• Interactions of diploid embryonal carcinoma cells and normal cell mass cells with 2.5 day mouse embryos following aggregation

      Waters, Barbara Kathleen.; Department of Biological Sciences (Brock University, 1984-07-09)
      Spontaneous teratocarcinomas are ovarian or testicular tumors which have their origins in germ cells. The tumors contain a disorganized array of benign differentiated cells as well as an undifferentiated population of malignant stem cells, the embryonal carcinoma or EC cells. These pluripotent stem cells in tissue culture share many properties with the transient pluripotent cells of the early embryo, and might therefore serve as models for the investigation of developmental events ill vitro. The property of EC cells of prime interest in this study is an in vivo phenomenon. Certain EC cell lines are known to be regulated ill vivo and to differentiate normally in association with normal embryonic cells, resulting in chimeric mice. These mice have two genetically distinct cell populations, one of which is derived from the originally malignant EC cells. This has usually been accomplished by injection of the EC cells into the Day 3 blastocyst. In this study, the interactions between earlier stage embryos and EC cells have been tested by aggregating clumps of EC cells with Day 2 embryos. The few previous aggregation studies produced a high degree of abnormality in chimeric embryos, but the EC cells employed had known chromosomal abnormalities. In this study, two diploid EC cell lines (P19 and Pi0) were aggregated with 2.5 day mouse embryos, and were found to behave quite differently in the embryonic environment. P19 containing aggregates generally resorbed early, and the few embryos recovered at midgestation were normal and non-chimeric. Pi0 containing aggregates survived in high numbers to midgestation, and the Pi0 cells were very successful in colonizing the embryo. All these embryos were chimeric, and the contribution by the EC cells to each chimera was very high. However, these heavily chimeric embryos were all abnormal. Blastocyst injection had previously produced some abnormal embryos with high Pl0 contributions in addition to the live born mice, which had lower EC contributions. This study now adds more support to the hypothesis that high EC contributions may be incompatible with normal development. The possibility that the abnormalities were due to the mixing of temporally asynchronous embryonic cell types in the aggregates was tested by aggregating normal pluripotent cells taken from 3.5 day embryos with 2.5 day embryos. Early embryo loss was very high, and histological studies showed that the majority of these embryos died by 6.5 days development. Some embryos escaped this early death such that some healthy chimeras were recovered, in contrast to recovery of abnormal chimeric embryos following Pl0-morula aggregations, and non-chimeric embryos following P19-morula aggregations. This somewhat surprising adverse effect on development following aggregation of normal cell types suggests that there are developmental difficulties associated with the mixing of asynchronous cell types in aggregates. However, the greater magnitude of the adverse effects when the aggregates contained tumor derived cells suggests that EC cells should not be considered the complete equivalent of the pluripotent cells of the early embryo.
    • Interactions of onion (Allium cepa) and yellow wax bean (Phaseolus vulgaris) in monoculture and intercropping with weeds, Chenopodium album and Amaranthus hybridus

      Lin, Yi An; Department of Biological Sciences
      Intercropping systems are seen as advantageous as they can provide higher crop yield and diversity along with fewer issues related to pests and weeds than monocultures. However, plant interactions in intercropped crop species and between crops and weeds in these systems are still not well understood. The main objective of this study was to investigate interactions between onion (Allium cepa) and yellow wax bean (Phaseolus vulgaris) in monocultures and intercropping with and without the presence of a weed species, either Chenopodium album or Amaranthus hybridus. Another objective of this study was to compare morphological traits of C. album from two different populations (conventional vs. organic farms). Using a factorial randomized block design, both crop species were planted either in monoculture or intercropped with or without the presence of one of the two weeds. The results showed that intercropping onion with yellow wax bean increased the growth of onion but decreased the growth of yellow wax bean when compared to monocultures. The relative yield total (RYT) value was 1.3. Individual aboveground dry weight of both weed species under intercropping was reduced about 5 times when compared to the control. The poor growth of weeds in intercropping might suggest that crop diversification can help resist weed infestations. A common garden experiment indicated that C. album plants from the conventional farm had larger leaf area and were taller than those from the organic farm. This might be associated with specific evolutionary adaptation of weeds to different farming practices. These findings contribute to the fundamental knowledge of crop-crop interactions, crop-weed competition and adaptation of weeds to various conditions. They provide insights for the management of diversified cropping systems and integrated weed management as practices in sustainable agriculture.
    • Interactions with bioenergetics by the mitochondria-targeted anti-apoptotic imidazole fatty acid derivative ‘TPP-IOA’.

