immense region of Latin America has made little progress. Recognising this, a group of scientists working on plant invasions in Latin America met in Chile in late 2010 to develop a research agenda for the region based on lessons learned elsewhere. Our three main findings are as follows. (1) Globalisation is inevitable, but the resultant plant introductions can be slowed or prevented by effective quarantine and early intervention. Development of spatially explicit inventories, research on the invasion process and weed risk assessments can help prioritise and streamline action. (2) Eradication has limited application for plants and control is expensive and requires strict prioritisation and careful planning and evaluation. (3) Accepting the concept of novel ecosystems, new combinations of native and introduced species that no longer depend on human intervention, may help optimise invasive species management. Our vision of novel ecosystem management is through actions that: (a) maintain as much native biodiversity and ecosystem functionality as possible, (b) minimise management intervention to invasives with known impact, and (c) maximise the area of intervention. We propose the creation of a Latin American Invasive Plants Network to help focus the new research agenda for member countries. The network would coordinate research and training and establish funding priorities, develop and strengthen tools to share knowledge, and raise awareness at the community, governmental and intergovernmental levels about the social, economic and environmental costs of plant invasions.

Responsiveness to change for the Brazilian Scale of Oral Health Outcomes for 5-year-old children (SOHO-5)

Abstract Background The responsiveness of oral health-related quality of life (OHRQoL) instruments has become relevant, given the increasing tendency to use OHRQoL measures as outcomes in clinical trials and evaluations studies. The purpose of this study was to assess the responsiveness of the Brazilian Scale of Oral Health Outcomes for 5-year-old children (SOHO-5) to dental treatment. Methods One hundred and fifty-four children and their parents completed the child self- and parental’ reports of the SOHO-5 prior to treatment and 7 to 14 days after the completion of treatment. The post-treatment questionnaire also included a global transition judgment that assessed subject’s perceptions of change in their oral health following treatment. Change scores were calculated by subtracting post-treatment SOHO-5 scores from pre-treatment scores. Longitudinal construct validity was assessed by using one-way analysis of variance to examine the association between change scores and the global transition judgments. Measures of responsiveness included standardized effect sizes (ES) and standardized response mean (SRM). Results The improvement of children’s oral health after treatment are reflected in mean pre- and post-treatment SOHO-5 scores that declined from 2.67 to 0.61 (p < 0.001) for the child-self reports, and 4.04 to 0.71 (p < 0.001) for the parental reports. Mean change scores showed a gradient in the expected direction across categories of the global transition judgment, and there were significant differences in the pre- and post-treatment scores of those who reported improving a little (p < 0.05) and those who reported improving a lot (p < 0.001). For both versions, the ES and SRM based on change scores mean for total scores and for categories of global transitions judgments were moderate to large. Conclusions The Brazilian SOHO-5 is responsive to change and can be used as an outcome indicator in future clinical trials. Both the parental and the child versions presented satisfactory results.

Cross-cultural adaptation and psychometric properties of the Brazilian version of the scale of oral health outcomes for 5-year-old children (SOHO-5)

Abstract Background Most of the instruments available to measure the oral health-related quality of life (OHRQoL) in paediatric populations focus on older children, whereas parental reports are used for very young children. The scale of oral health outcomes for 5-year-old children (SOHO-5) assesses the OHRQoL of very young children through self-reports and parental proxy reports. We aimed to cross-culturally adapt the SOHO-5 to the Brazilian Portuguese language and to assess its reliability and validity. Findings We tested the quality of the cross-cultural adaptation in 2 pilot studies with 40 children aged 5–6 years and their parents. The measurement was tested for reliability and validity on 193 children that attended the paediatric dental screening program at the University of São Paulo. The children were also clinically examined for dental caries. The internal consistency was demonstrated by a Cronbach's alpha coefficient of 0.90 for the children’s self-reports and 0.77 for the parental proxy reports. The test-retest reliability results, which were based on repeated administrations on 159 children, were excellent; the intraclass correlation coefficient was 0.98 for parental and 0.92 for child reports. In general, the construct validity was satisfactory and demonstrated consistent and strong associations between the SOHO-5 and different subjective global ratings of oral health, perceived dental treatment need and overall well-being in both the parental and children’s versions (p < 0.001). The SOHO-5 was also able to clearly discriminate between children with and without a history of dental caries (mean scores: 5.8 and 1.1, respectively; p < 0.001). Conclusion The present study demonstrated that the SOHO-5 exhibits satisfactory psychometric properties and is applicable to 5- to 6-year-old children in Brazil.

Non-normal modal logics, quantification, and deontic dilemmas. A study in multi-relational semantics

This dissertation is devoted to the study of non-normal (modal) systems for deontic logics, both on the propositional level, and on the first order one. In particular we developed our study the Multi-relational setting that generalises standard Kripke Semantics. We present new completeness results concerning the semantic setting of several systems which are able to handle normative dilemmas and conflicts. Although primarily driven by issues related to the legal and moral field, these results are also relevant for the more theoretical field of Modal Logic itself, as we propose a syntactical, and semantic study of intermediate systems between the classical propositional calculus CPC and the minimal normal modal logic K.

Introduzione alla logica e alla matematica intuizioniste

L'elaborato propone un'introduzione alla matematica intuizionista e alla sua formalizzazione. Dopo aver inquadrato storicamente il movimento e averne tratteggiato il pensiero, si approfondiscono la sintassi e la semantica ad esso associate grazie ai lavori di Heyting e Kripke, si confronta la teoria con quella classica, e si indica il legame tra la logica intuizionista e quella modale. Il capitolo conclusivo è dedicato alla costruzione dei numeri naturali e reali, con particolare attenzione alle conseguenze metodologiche del pensiero intuizionista, ai concetti originali e alle nozioni più fini in cui si suddividono alcune nozioni fondamentali della matematica tradizionale.