mPIP is a mouse homologue of human PIP/GCDFP-15 which is an established marker of both malignant and benign pathological conditions of the mammary gland. mPIP gene expression has been identified in both lacrimal and salivary glands of healthy mice and the mPIP protein has been detected in saliva. The mPIP protein has been found to bind oral bacteria, showing the highest affinity for streptococci, suggesting a potential function of mPIP in the non-immune host defense in the mouse oral cavity. Since the exact functions of mPIP are still unknown, we examined the roles of mPIP through both in vitro and in vivo studies, specifically to address the possible role of this protein in non-immune host response through modulating the oral flora. The in vitro studies were primarily focused on elucidation of the consequences of interaction between mPIP and oral bacteria, in particular to examine whether mPIP plays a role in bacterial aggregation. The in vivo studies addressed the roles of mPIP through the analysis of an mPIP knockout mouse model generated in our laboratory. Following confirmation of the null mutation, the delineating the phenotype of this model was pursued through morphopathological analysis as well as examination of the impact of the lack of mPIP on the mouse oral flora. The null mutation in the mPIP knockout mice was confirmed by both the gene and protein analysis. Histological analysis revealed lymphocytic proliferation in both the submaxillary and prostate glands of the mPIP knockout mice. In addition, both quantitative and composition differences in the oral flora of mPIP knockout mice were identified when compared with wild-type controls. Specifically, a higher proportion of the oral bacteria of mPIP knockout mice were found to belong to genus Streptococcus and certain genera were found to be absent from the oral cavity of these mice. The effect of knockout mouse saliva, which lacks mPIP, on the aggregation of oral bacteria was compared to wild-type mouse saliva. Our data suggests that mPIP contributes to saliva-induced bacterial aggregation. While oral flora has multiple functions, including protection against infection, mPIP might play a role in the non-innate host defense through modulating the resident oral flora in the mouse. The identification of lymphocytic proliferation in submaxillary and prostate glands of mPIP knockout mice suggests that mPIP might also interfere with lymphocyte activity, playing a possible immunomodulatory role.

The primary objective of this study was to determine the role of the host community in the marketing of arts festivals with reference to the Grahamstown National Arts Festival. This goal was achieved firstly by analysing the relationship between the events industry and the community. Secondly, a literature study was conducted to explore the role of the community in tourism marketing. Thirdly, a survey was done to determine the perceptions of the community in terms of the marketing of the Festival as well as the social impact thereof. From previous studies it was found that the local community can be seen as a group of people of different gender, race, age and different race and social groups living together in a certain geographic area. The local community is the main suppliers of an event; they provide many of the businesses, public places and hospitality services used by tourists and visitors at an event. Staff, volunteers and many other resources come from the local community. The attitude of the local community towards the Festival is proven to be important. Positive attitudes are associated with economic and social benefits gained from the event. The attitude of the local community can influence their behaviour towards tourists as well as their message spread concerning the event, through word-of-mouth communication. They can influence the buying behaviour of tourists and visitors. Research was undertaking at Grahamstown National Arts Festival to determine the perceptions and attitudes of the local community towards the Festival and the marketing thereof. This was determined by means of a questionnaire as adopted from Fredline, Jago and Deery. It was distributed among the local community according to a stratified random sampling procedure. A total of 265 questionnaires were completed by residents. Firstly, the data was used to compile tables and graphs with a view to interpret the descriptive data: demographic information as well as perceptions and attitudes of the local community towards the Festival and the marketing thereof. It was found that the Festival has a positive impact on their personal lives as well as on the community as a whole. Positive and negative social impacts were identified, which indicates that the community are aware of the impacts: positive, including the range of things to do in Grahamstown has improved (80%), the number of people in the area has increased (80%) and there are more opportunities to meet new people (80%). The highest rated negative social impacts include the fact that the availability of parking has decreased (79%), that the public money spent on the Festival would be better spent on other things (43%) and that ordinary residents