Diatoms as indicators of lake trophic status

Relationships between surface sediment diatom assemblages and lake trophic status were studied in 50 Canadian Precambrian Shield lakes in the Muskoka-Haliburton and southern Ontario regions. The purpose of this study was to develop mathematical regression models to infer lake trophic status from diatom assemblage data. To achieve this goal, however, additional investigations dealing with the evaluation of lake trophic status and the autecological features of key diatom species were carried out. Because a unifying index and classification for lake trophic status was not available, a new multiple index was developed in this study, by the computation of the physical, chemical and biological data from 85 south Ontario lakes. By using the new trophic parameter, the lake trophic level (TL) was determined: TL = 1.37 In[1 +(TP x Chl-a / SD)], where, TP=total phosphorus, Chl-a=chlorophyll-a and SD=Secchi depth. The boundaries between 7 lake trophic categories (Ultra-oligotrophic lakes: 0-0.24; Oligotrophic lakes: 0.241-1.8; Oligomesotrophic lakes: 1.813.0; Mesotrophic lakes: 3.01-4.20; Mesoeutrophic lakes: 4.21-5.4; Eutrophic lakes: 5.41-10 and Hyper-eutrophic lakes: above 10) were established. The new trophic parameter was more convenient for management of water quality, communication to the public and comparison with other lake trophic status indices than many of the previously published indices because the TL index attempts to Increase understanding of the characteristics of lakes and their comprehensive trophic states. It is more reasonable and clear for a unifying determination of true trophic states of lakes. Diatom specIes autecology analysis was central to this thesis. However, the autecological relationship of diatom species and lake trophic status had not previously been well documented. Based on the investigation of the diatom composition and variety of species abundance in 30 study lakes, the distribution optima of diatom species were determined. These determinations were based on a quantitative method called "weighted average" (Charles 1985). On this basis, the diatom species were classified into five trophic categories (oligotrophic, oligomesotrophic, mesotrophic, mesoeutrophic and eutrophic species groups). The resulting diatom trophic status autecological features were used in the regressIon analysis between diatom assemblages and lake trophic status. When the TL trophic level values of the 30 lakes were regressed against their fi ve corresponding diatom trophic groups, the two mathematical equations for expressing the assumed linear relationship between the diatom assemblages composition were determined by (1) uSIng a single regression technique: Trophic level of lake (TL) = 2.643 - 7.575 log (Index D) (r = 0.88 r2 = 0.77 P = 0.0001; n = 30) Where, Index D = (0% + OM% + M%)/(E% + ME% + M%); 4 (2) uSIng a' multiple regressIon technique: TL=4.285-0.076 0%- 0.055 OM% - 0.026 M% + 0.033 ME% + 0.065 E% (r=0.89, r2=0.792, P=O.OOOl, n=30) There was a significant correlation between measured and diatom inferred trophic levels both by single and multiple regressIon methods (P < 0.0001, n=20), when both models were applied to another 20 test lakes. Their correlation coefficients (r2 ) were also statistically significant (r2 >0.68, n=20). As such, the two transfer function models between diatoms and lake trophic status were validated. The two models obtained as noted above were developed using one group of lakes and then tested using an entirely different group of lakes. This study indicated that diatom assemblages are sensitive to lake trophic status. As indicators of lake trophic status, diatoms are especially useful in situations where no local trophic information is available and in studies of the paleotrophic history of lakes. Diatom autecological information was used to develop a theory assessing water quality and lake trophic status.

Postglacial water levels in the Great Lakes Region in relation to Holocene climate change : Thecamoebian and Palynological evidence /

