Stabilization of troponin I for applications in clinical chemistry and forensic medicine. Cardiac troponin I: A time of death marker

Cardiac troponin I (cTnI) is one of the most useful serum marker test for the determination of myocardial infarction (MI). The first commercial assay of cTnI was released for medical use in the United States and Europe in 1995. It is useful in determining if the source of chest pains, whose etiology may be unknown, is cardiac related. Cardiac TnI is released into the bloodstream following myocardial necrosis (cardiac cell death) as a result of an infarct (heart attack). In this research project the utility of cardiac troponin I as a potential marker for the determination of time of death is investigated. The approach of this research is not to investigate cTnI degradation in serum/plasma, but to investigate the proteolytic breakdown of this protein in heart tissue postmortem. If our hypothesis is correct, cTnI might show a distinctive temporal degradation profile after death. This temporal profile may have potential as a time of death marker in forensic medicine. The field of time of death markers has lagged behind the great advances in technology since the late 1850's. Today medical examiners are using rudimentary time of death markers that offer limited reliability in the medico-legal arena. Cardiac TnI must be stabilized in order to avoid further degradation by proteases in the extraction process. Chemically derivatized magnetic microparticles were covalently linked to anti-cTnI monoclonal antibodies. A charge capture approach was also used to eliminate the antibody from the magnetic microparticles given the negative charge on the microparticles. The magnetic microparticles were used to extract cTnI from heart tissue homogenate for further bio-analysis. Cardiac TnI was eluted from the beads with a buffer and analyzed. This technique exploits banding pattern on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by a western blot transfer to polyvinylidene fluoride (PVDF) paper for probing with anti-cTnI monoclonal antibodies. Bovine hearts were used as a model to establish the relationship of time of death and concentration/band-pattern given its homology to human cardiac TnI. The final concept feasibility was tested with human heart samples from cadavers with known time of death. ^

Managing small-medium cities in a time of globalization: Experiences and evidence from Florida's public managers

This study examines how public management practitioners in small and medium-sized Florida cities perceive globalization and its impact on public management practice. Using qualitative analysis, descriptive statistics and factor analysis methods, data obtained from a survey and semi-structured interviews were studied to comprehend how public managers view the management and control of their municipalities in a time of globalization. The study shows that the public managers’ perceptions of globalization and its impact on public management in Florida’s small-medium cities are nuanced. Whereas some public managers feel that globalization has significant impacts on municipalities’ viability, others opine that globalization has no local impact. The study further finds that globalization processes are perceived as altering the public management functions of decision-making, economic development and service delivery in some small-medium cities in Florida as a result of transnational shifts, rapidly changing technologies, and municipalities’ heightened involvement in the global economy. The study concludes that the globalization discourse does not resonate among some public managers in Florida’s small-medium cities in ways implied in extant literature.

Using soil profiles of seeds and molecular markers as proxies for sawgrass and wet prairie slough vegetation in Shark Slough, Everglades National Park

We measured the abundance of Cladium jamaicense (Crantz) seeds and three biomarkers in freshwater marsh soils in Shark River Slough (SRS), Everglades National Park (ENP) to determine the degree to which these paleoecological proxies reflect spatial and temporal variation in vegetation. We found that C. jamaicense seeds and the biomarkers Paq, total lignin phenols (TLP) and kaurenes analyzed from surface soils were all significantly correlated with extant aboveground C. jamaicense biomass quantified along a vegetation gradient from a C. jamaicense to a wet prairie/slough (WPS) community. Our results also suggest that these individual proxies may reflect vegetation over different spatial scales: Paq and kaurenes correlated most strongly (R 2 = 0.88 and 0.99, respectively) with vegetation within 1 m of a soil sample, while seeds and TLP reflected vegetation 0–20 m upstream of soil samples. These differences in the spatial scale depicted by the different proxies may be complementary in understanding aspects of historic landscape patterning. Soil profiles of short (25 cm) cores showed that downcore variation in C. jamaicense seeds was highly correlated with two of the three biomarkers (Paq, R 2 = 0.84, p<0.005; TLP, R 2 = 0.97, p<0.0001), and all four of the proxies indicated a recent increase in C. jamaicense biomass at the site. Using a preliminary depth-to-age relationship based on matching charcoal peaks with available ENP fire records (1980-present) specific to our coring site, we found that peak-depths in C. jamaicense seed concentration appeared to correspond to recent minimum water levels (e.g., 1989 and 2001), and low seed abundance corresponded to high water levels (e.g., 1995), consistent with the known autecology of C. jamaicense. In summary, the combination of C. jamaicense seeds and biomarkers may be useful for paleoecological reconstruction of vegetation change and ultimately in guaging the success of ongoing efforts to restore historic hydrologic conditions in the South Florida Everglades.