      Maddalena, Lucas; Department of Biological Sciences
      The mitochondrial pathway of apoptosis contributes to cell death and tissue degeneration in a variety of human diseases. Targeting the molecular events of this pathway represents a promising therapeutic strategy under pathological scenarios, particularly in tissues with limited regenerative capability (e.g. brain, heart). Recently, the small molecule ‘TPP-IOA’ was designed to target mitochondria and therein inhibit the peroxidase activity of cytochrome c that promotes activation of the apoptotic death pathway (Atkinson et al., Nat. Commun. 2011; 2:497). Therefore, TPP-IOA holds promise as a potential therapeutic agent in many pathological scenarios of cell loss. However, TPP-IOA’s target protein and organelle perform critical functions in aerobic ATP production via oxidative phosphorylation, and yet little is known about TPP-IOA’s interaction with mitochondrial energetics. This is an important consideration for TPP-IOA’s potential future therapeutic utility, as avoiding interference to mitochondrial energetics will be essential for many target cell types that feature a high rate of oxidative phosphorylation (e.g. neurons, cardiomyocytes). Using the purified protein target, isolated organelle target, and live cultured cells, this thesis investigated TPP-IOA’s interaction with fundamental components of energetics-related mitochondrial biology. Assessments with pure cytochrome c revealed that TPP-IOA can inhibit the respiration-associated reduction activity of cytochrome c, and that this occurred at concentrations marginally higher than those required to inhibit the protein’s pro-apoptotic peroxidase activity. In isolated rat liver mitochondria, TPP-IOA impaired oxidative phosphorylation via inhibition of both ADP-phosphorylating-respiration and FCCP-uncoupled respiration, and stimulation of non-phosphorylating respiration. Critically, TPP-IOA was unable to inhibit pro-apoptotic peroxidase activity at concentrations that avoided interference to oxidative phosphorylation, suggesting it may be unable to perform its desired pharmacological activity without causing toxicity to mitochondrial energetics. In live cultured cells, TPP-IOA caused effects associated with impaired oxidative phosphorylation, including a collapsed mitochondrial membrane potential, fragmented mitochondrial network morphology, and apparent loss of mitochondria. Furthermore, TPP-IOA effectively inhibited apoptotic death under lethal conditions in cells that preferentially utilized anaerobic glucose metabolism, but not in cells manipulated to be heavily reliant on oxidative phosphorylation and thus more sensitive to the perturbations to mitochondrial energetics. Altogether, these findings indicate that relative cellular dependence on oxidative phosphorylation is a critical factor influencing TPP-IOA’s pharmacological efficacy. Furthermore, potential therapeutic applications may be limited to pathologies involving predominantly less aerobic tissues/cell types or depressed aerobic metabolism. Additionally, these findings have broader implications for mitochondrial medicine based on similar mitochondrial drug targeting strategies.
    • Intraspecific and interspecific social variation in the sweat bee Lasioglossum Malachurum and other members of the subgenus Evylaeus

      Wyman, Laura M.; Department of Biological Sciences (Brock University, 2002-10-02)
      Sweat bees exhibit a range of social behaviours, from solitary nesting, in which no workers are produced, to strong eusociality, in which workers exhibit a high degree of altruism, behaviour that is measured by the degree of personal reproductive sacrifice. Field studies were carried out for seven weeks during May-June 2000 in southern Greece in order to investigate intraspecific social variation, and test the hypothesis of a north-south cline of decreasing eusociality in the obligately eusocial sweat bee L. (E.) malachurum. A comparative study, using principal components analysis, was performed to determine if patterns of intraspecific social variation in L. malachurum reflect the patterns of social variation within the subgenus, Evylaeus, as a whole. The results of the field study reveal that, in Greece, two worker broods were produced followed by a third brood consisting of gynes, males and some workers, indicating that there was an overlap in worker and gyne production. There was strong caste distinction between queens and workers. Workers actively foraged and participated in nest construction as most workers (58%, n=303) had a high degree of mandibular wear. Workers did not participate in the oviposition of Brood 3 gynes since only 0.7% (n=278) of workers were mated. Furthermore, queen survival until the end of Brood 3 and a substantial size differential of 10.6% between queens and workers suggested that queen domination over worker behaviour during the early to mid-part of the colony cycle was plausible. Male production in Brood 3 by some workers was likely, since the timing of worker ovarian development corresponded with the timing of male production. These findings suggest that workers of the first two broods were primarily altruistic, but some (28%) Brood 1 (9%) and Brood 2 (19%) workers produced males, indicating that the degree of altruistic behaviour declined during the lifetime of the colony. In comparison with other L. malachurum populations in Europe, the Greek population of L. malachurum had a weaker social level as a result of the higher proportion of workers potentially involved in male production, thus 3 supporting the hypothesis of a southerly cline of decreasing eusociality. Furthermore, intraspecific variation in social level across Europe appears to be due to longer breeding seasons in more southerly locations that would promote the production of larger colonies and provide opportunities for workers to evade queen control. The comparative study using principal components analysis on 20 solitary (of the subgenera Evylaeus and Lasioglossum), eusocial and socially polymorphic Evylaeus species and populations reveals that six traits are closely associated with stronger eusociality in Evylaeus. These traits are: (1) a reduction in the proportion of males in the early brood(s); (2) a reduction in the proportion of females that mate; (3) an increase in the mean number of first brood workers; (4) a reduction in the proportion of females with developed ovaries; (5) an increase in size dimorphism between castes, and (6) nest guarding. These are traits that most significantly define principal component one and therefore distinguish social type as indicated by a clear separation of the eusocial and the solitary populations, with a socially polymorphic species falling in between. Furthermore, most of these traits are under foundress control and may suggest that the evolutionary loss or gain of eusociality is based on selection pressures on a founding female. Colony size and female ovarian development are common factors distinguishing social variation in L. malachurum and within the subgenus as a whole. The principal components analysis excluding the solitary species and the socially aberrant L. marginatum populations show the L. malachurum populations separated based on an increasing proportion of workers with developed ovaries as populations are found more south, lending further support to the hypothesis of a north-south cline of decreasing eusociality.
    • Introduced taxa in disturbed ecosystems