get no say in the planning and management of the Festival (50%). In terms of the marketing, the local community markets the Festival verbally by telling friends and family about it and also by attending the Festival themselves. However, the local community feel that they should be more involved in the planning and marketing of the Festival. Secondly, a factor analysis was performed with a view to synthesise the large amount of data concerning the perceptions and marketing of the Festival in smaller, more descriptive factors. These factors were used to determine the role of the community in the marketing of the Festival. The factor analysis yielded a 2-factor solution and the factors were labelled: Factor 1: Verbal marketing of the Festival; Factor 2: Action marketing of the Festival. Thirdly, a one-way analysis of variance (ANOVA) was conducted to explore the impact of the variables: age, gender, race, income, level of education, attitudes towards continuation of the Festival, attendance of the current festival, attendance of previous festivals, level of interest in the Festival, length of stay in Grahamstown, attitudes towards living in Grahamstown on the dimensions Verbal and Active marketing, as determined in the factor analysis. It was found that a medium effect exists between the middle-aged group and the older-age group on the Verbal marketing dimension. A medium effect also exists between the black and Asian respondents on the dimension Verbal marketing. There is a large effect between attendance of the Festival and Verbal marketing but a medium effect on the dimension Action marketing. A large effect was determined between avid fans and those that are not interested but attend occasionally, between avid fans and those who have no interest in the Festival, between those that are interested and attend some aspects of the Festival and those that are not interested but attend, those that are interested and attend and those that are not interested, on the Verbal marketing dimension. A medium effect was discovered between avid fans of the Festival and those that attended some of the aspects of the Festival, and those that are not interested but attend some aspects, and those that are not interested at all, on the Verbal marketing dimension. On the Verbal marketing dimension a medium effect was determined between the following: A medium effect was determined between avid fans of the Festival and those that attended some of the aspects of the Festival, avid fans and those that are not interested but attend occasionally, those that are interested and attend and those that are not interested, on the Action marketing dimension. This study contributes to the information that already exists concerning the role of the community in the marketing of arts festivals and on marketing communication research. / Thesis (M.Com. (Tourism))--North-West University, Potchefstroom Campus, 2009.

Studying how SN Ia rates (SNR) correlate with host galaxy properties is an important step in understanding the exact nature of SN Ia. Taking a sample of SNe and galaxies from the SDSS, we obtain the optimum parameter values for the A+B model for SNR, which states that SNR scale linearly with mass and star formation rate of the host, and compare them with previous work. We then proceed to show that the A+B model deviates very significantly from the SNR behaviour in our sample, demonstrate that no reasonable values for A and B could possibly match the observations, and investigate the possibility of a third-parameter correction to the generic A+B model. We find that several hypothesised models seem to match the distribution of SNRs in our sample; however, discriminating between them is a difficult task. We interpret the above to be an indicator that a new parameter may need to be taken into account when modelling SNR, and we present metallicity as a possible candidate for the new parameter. Also, by investigating decomposed bulge + disk components of the host galaxies, we find that the spatial positions of SNe Ia are correlated with bulge luminosity, but not with galaxy total luminosity or disk luminosity. It is also shown that SNe do not preferentially occur in bulge-dominated galaxies. Our interpretation of these results is that SNe arise from a population having a spatial distribution which correlates very well with bulge luminosity, but does not usually contribute to bulge luminosity. / Graduate

