Purification and physical properties of the male and female double sex proteins of Drosophila.

The double sex gene (dsx) encodes two proteins, DSX(M) and DSX(F), that regulate sex-specific transcription in Drosophila. These proteins bind target sites in DNA from which the male-specific DSX(M) represses and the female-specific DSX(F) activates transcription of yolk protein (Yp) genes. We investigated the physical properties of these DSX proteins, which are identical in their amino-terminal 397 residues but are entirely different in their carboxyl-terminal sequences (DSX(F), 30 amino acids; DSX(M), 152 amino acids). DSX(M) and DSX(F) were overexpressed in cultured insect cells and purified to near homogeneity. Gel filtration chromatography and glycerol gradient sedimentation showed that at low concentrations both proteins are dimers of highly asymmetrical shape. The axial ratios are approximately 18:1 (DSX(M), 860 X 48 angstroms; DSX(F), 735 X 43 angstroms). At higher concentrations, the proteins form tetramers. Through use of a novel, double crosslinking assay (protein-DNA plus protein-protein), we demonstrated that a DNA regulatory site binds to both monomers of the DSX dimer and to only two monomers of the tetramer. Furthermore, binding another DNA molecule to what we presume is the second and identical site in the tetramer dramatically shifts the equilibrium from tetramers to dimers. These oligomerization and DNA binding properties are indistinguishable between the male and female proteins.

Efficient high-resolution genetic mapping of mouse interspersed repetitive sequence PCR products, toward integrated genetic and physical mapping of the mouse genome.

The ability to carry out high-resolution genetic mapping at high throughput in the mouse is a critical rate-limiting step in the generation of genetically anchored contigs in physical mapping projects and the mapping of genetic loci for complex traits. To address this need, we have developed an efficient, high-resolution, large-scale genome mapping system. This system is based on the identification of polymorphic DNA sites between mouse strains by using interspersed repetitive sequence (IRS) PCR. Individual cloned IRS PCR products are hybridized to a DNA array of IRS PCR products derived from the DNA of individual mice segregating DNA sequences from the two parent strains. Since gel electrophoresis is not required, large numbers of samples can be genotyped in parallel. By using this approach, we have mapped > 450 polymorphic probes with filters containing the DNA of up to 517 backcross mice, potentially allowing resolution of 0.14 centimorgan. This approach also carries the potential for a high degree of efficiency in the integration of physical and genetic maps, since pooled DNAs representing libraries of yeast artificial chromosomes or other physical representations of the mouse genome can be addressed by hybridization of filter representations of the IRS PCR products of such libraries.

Bioactive compounds and physical and chemical characteristics of mini tomatoes: influence of postharvest treatments