      Stirling, Ana Lorraine.; Department of Biological Sciences (Brock University, 1990-07-09)
      In the literature, introduced taxa are assumed to be present, more abundant, and occupy greater physical space in portions of ecosystems disturbed by human activity. This study tested this principle in two sites, Short Hills provincial Park ("SHU) and Backus Woods ("B~l"). spatial distribution of introduced taxa of vegetation, isopods, and earthworms was determined with the runs test along 300m transects encompassing gradients of anthropogenic disturbance severity. The hypothesis was that introduced taxa would be aggregated along these transects; the null hypothesis was that they would not be aggregated. The null hypothesis was rejected for the introduced taxa as a unit, and vegetation and earthworms individually. Introduced taxa were aggregated along 53.33% (N~30) and 57.14% (N~21) of the transects in SH and BW (respectively). Introduced vegetation (90.00%, N~10 and 100.00%, N~7) and earthworms (50.00%, N~10 and 50.00%, N~8) were also significantly aggregated within the sites. Introduced isopods, however, were not significantly aggregated at either place (20. 00%, N-=10 and 16. 67%, ~J~6). This study demonstrated that introduced taxa are aggregated within ecosystems disturbed by human activity. However, since introduced isopods were not significantly aggregated it was also shown that taxa respond differently.
    • Investigating how Notch, and JAK/STAT Signaling Synergistically Regulate Intestinal Stem Cell Homeostasis Using Engineered Optogenetic Alleles

      Lidster, Taylor; Department of Biological Sciences
      Stem cells are a class of undifferentiated cells that have the unique ability to give rise to a variety of specialized cell types during tissue development and growth. These cells communicate amongst one another by sending and receiving signals from multiple pathways that regulate cell fate decisions by promoting either self-renewal or differentiation. The Notch, and JAK/STAT signaling pathways are central regulators of multicellular development and are vital for tissue maintenance. The interplay between the Notch, and JAK/STAT pathways required for tissue homeostasis has not been fully elucidated, particularly as it relates to the intestinal epithelium. I utilize the Drosophila melanogaster midgut as a model system to study stem cell dynamics and more specifically, how Notch and JAK/STAT signaling cooperatively regulate intestinal stem cell turnover. In order to accomplish this, I employed both traditional and optogenetic methods to elicit Notch blockade and ectopic activation of JAK/STAT signaling in the midgut. First, I demonstrated that ectopic JAK/STAT signaling paired with Notch knockdown causes substantial ISC overproliferation, leading to the formation of large ISC tumors spanning the entire midgut. Quantitative assessment of ISC pools confirmed that Notch and JAK/STAT signaling work in a synergistic manner, rather than an additive manner, to regulate ISC homeostasis. I also utilized two optogenetic alleles, Opto-Delta and Opto-Domeless, to recapitulate Notch blockade and JAK/STAT hyperactivation using light. Opto-Delta was tested in the intestinal epithelium and demonstrated the ability to block Notch signaling in the ISCs, resulting in the formation of stem cell clusters. Subsequently, an Opto-Domeless construct was created and expressed in the nervous system during embryogenesis to assess its efficacy to activate JAK/STAT signaling. The embryos that expressed Opto-Domeless exhibited a significant decrease in viability when subjected to the light condition, suggesting possible light-responsiveness. Expression of Opto-Domeless in the midgut was also achieved, however, ISCs expressing Opto-Domeless appeared to have JAK/STAT activity in the absence of light exposure, suggesting that Opto-Domeless is not behaving in a binary fashion and will require further validation. Altogether, these findings reveal great potential for light-gated proteins, as they provide a powerful approach to disentangle dynamic cellular signals by using light.
    • Investigating the Modulation of Drosophila melanogaster Body-wall Muscle Contraction by the Neuropeptide DPKQDFMRFamide