Various lake phases have developed in the upper Great Lakes in response to isostatic adjustment and changes in water supply since the retreat of the Laurentide Ice Sheet. Georgian Bay experienced a lowstand that caused a basin wide unconformity approximately 7,500 years ago that cannot be explained by geological events. Thecamoebians are shelled protozoans abundant in freshwater environments and they are generally more sensitive to changing environmental conditions than the surrounding vegetation. Thecamoebians can be used to reconstruct the paleolimnology. The abundance of thecamoebians belonging to the genus Centropyxis, which are known to tolerate slightly brackish conditions (i.e. high concentrations of ions) records highly evaporative conditions in a closed basin. During the warmer interval (9000 to 700 yBP), the Centropyxis - dominated population diminishes and is replaced by an abundant and diverse Difflugia dominate population. Historical climate records from Tobermory and Midland, Ontario were correlated with the Lake Huron water level curve. The fossil pollen record and comparison with modem analogues allowed a paleo-water budget to be calculated for Georgian Bay. Transfer function analysis of fossil pollen data from Georgian Bay records cold, dry winters similar to modem day Minneapolis, Minnesota. Drier climates around this time are also recorded in bog environments in Southem Ontario - the drying of Lake Tonawanda and inception of paludification in Willoughby Bog, for instance, dates around 7,000 years ago. The dramatic impact of climate change on the water level in Georgian Bay underlines the importance of paleoclimatic research for predicting future environmental change in the Great Lakes.

Autonomic regulation and blood pressure in children

Vagal baroreflex sensitivity (BRS) is a measure of short term blood pressure (BP) regulation through alterations in heart rate. Low BRS reflects impaired autonomic system regulation and has been found to be a surrogate marker for cardiovascular health. In particular, it has found to be associated with the pathogenesis of adult hypertension. However, only limited information exists as to the negative consequences of childhood BP on baroreflex function. The objective of this study was to investigate BRS in children with 2 different BP profiles while controlling for the effects of age, maturation, sex, and body composition. A preliminary subsample of 11-14 year-old children from the HBEAT (Heart Behavioural Environmental Assessment Team) Study was selected. The children were divided into 2 BP groups; high BP (HBP; 2:95tl1 percentile, n=21) and normal BP (NBP; <90th percentile, n=85). Following an initial 15 minutes of supine rest, 5 minutes of continuous beat-to-beat BP (Finapres) and RR interval (RRI) were recorded (standard ECG). Spectral indices were computed using Fast Fourier Transform and transfer function analysis was used to compute BRS. High frequency (HF) and low frequency (LF) power spectral areas were set to 0.15-0.4 Hz and 0.04-0.15 Hz, respectively. Body composition was measured using body mass index. After adjusting for body composition, maturation, age and sex ANCOV A results were as follows; LF and HF BRS, LF and HF RRI, and RRI total power were lower in the HBP versus NBP participants (p<0.05). As well, LF IHF SBP ratio was significantly higher in the HBP compared to the NBP group (p<0.05). The regression coefficients (unstandardized B) indicated that in changing groups (NBP to HBP) LF and HF BRS decreases by 4.04 and 6.18 ms/mmHg, respectively. Thus, as BP increases, BRS decreases. These data suggest that changes in autonomic activity occur in children who have HBP, regardless of age, sex, maturation, and body composition. Thus, despite their young age and relatively short amount of time having high BP compared with adults, these children are already demonstrating poor BP regulation and reduced cardiovagal activity. Given that childhood BP is associated with hypertension in adulthood, there is a growing concern in regards to the current cardiovascular health of our children and future adults.

Baroreflex sensitivity and developmental coordination disorder

Developmental coordination disorder (DCD) is a motor coordination disorder that is characterized by impairment of motor skills which leads to challenges with performing activities of daily living. Children with DCD have been shown to be less physically active and have increased body fatness. This is an important finding since a sedentary lifestyle and obesity are risk factors for cardiovascular disease. One indicator of cardiovascular health is baroreflex sensitivity (BRS), which is a measure of short term BP regulation that is accomplished through changes in HR. Diminished BRS is predictive of cardiovascular morbidity and mortality. The purpose of this study was to investigate BRS in 117 children aged 12 to 13 years with probable DCD (pOCO) and their matched controls with normal coordination. Following 15 minutes of supine rest, five minutes of continuous beat-by-beat blood pressure (Finapres) and RR interval were recorded (standard ECG). Spectral indices were computed using Fast Fourier Transform and transfer function analysis was used to compute BRS. High frequency and low frequency power spectral areas were set to 0.15-0.6 Hz and 0.04-0.15 Hz, respectively. BRS was compared between groups with an independent t-test and the difference was not significant. It is likely that a difference in BRS was not seen between groups since the difference in BMI between groups was small. As well, differences in BRS may not have manifested yet at this early age. However, the cardiovascular health of this population still deserves attention since differences in body composition and fitness were found between groups.

