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.

Effect of pH on non-photochemical quenching in spinach photsystem 2 particles as measured by picosecond fluorescence decay kinetics

Single photon timing was used to study picosecond chlorophyll a fluorescence decay kinetics of pH induced non-photochemical quenching in spinach photosystem 2 particles. The characteristics of this quenching are a decrease in chlorophyll a fluorescence yield as well as a decrease in photochemistry at low pH. Picosecond kinetics of room temperature fluorescence temporally resolve the individual components of the steady state fluorescence yield into components that are related to primary energy conversion processes in photosystem 2. Four components were resolved for dark adapted (Fo), light saturated (Fm), and chemically reduced (Nadithionite) photosystem 2 reaction centres. The fastest and slowest components, indicative of energy transfer to and energy capture by the photosystem 2 reaction centre and uncoupled ("dead") chlorophyll, respectively, were not affected by changing pH from 6.5 to 4.0. The two intermediate components, indicative of electron transfer processes within the reaction centre of photosystem 2, were affected by the pH change. Results indicate that the decrease in the steady state fluorescence yield at low pH was primarily due to the decrease in lifetime and amplitude of the slower of the intermediate components. These results imply that the decrease in steady state fluorescence yield at low pH is not due to changes in energy transfer to and energy capture by the photosystem 2 reaction centre, but is related to changes in charge stabilization and charge recombination in the photosystem 2 reaction centre.

An investigation into the functional role of the D1:1 and D1:2 polypeptides in photosystem II in cyanobacteria : the effect of changing PSI/PSII ratio on photoinhibition in Synechococcus sp. PCC7942 /

The cyanobacterium Synechococcus sp. PCC 7942 (Anacystis nidulans R2) adjusts its photosynthetic function by changing one of the polypeptides of photosystem II. This polypeptide, called Dl, is found in two forms in Synechococcus sp. PCC 7942. Changing the growth light conditions by increasing the light intensity to higher levels results in replacement of the original form of D 1 polypeptide, D 1: 1, with another form, D 1 :2. We investigated the role of these two polypeptides in two mutant strains, R2S2C3 (only Dl:l present) and R2Kl (only Dl:2 present) In cells with either high or low PSI/PSII. R2S2C3 cells had a lower amplitude for 77 K fluorescence emission at 695 nm than R2Kl cells. Picosecond fluorescence decay kinetics showed that R2S2C3 cells had shorter lifetimes than R2Kl cells. The lower yields and shorter lifetimes observed in the D 1 and Dl:2 containing cells. containing cells suggest that the presence of D 1: 1 results in more photochemical or non-photochemical quenching of excitation energy In PSII. One of the most likely mechanisms for the increased quenching in R2S2C3 cells could be an increased efficiency in the transfer of excitation energy from PSII to PSI. However, photophysical studies including 77 K fluorescence measurements and picosecond time resolved decay kinetics comparing low and high PSI/PSII cells did not support the hypothesis that D 1: 1 facilitates the dissipation of excess energy by energy transfer from PSII to PSI. In addition physiological studies of oxygen evolution measurements after photoinhibition treatments showed that the two mutant cells had no difference in their susceptibility to photoinhibition with either high PSI/PSII ratio or low PSI/PSII ratio. Again suggesting that, the energy transfer efficiency from PSII to PSI is likely not a factor in the differences between Dl:l and Dl:2 containing cells.

The effect of moisture on decomposition processes in the disturbed peatlands of the Wainfleet bog /

The purpose of this study was to determine the effect of increased soil moisture levels on the decomposition processes in a peat-extracted bog. Field experiments, in which soil moisture levels were manipulated, were conducted using 320 microcosms in the Wainfleet Bog from May 2002 to November 2004. Decomposition was measured using litter bags and monitoring the abundance of macro invertebrate decomposers known as Collembola. Litter bags containing wooden toothpicks (n=2240), filter paper (n=480) and Betula pendula leaves (n=40) were buried in the soil and removed at regular time intervals up to one year. The results of the litter bag studies demonstrated a significant reduction of the decomposition of toothpicks (p<0.001), filter paper (p<0.001), and Betula pendula leaves (peffect of increased moisture levels on the abundance of Collembola was more variable and difficult to interpret. The conclusions of this study indicate that the Wainfleet Bog is a highly disturbed peatland and that the greatest reductions in decomposition can be obtained by restoring the soil moisture levels near those of undisturbed conditions.