      Wasilewicz, Luc; Department of Biological Sciences
      The chemical synapse is the site of communication between a neuron and its target cell, where an electrical impulse depolarizes the presynaptic cell causing chemical release. The chemicals released at the synapse are signaling molecules referred to as transmitters and co-transmitters that exert effects on the target cell and can sometimes modulate the effects of each other. A class of signaling molecules, known as neuropeptides, can act as transmitters or can be released as hormones that can modulate chemical synapses and ultimately affect many physiological functions. The neuropeptide, DPKQDFMRFamide, is an important neuromodulator of neuromuscular junctions in the fruit fly, Drosophila melanogaster. DPKQDFMRFamide has previously been shown to enhance excitatory junctional potentials (EJPs) elicited by specific neurons, to enhance nerve-evoked contractions, and to induce contractions directly in Drosophila 3rd instar larval body-wall muscles. This thesis investigated how the DPKQDFMRFamide peptide modulates muscle contractions elicited by the excitatory transmitter of the neuromuscular junction, L-glutamate, in D.melanogaster 3rd instar larvae. Effects were assessed by co-applying peptide with L-glutamate after removing the central nervous system. The results indicate that DPKQDFMRFamide enhances glutamate-evoked contractions in a dose-dependent manner, and there was synergy between the effects of L-glutamate and DPKQDFMRFamide on muscle contraction. DPKQDFMRFamide increased membrane depolarization in muscle when co-applied with glutamate, and it enhanced contractions induced by caffeine in the absence of extracellular calcium. Thus, the peptide appears to act at the cell membrane to increase depolarization and at, or downstream of the sarcoplasmic reticulum (SR) to enhance caffeine-induced contractions. However, the effects of DPKQDFMRFamide do not appear to involve the 2nd messenger nitric oxide or the calcium/calmodulin activated protein kinase, CaMKII.
    • An investigation into differences between indoleacetic acid and fusicoccin in their influence on RNA synthesis, protein synthesis and growth in Avena coleoptile tissue

      Doo, Ahmed Cassin.; Department of Biological Sciences (Brock University, 1979-07-09)
      Growth stimulation of Avena coleoptile tissue by indoleacetic acid (IAA) and fusicoccin (FC) was compared by measuring both their influence on RNA and protein synthesis during IAA or FC stimulated growth. FC stimulated growth more than IAA during the initial four hour exposure, after which the growth rate gradually declined to the control rate. FC, but not IAA, increased the uptake of 3H-Ieucine into tissue and the specific radioactivity of extracted protein. Cycloheximide inhibited the incorporation of 3H-Ieucine into protein by approximately 60% to 70% in all cases. In the presence of cycloheximide 3H-radioactivity accumulated in FC-treated tissue, whereas IAA did not seem to influence 3H-accumulation. These results suggest that FC stimulated leucine uptake into the tissue and that increased specific activity of coleoptile protein is due to increased leucine uptake, not an increased rate of protein synthesis. There was no measurable influence of IAA and/or FC on RNA and protein synthesis during the initial hours of a growth stimulation. Inhibitors of RNA and protein synthesis, actinomycin D and cycloheximide, respectively, severely inhibited IAA enhanced growth but only partially inhibited FC stimulated growth. The data are consistent with suggestions that a rapidly turning over protein participates in IAA stimulated growth, and that a continual synthesis of RNA and proteins is an absolute requirement for a long term growth response to IAA. On the contrary, FC-stimulated growth exhibited less dependency on the transcription and translation processes. The data are consistent with proposals suggesting different sites of action for FC and IAA stimulated growth. l?hen compared to CO2-free air, CO2 at 300 ppm had no significant influence on coleoptile growth and protein synthesis in the presence or absence of lAA or FC. Also, I mM malate, pH 6.0 did not influence growth of coleoptiles in the presence or absence of lAA. This result was obtained despite reports indicating that 300 ppm CO2 or I mM malate stimulates growth and protein synthesis. This lack of difference between CO2-treated and untreated tissue could indicate either that the interstitial space CO2 concentration is not actually different in the two treatments due to significant endogenous respiratory CO2 or else the data would suggest a very loose coupling between dark CO2 fixation and growth. IAA stimulated the in vivo fixation of 14c-bicarbonate (NaHI4c03) by about 25% and the addition of cycloheximide caused an inhibition of bicarbonate fixation within 30 min. Cycloheximide has also been reported to inhibit IAA-stimulated H+ excretion. These data are consistent with the acid growth theory and suggest that lAA stimulated growth involves dark CO2 fixation. The roles of dark CO2 fixation in lAA-stimulated growth are discussed.
    • An investigation into the effects of homopteran honeydew sugars versus floral nectar sugars on black fly longevity, flight performance and digestion /

      Stanfield, Trudy K.; Department of Biological Sciences (Brock University, 2003-07-14)
      Floral nectar is thought to be the primary carbohydrate source for most dipteran species. However, it has been shown that black flies (Burgin & Hunter 1997 a,b,c), mosquitoes (Foster 1995; Burkett et al. 1999; Russell & Hunter 2002), deer flies (Magnarelli & Burger 1984; Janzen & Hunter 1998; Ossowski & Hunter 2000), horse flies (Schutz & Gaugler 1989; Hunter & Ossowski 1999) and sand flies (MacVicker et al. 1990; Wallbanks et al. 1990; Cameron et al. 1992, 1995; Schlein & Jacobson 1994, 1999; Hamilton & EI Naiem 2000) feed on homopteran honeydew as well as floral nectar. Prior to 1997 floral nectar was thought to be the main source of carbohydrates for black flies. However, Burgin & Hunter (1 997a) demonstrated that up to 35% of black flies had recently consumed meals of homo pte ran honeydew. This information has necessitated a re-assessment of many life history aspects of black flies. Attempts are being made to examine the effects of nectar versus honeydew on black fly fecundity and parasite transmission (Hazzard 2003). Recently, Stanfield and Hunter (unpublished data) have shown that in female black flies, honeydew sugars produce flights of longer distance and duration than do nectar sugars. This thesis examines two aspects of black fly biology as it relates to sugar meal consumption. First, the effects of honeydew and nectar on black fly longevity are examined. Second, the proximate causation behind longer flight performances in honeydew-fed flies will be examined. The comparison between these two sources is important because nectar is composed of mainly simple sugars (monosaccharides and disaccharides) whereas honeydew is composed of both simple and complex sugars (including trisaccharides and tetrasaccharides ).
    • 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 /

