[Truncated abstract] This thesis examines the potential role of dietary intake in the development of two common conditions affecting the prostate gland; prostate cancer and benign prostatic hyperplasia (BPH). Diet is of interest as a potential risk factor for prostate cancer because of geographical variations in prostate cancer incidence and increased prostate cancer risks associated with migration from Asian to western countries. Some geographical variation has been suggested for BPH, but this is less certain. However, both prostate cancer and BPH have potential links with diet through their positive associations with sex hormone levels, metabolic syndrome, increased insulin levels and chronic inflammation. In addition, zinc is an essential dietary micronutrient required for semen production in the prostate gland. The original work for this thesis is presented in six manuscripts of which, four have been published in peer-reviewed journals (at the time of thesis completion). BPH investigated in this thesis is defined as surgically-treated BPH. The following hypotheses were investigated. Regarding foods, nutrients and the risk of prostate cancer and BPH: 1. Increasing intakes of fruits, vegetables and zinc are inversely associated with the risk of prostate cancer and BPH 2. Increasing intakes of total fat and calcium are positively associated with the risk of prostate cancer and BPH. 3. Dietary patterns characterised by high meat, processed meat, calcium and fat content are positively associated with the risk of prostate cancer and BPH. 4. Dietary patterns characterised by high fruit and vegetable and low meat content are inversely associated with the risk of prostate cancer and BPH. v Regarding methodological issues related to the study of diet-disease relationships: 5. Dietary patterns (overall diet) elicited from principal components analysis yield stronger diet-disease associations than when studying isolated nutrients. 6. Remotely recalled dietary intake is reliable enough to be used in studies of chronic disease with long latency periods, such as prostate cancer and BPH. Methods: Data from two studies was used to address the hypotheses above. ... Based on the literature reviewed and the original work for this thesis, the most important dietary risk factors for prostate cancer and BPH appear to be those common to western style diets, i.e. diets high in red meat, processed meat, refined grains, dairy products, and low in fruit and vegetables. This type of diet is likely to result in marginal intakes of antioxidants and fibre, excess intakes of fat and possibly, moderate intakes of carcinogens associated with processed meat and meat cooked at high temperatures. These dietary factors have been linked with biomarkers of inflammation, and they support the hypotheses that chronic inflammation is involved in the development of both prostate cancer and BPH. In addition, this work builds on evidence that zinc is an important factor in prostate health. There is scope for more investigation into the reliability of dietary patterns and the use of nutrient patterns as an alternative to focussing on single food components. Further studies on the reliability of remote dietary intake would also be useful. Because of the latency of chronic disease, it can be theorised that remote dietary recall may uncover more robust diet-disease relationships.

Androgens are critical for the development and maintenance of adult male characteristics such as muscle mass and sexual function. Consequently, the established decline with age of serum testosterone (T) in males has major health implications. While the androgen receptor (AR) is the major mediator of genomic androgen action and is required for the development of the male phenotype, reproductive organs and the maintenance of male secondary sexual characteristics, it is the entrance of androgens into the cell that mediates the activation of the AR and the subsequent modulation of expression of androgen regulated genes. Testosterone, biologically the most important androgen in male serum, circulates either free, loosely bound to albumin or tightly bound to sex hormone binding globulin (SHBG). Each of these forms of serum T have different abilities to enter cells, and which proportion of serum T is capable of entering cells and initiating the androgen signalling cascade, thereby leading to the activation of the AR has not been precisely defined. The AR amino terminal domain (NTD) is responsible for the majority of the ability of the AR to activate genes but the relative roles of the two activation functions in the AR NTD (activation functions 1 and 5; AF1 and 5) have not been precisely defined while the role of the AF2 surface which forms in the ligand binding domain upon agonist binding is responsible for interactions with key coregulators and also