Programmed function of the tammar mammary gland; the role of milk proteins in gut development

 This thesis aimed to exploit the unique reproductive strategy of marsupials such as the tammar wallaby to prove that milk may regulate postnatal growth and development of organs such as the stomach.

Comparative analysis of caveolins in mouse and tammar wallaby: role in regulating mammary gland function.

Recent studies using the mouse showed an inverse correlation between the Caveolin 1 gene expression and lactation, and this was regulated by prolactin. However, current study using mammary explants from pregnant mice showed that while insulin (I), cortisol (F) and prolactin (P) resulted in maximum induction of the β-casein gene, FP and IFP resulted in the downregulation of Caveolin 1. Additionally, IF, FP and IFP resulted in the downregulation of Caveolin 2. Immunohistochemistry confirmed localisation of Caveolin 1 specific to myoepithelial cells and adipocytes. Comparative studies with the tammar wallaby showed Caveolin 1 and 2 had 70-80% homology with the mouse proteins. However, in contrast to the mouse, Caveolin 1 and 2 genes showed a significantly increased level of expression in the mammary gland during lactation. The regulation of tammar Caveolin 1 and 2 gene expression was examined in mammary explants from pregnant tammars, and no significant difference was observed either in the absence or in the presence of IFP.

Innate immunity in the mammary gland and the role of the milk peptidome

 This research investigated how the mammary gland responds to disease states such as mastitis and how the milk plays a protective role. An in vitro mammary model was developed and shown to replicate the same responses to the breast when challenged with bacterial proteins demonstrating the usefulness of this model for future investigation of therapeutic interventions.

Altered expression of two zinc transporters, SLC30A5 and SLC30A6, underlies a mammary gland disorder of reduced zinc secretion into milk

Two cases of zinc deficiency in breastfed neonates were investigated where zinc levels in the mothers' milk were reduced by more than 75 % compared to normal. The objective of this study was to find the molecular basis of the maternal zinc deficiency condition. Significant reductions in mRNA expression and protein levels of the zinc transporters SLC30A5 and SLC30A6 were found in maternal tissue, suggesting a causal link to the zinc-deficient milk. Novel splice variants of the SLC30A6 transcript were detected. No modifications were found in coding regions, or in transcription binding sites of promoter regions or in 5' and 3' untranslated regions of both transporters in lymphoblasts and fibroblasts isolated from both mothers. Altered DNA methylation in SLC30A5 at two CpG sites was detected and may account for the reduced levels of SLC30A5 mRNA and protein in lymphoblasts. Reduced SLC30A6 mRNA and protein levels in lymphoblasts may be secondary to reduced SLC30A5 expression, as they function as a heterodimer in zinc transport. In conclusion, two cases of zinc deficiency are linked to low levels of the SLC30A5 and SLC30A6 zinc transporters. These two zinc transporters have not been previously associated with zinc deficiency in milk.

A colostrum trypsin inhibitor gene expressed in the Cape fur seal mammary gland during lactation

The colostrum trypsin inhibitor (CTI) gene and transcript were cloned from the Cape fur seal mammary gland and CTI identified by in silico analysis of the Pacific walrus and polar bear genomes (Order Carnivora), and in marine and terrestrial mammals of the Orders Cetartiodactyla (yak, whales, camel) and Perissodactyla (white rhinoceros). Unexpectedly, Weddell seal CTI was predicted to be a pseudogene. Cape fur seal CTI was expressed in the mammary gland of a pregnant multiparous seal, but not in a seal in its first pregnancy. While bovine CTI is expressed for 24-48h postpartum (pp) and secreted in colostrum only, Cape fur seal CTI was detected for at least 2-3months pp while the mother was suckling its young on-shore. Furthermore, CTI was expressed in the mammary gland of only one of the lactating seals that was foraging at-sea. The expression of β-casein (CSN2) and β-lactoglobulin II (LGB2), but not CTI in the second lactating seal foraging at-sea suggested that CTI may be intermittently expressed during lactation. Cape fur seal and walrus CTI encode putative small, secreted, N-glycosylated proteins with a single Kunitz/bovine pancreatic trypsin inhibitor (BPTI) domain indicative of serine protease inhibition. Mature Cape fur seal CTI shares 92% sequence identity with Pacific walrus CTI, but only 35% identity with BPTI. Structural homology modelling of Cape fur seal CTI and Pacific walrus trypsin based on the model of the second Kunitz domain of human tissue factor pathway inhibitor (TFPI) and porcine trypsin (Protein Data Bank: 1TFX) confirmed that CTI inhibits trypsin in a canonical fashion. Therefore, pinniped CTI may be critical for preventing the proteolytic degradation of immunoglobulins that are passively transferred from mother to young via colostrum and milk.

The relationship between milk, mammary adipocytes and ECM in regulating murine mammary gland involution

 This thesis investigated the role of milk, extracellular matrix and mammary adipocytes in regulating mammary gland function during involution in mice and explored the use of an in vitro culture model, the mammosphere model system to study the same.