      Al-Hazmi, Abdul Aziz.; Department of Biological Sciences (Brock University, 1999-05-21)
      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.
    • Investigation into the relationships between molecular chaperones, mitochondrial antioxidant enzymes, and endothermic vertebrate longevity

      Christoff, Casey; Department of Biological Sciences (Brock University, 2014-04-21)
      The maximum lifespan (MLSP) of endothermic vertebrates can range from as little as a year to over two centuries, yet the underlying phenotype of aging is very similar amongst this group of organisms. One organelle that may be important in the phenotype of aging is the mitochondrion. When damaged, this organelle is thought to contribute to many of the neurodegenerative diseases of aging. For this thesis, mitochondria from brain tissues of 7 mammalian and 2 avian species were isolated to assess whether the antioxidant glutathione system and major molecular chaperone, HSP60, is correlated to species MLSP. Furthermore, HSP60, and the major endoplasmic reticulum chaperone, GRP78, were measured under basal conditions, and following the introduction of an oxidative stress (hydrogen peroxide) in cultured mammalian myoblasts from 10 different species. My results indicate that the enzymes involved in the glutathione defense system are not correlated to species MLSP in brain mitochondria; however HSP60 levels are indeed higher in the longer-lived species. HSP60 levels are also higher at the basal level in cultured mammalian myoblasts and after 1 hour of hydrogen peroxide exposure. GRP78 induction is not correlated to species MLSP at the basal level or following hydrogen peroxide exposure. Therefore, these results suggest that HSP60 is a correlate of longevity in endothermic vertebrate species, but neither the glutathione antioxidant defense system, nor GRP78, correlates to species longevity.
    • The investigation of edge effects within ragmented forest islands in Short Hills Provincial Park

      Jonsson-Ninniss, Susan.; Department of Biological Sciences (Brock University, 1994-10-02)
      It has been well documented, within the field of landscape ecology, that terrestrial fragmentation contributes to increased heterogeneity at the landscape level. It has also been observed that elevated areas of edge habitat occur within fragmented landscapes. Spatial and temporal edge effects were investigated in four areas designated as Nature Reserve Zones within Short Hills Provincial Park, near St. Catharines, Ontario. Random sampling along exposed edges was performed on trees and saplings, at 5 and 25 ill edge depths, using the point-centred quarter method. Diameter at breast height (dbh) and distance from point measurements were used to establish relative density, dominance, frequency and importance value. One-way analyses of variance were used on dbh measurements of tree species and Chi-Square contingency tables were used on size class distributions of saplings species to determine significant differences between 5 and 25 metres. Qualitative comparisons of importance values were also used to determine differences between 5 and 25 metres as well as between trees and saplings. These statistical and qualitative comparisons suggest that a significant overall spatial edge effect is currently exhibited by fragmented wooded islands within the park. The major species of the park, Acersaccharuln, may be exhibiting a temporal edge effect. The heterogeneous nature of the park may be of importance in understanding this area as a complex, ecological system. It is possible that the remaining forest tracts of the park have been affected, and continue to be affected by previous disturbances. Based on these findings, recommendations are made to the Ontario Ministry of Natural Resources concerning the management of Short Hills Provincial Park in accordance with their 1990 proposed Management Plan.
    • An investigation of ethylene inhibition of growth in etiolated Avena sativa coleoptiles /

      Dymock, Ian James.; Department of Biological Sciences (Brock University, 1975-06-09)
      Growth rates of etiolated Avena sativa coleoptiles in pH 7.0 buffered medium are stimulated in a synergistic manner by IAA and 320 ~l/l carbon dioxide. The suggestion that carbon dioxide stimulated growth involves dark fixation is supported by the ability of 1 mM malate to replace carbon dioxide, with neither factor able to stimulate growth in the presence of the other (Bown, Dymock and Aung, 1974). The regulation of Avena coleoptile growth by ethylene has been investigated in the light of this data and the well documented antagonism between carbon dioxide and ethylene in the regulation of developmental processes. The influence of various permutations of ethylene, IAA, carbon dioxide and malate on the rates of growth, l4c-bicarbonate incorporation, l4C-bicarbonate fixation, and malate decarboxylation have been investigated. In the presence of 320 ~l/l carbon dioxide, 10.8 ~l/l ethylene inhibited growth both in the absence and presence of 20 ~M IAA with inhibition times, of 8-10 and 12-13 minutes respectively. In contrast ethylene inhibition of growth was not significant in the absence of growth stimulation by CO2 or 1 mM malate, and the normal growth increases in response to CO2 and malate were blocked by the simultaneous application of ethylene. The rates of incorporation and dark fixation of l4C-bicerbonate were not measurably. influenced by ethylene, IAA or malate, either prior to or during the changes in growth ,ates induced by these agents. The data does not support the hypothesis that ethylene inhibition of growth results from an inhibition of dark fixation, but suggests that ethylene may inhibit a process which is subsequent to fixation.
    • An investigation of sugar feeding by black flies (Diptera: Simuliidae)