with the NTD in the amino-carboxyl (N/C) interaction. Our laboratory has recently identified a region within AF5 between amino acids 500-535 to which somatic mutations in castrate resistant prostate tumour samples collocate. Due to the lack of functional information on the AF5 region and the NTD in general, the function of this region and the functional consequences of the mutations remain to be defined. The objectives of this thesis were to develop a specific mammalian cell based bioassay capable of reliable measuring T in serum and to determine the ability of this bioassay to measure a physiologically relevant fraction of T in serum. Additionally, this thesis aimed to determine the relative contributions and roles of the activation functions of the AR to overall AR transcriptional activity along with the functional consequences for AR signalling of prostate cancer mutations which have previously been identified in the AF5 region of the AR NTD. The mammalian-cell based bioassay developed in this thesis is capable of sensitively and reliably measuring serum T. However, evaluation of this bioassay utilising approximately 1000 serum samples from the Florey Adelaide Male Aging Study reveals that this bioassay measures a fraction of T in serum that most closely relates to serum T. Furthermore, this measure does not correlate more strongly with grip strength, sexual function or waist circumference than the existing immunoassay-based measures of serum T, highlighting the limitations of utilising a static mammalian cell-based androgen bioassay to measure physiological levels of serum T in males. The investigation of the roles of the activation functions in the AR in this thesis have revealed that while the AF1 domain is responsible for the majority of the transactivation activity of the AR, AF5 and AF2 govern the sensitivity and cellular response of the AR to androgens by providing protein and interdomain interaction interfaces. Furthermore, the evidence in this thesis demonstrates that the AR requires interdomain communication for sensitive AR signalling. Finally, the findings in this thesis demonstrate that the AF5 surface is required for the N/C interaction and coregulator interactions while advanced prostate cancer mutations identified within this region confer increased transactivation activity of the AR in the presence of high cellular levels of coregulators. Collectively, the findings in this thesis provide several novel insights into the mechanism of action of serum androgens and challenges several long held assumptions of androgenic action in males. These findings also delineate a mechanism of treatment failure in advanced prostate cancer, provide a novel model for the events leading to sensitive AR transactivation and contribute to the understanding of physiologically relevant levels of serum T. / Thesis (Ph.D.) -- University of Adelaide, School of Medicine, 2008

Objectifs : Rapporter le développement et les expérimentations d'un nouveau système robotisé destiné à la curiethérapie prostatique possédant un système de suivi de la prostate et une possibilité de fusion écho-IRM. Matériel et méthodes : Un robot d'implantation d'aiguilles transpérinéales guidé par échographie transrectale avec suivi per-opératoire des mouvements et de la déformation de la prostate a été créé. Les expériences ont été conduites sur 90 cibles réalisées dans 9 fantômes conçus pour être mobiles et déformables. Les expériences ont été ensuite conduites chez 2 cadavres. Le robot a cherché à déposer des billes de verre simulant des grains de curiethérapie aussi près que possible des cibles dans des fantômes évaluables par différentes modalités d'imagerie dont le scanner et dans des prostates de cadavre. Les résultats étaient mesurés en segmentant les cibles et les billes de verre sur des volumes tomodensitométriques des fantômes et des cadavres. Résultats : Le robot était capable d'atteindre les cibles choisies dans les fantômes avec une précision médiane de 2.73 mm, avec un déplacement médian de la prostate de 5.46 mm. La précision était meilleure à la base qu'à l'apex (2.28 mm vs 3.83 mm, p<0.01) et n'était pas significativement différente pour les implantations horizontales et obliques (2.7 vs 2.82 mm, p=0.18). Les tests sur cadavre ont montré la faisabilité et l'ergonomie du robot en salle d'opération mais des expérimentations plus poussées sont nécessaires. Conclusion : Ce robot destiné à la curiethérapie prostatique est le premier système utilisant le suivi de la prostate per-opératoire pour guider des aiguilles dans la prostate. Les expériences préliminaires montrent sa capacité à atteindre des cibles malgré les mouvements de la prostate. Les applications pourraient être élargies à la thérapie focale et aux biopsies guidées compte-tenu de sa possibilité à fusionner l'imagerie IRM et l'échographie.