Autonomic Cardiovascular Control in Children and Adolescents

This thesis investigated the impact of pubertal maturation and sex on cardiovagal baroreflex sensitivity (BRS) and arterial properties of the common carotid artery (CCA), and the relationship between CCA arterial properties and BRS. This thesis also investigated the effect of orthostatic stress on arterial properties of the CCA and carotid sinus (CS), as well as their impact on BRS in response to orthostatic stress. Children and adolescents between the ages of 8-18 years were examined. To assess pubertal maturation participants were organized into five pubertal groups based on the criteria of Tanner. BRS was assessed by transfer function analysis in the low frequency range (0.05 – 0.15Hz). Pulse pressure (PP) was measured at the CCA (PPCCA) and CS (PPCS) using applanation tonometry, and at the finger (PPFinger) using photoplethysmography. Ultrasound sonography and applanation tonometry were used to determine the distensibility coefficient (DC) at the CCA (DCCCA) and CS (DCCS). A moderate posture stimulus was implemented by passively moving participants into a 50° seated-recumbent (SR) position. The results demonstrated a sex-by-maturation interaction on BRS (p= 0.019). BRS decreased from early- to post-puberty in males (p<0.01), and remained unchanged in females. Females demonstrated greater BRS compared to males post-puberty (p<0.05). CCA distensibility was not affected by sex or maturation and was not related to BRS. PPCS was greater than PPCCA (p<0.001), while PPFinger was greater than both PPCCA (p<0.001) and PPCS (p<0.001). In response to SR, the relative change in PPFinger was significantly different than the relative change in PPCCA (p<0.001) and PPCS (p<0.001), while the relative change between PPCCA and PPCS were not different. Finally, in response to SR there was a significant decrease in DCCS (p=0.001), but not DCCCA. The relative change in BRS in response to SR was significantly correlated to the relative change in DCCS (p=0.004), but not DCCCA. The findings demonstrated an important sex-dependent maturation effect on BRS in children and adolescents that was not explained by CCA distensibility. Also, the CS and CCA responded differently to orthostatic stress. The CS was more suitable to evaluate the effect of arterial distensibility on BRS in response to posture change.

Reactivity characteristics of cytochrome c, cytochrome oxidase and the electrostatic cytochrome c - cytochrome oxidase complex /

The mechanism whereby cytochrome £ oxidase catalyses elec-. tron transfer from cytochrome £ to oxygen remains an unsolved problem. Polarographic and spectrophotometric activity measurements of purified, particulate and soluble forms of beef heart mitochondrial cytochrome c oxidase presented in this thesis confirm the following characteristics of the steady-state kinetics with respect to cytochrome £: (1) oxidation of ferrocytochrome c is first order under all conditions. -(2) The relationship between sustrate concentration and velocity is of the Michaelis-Menten type over a limited range of substrate. concentrations at high ionic strength. (3) ~he reaction rate is independent from oxygen concentration until very low levels of oxygen. (4) "Biphasic" kinetic plots of enzyme activity as a function of substrate concentration are found when the range of cytochrome c concentrations is extended; the biphasicity ~ is more apparent in low ionic strength buffer. These results imply two binding sites for cytochrome £ on the oxidase; one of high affinity and one of low affinity with Km values of 1.0 pM and 3.0 pM, respectively, under low ionic strength conditions. (5) Inhibition of the enzymic rate by azide is non-c~mpetitive with respect to cytochrome £ under all conditions indicating an internal electron transfer step, and not binding or dissociation of £ from the enzyme is rate limiting. The "tight" binding of cytochrome '£ to cytochrome c oxidase is confirmed in column chromatographic experiments. The complex has a cytochrome £:oxidase ratio of 1.0 and is dissociated in media of high ionic strength. Stopped-flow spectrophotometric studies of the reduction of equimolar mixtures and complexes of cytochrome c and the oxidase were initiated in an attempt to assess the functional relevance of such a complex. Two alternative routes -for reduction of the oxidase, under conditions where the predominant species is the £ - aa3 complex, are postulated; (i) electron transfer via tightly bound cytochrome £, (ii) electron transfer via a small population of free cytochrome c interacting at the "loose" binding site implied from kinetic studies. It is impossible to conclude, based on the results obtained, which path is responsible for the reduction of cytochrome a. The rate of reduction by various reductants of free cytochrome £ in high and low ionic strength and of cytochrome £ electrostatically bound to cytochrome oxidase was investigated. Ascorbate, a negatively charged reagent, reduces free cytochrome £ with a rate constant dependent on ionic strength, whereas neutral reagents TMPD and DAD were relatively unaffected by ionic strength in their reduction of cytochrome c. The zwitterion cysteine behaved similarly to uncharged reductants DAD and TI~PD in exhibiting only a marginal response to ionic strength. Ascorbate reduces bound cytochrome £ only slowly, but DAD and TMPD reduce bound cytochrome £ rapidly. Reduction of cytochrome £ by DAD and TMPD in the £ - aa3 complex was enhanced lO-fold over DAD reduction of free £ and 4-fold over TMPD reduction of free c. Thus, the importance of ionic strength on the reactivity of cytochrome £ was observed with the general conclusion being that on the cytochrome £ molecule areas for anion (ie. phosphate) binding, ascorbate reduction and complexation to the oxidase overlap. The increased reducibility for bound cytochrome £ by reductants DAD and TMPD supports a suggested conformational change of electrostatically bound c compare.d to free .£. In addition, analysis of electron distribution between cytochromes £ and a in the complex suggest that the midpotential of cytochrome ~ changes with the redox state of the oxidase. Such evidence supports models of the oxidase which suggest interactions within the enzyme (or c - enzyme complex) result in altered midpoint potentials of the redox centers.

