Investigation of the Time Dependent Influence of Extracellular Osmotic Stress on Protein Turnover in Skeletal Muscle Cells

Acute alterations in cell volume can substantively modulate subsequent metabolism of substrates. However, how such alterations in skeletal muscle modulate protein metabolism is limited. The purpose of this study was to determine the time dependent influence of extracellular osmotic stress on protein turnover in skeletal muscle cells. L6 cells were incubated in hyperosmotic (HYPER; 425.3 ± 1.8mmol/kg), hypo-osmotic (HYPO; 235.4 ± 1.0mmol/kg) or control (CON; 333.5 ± 1.4mmol/kg) media for 4, 8, 12, or 24hrs. During the final 4hrs, incorporation of L-[ring-3,5-3H]-tyrosine was measured to estimate protein synthesis. Western blotting measured markers of protein synthesis and degradation. No differences were observed in any outcomes except p70S6K phosphorylation whereby HYPO was lower (p<0.05) than CON and HYPER; which remained similar except for a large increase at 8hrs for HYPER. These findings suggest that regardless of duration, extracellular osmotic stress does not significantly affect protein metabolism in L6 cells.

Expression and function of endothelin and its receptors in vascular adventitial fibroblasts

Objective: The adventitia has been recognized to play important roles in vascular oxidative stress, remodelling and contraction. We recently demonstrated that adventitial fibroblasts are able to express endothelin-1 (ET-1) in response to angiotensin II (ANG II). However, the mechanisms by which ANG II induces ET-1 expression are unknown. It is also unclear whether the ET-1 receptors are expressed in the adventitia. We therefore examined the role of oxidative stress in the regulation of ET-1. We also investigated the expression of both the ETA and ETB receptors and the roles of these two types of receptors in collagen synthesis and ET-1 clearance in adventitial fibroblasts. Methods and Results: Adventitial fibroblasts were isolated and cultured from the thoracic mouse aorta. Cells were treated with ANG II (lOOnM), ET-1 (lOpM), NADPH oxidase inhibitor apocynin (lOOfiM), the superoxide anion scavenger tempol (lOOfiM), the ANG II receptor antagonists (100[aM), losartan (AT| receptor) and PD 1233 19 (AT2 receptor), the ET-1 receptor antagonists (lOOuM), BQ123 (ETA receptor) and BQ788 (ETB receptor), and the ETB receptor agonist (lOOnM) Sarafotoxin 6C. ET-1 peptide levels were determined by ELISA, while ETA ,ETB and collagen levels were determined by Western blot. ANG II increased ET-1 peptide levels in a time-dependent manner reaching significance when incubated for 24 hours. NAD(P)H oxidase inhibitor, apocynin, as well as the superoxide scanverger, tempol, significantly reduced ANG Il-induced ET-1 peptide levels while over-expression of SOD1 (endogenous antioxidant enzyme) significantly decreased ANG Il-induced collagen I expression, therefore implicating reactive oxygen species in the mediation of ET-1. ANG II increased ETA receptor protein as well as collagen in a similar fashion, reaching significance after 4, 6, and 24 hours treatment. ANG II induced collagen was reduced while in the presence of the ETA receptor antagonist suggesting the role of the ETa receptor in the regulation of the extracellular matrix. ANG II treatment also increased ETB receptor protein levels in a time-dependent manner. ANG II treatment in the presence of the ETB receptor antagonist significantly increased ET-1 peptide levels. On another hand, the ETB receptor agonist, Sarafotoxin 6C, significantly decreased ET-1 peptide levels. These data implicate the role of the ETb receptor in the clearance of the ET-1 peptide. Conclusion: ANG II-induced increases of ET-1 peptide appears to be mediated by reactive oxygen species derived from NAD(P)H oxidase. Both the ETA and ETB receptors are expressed in adventitial fibroblasts. The ETA receptor subtype mediates collagen I expression, while the ETB receptor may play a protective role through increasing the clearance of the ET- 1 peptide.