An electrophysiological examination of intentional and inadvertent sleep onset : the effect of intention on the sleep onset process

The main purpose ofthis study was to examine the effect ofintention on the sleep onset process from an electrophysiological point ofview. To test this, two nap conditions, the Multiple Sleep Latency Test (MSLT) and the Repeated Test of Sustained Wakefulness (RTSW) were used to compare intentional and inadvertent sleep onset. Sixteen female participants (aged 19-25) spent two non-consecutive nights in the sleep lab; however, due to physical and technical difficulties only 8 participants produced compete sets of data for analysis. Each night participants were given six nap opportunities. For three ofthese naps they were instructed to fall asleep (MSLT), for the remaining three naps they were to attempt to remain awake (RTSW). These two types of nap opportunities represented the conditions ofintentional (MSLT) and inadvertent (RTSW) sleep onset. Several other sleepiness, performance, arousal and questionnaire measures were obtained to evaluate and/or control for demand characteristics, subjective effort and mental activity during the nap tests. The nap opportunities were scored using a new 9 stage scoring system developed by Hori et al. (1994). Power spectral analyses (FFT) were also performed on the sleep onset data provided by the two nap conditions. Longer sleep onset latencies (approximately 1.25 minutes) were obseIVed in the RTSW than the MSLT. A higher incidence of structured mental activity was reported in the RTSW and may have been reflected in higher Beta power during the RTSW. The decent into sleep was more ragged in the RTSW as evidenced by an increased number shifts towards higher arousal as measured using the Hori 9 stage sleep scoring method. 1ll The sleep onset process also appears to be altered by the intention to remain awake, at least until the point ofinitial Stage 2 sleep (i.e. the first appearance of spindle activity). When only examining the final 4.3 minutes ofthe sleep onset process (ending with spindle activity), there were significant interactions between the type ofnap and the time until sleep onset for Theta, Alpha and Beta power. That is to say, the pattern of spectral power measurements in these bands differed across time as a function ofthe type ofnap. The effect ofintention however, was quite small (,,2 < .04) when compared to the variance which could be accounted for by the passage oftime (,,2 == .10 to .59). These data indicate that intention alone cannot greatly extend voluntary wakefulness if a person is sleepy. This has serious implications for people who may be required to perform dangerous tasks while sleepy, particularly for people who are in a situation that does not allow them the opportunity to engage in behavioural strategies in order to maintain their arousal.

Effect of age on female choice in the field cricket, Gryllus interger

Female choice is an important element of sexual selection that may vary among females of the same species. Few researchers have investigated the causes of variation in selectivity with respect to potential mates and overall level of motivation toward a stimulus source representative of a mate. This study demonstrates that female age may be one cause of variation in female choice. Females of different ages may have different mate preferences. As females age, they have less time left to reproduce, and their residual reproductive value decreases. This should correspond to a higher reproductive effort which may be represented as increased motivation and/or decreased selectivity. The effect of age on mate choice in Gryllus integer was investigated by using a non-compensating treadmill, called the Kugel, to measure female phonotaxis. Artificially generated male calling songs of varying pulse rates were broadcast in either a singlestimulus or a three-stimulus experimental design. The pulse rates used in the calling song stimuli were 70, 64, 76, 55 and 85 pulses per second. These corresponded to the documented mean pulse rate for the species at the experimental temperature, one standard deviation below and above the mean, and 2.5 standard deviations below and above the mean, respectively. Test females were either 11-14 days or 25-28 days post-ecdysis. Trials usually were conducted two to seven hours into the scotophase. In the single-stimulus experiment, females were presented with stimuli with only one pulse rate. Older females achieved higher vector scores than younger females, indicating that older females are more motivated to mate. Both groups showed little phonotactic response towards 55 or 85 pIs, both of which lie outside the natural range of G. integer calling song at the experimental temperature. Neither group discriminated among the three pulse rates that fell within the natural range of calling song. In the three-stimulus experiment, females were presented with stimuli with one of three pulse rates, 64, 70 or 76 pIs, In alternation. Both age groups had reduced responsiveness in this experiment, perhaps due to an increase in perceived male density. Additionally, younger females responded significantly more to 64 and 70 pIs than to the higher pulse rate, indicating that they are selective with respect to mate choice. Older females did not discriminate among the three pulse rates. Therefore, it was concluded that selectivity decreases with age. A further study was conducted to determine that these effects were due to age and not due to the differing periods without a mating between the two age groups. Again, stimuli were presented in a three-stimulus experimental design. Age was held constant at 28 days and time since last mating varied from 11 to 25 days. Females varyIng in time since last mating did not differ in their responses to the calling song pulse rates. This indicated that the increased motivation and decreased selectivity exhibited In the initial experiments were due to age and not to time without a mating. Neither time of trial nor female weight had an effect upon female phonotaxis. Data are discussed in terms of mate choice, residual reproductive value, and costs of choice.

