Made available in DSpace on 2014-06-11T19:30:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-10-04Bitstream added on 2014-06-13T20:40:53Z : No. of bitstreams: 1 ortiz_g_dr_rcla.pdf: 3017128 bytes, checksum: d70cec429499f79c59df0214adaddbed (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A espermateca dos insetos estoca os espermatozóides desde a cópula até a fecundação dos ovos, e consiste de uma bolsa que freqüentemente possui glândulas associadas. Considerações sobre a anatomia, histologia e ultra-estrutura de espermatecas tornam-se muito mais complexas quando são considerados os insetos sociais, que apresentam diferenças no tamanho e na forma entre rainhas e operárias, e ainda, entre as espécies. Apesar desta complexidade, os resultados morfológicos, histoquímicos e ultra-estruturais aqui obtidos mostraram as diferenças entre as espermatecas das espécies: Atta sexdens rubropilosa, Atta laevigata, Atta sexdens sexdens, Atta sexdens piriventris, Atta capiguara, Acromyrmex balzani, Acromyrmex landolti, Acromyrmex landolti balzani, Acromyrmex coronatus e Cyphomyrmex muelleri. As espécies Atta sexdens rubropilosa, Atta laevigata, Acromyrmex coronatus e Cyphomyrmex muelleri apresentaram apenas a glândula espermatecal (porção secretora externa à espermateca). Atta sexdens sexdens, Atta capiguara e Acromyrmex landolti balzani apresentaram tanto a glândula espermatecal como um epitélio secretor interno. Atta sexdens piriventris, Acromyrmex balzani e Acromyrmex landolti, no entanto, apresentaram apenas epitélio secretor interno, sugerindo que nestas espécies a espermateca seja mais independente na manutenção dos espermatozóides. As formigas cortadeiras vêm sendo cada vez mais estudadas do ponto de vista filogenético, tendo como principal marcador o DNA mitocondrial. Estabelecer a filogenia das formigas atines auxiliaria na resposta de algumas questões evolutivas sobre a relação destas com seus fungos. Realizou-se aqui a correlação das informações obtidas através das seqüências de DNA e do estudo morfológico das espermatecas de rainhas Atta sexdens rubropilosa, Atta laevigata, Atta sexdens sexdens, Atta sexdens piriventris... / The insects spermatheca stores the spermatozoa from the copulation until the egg fecundation, and consists of a sac frequently associated with glands. Considerations about anatomy, histology and ultrastructure of spermathecae become much more complex in social insects, because they have queens and workers with different sizes and shapes, as well as among the species. Besides this complexity, the morphological, histochemical and ultrastructural results show the differences between the spermathecae of the following species: Atta sexdens rubropilosa, Atta laevigata, Atta sexdens sexdens, Atta sexdens piriventris, Atta capiguara, Acromyrmex balzani, Acromyrmex landolti, Acromyrmex landolti balzani, Acromyrmex coronatus and Cyphomyrmex muelleri. The species Atta sexdens rubropilosa, Atta laevigata, Acromyrmex coronatus and Cyphomyrmex muelleri presented only the spermathecal gland (external secretory portion). Atta sexdens sexdens, Atta capiguara and Acromyrmex landolti balzani presented both the spermathecal gland and an internal secretory epithelium. Atta sexdens piriventris, Acromyrmex balzani and Acromyrmex landolti, however, presented only the internal secretory epithelium, suggesting that the spermatheca in these species is more independent in spermatozoa maintenance. The leaf-cutter ants are being studied more from a phylogenetic point of view, having as a principal marker mitochondrial DNA. The establishment of the phylogeny of the attines ants would help in the response of some evolutionary questions about their relation with their fungus. The present study correlated the information obtained from the DNA sequences and morphological studies of spermathecae of Atta sexdens rubropilosa, Atta laevigata, Atta sexdens sexdens, Atta sexdens piriventris, Atta capiguara, Acromyrmex balzani, Acromyrmex landolti, Cyphomyrmex muelleri, Apterostigma sp. and Mycocepurus goeldii queens... (Complete abstract click electronic access below)