An exploration of women elementary school teachers' perceptions of how their decision to return to teaching part-time from a maternity leave influences their professional and personal lives /

The purpose of this research was to explore women elementary teachers' perceptions of how their decision to return to teaching part-time from a maternity leave influences their professional and personal lives. The investigation focused on the decisions surrounding a mother's choice to reenter the teaching profession parttime in a field where each mother had previously been employed full-time. A collective case study was undertaken based on an in-depth interview with five mothers who had made the choice to return to the classroom part-time. The data collected in this study were analyzed and interpreted using qualitative methods. The following four major themes emerged from the interviews: decisionmaking process, challenges faced by mothers who teach part-time, the importance of support, and the enhancement of instructional practice from parenthood. Using these four themes, an analysis was conducted to examine the similarities and differences among the experiences of the participants. The mothers' reflections, my analysis, and the related literature were used at the conclusion of this report to compile implications for teaching practice, theory, and further research.

Nursing staffs' perceptions of a generic service manager position

The recent reengineering within the health care industry has challenged many assumptions regarding traditional structures and roles. Within a product-line management structure, the traditional viewpoint that those who manage patient care areas must have a nursing background, is an example of one such assumption being challenged. The nursing profession is often seen as the greatest obstacle to the implementation of a product-line management structure and generic manager positions (does not require a nursing background), due to the perceived loss of professional identity. This qualitative study focused on how nursing staff within a chronic care and rehabilitation facility perceived a generic service manager position. Focus groups were conducted in three phases, over a 14 month period of time. The data collected from the focus groups were then coded according to common themes. Each phase was analyzed independently, with the study concluding with an analysis and interpretation of the collective results. The results of this study revealed a significant shift in how the nursing staff perceived their professional identity and accountability in light of the implementation of the generic Service Manager position. Initial reactions of personal and professional vulnerability and resentment were seen to transform into an increased ability to explicitly articulate the role of nursing. Changes in behavior that were described included: increased consultation and collaboration with other

Temporal variation in the coral reefs of the east coast of the inner gulf of Thailand

A series of permanent line transects established on fourteen reefs on the eastern seaboard of the Gulf of Thailand were monitored through a three-year period (1995- 1998) using a video transect method. Hierarchical cluster analysis shows three distinctive reef community types dominated by 1) Porites, 2) Acropora and 3) zoantharians. The reefs are developed under naturally turbid conditions and relatively low salinity due to the proximity of four major river outlets located in the uppermost area of the gulf. The number of Acroporid species on the reefs is positively correlated with distance from the major flver outlets. Eighty-seven species of scleractinian coral were found on the transects. Over the three-year period, the comparison of 1995-97-98 matched stations using Repeated Measures ANOV A reveals no significant time-dependent change in percent area cover of reef components except for an overall significant reduction in the faviid coral component. In the 1997-98 matched station comparison, statistical tests reveal significant increases in both Acropora and Porites components that translated into an overall increase in total living coral cover. These findings indicate that the overall environmental conditions have been favorable for coral growth. Outcompetition of massive corals by faster growing corals on several reefs also indicates conditions favorable for reef expansion. Growth of newlyformed Porites colonies over primary rock substrate and dead coral skeleton was presumably responsible for its rapid increase. Although these reefs are in an area of rapid industrialization and population growth, resultant anthropogenic effects have not yet stopped active coral accretion.

The influence of familial smoking behaviours on adolescent smoking behaviours

The ability to identify adolescents who are at risk for becoming smokers may prove useful in developing effective smoking prevention programs. The purpose of this stUdy was to assess the importance of familial smoking behaviours on adolescent smoking patterns. The results were based on responses to The Grade 7 Lincoln County Smoking Survey designed by Chudzik and Partington (1994), and are a part of the "Peer Assisted Learning Program· (PAL) presented by the Niagara Regional Health Services Department, with the cooperation of a local Board of Education (Region of Niagara). The results indicate that 12% of the total group of 450 Grade 7 student respondents were current smokers at the time the data were collected (13% males and 11% females), while more than 37% of individuals indicated that they had tried smoking previously. Of the individuals who were classified as smokers, 11% reported that they smoked because their parents smoked, but only 6% reported that they smoked because their siblings smoked. More concerning, however, is the finding that 4% of smokers reported that they felt pressured to smoke by their relatives. In a society that is becoming increasingly concerned about health, it is also alarming to observe that only 50% of the respondents within this sample reported that there were no smokers (parents/siblings) in their homes. The results also indicate that 33% percent of respondents had grandparents who continued to smoke, and 53% of respondents indentified other relatives who continued to smoke.