Viruses are common causative agents of ocular infection among humans. Epidemic keratoconjuntivitis (EKC) is a severe and contagious ocular disease with reported outbreaks worldwide. It is estimated that this disease affects 20-40 million individuals every year, which leads to huge socioeconomic costs for the affected countries. EKC is characterized by keratitis and conjunctivitis but is also associated with pain, edema, lacrimation, and decreased vision that can prolong for months after the infection and in rare cases years. This disease is caused by human adenoviruses (HAdVs), which belong to the family of Adenoviridae. Currently, there is no available treatment against EKC. EKC is mainly caused by HAdV-8, HAdV-19, HAdV-37, HAdV-53, HAdV-54, and HAdV-56, which belong to species D HAdVs. HAdV-8, HAdV-19 and HAdV-37 have previously been shown to use sialic acid (SA)-containing glycans as cellular receptors to bind to and infect human corneal epithelial (HCE) cells. To characterize the receptor in more detail, we performed a glycan array, which included SA-containing glycans. A branched hexasaccharide terminating with SA in each arm was identified as a candidate receptor. This glycan corresponds to the glycan motif found on a ganglioside, GD1a. By performing a series of biological and biochemical experiments we confirmed the function of the GD1a glycan as a cellular receptor for EKC-causing HAdVs. However, the glycan used as a receptor was linked to plasma membrane protein(s) through O-glycosidic bonds, rather than to a lipid (as in the ganglioside). X-ray crystallography analysis showed that the two terminal SA:s interacted with two of the three previously identified SA-binding sites on the knob domain of the HAdV-37 capsid protein known as the fiber. Based on the structural features of the GD1a:HAdV-37 knob interaction, we assumed that a three-armed molecule with each arm terminating with SA would be an efficient inhibitor. Such molecules were designed, synthesized and found to efficiently prevent HAdV-37 binding to and infection of corneal cells. These results indicate that trisialic acids-containing compounds may be used for treatment of EKC. After binding to its primary receptor, most HAdVs have been shown to interact with αVβ3 and αVβ5 integrins to enter human cells. This interaction occurs through the RGD (arginine-alanine-aspartic acid) motif in the capsid protein known as the penton base. However, it was not clear if corneal epithelial cells express αVβ3 and αVβ5 integrins. Thus, to better understand additional early steps of infection by EKC-causing HAdVs, we performed binding and infection competition experiments using human corneal epithelial cells and siRNA, integrin specific antibodies, peptides and RGD-containing ligands indicating that α3, αV, β1 affected HAdV-37 infection of but not binding to HCE cells. We could also see that HAdV-37 co-localize with α3 and αV at after entry into HCE cells. In situ histochemistry confirmed that the expression of α3 and αV in human corneal tissue. Overall, our results suggest that αV and α3 integrins are important for HAdV-37 infection of corneal cells. Altogether, these results provide further insight into the biology of HAdVs and open up for development of novel antiviral drugs.

This study is concerned with two strigeid genera which utilise fish as their second intermediate host and piscivorous birds as a definitive host, i.e. Apatemon (Apatemon) Sudarikov, 1959 and Ichthyocotylurus Odening, 1969. Although the lifecycle has been ascertained for most Ichthyocotylurus spp., confusion and disagreement still exist as to the constituent species, while all of the life-stages have been described for only a single member of the subgenus Apatemon (Apatemon). In order to clarify species membership to these taxa and indeed the taxonomic position of the subgenus Apatemon (Apatemon) further information was required on the life-cycles and life-stages of these strigeids. Although, metacercariae from this family have been recorded from a variety of British fishes, confirmed records, i.e. those supported with life-cycle data, are limited to a single species. It was this lack of confidence in identifying metacercariae recovered from fishes and the lack of known good criteria for distinguishing the adults that prompted the present study. Collections of metacercariae from a variety of hosts and locations were made, from which all subsequent life-cycle stages were obtained. The project aims were to establish the identity of the forms occurring in British fishes, by applying discriminatory techniques to the experimentally reared life-stages. In addition to traditional methods, techniques with little previous application to these genera were used and included, scanning electron microscopy (SEM), chaetotaxy, principal components analysis (PCA), and karyology. Furthermore, behavioural aspects such as the release patterns of cercariae from their molluscan hosts were studied to investigate whether they would prove to be of diagnostic value. Metacercariae obtained from the sampling survey were tentatively identified, using all currently employed methods for their determination, i.e. morphology, nature of cyst, host and site specificities, as Ichthyocotylurus erraticus (Rudolphi, 1809), I. variegatus (Creplin, 1825), Apatemon gracilis (Rudolphi, 1819) and A. annuligerum (Nordmann, 1832). Material collected from Finland was considered to contain both Ichthyocotylurus spp. recovered in the U.K., as well as I. platycephalus (Creplin, 1825) and I. pileatus (Rudolphi, 1802). The Ichthyocotylurus spp. were found to be more host specific than A. gracilis, although A. annuligerum was considered oioxenic to perch Perea fluiatilis L. Records of I. erraticus from gwyniad Coregonus lavaretus (L.) and grayling Thymallus thymallus (L.), and A. gracilis from arctic charr Salvelinus alpinus (L.) constitute first listings from Britain. The large number of sensilla present