Medição de deslocamento através de sensores magnetoresistivos aplicada ao movimento mandibular

Neste trabalho é apresentado um método para medição de deslocamentos sem contato, utilizando sensores magnetoresistivos, os quais são sensibilizados pela variação do campo magnético produzido por um imã permanente em deslocamento no espaço. Os sensores magnetoresistivos possuem, internamente, uma ponte de Wheathestone, onde a resistência varia conforme varia o campo magnético, de modo que o circuito mais indicado para este caso é um amplificador e um filtro para cada sensor. O principal objetivo do trabalho é a obtenção de uma técnica para medir deslocamentos sem contato, e estender os resultados para medida de movimentos mandibulares. A montagem consiste de duas placas de celeron, distantes 30mm uma da outra e unidas por parafusos de polietileno. Em cada uma destas placas foram dispostos quatro sensores, num total de oito, sendo que para cada um deles existe um circuito de amplificação e filtragem do sinal de saída. Sob uma chapa de alumínio foi fixado este equipamento e uma mesa de calibração em 3D, a qual, após a obtenção da matriz de calibração, foi substituída por um emulador de movimento mandibular. Os parâmetros do modelo foram estimados através do método dos mínimos quadrados com auxílio do software Matlab, Release 12. Este software também foi utilizado para o sistema de aquisição de dados durante a realização dos experimentos. A imprecisão dos resultados obtidos na determinação dos deslocamentos, está na ordem de décimos de milímetros. O trabalho apresenta, também, o mapeamento do campo magnético do magneto utilizado nos experimentos através do software FEM2000 – Método de elementos finitos aplicado ao eletromagnetismo.

Estudo comparativo em radiografias panorâmicas do grau de inclinação dos terceiros molares inferiores e da direção de crescimento mandibular segundo os traçados de Altonen, Haavikko, Mattila e de Puricelli

Foi realizado um estudo comparativo entre os resultados das medidas obtidas segundo dois traçados (Altonen, Haavikko e Mattila x Puricelli), efetuadas sobre radiografias panorâmicas, verificando-se a presença ou ausência de compatibilidade entre os mesmos. Avaliou-se também a direção de crescimento mandibular (vertical ou sagital) na amostra composta por pacientes com maloclusão de Classe II esquelética e ainda a existência de uma relação desta direção com o grau de inclinação dos terceiros molares. Desta forma, as seguintes medidas foram comparadas: ângulo β de Altonen, Haavikko e Mattila correspondente ao γ de Puricelli, o qual indicou o relacionamento dos terceiros molares inferiores em relação aos segundos molares; ângulo γA de Altonen, Haavikko e Mattila correspondente ao M2L3 de Puricelli, o qual indicou o ângulo de inclinação do segundo molar com uma linha de base e ângulo α de Altonen, Haavikko e Mattila correspondente ao Pc-Go-B de Puricelli, proporcionando o ângulo mandibular. Baseando-se nos resultados obtidos, apresentados e discutidos no presente trabalho e de acordo com a metodologia estudada, pode-se concluir que: 1) Os traçados diferem entre si, com relação a todas as medidas. Entretanto, apresentam uma correlação positiva para todas as medidas estudadas. 2) Há um crescimento mandibular predominantemente vertical (condilar) segundo o traçado proposto por Puricelli. 3) O grau de inclinação do terceiro molar não se correlaciona com a direção do crescimento mandibular indicada através do ângulo mandibular em ambos os traçados. Desta forma, é imprescindível que ao se compararem os resultados de diferentes estudos analisem-se apenas aqueles que utilizaram o mesmo traçado.

Structural and ultrastructural characteristics of interrenal gland and chromaffin cell of matrinxa, Brycon cephalus Gunther 1869 (Teleostei-Characidae)

This work presents the structure and ultrastructure of the interrenal gland and chromaffin cells, as well as the morphology of the head kidney of Brycon cephalus, the head kidney is composed of fused bilateral lobes located anterior to the swim bladder and ventrolateral to the spinal column, the parenchyma revealed lympho-haematopoietic tissue, melano-macrophage centres, interrenal gland and chromaffin cells. The interrenal gland consisted of cords or strands of cells grouped around the posterior cardinal vein and their branches. Chromaffin cells are found in small groups, closely associated with the interrenal gland and/or under the endothelium of the posterior cardinal vein. So far, the ultrastructural analysis has revealed only one interrenal cell type which contained abundant smooth endoplasmic reticulum and numerous mitochondria with tubulo-vesicular cristae, characteristic of steroid-producing cells. Two types of chromaffin cells were observed. The first type was characterized by the presence of vesicles with round, strongly electron-dense granules, which were eccentrically located, Such cells were interpreted as noradrenaline cells, Meanwhile, cells which contained smaller vesicles and electron-lucent granules, with a small halo separating the granule from the vesicular limiting membrane, were identified as adrenaline cells.

Third eyelid gland protrusion in dogs: an experimental model proposal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of abutment's height and framework alloy on the load distribution of mandibular cantilevered implant-supported prosthesis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of cantilever length and framework alloy on the stress distribution of mandibular-cantilevered implant-supported prostheses

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

TLR4 and CD14 receptors expressed in rat pineal gland trigger NFKB pathway

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)