Evolutionary synthesis of stochastic gene network models using feature-based search spaces

A feature-based fitness function is applied in a genetic programming system to synthesize stochastic gene regulatory network models whose behaviour is defined by a time course of protein expression levels. Typically, when targeting time series data, the fitness function is based on a sum-of-errors involving the values of the fluctuating signal. While this approach is successful in many instances, its performance can deteriorate in the presence of noise. This thesis explores a fitness measure determined from a set of statistical features characterizing the time series' sequence of values, rather than the actual values themselves. Through a series of experiments involving symbolic regression with added noise and gene regulatory network models based on the stochastic 'if-calculus, it is shown to successfully target oscillating and non-oscillating signals. This practical and versatile fitness function offers an alternate approach, worthy of consideration for use in algorithms that evaluate noisy or stochastic behaviour.

Examining the role of goals and motives for physical activity and eating behaviour in commercial weight-loss program participants

Background: The purpose of this study was to examine the relationships between physical activity and healthy eating behaviour with the participant's motives and goals for each health behaviour. Methods: Participants (N 121; 93.2% female) enrolled in commercial weightloss programs at the time of data collection, completed self-reported instruments using a web-based interface that were in accordance with Deci and Ryan's (2002) Self-Determination Theory (SDT). Results: Multiple linear regression models revealed that motivation and goals collectively accounted for between 0.21 to 0.29 percent and 0.03 to 0.16 percent of the variance in physical and healthy eating behaviours in this sample. In general, goals regarding either behaviour did not appear to have strong predictive relationships with each health behaviour beyond the contributions of motives. Discussion: Overall, findings from this study suggest that motives seem to mattermore than goals for both physical activity and healthy eating behaviour in clientele of commercial weight-loss programs. Therefore commercial weight-loss program implementers may want to consider placing more attention on motives I than goals for their clientele when designing weight-loss and weight-maintenance initiatives.

Effects of naringenin on glucose uptake in L6 skeletal muscle cells

Diabetes mellitus is a disorder of inadequate insulin action and consequent high blood glucose levels. Type 2 diabetes accounts for the majority of cases of the disease and is characterized by insulin resistance and relative insulin deficiency resulting in metabolic deregulation. It is a complex disorder to treat as its pathogenesis is not fully understood and involves a variety of defects including ~-cell failure, insulin resistance in the classic target tissues (adipose, muscle, liver), as well as defects in a-cells and kidney, brain, and gastrointestinal tissue. Present oral treatments, which aim at mimicking the effects of insulin, remain limited in their efficacy and therefore the study of the effects of novel compounds on insulin target tissues is an important area of research both for potentially finding more treatment options as well as for increasing our knowledge of metabolic regulation in health and disease. In recent years the extensively studied polyphenol, resveratrol, has been reported to have antidiabetic effects showing that it increases glucose uptake by skeletal muscle cells and prevents fatty acid-induced insulin resistance in vitro and in vivo. Naringenin, a citrus flavonoid with structural similarities to resveratrol, is reported to have antioxidan.t, antiproliferative, anticancer, and anti-inflammatory properties. Effects on glucose and lipid metabolism have also been reported including blood glucose and lipid lowering effects. However, whether naringenin has insulinlike effects is not clear. In the present study the effects of naringenin on glucose uptake in skeletal muscle cells are examined and compared with those of insulin. Naringenin treatment of L6 myotubes increased glucose uptake in a dose- and time dependent manner and independent of insulin. The effects of naringenin on glucose uptake achieved similar levels as seen with maximum insulin stimulation and its effect was additive with sub-maximal insulin treatment. Like insulin naringenin treatment did not increase glucose uptake in myoblasts. To elucidate the mechanism involved in naringenin action we looked at its effect on phosphatidylinositol 3-kinase (PI3K) and Akt, two signalling molecules that are involved in the insulin signalling cascade leading to glucose uptake. Naringenin did not stimulate basal or insulinstimulated Akt phosphorylation but inhibition of PI3K by wortmannin partially repressed the naringenin-induced glucose uptake. We also examined naringenin's effect on AMP-activated protein kinase (AMPK), a molecule that is involved in mediating glucose uptake by a variety of stimuli. Naringenin stimulated AMPK phosphorylation and this effect was not inhibited by wortmannin. To deduce the nature of the naringenin-stimulated AMPK phosphorylation and its impact on glucose uptake we examined the role of several molecules implicated in mod.ulating AMPK activity including SIRTl, LKB 1, and ca2+ Icalmodulin-dependent protein kinase kinase (CaMKK). Our results indicate that inhibition of SIRTI did not prevent the naringeninstimulated glucose uptake Of. AMPK phosphorylation; naringenin did not stimulate LKB 1 phosphorylation; and inhibition of CaMKK did not prevent naringeninstimulated glucose uptake. Inhibition of AMPK by compound C also did not prevent naringenin-stimulated glucose uptake but effectively inhibited the phosphorylation of AMPK suggesting that AMPK may not be required for the naringenin-stimulated glucose uptake.