The effect of diet and parasite load on female phonotaxis in the Texas field cricket, Gryllus integer

Female crickets respond selectively to variations in species-specific male calling songs. This selectivity has been shown to be age-dependent; older females are less choosy. However, female quality should also affect female selectivity. The effect of female quality on mate choice was examined in Gryllus integer by comparing the phonotactic responses of females on different diets and with different parasite loads to various synthetic models of conspecific calling song. Test females were virgin, 11-14 days old, and had been maintained on one of five diets varying in protein and fat content. Phonotaxis was quantified using a non-compensating Kugel treadmill which generates vector scores incorporating the speed and direction of movement of each female. Test females were presented with four calling song models which differed in pulse rate, but were still within the natural range of the species for the experimental temperature. After testing, females were dissected and the number of gregarine parasites within the digestive tract counted. There were no significant effects of either diet or parasitism on female motivation to mate although the combined effects of these variables seem to have an effect with no apparent trend. Control females did not discriminate among song types, but there was a trend of female preferences for lower pulse rates which are closest to the mean pulse rate for the species. Heavily parasitized females did not discriminate among pulse rates altho~gh there was a similar trend of high vector scores for low pulse rates. Diet, however, affected selectivity with poorly-fed females showing significantly high vector scores for pulse rates near the species mean. Such findings raise interesting questions about energy allocation and costs and risks of phonotaxis and mate choice in acoustic Orthoptera. These results are discussed in terms of sexual selection and female mate choice.

"Re-huminization": the role of human-animal similarity in predicting prejudice towards immigrants and non-human animals

Research implies that there ~ay be an association between attitudes toward margil1alized human outgroups and non-human animals. Very few studies, however, have specifically tested this relation empirically. The general purpose of the present research was to determine if such a relation exists and if perceptions of human-animal similarity avail as a common predictor of both types of attitudes. Ideological orientations associated with prejudiced attitudes (Social Dominance Orientation, Right-Wing Authoritarianism, and Universal Orientation) were also examined as individual differences in predicting perceptions of human-animal similarity. As predicted, people who endorsed prejudiced attitudes toward human outgroups (Study 1) and immigrants in particular (Studies 2 and 3), were more likely to endorse prejudiced attitudes toward non-human animals. In Study 2, perceptions that humans are superior (versus similar) to other animals directly predicted higher levels of prejudice toward non-human animals, whereas the effect of human superiority beliefs on immigrant prejudice was mediated by dehumanization. In other words, greater perceptions of humans as superior (versus similar) to other animals "allowed for" greater dehumanization of immigrants, which in turn resulted in heightened immigrant prejudice. Furthermore, people higher in Social Dominance Orientation or Right-Wing Authoritarianism were particularly likely to perceive humans as superior (versus similar) to other animals, whereas people characterized by a greater Universal Orientation were more likely to perceive humans and non-human animals as similar. Study 3 examined whether inducing perceptions of human-animal similarity through experimental manipulation would lead to more favourable attitudes toward non-human animals and immigrants. Participants were randomly assigned to read one of four 11 editorials designed to highlight either the similarities or differences between humans and other animals (i.e., animals are similar to humans; humans are similar to animals;~~nimals are inferior to humans; humans are superior to animals) or to a neutral control condition. Encouragingly, when animals were described as similar to humans, prejudice towards non-human animals and immigrants was significantly lower, and to some extent this finding was also true for people naturally high in prejudice (i.e., high in Social Dominance Orientation or Right-Wing Authoritarianism). Inducing perceptions that nonhuman animals are similar to humans was particularly effective at reducing the tendency to dehumanize immigrants ("re-humanization"), lowering feelings of personal threat regarding one's animal-nature, and at increasing inclusive intergroup representations and empathy, all of which uniquely accounted for the significant decreases in prejudiced attitudes. Implications for research, theory and prejudice interventions are considered.