Tomatoes are among the most cultivated and used vegetables in the world. They are very succeptible to post harvest losses due to high perishability, therefore the use of postharvest treatments may contribute to conservation of this fruit, however the treatments might affect significantly physico-chemical, sensory and nutritional characteristics of tomatoes. Given the perishability of tomato and the economic importance of small tomato fruits, the purpose of the present study was to determine the effect of gamma radiation, carnauba coating and 1-MCP treatments on tomato fruit quality during storage. The study may be divided into two parts. In the first, mini tomatoes cv. Sweet Grape were harvested at breaker stage, divided into 4 grous and treated with gamma radiation (0.6 kGy), carnauba coating (1 L 1000 kg-1) and 1-MCP (500 nL L-1) and then stored at 25±2°C for 30 days with a control group of tomatoes. In the seconnd part, tomatoes harvested at light-red stage were submitted to the same treatments and storage period. Every 6 days tomatoes were evaluated for color modifications, fruit firmness, souble and total pectin (only for light-red tomatoes), mass loss, titratable acidity (TA), soluble solids (SS), SS/TA ratio, carotenoids profile, formation of lycopene isomers, total phenolic compounds, ascorbic acid and antioxidant capacity. For tomatoes harvested at breaker stage and submitted to the treatments the results showed mass loss was delaying mainly by carnauba wax, and to a lesser extend by 1-MCP. Fruit firmness were better retained for 1-MCP treated fruits and carnauba treatment showed a transient effect in preserving fruit firmness. SS/TA of tomatoes treated with gamma radiation and carnauba presented no differences from control values, and were lower with the application of 1-MCP. Color was negatively affected by 1-MCP and earlier changed (6th day) when gamma radiation was applied. In relation to bioactive compounds of tomatoes harvest at breaker stage, results indicated gamma radiation and 1-MCP decreased the final content of lycopene and produced more (Z)-isomers of lycopene. Gamma radiation also induced a decreased in ?-carotene and an increased in phenolic compounds by the end of storage period. 1-MCP treatment promoted a slow down increase in ascorbic acid content during storage. Antioxidant capacity of the hydrophilic fraction was not dramatically affected by treatments and the lipophilic fraction was lower, especially for 1-MCP fruits. In addition, contents of ?-carotene, lycopene, (Z)-isomers of lycopene, ascorbic acid and antioxidant capacity increased during the period of storage while contents of lutein and phenolic compounds tended to decrease. Regarding tomatoes harvest at light-red stage, the most effective treatments for delaying fruit firmness and mass loss was carnauba and 1-MCP, while gamma radiation was the treatment with higher mass loss and the less fruit firmness, which could be associated with the higher solubilization of pectins promoted by radiation treatment. Color (L* and Hue) was mainly affected by 1-MCP treatment which delayed color development, however, by the end of storage, the values were not different from the other treatments. SS/TA ratio was lower for fruits treated with 1-MCP and TA was not so dramatically affected by treatments. Furthermore, mini tomatoes harvested at light-red stage, demonstrated irradiation induced changes in the final content of lycopene, increasing it, and formed less (13Z)-lycopene, while 1-MCP and carnauba coating slow down the increase in lycopene and slown down the decrease of ascorbic acid and phenolic compounds. Antioxidant capacity of lipophilic fraction was not affected by treatments and the hydrophilic fraction was lower for irradiated fruits only on day 0 as well as phenolic compounds. In the other days, no differences among treatments were observed for hydrophilic antioxidant capacity. Considering the results, the best combination of SS and TA and fruit preservation for mini tomatoes harvest at breaker stage was promoted by carnauba coating, which seems to be the treatment that causes fewer changes in bioactive compounds of breaker tomatoes. However, when mini tomatoes were harvested at light-red stage, SS/TA ratio and color were better and, to preserve the quality of these fruits, besides carnauba coating, 1-MCP also could be indicated

Orbital and Physical Properties of the σ Ori Aa, Ab, B Triple System

We provide a complete characterization of the astrophysical properties of the σ Ori Aa, Ab, B hierarchical triple system and an improved set of orbital parameters for the highly eccentric σ Ori Aa, Ab spectroscopic binary. We compiled a spectroscopic data set comprising 90 high-resolution spectra covering a total time span of 1963 days. We applied the Lehman-Filhés method for a detailed orbital analysis of the radial velocity curves and performed a combined quantitative spectroscopic analysis of the σ Ori Aa, Ab, B system by means of the stellar atmosphere code FASTWIND. We used our own plus other available information on photometry and distance to the system for measuring the radii, luminosities, and spectroscopic masses of the three components. We also inferred evolutionary masses and stellar ages using the Bayesian code BONNSAI. The orbital analysis of the new radial velocity curves led to a very accurate orbital solution of the σ Ori Aa, Ab pair. We provided indirect arguments indicating that σ Ori B is a fast-rotating early B dwarf. The FASTWIND+BONNSAI analysis showed that the Aa, Ab pair contains the hottest and most massive components of the triple system while σ Ori B is a bit cooler and less massive. The derived stellar ages of the inner pair are intriguingly younger than the one widely accepted for the σ Orionis cluster, at 3 ± 1 Ma. The outcome of this study will be of key importance for a precise determination of the distance to the σ Orionis cluster, the interpretation of the strong X-ray emission detected for σ Ori Aa, Ab, B, and the investigation of the formation and evolution of multiple massive stellar systems and substellar objects.