on the body surface of these metacercariae, observed by SEM and chaetotaxy, precluded their diagnostic use. PCA was, however, found to be of value for distinguishing between species and determining morphological variation within a species. I. erraticus, I. variegatus and A. gracilis adults were successfully reared in experimental hosts using metacercariae from a variety of fish hosts, sites within a single fish host and geographical sites. The adults obtained enabled clarification of the identities assigned to the metacercariae. Those metacercariae believed to represent I. pileatus and A. annuligerum failed to establish in experimental hosts. Herring gulls and lesser black-backed gulls proved to be extremely good experimental hosts for both Ichthyocotylurus spp., with the vast majority of infections establishing and providing high yields of eggs and adults. These infections yielded information on the establishment, development, fecundity, site specificity, longevity and morphological variability of the adults. Aspects of the morphology and biology of I. variegatus adults recorded were found to support its validity as a species discrete from I. platycephalus which was in some doubt. The experimental hosts used for A. gracilis infections, domestic and mallard ducklings, were found to be less satisfactory. Challenges were performed with A. gracilis metacercariae from three sources, rainbow trout, salmon parr and stone loach. The latter source was the only one to result in egg producing adults, with specimens exhibiting normal morphology and demonstrating an increased longevity over adults raised from salmonid metacercariae. These findings suggest that the metacercarial host may affect the successful completion of the life-cycle. Eggs of known origin were collected for all three cultured strigeid species, enabling further life-cycle studies, these were incubated and miracidia successfully raised. Developmental periods were found to be temperature dependent and differed for the three species at 20°C: A. gracilis < I. erraticus < I. variegatus. Light microscopy revealed the morphology of all three species to be identical, as were the epidermal plate formulae and chaetotaxy, indicated by silver-staining. The nomenclature for the distribution of miracidial sensilla derived by Dimitrov et al. (1989) was amended to enable a full description of these species. Osmotic shock resulted in an improved deciliation of the miracidia compared to sonication and subsequent SEM observation confirmed the arrangement of body surface structures, while revealing sensilla forms. Behavioural aspects of I. variegatus miracidia were examined, with a maximum longevity (< 11 hours) recorded at the lowest temperature studied (l0 degrees C), and host finding demonstrated to occur by an increased turning response in the presence of substances emitted from the susceptible snail host, following an initial unresponsive dispersal phase. Ichthyocotylurus cercariae were found in naturally infected Valvata piscinalis which constitutes the first record in Britain of cercariae of this genus. Cercariae of I. erraticus and I. variegatus were successfully raised experimentally from miracidia of known identity and origin within naive, experimentally raised V. piscinalis hosts, while A. gracilis cercariae were obtained from laboratory reared Lymnaea peregra. Cercarial developmental periods within the molluscan host were found to be temperature dependent and markedly different for the strigeid genera investigated, as were their behaviour and morphology. The Ichthyocotylurus spp. exhibit a distinct diurnal emergence rhythm from their molluscan host, being shed during the hours of daylight, while A. gracilis cercariae demonstrate a reciprocal pattern, emerging during the hours of darkness. Behavioural contrasts were also observed in longevities, emergence strategies (route of exit) and swimming behaviour. The two Ichthyocotylurus spp. were extremely similar, the only cercarial features found to be of diagnostic use were: the presence or absence of eye-spots; their differing developmental periods from miracidium to cercaria; the number and distribution of sensilla when compared by PCA; and their differing longevities at 20 degrees C. Characters considered to be of value in differentiating between strigeid cercariae at the species level, including the armature, chaetotaxy pattern and resting posture, did not differ between these two species. SEM observations enabled descriptions of the variety and structure of sensilla present on different life-stages, while transmission electron microscopy revealed the internal structure of cercarial sensory structures. Experimentally raised cercariae were found to be infective and the life-cycle was completed for the three strigeid species. Host specificities were observed for I. erraticus and A. gracilis, being particularly stringent for the latter species, while site specificities recorded were as observed in natural infections. Metacercarial maturation periods (for encystment) were highly temperature dependent, being comparable for the two Ichthyocotylurus spp. and more rapid than for A. gracilis specimens.

No description available.