The study of ligand binding specificities of the lipid binding proteins : recombinant human a-tocopherol transport protein (a-ttp), supernatant protein factor (spf) and S. cerevisiae Sec 14p for vitamin e (rrr-a-tocopherol) and other hydrophobic ligands.

One of the various functions of proteins in biological systems is the transport of small molecules, for this purpose proteins have naturally evolved special mechanisms to allow both ligand binding and its subsequent release to a target site; a process fundamental to many biological processes. Transport of Vitamin E (a-tocopherol), a lipid soluble antioxidant, to membranes helps in the protection of polyunsaturated fatty acids against peroxidative damage. In this research, the ligand binding characteristics of several members of the CRALTRIO family of lipid binding proteins was examined; the recombinant human a-Tocopherol Transfer Protein (a-TIP), Supernatant Protein Factor (SPF)ffocopherol Associated Protein (TAP), Cellular Retinaldehyde Binding Protein (CRALBP) and the phosphatidylinositol transfer protein from S. cerevisiae Sec 14p. Recombinant Sec 14p was expressed and purified from E. coli for comparison of tocopherol binding to the two other recombinant proteins postulated to traffic a-tocopherol. Competitive binding assays using [3H]-a-tocopherol and Lipidex-l000 resin allowed determination of the dissociation constants ~) of the CRAL-TRIO proteins for a-tocopherol and - 20 hydrophobic ligands for evaluation of the possible biological relevance of the binding interactions observed. The KIs (nM) for RRR-a-tocopherol are: a-TIP: 25.0, Sec 14p: 373, CRALBP: 528 and SPFffAP: 615. This indicates that all proteins recognize tocopherol but not with the same affinity. Sec 14p bound its native ligand PI with a KI of381 whereas SPFffAP bound PI (216) and y-tocopherol (268) similarly in contrast to the preferential binding ofRRR-a-tocopherol by a-TIP. Efforts to adequately represent biologically active SPFff AP involved investigation of tocopherol binding for several different recombinant proteins derived from different constructs and in the presence of different potential modulators (Ca+2, Mg+2, GTP and GDP); none of these conditions enhanced or inhibited a-tocopherol binding to SPF. This work suggests that only aTTP serves as the physiological mediator of a-tocopherol, yet structural homology between proteins allows common recognition of similar ligand features. In addition, several photo-affmity analogs of a-tocopherol were evaluated for their potential utility in further elucidation of a-TTP function or identification of novel tocopherol binding proteins.