      Burgin, Steve G.; Department of Biological Sciences (Brock University, 1995-07-09)
      Although much research has been conducted on blood-meal acquisition in adult female black flies (Diptera: Simuliidae), the same cannot be said for sugarmeals. Both sexes feed on sugar which provides energy for flight and it has been commonly held that nectar is the major carbohydrate source. This thesis addresses the question of whether a non-floral carbohydrate source, specifically homopteran honeydew, is ingested by male and female black flies. Black flies reared in the laboratory have been observed to readily ingest freshly excreted and older (dry) honeydew when presented with honeydew coated tamarack branches. Field work was conducted in Algonquin Park, Ontario in the spring and summer of 1993. Three separate studies were designed to test whether homopteran honeydew is an important carbohydrate source for black flies and whether flies from different habitats utilize different sugar sources. The sugars melezitose and / or stachyose are known to occur in a variety of homopteran honeydews and therefore were used as indicators of honeydew feeding by black flies. In the first study, black flies were collected with insect nets from a stand of Larix larcina heavily infested with honeydew - producing homopterans (Adelges lariciatus). Six black fly species were captured: Simulium venustum, S. rostra tum, S. vittatum, Stegopterna mutata, S. aureum and S. quebecense. Samples of honeydew and individual black flies were tested using thin layer chromatography (T. L. C.) with fructose, glucose, sucrose, turanose, melezitose, raffinose and stachyose as standards. All sugars except turanose and melezitose were found in the adelgid honeydew samples. Since the sugar melezitose was absent from ~ honeydew samples, stachyose was used to indicate that black flies were feeding from this particular honeydew source. Of the 201 black flies tested, 194 contained sugars which occurred in 16 combinations. Stachyose combinations excluding melezitose, present in 45.9 % of flies, were used to indicate that black flies had been feeding on the adelgid honeydew. In the second study, black flies were collected in the morning and evening on 8 collection dates, using a vehicle mounted insect net. The crops and midguts of 10 male and 10 female Simulium venustum were dissected on each sample date. In total the gut contents of 320 individual flies were analysed by T. L. C. The sugars identified from these flies were present in the following proportions: fructose (100.0%), glucose (100.0%), sucrose/turanose (50.4%), melezitose (30.3%), raffinose (18.8%) and stachyose (8.7%). These sugars occurred in fourteen different combinations. It is argued that the presence of melezitose and / or stachyose indicates that black flies had fed on homopteran honeydew. Significantly more female flies (40.0%) than male flies (27.5%) had fed on honeydew. In the third study, adult black flies were sampled by sweep netting vegetation in four habitats in the morning and evening on 8 collection dates. The habitats are as follows: (1) Davies Bog, (2) Abandoned Air Field (dominated by blueberries, Vaccinium spp.), (3) Deciduous Habitat and (4) Coniferous Habitat. Sugars in the crops and midguts of female flies were tested by T. L. C. and, for S. venustum, it was found that significantly fewer flies (18.8%) from the Air Field contained honeydew than from the other three sites (Davies Bog, 34.4%; Deciduous Habitat, 36.2%; Coniferous Habitat, 25.0%). Of the 1287 black flies tested individually by T. L. C. 441 (34.3%) contained melezitose and / or stachyose sugars indicating that this proportion of the population were feeding from Homopteran honeydew. It is therefore clear that floral (nectar) sugars are not the only source of carbohydrates available to black flies.
    • Investigation of the mechanism of transfer of a-tocopherol by the human a-tocopherol transfer protein (H-a-TTP) /