Some parasites may modify the behavior of their hosts. Altered behaviors may: 1) benefit the host in that they defend against the pathogen, 2) benefit the pathogen and represent manipulations of the host response, and 3) benefit neither the host or the pathogen and simply be a product of the host response to infection. In this thesis I examine four host/parasite systems. For each system, I explore host/parasite behavioral interactions, and examine them with regard to selective pressures that may be acting on both the host and the parasite. I test the Hamilton and Zuk hypothese in 26 species of lizards. I find an inverse relationship between a lizard species' brightness and parasite prevalence. My result lend credence to criticisms of the Hamilton and Zuk Hypothesis. If infection does occur, animals may alter their behavior to impair the growth and reproduction of the parasite. To test this prediction, I examine behavioral thermoregulation in two strains of the snail Biomphalaria glabrata, one resistant to, and one susceptible to, the parasite Schistosoma mansoni. The preferred temperature of infected snails drops five weeks after exposure to the parasite. I propose the hypothesis that pathogen-induced host defense responses result in altered host behaviors and enhanced predation. In particular, I examine the effects of the acute phase response (a physiological response whose symptoms include fever, reduced activity and malaise) on antipredatory behavior in bullfrog (Rana catesbeiana) tadpoles. This host response is associated with the preliminary stages of infection with many pathogens yet its behavioral effects have received little attention. I find that the stereotypical effects of the acute phase response can lead to increased predation. I suggest that altered behaviors may afford some parasites a potential pathway to their next host. I examine the behavioral effects of a yeast, Candida spp., a single-host parasite species in its natural host, the red-legged frog (Rana aurora). Infected tadpoles exhibit the same behavioral modifications that are noted in bacteria injected bullfrog tadpoles. These results suggest that some altered behaviors may occur due to a host response to infection and not due to parasitic manipulation. / Graduation date: 1993

The cationic host defence peptide LL-37 is an immunomodulatory agent that plays an important role in epithelial innate immunity. Previously, concentrations of LL-37 thought to represent levels present during inflammation have been shown to elicit the production of cytokines and chemokines by epithelial cells. To investigate the potential of lower concentrations of LL-37 to alter epithelial cell responses, normal primary keratinocytes and bronchial epithelial cells were treated with pro-inflammatory stimuli in the presence or absence of 1 – 3 μg/ml LL-37. Low, physiologically relevant concentrations of LL-37 synergistically increased IL-8 production by both proliferating and differentiated keratinocytes in response to IL-1β and the TLR5 agonist flagellin, and synergistically increased IL-8 production by bronchial epithelial cells in response to IL-1β, flagellin, and the TLR2/1 agonist PAM3CSK4. Treatment of bronchial epithelial cells with LL-37 and the TLR3 agonist poly(I:C) resulted in synergistic increases in IL-8 release and cytotoxicity. The synergistic increase in IL-8 production observed when keratinocytes were co-stimulated with flagellin and LL-37 was suppressed by pretreatment with inhibitors of Src-family kinase signalling and NF-κB translocation. These data suggest that low concentrations of LL-37 may alter epithelial responses to microbes in vivo. Microarray analysis of keratinocyte transcriptional responses after LL-37 treatment suggest that LL-37 may alter the expression of growth factors and a number of genes important to innate immune responses. LL-37 may thus play a more important role than previously suspected in the regulation of epithelial inflammation; an improved understanding of the mechanisms by which LL-37 alters chemokine responses could lead to the development of novel anti-infective and anti-inflammatory therapeutics.