Implementing the Physical Literacy Component of the 2015 Ontario Health and Physical Education Curriculum: Is the Stage Set?

Introduction: Current physical activity levels among children and youth are alarmingly low; a mere 7% of children and youth are meeting the Canadian Physical Activity Guidelines (Colley et al., 2011), which means that the vast majority of this population is at risk of developing major health problems in adulthood (Janssen & Leblanc, 2010). These high inactivity rates may be related to suboptimal experiences in sport and physical activity stemming from a lack of competence and confidence (Lubans, Morgan, Cliff, Barnett, & Okely, 2010). Developing a foundation of physical literacy can encourage and maintain lifelong physical activity, yet this does not always occur naturally as a part of human growth (Hardman, 2011). An ideal setting to foster the growth and development of physical literacy is physical education class. Physical education class can offer all children and youth an equal opportunity to learn and practice the skills needed to be active for life (Hardman, 2011). Elementary school teachers are responsible for delivering the physical education curriculum, and it is important to understand their will and capacity as the implementing agents of physical literacy development curriculum (McLaughlin, 1987). Purpose: The purpose of this study was to explore the physical literacy component of the 2015 Ontario Health and Physical Education curriculum policy through the eyes of key informants, and to explore the resources available for the implementation of this new policy. Methods: Qualitative interviews were conducted with seven key informants of the curriculum policy development, including two teachers. In tandem with the interviews, a resource inventory and curriculum review were conducted to assess the content and availability of physical literacy resources. All data were analyzed through the lens of Hogwood and Gunn’s (1984) 10 preconditions for policy implementation. Results: Participants discussed how implementation is affected by: accountability, external capacity, internal capacity, awareness and understanding of physical literacy, implementation expertise, and policy climate. Discussion: Participants voiced similar opinions on most issues, and the overall lack of attention given to physical education programs in schools will continue to be a major dilemma when trying to combat such high physical inactivity levels.

Structured Sports and Physical Activities: Their Critical Role

Even though one recognizes the physical and psychological kindness of h more and more involvement of the children to the physical and sporty activities, these programs seem more and more inaccessible to some. This article underlines the importance school déprogrammes centered on structured physical and sporty activities. In the setting of a recent survey, one asked youngsters to 5th year to take in note all their activities (as diary calendar) during two days of consecutive week. L·s children that participated in structured physical and sporty activities before and after the hours class sembkient to pull a lot more phisir of their day that those that didn't participate in these activities. Besides, seuk the young that took to structured physical and sporty activities reached the rates recommended of daily physical activity. The article proposes to the school administrators, to the parents, to the children and to the collectivities various measures encouraging the adoption of additional programs centered on the physical and sporty activities in school environment.

BMI and physical activity in diabetic adolescents followed Hospital Barcelos

Introduction: Physical activity is related to health and lifestyle and should be part of the daily routine of all individuals since it brings many benefits to the body. Ains: To study the adolescent population‘s body mass index (BMI). To study the relation between physical activity and gender. Materials and Methods: We performed a quantitative, observational, analytic and cross-sectional study. After the use of exclusion criteria, a sample of 36 individuals was selected from a population of diabetic adolescents. A validated questionnaire was applied to collected physical activity and personal data. SPSS 22.0 was used to treat the data. The chi-square test was applied to study the relation between the level of physical activity and gender; Fisher’s exact test was applied to study the relation between level of physical activity and BMI. Results and discussion: The chi-square test showed a significant relation between the level of physical activity and gender (p-value = 0.018) with moderate intensity (phi = .4), which corroborates other national and worldwide studies. Fisher’s exact test showed no relation between BMI and the level of physical activity (p-value=0,646). Conclusion: 86,1% of the sampled diabetic adolescents are eutrophic, and 66,7% are considered insufficient active. A significant relation was found between gender and the level of physical activity. No relation was found between BMI and level of physical activity.