Prostate cancer is the most frequent type of cancer in men in Europe and the USA. The methods presently used to detect and diagnose prostate cancer are inexact, and new techniques are needed. Prostate tumours can be regarded as harder than the surrounding normal healthy glandular tissue, and therefore it is of interest to be able to reliably measure prostate tissue stiffness. In this dissertation the approach was to evaluate tactile resonance sensor technology and its ability to measure mechanical properties and to detect cancer in human prostate tissue. The tactile resonance sensor is based on a piezoelectric transducer element vibrating at its resonance frequency through a feedback circuit. A change in the resonance frequency is observed when the sensor contacts an object. This feature has been utilized to measure tissue stiffness variations due to various pathophysiological conditions. An impression-controlled tactile resonance sensor system was first used to quantify stiffness and evaluate performance on silicone. Then the sensor system was used on fresh human prostate tissue in vitro to measure stiffness using a combination of frequency change and force measurements. Significant differences in measured stiffness between malignant and healthy normal tissue were found, but there were large variations within the groups. Some of the variability was explained by prostate tissue histology using a tissue stiffness model. The tissue content was quantified at four depths in the tissue specimens with a microscope-image-based morphometrical method involving a circular grid. Numerical weights were assigned to the tissue data from the four depths, and the weighted tissue proportions were related to the measured stiffness through a linear model which was solved with a least-squares method. An increase in the proportion of prostate stones, stroma, or cancer in relation to healthy glandular tissue increased the measured stiffness. Stroma and cancer had the greatest effect and accounted for 90 % of the measured stiffness (45% and 45%, respectively). The deeper the sensor was pressed, the greater, i.e., deeper, volume it sensed. A sensing depth was extrapolated from the numerical weights for the measurements performed at different impression depths. Horizontal surface tissue variations were studied by altering the circular grid size relative to the contact area between the sensor tip and the tissue. The results indicated that the sensing area was greater than the contact area. The sensor registered spatial tissue variations. Tissue density-related variations, as measured by the frequency change, were weakly significant or non-significant. The measured force registered elastic-related tissue variations, to which stroma and cancer were the most important variables. A theoretical material-dependent linear relation was found between frequency change and force from theoretical models of frequency change and force. Tactile resonance sensor measurements on prostate tissue verified this at small impression depths. From this model, a physical interpretation was given to the parameters used to describe stiffness. These results indicate that tactile resonance sensor technology is promising for assessing soft tissue mechanical properties and especially for prostate tissue stiffness measurement with the goal of detecting prostate cancer. However, further studies and development of the sensor design must be performed to determine the full potential of the method and its diagnostic power. Preferably, measurements of tissue mechanical properties should be used in combination with other methods, such as optical methods, to increase the diagnostic power.

The objectives were to 1) conduct a meta-analysis to assess the association between alpha-linolenic acid (ALA) and prostate cancer; 2) analyze a trial of ALA on coronary heart disease with PSA as a post hoc outcome; 3) assess the effect of trial serum and also ALA directly on LNCaP cell growth. 1) The ALA meta-analysis of prospective and case-control studies showed no overall effect on prostate cancer. However, removal of one study from the analysis of prospective studies changed the result to a significant protective effect (RR=0.91; 95%CI:0.83,0.99). 2) No significant treatment difference was seen in the change in PSA in the randomized controlled trial. 3) The ALA treatment serum from the clinical trial did not affect LNCaP cell growth. However, ALA decreased LNCaP cell growth in a dose dependent manner when added to cell culture. The results provide no positive evidence for an effect of ALA on prostate cancer.

The objectives were to 1) conduct a meta-analysis to assess the association between alpha-linolenic acid (ALA) and prostate cancer; 2) analyze a trial of ALA on coronary heart disease with PSA as a post hoc outcome; 3) assess the effect of trial serum and also ALA directly on LNCaP cell growth. 1) The ALA meta-analysis of prospective and case-control studies showed no overall effect on prostate cancer. However, removal of one study from the analysis of prospective studies changed the result to a significant protective effect (RR=0.91; 95%CI:0.83,0.99). 2) No significant treatment difference was seen in the change in PSA in the randomized controlled trial. 3) The ALA treatment serum from the clinical trial did not affect LNCaP cell growth. However, ALA decreased LNCaP cell growth in a dose dependent manner when added to cell culture. The results provide no positive evidence for an effect of ALA on prostate cancer.