Muscle strength and activation characteristics of power- trained and non-athlete boys and men

During maturation, muscle strength is enhanced through muscle growth, although neuro-muscular factors are also believed to be involved. In adults, training for power sports has been shown to enhance muscle strength and activation. The purpose of this study was to examine muscle strength and activation in power-trained athletes (POW) compared with non-athletes (CON), in boys and in adults. After familiarization subjects performed ten 5-s explosive maximal voluntary contractions for elbow and knee flexion and extension. The adults were stronger then the boys and the adult POW were stronger then the adult CON, even after correction for muscle size. Normalized rate of torque development was higher in the adults then in the boys and higher in the POW then CON boys. The rate of muscle activation was higher in the adults and POW groups. The results suggest that maturation and power-training have an additive effect on muscle activation.

The effect of acute exercise type on body image attitudes

The current study examined the effect of two types of acute exercise (resistance and mind-body yoga) on state body image attitudes, to see which, was most effective at reducing body image concerns. I t was hypothesized that both types of exercise would lead to improvements in body image, with yoga showing the biggest benefits. Collegeaged female non/infrequent exercisers (n = 40) completed state measures of body satisfaction, social physique anxiety (SPA), and appearance orientation prior to and following participation in a single yoga and resistance class. Participation in the yoga class was associated with decreases in SPA and increases in body satisfaction. However, participation in the resistance class was associated with no changes in any of the study variables. These findings indicate participation in a single yoga class may have positive effects on body image attitudes, which may encourage non-exercisers to become more active.

Extending the irrelevant sound effect beyond serial recall

The finding that serial recall perfonnance for visually presented items is impaired by concurrently presented speech or sounds is referred to as the irrelevant sound effect (lSE). The foremost explanation for the effect is based on interference with rehearsal and seriation processes. The present series of experiments demonstrates tha t neither rehearsal nor seriation processes is necessary to observe the ISE. Evidence comes from three experiments that a) allow participants to report to-be-remembered items in any order, b) eliminate rehearsal by engaging participants in a cover task and surprising them with a memory test, and c) show that surprise non-serial recognition is immune to rehearsal-based experimental manipulations that modulate the ISE in more typical serial recall tasks. Together,the results show that models that rely on rehearsal or seriation processes to account for the ISE need to be reconsidered. Results are discussed in tenns of interference with encoding of to-be-remembered material.

Hoosiers on the Hardwood: A Critical Examination of Indiana Basketball Culture and its Effect on Identity Formation

The purpose of this research was to examine the nexus at which Indiana basketball and the state’s ‘hoosier’ identity meet. More specifically, this thesis interrogates the romanticization of this sporting culture for its pedagogical role in the creation of twenty-first century ‘hoosier’ bodies. Adopting a theoretical orientation rooted in critical race theory, I argue that Indiana’s basketball culture represents a normalized / normalizing structure underneath which Otherness is reified to produce hypervisible “different” outsiders (‘non-hoosiers’), and invisible “disciplined” insiders (i.e. ‘hoosiers’). Utilizing data gleaned over a two-month period spent conducting fieldwork in the “hoosier state” (document analysis, unstructured interviewing, and participant observation), I specifically tailor my analysis to uncover people’s understanding, negotiation, and performance of this regional and national subject position. From this point of inquiry, authentic ‘hoosierness’ comes to be represented, known, practiced, and felt in relation to hierarchies of power that privilege white, hypermasculine, rural, and conservative bodies.