      Frahm, Grant E.; Department of Biological Sciences (Brock University, 2007-06-01)
      The human a-tocopherol transfer protein (h-a-TTP) is understood to be the entity responsible for the specific retention of a-tocopherol (a-toc) in human tissues over all other forms of vitamin E obtained from the diet. a-Tocopherol is the most biologically active form of vitamin E, and to date has been studied extensively with regard to its antioxidant properties and its role of terminating membrane lipid peroxidation chain reactions. However, information surrounding the distribution of a-tocopherol, specifically its delivery to intracellular membranes by a-TTP, is still unclear and the molecular factors influencing transfer remain elusive. To investigate the mechanism of ligand transfer by the h-a-TTP, a fluorescent analogue of a-toc has been used in the development of a fluorescence resonance energy transfer (FRET) assay. (/?)-2,5,7,8-tetramethyl-2-[9-(7-nitro-benzo[l,2,5]oxdiazol-4-ylamino)-nonyl]- chroman-6-ol (NBD-toc) has allowed for the development of the FRET-based ligand transfer assay. This ligand has been utilized in a series of experiments where changes were made to acceptor lipid membrane concentration and composition, as well as to the ionic strength and viscosity of the buffer medium. Such changes have yielded evidence supporting a collisional mechanism of ligand transfer by a-TTP, and have brought to light a new line of inquiry pertaining to the nature of the forces governing the collisional transfer interaction. Through elucidation of the transfer mechanism type, a deeper understanding of the transfer event and the in vivo fate of a-tocopherol have been obtained. Furthermore, the results presented here allow for a deeper investigation of the forces controlling the collisional protein-membrane interaction and their effect on the transfer of a-toc to membranes. Future investigation in this direction will raise the possibility of a complete understanding of the molecular events surrounding the distribution of a-toc within the cell and to the body's tissues.
    • Investigations on the host-parasite interface of a mycoparasite system /

      Letourneau, Denis Raymond.; Department of Biological Sciences (Brock University, 1977-09-23)
      The cell wall composition of Choanephora cucur - bitarum and the host-parasite interface, after infection with Piptocephalis virginiana , were examined in detail. The cell walls of C_. cucurbitarum were determined to be composed of chitin (17%), chitosan (28.4%), neutral sugars (7.2%),uronic acid (2.4%), proteins (8.2%) and lipids (13.8%). The structure of hyphal walls investigated by electron microscopy of shadowed replicas before and after alkali-acid hydrolysis, showed two distinct regions: microfibrillar and amorphous. The microfibrils which were composed of mainly chitin, were organized into two distinct layers: an outer, thicker layer of randomly orientated microfibrils and an inner, thin layer of parallel microfibrils.Electronmicrographs of the host-parasite interface of C_. cucurbitarum and the mycoparasite , P_. virginiana , 30 h following inoculation, showed that the sheath zone has a similar electron density to that of the host cell wall. The sheath was not present around the young (18 h old) haustorium. High-resolution autoradiographs of infected host hyphae showed that radioactive N-acetyl-D-glucosamine , a precursor of chitin, was incorporated preferentially in the host cell wall and sheath zone. Cell fractionation of label fed hyphae showed that 84% of the label was present in the cell wall and specifically in the chitin portion of the wall. The antifungal antibiotic, Polyoxin D, a specific inhibitor of the enzyme, chitin synthetase, suppressed the incorporation of the label in the cell wall and sheath zone and resulted in a decrease in electron density of the developing sheath. The significance of these results is discussed in the light of host resistance.
    • Investigations on the production of long chain fatty acids in Choanephora cucurbitarum (Berk.

      Parmar, Yashpal Inder.; Department of Biological Sciences (Brock University, 1979-11-04)
      The fatty acid composition of the total cellular lipids of Choanephora cucurbitarum incubated for 96 hrs on either glucose-ammonium sulfate or malt-weast extract media was determined. The major fatty acids were palmitic, palmitoleic, stearic and linoleic acids. The saturated fatty acid possessing the longest acyl chain was stearate (C 18:0). The presence of glutamic acid (2.0 x 10-1% or 1.36 x la-2M) in either of the above growth media resulted in increase in percent of 1f-linolenic acid, decrease in percent of linoleic ~iCid and appearance of a new series of fatty acid> C ~8 e.g. C ",,,,'V' C2k:O, C26,O. The addition of glutamic acid had no effect on the lipid yield but slightly decreased the degree of unsaturation. Compounds which duplicated the effect of glutamic acid were acetate, malate, citrate, succinate, 0( -ketoglutarate, prOline, -y -aminobutyric acid and glucose (3%) but not aspartic acid or alanine. ~o correlation was found between glutamic acid pool concentration and the presence in the growth medium of those compounds which stimulate long chain fatty acid production. Four hours of incubation with 27 JJ 1-1 glutamate supported the production of long chain fatty acids. This stimulation is inhibited if 272 .u M isophthalic acid is added with 27 AJ M glutamate. But, long chain fatty acids were detected when 27 JJ M eX -ketoglutarate is also present in the incubation mixture. Five hours of incubation with 100 ,Mg/ml of cycloheximide resulted in over 9CY/o inhibition of cytoplasmic :protein synthesise Glutamate (27 .uM) enhanced the synthesis of long chain fatty acids under these conditions. These findings are discussed in an attempt to provide a plausible explanation COmmon to compounds that support the production of long chain fatty acids.
    • An investigative study on the effects of black fly (diptera: simuliidae) sugar meals on reproductive success and parasite transmission /