Ο καρκίνος του προστάτη αποτελεί σημαντικό πρόβλημα της δημόσιας υγείας ιδιαίτερα στο δυτικό κόσμο. Είναι ο πιο συχνός σπλαχνικός καρκίνος και ο δεύτερος πιο θανατηφόρος μετά τον καρκίνο του πνεύμονα. Η εισαγωγή και χρήση του PSA από τα τέλη της δεκαετίας του ’80 προσέθεσε ένα ισχυρό διαγνωστικό εργαλείο στα χέρια των κλινικών, το οποίο αναχαίτισε τη μέχρι τότε αυξητική τάση του αριθμού των θανάτων οφειλομένων στον καρκίνο του προστάτη. Το PSA ωστόσο χαρακτηρίζεται από χαμηλή ειδικότητα αλλά και ευαισθησία, χαρακτηριστικά που επιβάλλουν περιορισμούς στη χρήση του. Εξάλλου δεν προβλέπει το βαθμό της επιθετικότητας ενός αδενοκαρκινώματος του προστάτη, συμβάλοντας στο φαινόμενο της υπερδιάγνωσης αλλά κυρίως της υπερθεραπείας του καρκίνου του προστάτη. Κατά συνέπεια είναι απαραίτητη η ανάπτυξη μοριακών εργαλείων (δεικτών) οι οποίοι θα διαγιγνώσκουν τη νόσο με μεγαλύτερη ασφάλεια αλλά κυρίως θα μας δίνουν προγνωστικές πληροφορίες για τη βιολογική συμπεριφορά του όγκου, προλαμβάνοντας μια άσκοπη θεραπευτική παρέμβαση. Ενδιαφέρον πεδίο αναζήτησης τέτοιων δεικτών αποτελεί η χρωμοσωμική περιοχή 8q24. Ο μικροδορυφόρος DG8S737 εδράζεται στην περιοχή 8q24 και έχει δειχθεί, σε διαφορετικούς πληθυσμούς, ότι συγκεκριμένο αλληλόμορφό του σχετίζεται με επιθετικό, κλινικά σημαντικό καρκίνο του προστάτη. Στη συγκεκριμένη μελέτη ανιχνεύονται οι συχνότητας των αλληλομόρφων του μικροδορυφόρου DG8S737 σε μια ομάδα γενικού πληθυσμού (control) και σε μια ομάδα ασθενών οι οποίοι πάσχουν από επιθετικό καρκίνο του προστάτη. Τα αποτελέσματα επιβεβαιώνουν την τάση του αλληλομόρφου -8 να ανιχνεύεται συχνότερα σε ασθενείς παρά σε υγιείς. Εξάλλου παρατηρήθηκε για πρώτη φορά η ίδια τάση και για το αλληλόμορφο -10. Αυτά τα αποτελέσματα ενισχύουν το δυναμικό χρήσης του DG8S737 ως δείκτη για το μέτρο της επιθετικότητας του καρκίνου του προστάτη. / Prostate cancer represents a major public health issue in western world. It is the most frequently diagnosed visceral cancer whereas it is second in mortality. The use of PSA since the late 80s restrained the rising tendency of mortality of prostate cancer. PSA though, lacks in specificity. Besides it contributes to the phenomenon of overdiagnosis which leads to overtreatment of prostate cancer. Consequently it is necessary for novel biomarkers to emerge in order to diagnose more accurately and predict the aggressiveness of prostate cancer. The 8q24 region of chromosome 8 is a region which could harbor potential biomarkers for prostate cancer. The microsatellite DG8S737 in that region has a number of alleles, one of which has the tendency to be more often detected in patients with aggressive prostate cancer. We have studied the frequencies of alleles of DG8S737 in a group of patients with aggressive prostate cancer as well as in a group of control volunteers. The results confirm the findings of previous studies. The allele -8 of DG8S737 has been detected more often in patients than in healthy volunteers. A new finding is that allele -10 also is more frequently detected in the patients group rather than the control group. The results confirm previous findings and enforce the potential use of DG8S737 as novel biomarker for the aggressive prostate cancer.