Abstract not available

The aim of this project was to study the nature and relative proportion of the volatile components in the sternal-gland secretions obtained from a wide range of Australian marsupials.The results obtained were then used to investigate the evolution of semiochemicals in Australian marsupials by using the current phylogenetic tree as a template.The initial part of the study was dedicated to the investigation of some of the techniques available for the sampling and analysis of gland secretions. Individuals from 8 families within the Marsupialia and 1 family from the Monotremata were sampled over an 18 month period.The obtained results were then subjected to multivariate statistical analysis followed by cladistic analysis.In several species the secretion composition was found to be affected by the breeding status of individuals for both genders.Many other factors such as animal-age, hierarchical status, diet,and lifestyle were also observed to affect the secretion composition. Finally, cladistic analysis demonstrated the differences in the levels of divergence at the species, familial and ordinal levels and highlighted secretion components that could be used to differentiate between super families, species and even sexual status of individuals. / Master of Science (Hons)

The pituitary hormone prolactin (Prl) is essential for alveolar morphogenesis and plays a role in breast carcinogenesis, however the mechanism that underlies these actions remains to be defined. Alterations in serum Prl provide the primary endocrine signal regulating developmental events in the mammary gland in sexually mature mammals. Prl production and post-translational phosphorylation by the pituitary is regulated by the neuropeptide Galanin (Gal) in response to hypothalamic signals integrating neuronal and endocrine inputs. Prl exerts its effects on the mammary epithelium in two ways, indirectly by modulation of the systemic hormonal environment, for example the release of progesterone from the corpus luteum, and directly by binding to Prl receptors (Prlr) within the mammary epithelium. Prl binding to Prlr initiates signalling predominantly via activation of the Jak2/Stat5 pathway, leading to altered patterns of gene transcription. One of these target genes is the ets transcription factor Elf5, which is required by the epithelium for alveolar morphogenesis. This thesis aims to further our understanding of the mechanisms by which prolactin exerts its influence on the mammary gland during alveolar morphogenesis and carcinogenesis. Transcript profiling revealed a lactation signature of 35 genes in Prlr+/- mice, Gal-/- mice and mice treated with a Prl mutant (S179D) that mimics phosphorylated Prl. We discovered that the majority of changes in gene expression were produced by prolactin rather than by Gal. The action of Gal was predominantly via modulation of Prl phosphorylation and release, as its effects were very similar to that of S179D. Knockout of Elf5 phenocopied knockout of Prlr, resulting in failure of alveolar morphogenesis and reduced expression of milk and lipid synthesis genes. Forced Elf5 expression at puberty resulted in aberrant differentiation of the terminal end buds and milk protein synthesis during ductal morphogenesis. Re-expression of Elf5 in Prlr-/- mammary epithelial cells completely rescued alveolar morphogenesis. These observations indicate that Elf5 is a master regulator of alveolar morphogenesis downstream of the Prlr. Loss of mammary epithelial Prlr resulted in reduced proliferation of low-grade neoplastic lesions resulting in increased tumour latency in the C3(1)/SV40T model of mammary carcinogenesis. There was no change in the growth rate, proliferation nor the morphology of tumours in Prlr-/-/C3(1)/SV40T transplants, thus Prl acts early in carcinogenesis to drive the proliferation of pre-invasive lesions resulting in faster progression to cancer.