From simple beginnings, when only one host was required, numerous parasitic organisms have evolved complex life-cycles involving two or more host species. For example, trematode parasites reproduce in vertebrates, their definitive host, but their current life cycle also typically involves two intermediate hosts that were added during the course of evolution. Vertebrates are often considered to be the ancestral hosts of trematodes although other scenarios exist. While multi-host life cycles are observed in distantly related groups of parasites, their evolution remains largely unexplored. In trematodes, while recent phylogenetic studies have shed light on the sequence along which the different hosts were incorporated in the cycle, conditions that favoured the evolution of such complex life cycles can only be hypothesized. However, one opportunity to understand the force shaping the evolution of complex life cycles is provided by the few trematode species in which the classical three-host cycle facultatively reverts to a shorter cycle (i.e. life cycle abbreviation). In this study, the effects of different environmental factors on the life history strategy of the trematode Coitocaecum parvum were investigated using laboratory and field studies. C. parvum is able to abbreviate its life cycle from three to two hosts by maturing early (i.e. progenesis) and producing eggs inside the second intermediate host; both life history strategies occur simultaneously in C. parvum populations. Environmental factors such as predator densities should strongly influence parasite life history strategies. In fact, this study shows that laboratory reared Coitocaecum parvum adopt preferentially the normal three-host cycle when chemical cues from the definitive host are added to their environment, while the shorter cycle is favoured when these cues are absent. However, in nature, multiple environmental factors are likely to be perceived by parasites. Consequently, C. parvum�s ability to adapt its developmental strategy to definitive host densities may be confounded by the complex combination of various environmental parameters. Within-host competition between parasites sharing a common host is also likely to influence individual life history strategies. Parasites could then use alternative life strategies to adaptively respond to intraspecific and interspecific competition. Indeed, this study found that C. parvum preferentially adopts the abbreviated cycle in the presence of competitors. However, in interspecific competition, C. parvum�s strategy also depends upon the competitor species, possibly influenced by the other species� transmission route. Furthermore, intensity of intraspecific competition proved to constrain C. parvum�s ability to use the abbreviated life cycle. Finally, genetic relatedness between co-infecting C. parvum individuals seems to affect parasite life strategy through kin selection: closely related individuals are more likely to adopt the same developmental strategy, when they share a host, than unrelated ones. C. parvum individuals adopting the abbreviated cycle are enclosed within a cyst in their intermediate host and must produce eggs by self-fertilization, the most severe case of inbreeding. It was hypothesized that their offspring would have reduced fitness due to inbreeding depression, therefore selecting against the shorter cycle. However, this study found no difference in the survival and infection success of offspring produced through the abbreviated and normal cycles. Furthermore, no evidence for a genetic basis of life cycle abbreviation was detected: the same proportion of offspring from both reproductive strategies adopted the shorter life cycle. The work in this thesis provides evidence that although life cycle abbreviation provides Coitocaecum parvum with a viable alternative life strategy, numerous factors promote or restrict the adoption of this strategy. While this life history strategy has no detectable effect on parasite fitness, both environmental parameters and within-host competition affect C. parvum life-history strategies, alternatively selecting for either the shorter or normal life cycle. Overall, the complexity of the parasite environment could maintain both developmental strategies in C. parvum populations and, on a broader scale, could have influenced the evolution of complex life cycles in parasites.

CD4 is a co-receptor for binding of T cells to antigen-presenting cells (APC) and the primary receptor for human immunodeficiency virus-I. The disulfide bond in the second extracellular domain (D2) of CD4 is reduced on the cell surface, which leads to formation of disulfide-linked homodimers. A large conformational change must take place in D2 to allow for formation of the disulfide-linked dimer. Domain swapping of D2 is the most likely candidate for the conformational change leading to formation of two disulfide-bonds between Cysl30 in one monomer and Cysl59 in the other one (Cys133 and Cysl62 in the mouse CD4). Thus, we hypothesized that the domain swapping of D2 in CD4 regulates its co-receptor function of antigenspecific T cell activation. We found that mild reduction of the extracellular part of human CD4 resulted in formation of disulfide-linked dimers. We then tested the functional significance of dimer formation for co-receptor function using the engineered Jurkat T cell system by expressing wild-type or disulfide-bond mutant mouse CD4. Eliminating the D2 disulfide bond markedly impaired CD4's coreceptor function as assessed by antigen-specific IL-2 production. Exogenous wild type thioredoxin, but not redox-inactive thioredoxin, could inhibit the CD4-mediated IL-2 production, suggesting that the redox state of D2 disulfide bond is controlled by this oxidoreductase. Furthermore, structural modeling of the complex ofthe T cell receptor and domain-swapped CD4 dimer bound to class II major histocompatibility complex and antigen supports the domain-swapped dimer as the immune co-receptor. The known involvement of D4 residues Lys318 and Gln344 in dimer formation isalso accommodated by this model. These findings imply that disulfide-linked dimeric CD4 is the preferred functional co-receptor for binding to APC. Strategies to promote dimerisation of CD4 should, therefore, enhance the immune response, while inhibiting dimer formation is predicted to be immunosuppressive.