Synechococcus sp. PCC 7002 CpcB lyase null mutations alter phycocyanin chromophore function and, consequently, affect the redistribution of excitation energy via the light state transition /

Phycobilisomes are the major light harvesting complexes for cyanobacteria and phycocyanin is the primary phycobiliprotein of the phycobilisome rod. The phycocyanobilin lyases responsible for chromophorylating the phycocyanin p subunit (CpcB) have been recently identified in the cyanobacterium Synechococcus sp. PCC 7002. Surprisingly, mutants missing the CpcB lyases were nevertheless capable of producing pigmented phycocyanin. 10K absorbance measurements revealed that the energy states of the p phycocyanin chromophores were only subtly shifted; however, 77K steady state fluorescence emission spectroscopy showed excitation energy transfer involving the targeted chromophores to be highly disrupted. Such evidence suggests that phycobilin orientation within the binding domain is specifically modified. We hypothesized that alternate, less specific lyases are able to act on the p binding sites. A phycocyanin linker-polypeptide deficient mutant was similarly characterized. The light state transition, a short term adaptation of the photosynthetic light harvesting apparatus resulting in the redistribution of excitation energy among the photo systems, was shown to be dominated by the reallocation of phycocyanin-absorbed excitation energy. Treatment with a high M phosphate buffer effectively prevented the redistribution of both chlorophyll a- and phycobilisome- absorbed excitation energy, suggesting that the two effects are not strictly independent. The mutant strains required a larger redistribution of excitation energy between light states, perhaps to compensate for their loss in phycobilisome antenna function.

Transfection of newt blastema mesenchyme using the techniques of lipofection and direct gene transfer

The regenerating amphibian limb provides a useful system for studying genes involved in the establishment of positional information. While a number of candidate genes that may playa role in pattern formation have been identified, their function in vivo is unknown in this system. To better ascertain the role of these genes, it would be useful to be able to alter their normal patterns of expression in vivo and to assess the effects of this misexpression on limb pattern. In order to achieve this, a method of introducing a plasmid containing the eDNA of a gene of interest into a newt blastema (a growth zone of mesenchymal progenitor cells) is needed. Unfortunately, most commonly used transfection techniques cannot be used with newt blastema cells. In this study, I have used the techniques of lipofection and direct gene transfer to introduce plasmid DNA containing reporter genes into the cells of a regenerating newt limb. The technique of lipofection was most effective when the blastema cells were transfected in vitro. The optimal ratio for transfection was shown to be 1:3 DNA:Lipofectin (W/w) , and an increase in the amount of DNA present in the mixture (1:3 ratio maintained) resulted in a corresponding increase in gene expression. The technique of direct gene transfer was used to transfect newt blastema cells with and without prior complex formation with Lipofectin. Injection of plasmid DNA alone provided the most 3 promising results. It was possible to introduce plasmid DNA containing the reporter gene ~-galactosidase and achieve significant gene expression in cells associated with the injection site. In the future, it would be interesting to use this technique to inject plasmid DNA containing a gene which may have a role in pattern formation into specific areas of the newt blastema and to analyze the resulting limb pattern that emerges.

The role of daytime napping in sleepiness and cognitive function in 24-70 year olds /

Daytime napping improves well-being and performance for young adults. The benefits of napping in older adults should be investigated because they have fragmented nocturnal sleep, cognitive declines, and more opportunity to nap. In addition, experience with napping might influence the benefits of napping. Study 1 examined the role of experience with napping in young adults. Habitual (n = 23) and non-habitual nappers (n = 16) were randomly assigned to a 20-minute nap or a 20- minute reading condition. Both groups slept the same according to macro architecture. However, microarchitecture showed greater theta, alpha, and beta power during Stage 1, and greater delta, alpha, and sigma power during Stage 2 for habitual nappers, for the most part indicating better sleep. Both groups felt less sleepy after the nap. P2 latency, reflecting information processing, decreased after the nap for habitual nappers, and after the control condition for non-habitual nappers. In sum, both groups who slept felt better, but only the habitual nappers who napped gained a benefit in terms of information processing. Based on this outcome, experience with napping was investigated in Study 2. Study 2 examined the extent to which daytime napping enhanced cognition in older adults, especially frontal lobe function. Cognitive deficits in older adults may be due to sleep loss and age-related decline in brain functioning. Longer naps were expected to provide greater improvement, particularly for older adults, by reducing sleep pressure. Thirty-two adults, aged 24-70 years, participated in a repeated measures dose-response manipulation of sleep pressure. Twenty- and sixty-minute naps were compared to a no-nap condition in three age groups. Mood, subjective sleepiness, reaction time, working memory, 11 novelty detection, and waking electro physiological measures were taken before and after each condition. EEG was also recorded during each nap or rest condition. Napping reduced subjective sleepiness, improved working memory (serial addition / subtraction task), and improved attention (reduced P2 amplitude). Physiological sleepiness (i.e., waking theta power) increased following the control condition, and decreased after the longer nap. Increased beta power after the short nap, and seen with older adults overall, may have reflected increased mental effort. Older adults had longer latencies and smaller amplitudes for several event-related potential components, and higher beta and gamma power. Following the longer nap, gamma power decreased for older adults, but increased for young adults. Beta and gamma power may represent enhanced alertness or mental effort. In addition, Nl amplitude showed that benefits depend on the preceding nap length as well as age. Since the middle group had smaller Nl amplitudes following the short nap and rest condition, it is possible that they needed a longer nap to maintain alertness. Older adults did not show improvements to Nl amplitude following any condition; they may have needed a nap longer than 60 minutes to gain benefits to attention or early information processing. Sleep characteristics were not related to benefits of napping. Experience with napping was also investigated. Subjective data confirmed habitual nappers were happier to nap, while non-habitual nappers were happier to stay awake, reflecting self-identified napping habits. Non-habitual nappers were sleepier after a nap, and had faster brain activity (i.e., heightened vigilance) at sleep onset. These reasons may explain why non-habitual nappers choose not to nap.