Bovine mastitis is one of the most costly dairy-based diseases worldwide. Streptococcus uberis is a prevalent causative organism of mastitis and resides naturally in the environment of the dairy cow making prevention of the disease difficult. New strategies need to be developed to control this pathogen. However, a fundamental understanding of the complex relationships that exist between the cow, the pathogen and the environment are required in order to advance the development of prevention strategies. Microarray technology was used to evaluate the complex transcriptional changes which occur in the bovine mammary gland following the onset of clinical S. uberis mastitis. A 22,000 bovine cDNA microarray indicated that S. uberis mastitis led to the up-regulation of 1,283 genes and the down-regulation of 1,237 genes by greater than 1.5 fold. Gene ontology analysis demonstrated that S. uberis mastitis was typically associated with the up-regulation of genes that are involved in the immune response and homeostasis and a down-regulation of genes involved in lipid metabolism. Quantitative real-time analyses for a selection of genes associated with the immune response validated the microarray data. Mammary epithelial cell cultures did not show an increase in the expression of any of these immune factors in response to the same S. uberis strain used to induce clinical mastitis. This indicates that the expression of immune-related genes by mammary epithelial cells may be initiated by host factors and not S. uberis. The application of bacteriocins, proteinaceous antimicrobials produced by bacteria which typically inhibit the same or closely-related species to that of the producer organism, has been suggested as one possible approach in the control of mastitis. S. uberis have been previously found to commonly produce bacteriocin-like inhibitory substances (BLIS). The BLIS activities of a set of fifteen S. uberis and S. bovis strains were assessed. The results confirmed the prolific and varied nature of BLIS production by S. uberis and S. bovis and also indicated that these strains may commonly produce more than one inhibitory agent. This survey of BLIS production led to the detection and characterisation of a novel circular bacteriocin, uberolysin, produced by S. uberis strains 233 and 42. The structural gene of uberolysin was subsequently identified in nine (64%) of the fifteen test strains. Multiplex PCR analysis showed that 93% of 158 New Zealand S. uberis isolates contained the structural genes of at least one of the four known S. uberis bacteriocins (uberolysin, nisin U, ubericin A and ubericin 63). However, no apparent direct association was identified between any one of these bacteriocin-related loci and apparent ability to cause mastitis on New Zealand dairy farms. The uberolysin structural gene was detected in 91% of the isolates and this widespread distribution prompted the advancement and evaluation of a potential role for uberolysin in immunomodulation within the bovine mammary gland. Two different preparations of uberolysin were found to have different stimulatory effects on monocytes, neutrophils and epithelial cells. The less highly purified preparation appeared to diminish the production of TNF-α by monocytes in the presence of a bacterial stimulus and to decrease neutrophil phagocytosis. By contrast, the relatively more highly purified preparation of uberolysin itself induced a significant immune response by monocytes. Consistent with this, the purer preparation of uberolysin induced an increase in C3, IL-1β, IL-6, IL-8, the β-defensin LAP, the acute-phase protein MSAA, the calcium-binding protein S100A12 and TLR2 by quantitative real-time analysis. Although currently only two S. uberis bacteriocins (uberolysin and nisin U) have been fully characterised, the present study has shown that this species may be an important source of novel antimicrobials. Furthermore, bacteriocin production by S. uberis may have an immunomodulation role within the mammary gland. A better understanding of the complex immune response initiated at the onset of clinical S. uberis mastitis and of the role that bacteriocins have in S. uberis pathogenesis may lead to development of improved strategies to combat this disease.

Thesis (M.S.)--Baylor University, 2005. / Includes bibliographical references (p. 61-68).

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

This dissertation examines some important physiological and pheromonal factors regulating foraging and colony growth in honey bee colonies. The first study analyzed effects of soybean trypsin inhibitor (SBTI) on the development of hypopharyngeal gland, midgut enzyme activity and survival of the honey bee. In this study newly emerged caged bees were fed pollen diets containing three different concentrations of SBTI. Bees fed 1% SBTI had significantly reduced hypopharyngeal gland protein content. This study indicated that nurse bees fed a pollen diet containing at least 1% SBTI would be poor producers of larval food. In the second study nurse bee biosynthesis of brood food was manipulated using SBTI, and the resulting effects on pollen foraging were measured. Experimental colonies were given equal amounts of SBTI treated and untreated pollen. SBTI treatments had significantly lower hypopharyngeal gland protein content than controls. There was no significant difference in the ratio of pollen to non-pollen foragers and pollen load weights collected between the treatments. These results supported the pollen foraging effort predictions generated from the direct independent effects hypothesis. In the third study we tested whether brood pheromone (BP) regulated queen egg laying via modulation of worker-queen interactions and nurse bee rearing behaviors. This experiment had BP and control treatments. Queens in the BP treatment laid greater number of eggs, were fed for a greater amount of time and were less idle. Significantly more time was spent in cell cleaning by the bees in BP treatments. The results suggest that brood pheromone regulated queen egg-laying rate by modulating worker-queen interactions and nurse bee rearing behavior. The final study of this dissertation focused on how dose-dependent BP-mediated division of labor affected the partitioning of non-foraging and foraging work forces and the amount of brood reared. Triple cohort colonies were used and there were three treatments, Low BP, High BP and Control. Low BP treatments had significantly higher ratio of pollen to non-pollen foragers and greater pollen load weights. Low BP treatment bees foraged at a significantly younger age. This study has shown that BP elicits dose-dependent modulation of foraging and brood rearing behaviors.