Actuellement, les injections systémiques de médicaments atteignent faiblement les sites tumoraux et de fortes doses sont alors administrées provoquant des effets secondaires parfois lourds. Les possibilités offertes par les applications en médecine des nanoparticules permettent de nouvelles stratégies pour vectoriser des substances actives dans les cellules malades. Ces travaux de thèse portent sur le cancer de la prostate qui est le deuxième cancer le plus diagnostiqué et la cinquième cause de décès chez les hommes dans le monde.Les nanotubes de titanate (TiONts) sont synthétisés par voie hydrothermale et présentent une longueur moyenne de 170 nm, un diamètre extérieur de 10 nm et une cavité interne accessible de 4 nm. Leur morphologie tubulaire permet aux TiONts d’être internalisés plus facilement dans les cellules, sans induire de cytotoxicité, tout en créant un effet radiosensibilisant.Deux nanohybrides ont été développés dans cette thèse, pour lutter contre le cancer de la prostate par injection intratumorale (IT) et une attention particulière a été portée sur leur élaboration. Ces nouveaux nanomédicaments ont été pleinement caractérisés par différentes techniques (MET, ATG, potentiel zêta, XPS, spectroscopies UV visible, IR et Raman).La première approche consiste à combiner les TiONts avec un agent thérapeutique (le docétaxel, DTX), largement utilisé pour inhiber les tumeurs de prostate et un agent chélatant (le DOTA, radiomarqué avec l’111In) pour suivre la biodistribution des tubes par SPECT/CT. La surface des TiONts a été préalablement fonctionnalisée par l’APTES et le poly(éthylène) glycol (PEG3000) pour rendre les TiONts stables et biocompatibles. Afin d’évaluer l’efficacité de ce nanohybride, des tests in vitro ont montré que l’association entre les TiONts et le DTX permettait de maintenir une activité cytotoxique sur des lignées cellulaires de prostate (cellules 22Rv1 et PC 3) alors que les TiONts sans le DTX n’étaient pas toxiques. Les études in vivo ont montré, sur des souris Swiss nude mâles, que plus de 70% des nanovecteurs étaient retenus dans la tumeur, après injection IT, après 7 jours. De plus, un retard de croissance tumorale pour les souris ayant reçu le nanohybride avec la radiothérapie (RT) est observé, par rapport aux souris ayant reçues seulement le DTX. Après cette étude, d’autres molécules organiques ont été greffées avec succès à la surface des TiONts pour améliorer la stabilité colloïdale et la biocompatibilité des nanotubes : AHAMTES, catéchols (LDOPA, DHCA et NDOPA), phosphonates (PHA, ALD et un polymère hétérobifonctionnel de type phosphonate : (HO)2 (O)P-PEG NH2). De plus, le greffage de différentes longueurs de chaîne de PEG a été évalué par deux voies de synthèses. Le greffage de ces PEG en milieu organique (PyBOP) s’est avéré très prometteur pour améliorer leur taux de greffage et leur stabilité colloïdale.Dans une seconde approche, pour accroître l’effet radiosensibilisant, des nanoparticules d’or (AuNPs), elles-mêmes modifiées par le DTDTPA, ont été couplées avec les TiONts en présence ou non de DTX. Cette nouvelle combinaison a pour objectif le maintien des AuNPs, par les TiONts, dans la tumeur afin d’améliorer l’effet de la RT. Grâce aux AuNPs modifiées par le DTDTPA, le nanohybride est également détectable par imagerie X et par SPECT/CT. Les résultats in vitro ont démontré l’activité cytotoxique de l’édifice final. Des études de biodistribution et de croissance tumorale ont également été réalisées sur des tumeurs PC-3 xénogreffées sur des souris.Ces TiONts fonctionnalisés apparaissent comme un nouvel outil polyvalent dans le domaine médical, notamment pour lutter contre le cancer de la prostate. / Currently, the systemic injections of drugs reach very weakly tumor sites and large doses are thus administered causing adverse side effects. The new implementations of nanoparticles in the medical field offer new strategies to vectorize an active substance in diseased cells. This work is focused on the prostate cancer, which is the second most frequently diagnosed cancer and the fifth leading cause of cancer death in men worldwide.Titanate nanotubes (TiONts) are synthetized by a hydrothermal process and have average dimensions of about 170 nm in length, 10 nm in outer diameter and also have an internal cavity of 4 nm in diameter. Their needle-shaped morphology allows them to be internalized more easily into cells without inducing cytotoxicity while providing a radiosensitization effect.In the present manuscript are described two TiONts-based nanohybrids which were developed with a view to fight against prostate cancer by intratumoral (IT) injection and a particular attention