Mapping one university's response to internationalization : a case study

This study explored one university's response to the internationalization of higher education. Case study methodology was employed through a review of current and archival documents and interviews with key actors in the international spheres of the university. The historical, current, and future contexts were considered to situate the case study on a time line. Data analysis revealed that there were several points of division among the university community related to the response to internationalization, but also a major point of coherence in the centrality of inter-cultural understanding in efforts to internationalize. Other key findings included strengths, areas for improvement, and future directions of the university's response to internationalization. All of these findings were contextualized in findings related to the history of the university. In addition to these major findings, three themes in relation to the vision for internationalization at the institution were revealed: ( a) intercultural understanding, (b) the comprehensive status of the university, and (c) the financial benefits of internationalization. Recommendations are made for practice at the university in order to clarify this vision to develop a clear foundation from which to further build a response to internationalization that is solidly based on inter-cultural understanding, and recommendations for future research into the process of internationalization at the institutional level in Canada are suggested.

I Believe in me, not OCB: Evaluation of Group Function-Based CBT as a Treatment for Obsessive Compulsive Behaviour in Four Children with HFA

Researchers have conceptualized repetitive behaviours in individuals with Autism Spectrum Disorder (ASD) on a continuum oflower-Ievel, motoric, repetitive behaviours and higher-order, repetitive behaviours that include symptoms ofOCD (Hollander, Wang, Braun, & Marsh, 2009). Although obsessional, ritualistic, and stereotyped behaviours are a core feature of ASD, individuals with ASD frequently experience obsessions and compulsions that meet DSM-IV-TR (American Psychiatric Association, 2000) criteria for Obsessive-Compulsive Disorder (OCD). Given the acknowledged difficulty in differentiating between OCD and Autism-related obsessive-compulsive phenomena, the present study uses the term Obsessive Compulsive Behaviour (OCB) to represent both phenomena. This study used a multiple baseline design across behaviours and ABC designs (Cooper, Heron, & Heward, 2007) to investigate if a 9-week Group Function-Based Cognitive Behavioural Therapy (CBT) decreased OCB in four children (ages 7 - 11 years) with High Functioning Autism (HFA). Key treatment components included traditional CBT components (awareness training, cognitive-behavioural skills training, exposure and response prevention) as well as function-based assessment and intervention. Time series data indicated significant decreases in OCBs. Standardized assessments showed decreases in symptom severity, and increases in quality of life for the participants and their families. Issues regarding symptom presentation, assessment, and treatment of a dually diagnosed child are discussed.