Salt glands are important structural adaptations in some plant and animal species that are involved in the excretion of excess salts. Zoysia matrella is a highly salt tolerant turf grass that has salt glands. Two cultivars of Z. matrella, ‘Diamond’ and ‘Cavalier’, were examined in this study to look for salt gland related factors responsible for the differences in their degree of salt tolerance. In addition to the adaxial salt gland density being higher in ‘Diamond’, the salt glands in salt treated (300 mM NaCl) plants of this cultivar were bigger than the ones in ‘Cavalier’. ‘Diamond’, as well as some of the ‘Diamond’ x ‘Cavalier’ hybrid lines, showed a significant induction in salt gland density in response to salt treatment. Examination of salt gland density in ‘Diamond’ x ‘Cavalier’ hybrid lines showed that salt gland density was a highly heritable trait in the salt-treated population. Ultrastructural modifications in the salt glands observed with Transmission Electron Microscopy (TEM), coupled with Cl- localization studies, suggested a preference for symplastic transport of saline ions in Z. matrella. Salt glands have been studied in several plant species; however, no studies have tried to associate the role of ion transporters with the functioning of salt glands in plants. RNA in situ studies with Na+ transporters showed localization of ZmatHKT1 transcripts in the adaxial salt glands, leaf mesophyll and bundle sheath cells for both cultivars. ZmatSOS1 expression was observed in the xylem parenchyma cells for leaves from both cultivars, but the expression was markedly different around the cells bordering the vascular tissue. The strongest expression of ZmatSOS1 for ‘Diamond’ was seen in the bundle sheath cells and the phloem, while for ‘Cavalier’ the signal was strongest in the mestome sheath cells and in cells surrounding the phloem. No expression of ZmatSOS1 was seen in the salt glands for either cultivars. ZmatNHX1 expression in both cultivars was very low, and observed in the salt glands and neighboring epidermal cells. Three alleles of ZmatNHX1 were identified in Z. matrella, along with three alternatively-spliced forms of ZmatNHX1, the expression of which were cultivar and treatment specific. Together, these results provide a model for salt transport in Z. matrella and signify potential roles of salt glands and select ion transporters in the salt tolerance of this species.

Endometrial glands are required for adult uterine function and develop post-natally in mammalian species. Therefore, studies were conducted using neonatal ewes as a model to determine: 1) the roles of estradiol-17-alpha and estrogen receptor-alpha (ER-beta) in endometrial gland development; 2) the role of ovaries in endometrial gland development; 3) the role of prolactin in endometrial gland development; and 4) factors regulating prolactin receptor expression in endometrial glands. Study one determined the effects of neonatal exposure of ewes to estradiol-17-alpha valerate (EV); EM-800, an ER-beta antagonist; or CGS-20267, an aromatase inhibitor on endometrial gland development. Results indicate E2-17-alpha does not regulate endometrial gland differentiation or development. Additionally, ER-beta does not regulate primary differentiation of glandular epithelium, but does influence coiling and branching morphogenesis of endometrial glands. Study two determined the effects of ovariectomy on endometrial gland morphogenesis. Results suggest that the ovary and, thus, an ovarian-derived factor(s) regulate, in part, the coiling and branching of endometrial glands. Expression of subunits of activin, follistatin, and inhibin in the neonatal ovine ovary in addition to modulation of the components of the activin/follistatin system in the uterus of ovariectomized ewes supports the hypothesis that the ovarian factors that influence endometrial adenogenesis in the neonatal ewe may be activin, follistatin, and/or inhibin. Studies three and four determined the role of prolactin in endometrial adenogenesis in the neonatal ewe. Studies in which either hypoprolactinemia or hyperprolactinemia were induced indicate that prolactin regulates ovine endometrial adenogenesis in the neonatal ewe. The aim of study five was to determine transcription factors that regulate the glandular epithelium specific expression of prolactin receptor. Prolactin receptor exon 2 was cloned and sequenced, but no identifiable exon 1 or promoter was found. Additionally, many bovine contigs containing portions of the prolactin receptor gene were identified suggesting the bovine genome will be a useful tool as it becomes more complete. These results indicate ER-beta, prolactin and prolactin receptor, along with an unidentified ovarian factor(s), influence endometrial gland development in the neonatal ewe; however, exposure of the neonatal ewe to exogenous estradiol-17-alpha prevents differentiation and development of endometrial glands.