Individual differences in risk-taking and associated characteristics : a test of specificity in executive functions /

Multiple measures have been devised by clinicians and theorists from many different backgrounds for the purpose of assessing the influence of the frontal lobes on behaviour. Some utilize self-report measures to investigate behavioural characteristics such as risktaking, sensation seeking, impulsivity, and sensitivity to reward and punishment in an attempt to understand complex human decision making. Others rely more on neuroimaging and electrophysiological investigation involving experimental tasks thought to demonstrate executive functions in action, while other researchers prefer to study clinical populations with selective damage. Neuropsychological models of frontal lobe functioning have led to a greater appreciation of the dissociations among various aspects of prefrontal cortex function. This thesis involves (1) an examination of various psychometric and experimental indices of executive functions for coherence as one would predict on the basis of highly developed neurophysiological models of prefrontal function, particularly those aspects of executive function that involve predominantly cognitive abilities versus processes characterized by affect regulation; and (2) investigation of the relations between risk-taking, attentional abilties and their associated characteristics using a neurophysiological model of prefrontal functions addressed in (1). Late adolescence is a stage in which the prefrontal cortices undergo intensive structural and functional maturational changes; this period also involves increases in levels of risky and sensation driven behaviours, as well as a hypersensitivity to reward and a reduction in inhibition. Consequently, late adolescence spears to represent an ideal developmental period in which to examine these decision-making behaviours due to the maximum variability of behavioural characteristics of interest. Participants were 45 male undergraduate 18- to 19-year olds, who completed a battery of measures that included self-report, experimental and behavioural measures designed to assess particular aspects of prefrontal and executive functioning. As predicted, factor analysis supported the grouping of executive process by type (either primarily cognitive or affective), conforming to the orbitofrontal versus dorsolateral typology; risk-taking and associated characteristics were associated more with the orbitofrontal than the dorsolateral factor, whereas attentional and planning abilities tended to correlate more strongly with the dorsolateral factor. Results are discussed in light of future assessment, investigation and understanding of complex human decision-making and executive functions. Implications, applications and suggestions for future research are also proposed.

Orientation of Non-Native 2-Monosubstituted and 2,3-Disubstituted 1,4-Naphthoquinones in the A1 binding site of PSI and the effect on the rate of electron transfer : an electron paramagnetic resonance spectroscopy study