Elementary music and classroom teachers' time -use: A comparison by grade level, time of day, and day of the week

This study investigated time-use of elementary music teachers and elementary classroom teachers to determine: (1) whether there was a relationship between grade level, time of day, and day of the week and teachers' time-use in teaching, monitoring, and non-curricular, and (2) whether ethnicity, training, and years of experience affect teacher time-use. Sixty-nine music teachers and 55 classroom teachers participated. ^ A MANOVA was used to examine the hypothesized relationship. ANOVA results were significant for time spent teaching, monitoring, and non-curricular. An independent t test revealed a significance difference (t (302) = 5.20, p < .001) between the two groups of teachers. A significant difference was found for teaching, t (302) = 5.20, p < .001: music teachers spent more time actively teaching than did classroom teachers. There was a significant difference for monitoring (t (302) = 13.62, p < .001): classroom teachers allocated more time to monitoring than did music teachers. A significant difference was also found for non-curricular (t (302) = 7.03, p < .001): music teachers spent more time in this category of activities than did classroom teachers. ^ Analyses of the activities subsumed under the major categories indicated significant differences between elementary music teachers and elementary classroom teachers, overall, in subject matter (p < .001), discussion (p < .05), school-wide activities (p < .001), seatwork (p < .001), giving directions (p < .001), changing activities (p < .001), lunch (p < .05), planning (p < .001) and interruption (p < .001). Analyses of the relationship and ethnicity, training, degree, experience indicated significant difference for main effect, ethnicity (F(2, 116) = 4.22, p < .017). Time-use for black non-Hispanic teachers was higher than time-use for those who were Hispanic and white non-Hispanic. ^ Analyses of time-use by grade showed no increase for either group as grade level increased. A statistically significant Wilks Lambda ( F (1,294) = .917 p < .013) was found for the independent variable day of the week. ANOVA indicated that elementary classroom teachers monitored more on Thursdays and Fridays: music teachers allocated more time to non-curricular activities on Fridays. ^

Elementary music and classroom teachers' time-use: a comparison by grade level, time of day, and day of the week

This study investigated time-use of elementary music teachers and elementary classroom teachers to determine: (1) whether there was a relationship between grade level, time of day, and day of the week and teachers' time-use in teaching, monitoring, and non-curricular, and (2) whether ethnicity, training, and years of experience affect teacher time-use. Sixty-nine music teachers and 55 classroom teachers participated. A MANOVA was used to examine the hypothesized relationship. ANOVA results were significant for time spent teaching, monitoring, and non-curricular. An independent t test revealed a significance difference (t (302) = 5.20, p Analyses of the activities subsumed under the major categories indicated significant differences between elementary music teachers and elementary classroom teachers, overall, in subject matter ( p teachers was higher than time-use for those who were Hispanic and white non-Hispanic. Analyses of time-use by grade showed no increase for either group as grade level increased. A statistically significant Wilks Lambda ( F (1,294) = .917 p < .013 ) was found for the independent variable day of the week. ANOVA indicated that elementary classroom teachers monitored more on Thursdays and Fridays: music teachers allocated more time to non-curricular activities on Fridays.