      Hazzard, Christie-Lee.; Department of Biological Sciences (Brock University, 2003-05-21)
      Black flies are opportunistic sugar-feeders. They take sugar meals from Homopteran honeydew secretions or plant nectars, depending on availability. Homopteran honeydew secretions contain both simple and complex carbohydrates while plant nectars contain primarily simple carbohydrates. In order to determine whether honeydew secretions offer more energy than plant nectars to their insect visitors a study of wild-caught black flies was undertaken in Algonquin Provincial Park, Canada during the spring of 1 998 and 1 999. It was hypothesized that female black flies maintained on honeydew sugars will survive longer, produce more eggs and have a greater parasite vectoring potential than those maintained on artificial nectar or distilled water. Results demonstrated that: (1) host-seeking female Prosimulimfuscum/mixtum and Simulium venustum maintained on artificial honeydew did not survive longer than those maintained on artificial nectar when fed ad libitum; (2) fiiUy engorged S. venustum and Simulium rugglesi maintained on artificial honeydew did not produce more eggs than those maintained on artificial nectar when fed ad libitum; and (3) S. rugglesi did not have a greater vectoring potential of Leucocytozoon simondi when maintained on artificial honeydew as opposed to artificial nectar when fed ad libitum. However, all flies maintained on the two sugars (artificial honeydew and artificial nectar) survived longer, produce more eggs and had greater vectoring potential than those maintained on distilled water alone.
    • Involvement of fimbriae in host-mycoparasite recognition

      Rghei, Nezar A.; Department of Biological Sciences (Brock University, 1991-07-09)
      Extracellular, non-flagellar appendages, termed fimbriae are widespread among fungi. Fungal fimbriae range in diameter from 6-10 nm and exhibit lengths of up to 30 ~m. Fungal fimbriae have been implicated in several functions: adhesion, conjugation and flocculation. A possible role of fimbriae in host-mycoparasite interactions was the focus of this study . Using electron microscopy, fimbriae were observed on the surfaces of Mortiere lla cande labrum, Mortie re lla pusi lla and Phascolomyces articulosus with diameter means of 9.1±0.4 nm, 9.4±0.5 nm and 8.6±0.6 nm, respectively, and lengths of up to 25 ~m. Fimbriae were not observed on the surface of the mycoparasite, Piptocephalis virginiana. Polyclonal antiserum (AU) prepared against the fimbrial protein of Ustilago violacea cross-reacted with 60 and 57 kDa M. candelabrum proteins. In addition, AU cross-reacted with 64 kDa proteins from both M. pusilla and P. articulosus. The proteins that cross-reacted with AU were electroeluted from polyacrylamide gels and were shown to subsequently form fibrils. The diameter means for the electroeluted fibrils were: for M. candelabrum 9.7±0.3 nm, M. pusilla 8.4±0.6 nm and P articulosus 9.2±0.5 nm. Finally, to ascertain the role of fimbriae in host-mycoparasite interactions, AU was incubated with P. virginiana and M. pusilla (mycoparasite/susceptible host) and with P. virginiana and P . articulosus (mycoparasite/ resistant host). It was observed that AU decreased significantly the level of contact between P. virginiana and M. pusilla and between P. virginiana and P. articulosus in comparison to prelmmune serum treatments. Thus, it was proposed that fimbriae might play recognition and attachment roles in early events of mycoparasitism.

      Tolentino, Michael; Department of Biological Sciences
      Research into the molecular mechanisms of the psychoactive effects of cannabis has led to the discovery of the endocannabinoid system (ECS), a neuromodulatory system conserved throughout the animal kingdom. Little is known about its function in mammals, but there is evidence suggesting its contributions in the cellular processes that are important in CNS development and are conserved during CNS regeneration. However, these studies focussed primarily on mammals, which display limited abilities to regenerate after traumatic CNS injury. Furthermore, nothing is known regarding the role of endocannabinoids in CNS regeneration-competent species like the Mexican axolotl, one of the few vertebrates that can regenerate their spinal cord. The current study investigates the potential role of the ECS in influencing the pro-regenerative response observed in the axolotl spinal cord. I provide evidence that the main ECS receptor in the CNS (CB1) is upregulated in the regenerating caudal spinal cord and tail tissues of larval axolotls at 4 hours post amputation, lasting until 14 days post amputation. By performing immunofluorescence studies on these tissues, I demonstrate the expression of this receptor mainly in the ependymal region. In addition, bath application of the CB1 inverse agonist, AM251, significantly inhibited caudal growth of the spinal cord and tail by 7 days post amputation. The current study also identified an upregulation in a second ECS receptor, CB2, at 7- and 14-days post amputation. Immunofluorescence analysis revealed the localization of this receptor to the subependymal regions within the spinal cord. Furthermore, inhibition with the CB2 inverse agonist, AM630, similarly demonstrated an inhibition in spinal cord and tail regeneration by 7 days post amputation. An assessment of CB1 and CB2 expression was performed by identifying their localization in bromodeoxyuridine-positive (proliferating) and doublecortin-positive (differentiating neuronal) cells in 7-day regenerate tissue. These studies are the first to examine the role of the ECS during spinal cord regeneration in a regeneration-competent vertebrate and may aid in developing novel therapies for human nervous system injuries or pathologies.