The proce-ss ofoxygenic photosynthesis is vital to life on Earth. the central event in photosynthesis is light induced electron transfer that converts light into energy for growth. Ofparticular significance is the membrane bound multisubunit protein known as Photosystem I (PSI). PSI is a reaction centre that is responsible for the transfer of electrons across the membrane to reduce NADP+ to NADPH. The recent publication ofa high resolution X-ray structure of PSI has shown new information about the structure, in particular the electron transfer cofactors, which allows us to study it in more detail. In PSI, the secondary acceptor is crucial for forward electron transfer. In this thesis, the effect of removing the native acceptor phylloquinone and replacing it with a series of structurally related quinones was investigated via transient electron paramagnetic resonance (EPR) experiments. The orientation of non native quinones in the binding site and their ability to function in the electron transfer process was determined. It was found that PSI will readily accept alkyl naphthoquinones and anthraquinone. Q band EPR experiments revealed that the non-native quinones are incorporated into the binding site with the same orientation of the headgroup as in the native system. X band EPR spectra and deuteration experiments indicate that monosubstituted naphthoquinones are bound to the Al site with their side group in the position occupied by the methyl group in native PSI (meta to the hydrogen bonded carbonyl oxygen). X band EPR experiments show that 2, 3- disubstituted methyl naphthoquinones are also incorporated into the Al site in the same orientation as phylloquinone, even with the presence of a halogen- or sulfur-containing side chain in the position normally occupied by the phytyl tail ofphylloquinone. The exception to this is 2-bromo-3-methyl --.- _. -. - -- - - 4 _._ _ _ - _ _ naphthoquinone which has a poorly resolved spectrum, making determination of the orientation difficuh. All of the non-native quinones studied act as efficient electron acceptors. However, forward electron transfer past the quinone could only be demonstrated for anthraquinone, which has a more negative midpoint potential than phylloquinone. In the case of anthraquinone, an increased rate of forward electron transfer compared to native PSI was found. From these results we can conclude that the rate ofelectron transfer from Al to Fx in native PSI lies in the normal region ofthe Marcus Curve.

The association between blood pressure and vascular characteristics in children

Hypertension is thought to exist in up to five percent of children. A select number of studies have investigated the role elevated blood pressure plays in pediatric atherosclerotic progression. However these studies contain significant methodological flaws and fail to recognize important confounding factors. Therefore, the influence of elevated blood pressure on arterial health in children remains to be clearly understood. The purpose of this study was to investigate the association between blood pressure (BP) and arterial thickness and stiffuess in children. Common carotid artery (CCA) intima-media thickness (IMT) and distensibility (Dist), as well as systemic pulse wave velocity (PWV) were measured in 21 elevated blood pressure (EBP; BP ~ 95th percentile) and 83 normal blood pressure (NBP; BP < 90th percentile) children 11-14 years of age. Both EBP and NBP groups demonstrated BP within the normal clinical range, but EBP showed significantly elevated BP as compared to the NBP group. Independent t-tests failed to show significant differences between the EBP and NBP groups for CCA IMT (0.43 ± 0.05 mm and 0.42 ± 0.06 mm, respectively) and Dist (0.0058 ± 0.0024 mmHg-1 and 0.0064 ± 0.0019 mmHil respectively). In contrast, a significantly elevated PWV (pfunction as measured through CCA Dist and IMT, but do possess significantly altered systemic arterial stiffuess as measured through PWV. This may be the result of sympathetic predominance and its significant influence on the peripheral vasculature. More studies are needed to clearly illustrate the temporal sequence of pediatric atherosclerotic progression in response to elevated BP.

Behavioural and neural correlates of emotion recognition as a function of psychopathic personality traits

Psychopathy is associated with well-known characteristics such as a lack of empathy and impulsive behaviour, but it has also been associated with impaired recognition of emotional facial expressions. The use of event-related potentials (ERPs) to examine this phenomenon could shed light on the specific time course and neural activation associated with emotion recognition processes as they relate to psychopathic traits. In the current study we examined the PI , N170, and vertex positive potential (VPP) ERP components and behavioural performance with respect to scores on the Self-Report Psychopathy (SRP-III) questionnaire. Thirty undergraduates completed two tasks, the first of which required the recognition and categorization of affective face stimuli under varying presentation conditions. Happy, angry or fearful faces were presented under with attention directed to the mouth, nose or eye region and varied stimulus exposure duration (30, 75, or 150 ms). We found that behavioural performance to be unrelated to psychopathic personality traits in all conditions, but there was a trend for the Nl70 to peak later in response to fearful and happy facial expressions for individuals high in psychopathic traits. However, the amplitude of the VPP was significantly negatively associated with psychopathic traits, but only in response to stimuli presented under a nose-level fixation. Finally, psychopathic traits were found to be associated with longer N170 latencies in response to stimuli presented under the 30 ms exposure duration. In the second task, participants were required to inhibit processing of irrelevant affective and scrambled face distractors while categorizing unrelated word stimuli as living or nonliving. Psychopathic traits were hypothesized to be positively associated with behavioural performance, as it was proposed that individuals high in psychopathic traits would be less likely to automatically attend to task-irrelevant affective distractors, facilitating word categorization. Thus, decreased interference would be reflected in smaller N170 components, indicating less neural activity associated with processing of distractor faces. We found that overall performance decreased in the presence of angry and fearful distractor faces as psychopathic traits increased. In addition, the amplitude of the N170 decreased and the latency increased in response to affective distractor faces for individuals with higher levels of psychopathic traits. Although we failed to find the predicted behavioural deficit in emotion recognition in Task 1 and facilitation effect in Task 2, the findings of increased N170 and VPP latencies in response to emotional faces are consistent wi th the proposition that abnormal emotion recognition processes may in fact be inherent to psychopathy as a continuous personality trait.

Competing demands for a complex system : photosystem II repair, photoprotection and quantum yield

Photosynthesis in general is a key biological process on Earth and Photo system II (PSII) is an important component of this process. PSII is the only enzyme capable of oxidizing water and is largely responsible for the primordial build-up and present maintenance of the oxygen in the atmosphere. This thesis endeavoured to understand the link between structure and function in PSII with special focus on primary photochemistry, repair/photodamage and spectral characteristics. The deletion of the PsbU subunit ofPSII in cyanobacteria caused a decoupling of the Phycobilisomes (PBS) from PSII, likely as a result of increased rates of PSII photodamage with the PBS decoupling acting as a measure to protect PSII from further damage. Isolated fractions of spinach thylakoid membranes were utilized to characterize the heterogeneity present in the various compartments of the thylakoid membrane. It was found that the pooled PSIILHCII pigment populations were connected in the grana stack and there was also a progressive decrease in the reaction rates of primary photochemistry and antennae size of PSII as the sample origin moved from grana to stroma. The results were consistent with PSII complexes becoming damaged in the grana and being sent to the stroma for repair. The dramatic quenching of variable fluorescence and overall fluorescent yield of PSII in desiccated lichens was also studied in order to investigate the mechanism by which the quenching operated. It was determined that the source of the quenching was a novel long wavelength emitting external quencher. Point mutations to amino acids acting as ligands to chromophores of interest in PSII were utilized in cyanobacteria to determine the role of specific chromophores in energy transfer and primary photochemistry. These results indicated that the Hl14 ligated chlorophyll acts as the 'trap' chlorophyll in CP47 at low temperature and that the Q130E mutation imparts considerable changes to PSII electron transfer kinetics, essentially protecting the complex via increased non-radiative charge Photosynthesis in general is a key biological process on Earth and Photo system II (PSII) is an important component of this process. PSII is the only enzyme capable of oxidizing water and is largely responsible for the primordial build-up and present maintenance of the oxygen in the atmosphere. This thesis endeavoured to understand the link between structure and function in PSII with special focus on primary photochemistry, repair/photodamage and spectral characteristics. The deletion of the PsbU subunit ofPSII in cyanobacteria caused a decoupling of the Phycobilisomes (PBS) from PSII, likely as a result of increased rates of PSII photodamage with the PBS decoupling acting as a measure to protect PSII from further damage. Isolated fractions of spinach thylakoid membranes were utilized to characterize the heterogeneity present in the various compartments of the thylakoid membrane. It was found that the pooled PSIILHCII pigment populations were connected in the grana stack and there was also a progressive decrease in the reaction rates of primary photochemistry and antennae size of PSII as the sample origin moved from grana to stroma. The results were consistent with PSII complexes becoming damaged in the grana and being sent to the stroma for repair. The dramatic quenching of variable fluorescence and overall fluorescent yield of PSII in desiccated lichens was also studied in order to investigate the mechanism by which the quenching operated. It was determined that the source of the quenching was a novel long wavelength emitting external quencher. Point mutations to amino acids acting as ligands to chromophores of interest in PSII were utilized in cyanobacteria to determine the role of specific chromophores in energy transfer and primary photochemistry. These results indicated that the Hl14 ligated chlorophyll acts as the 'trap' chlorophyll in CP47 at low temperature and that the Q130E mutation imparts considerable changes to PSII electron transfer kinetics, essentially protecting the complex via increased non-radiative charge.