was paid on their elaboration. These new nanomedicines were extensively characterized by different techniques (TEM, TGA, ζ potential, XPS, UV visible, IR and Raman spectroscopies).The first approach that has been developed consists in combining TiONts with a therapeutic agent (docetaxel, DTX), widely used for the treatment of prostate cancer, and a chelating agent (DOTA) allowing the radiolabeling with 111In radionuclide to monitor TiONts biodistribution by SPECT/CT. The surface of TiONts was beforehand coated with a siloxane (APTES) and linked to a heterobifunctional polymer (PEG3000) to create well-dispersed and biocompatible TiONts. In vitro tests demonstrated that the association between TiONts and DTX had cytotoxic activity against prostate cancer cell lines (22Rv1 and PC-3 cells) whereas TiONts without DTX did not. The results of in vivo SPECT/CT imaging are also presented as well as first irradiation tests in Swiss nude mice after IT injection on PC-3 tumors. Biological tests showed that more than 70% of TiONts nanovectors were retained within the tumor for at least 7 days. In addition, tumor growth of mice receiving nanohybrids with radiotherapy was significantly slower than that of mice receiving free DTX. After this first study, other organic molecules were successfully grafted to the surface of TiONts to improve colloidal stability and biocompatibility of nanotubes: AHAMTES, catechols (LDOPA, DHCA and NDOPA) and phosphonates (PHA, ALD and a phosphonate heterobifunctional polymer-based: (HO)2 (O)P PEG NH2). Moreover, the influence of different PEG lengths has been considered on the nanomedicine efficacy by two different pathways. The grafting of these PEG in an organic medium (PyBOP) was very promising to improve their graft ratio and their colloidal stability.In a second approach and in order to improve the radiosensitizing effect, DTDTPA-modified gold nanoparticles (AuNPs) were coupled with TiONts in the presence of DTX. This novel combination aims at retaining these AuNPs into the tumor via the TiONts to enhance the radiotherapeutic effect. The nanohybrid was also detectable by X-ray and SPECT/CT imaging through AuNPs-DTDTPA. Preliminary in vitro results showed once again that our final nanohybrid had a satisfactory cytotoxic activity. Biodistribution and tumor growth studies were also realized on PC-3 xenografted tumors on mice.These functionalized-TiONts could thus become a new tool in the field of biomedicine to fight against prostate cancer and appear as versatile nanovectors.

Le cancer de la prostate est le cancer le plus fréquent chez l'homme en Europe, néanmoins il n'existe actuellement pas de technique d'imagerie permettant de détecter avec précision les tumeurs dans la glande. Sachant que les coupes histologiques contiennent la réalité de terrain concernant le diagnostic, il est nécessaire de recaler les images de chaque technique d'imagerie aux coupes histologiques afin de pouvoir les évaluer. De plus, comme il n'existe pas de méthode permettant de contrôler précisément le plan de coupe histologique, le recalage doit être considéré comme un problème 3D. Un dispositif permettant de réaliser, de manière rapide et standardisée, des marqueurs internes dans les coupes histologiques a été développé, de même qu'un algorithme permettant de détecter automatiquement ces marqueurs, de les identifier et d'aligner les coupes histologiques. La méthode a été testée sur 10 prostates, avec en moyenne 19.2 coupes par prostate, et a permis d'obtenir une précision de recalage moyenne de 0.18 ± 0.13 mm au niveau des marqueurs. Un deuxième algorithme a été développé pour recaler les coupes histologiques, une fois alignées, avec les images IRM. Ce recalage a été conçu pour être guidé par les canaux éjaculateurs, un repère anatomique présent dans chaque prostate et visible à la fois en histologie et dans les images IRM cliniques, acquises avec une résolution standard. L'algorithme a d'abord été testé en s'appuyant sur les marqueurs artificiels. La précision obtenue pour le recalage était en moyenne de 0.45±0.25 mm au niveau des marqueurs et de 1.04 ± 0.21 mm au niveau des canaux éjaculateurs. L'algorithme a enfin été testé en guidant le recalage à l'aide de la position des canaux éjaculateurs. La précision moyenne obtenue était alors de 0.16±0.05 mm au niveau des canaux éjaculateurs et de 2.82±0.41 mm au niveau des marqueurs. Ces résultats suggèrent une valeur du facteur de rétrécissement de l'ordre de 1.07±0.03 et une inclinaison vis à vis du plan de coupe histologique de l'ordre de 13.6◦ ± 9.61◦, avec une variance importante pour ces deux paramètres / Prostate cancer is the most frequently diagnosed cancer of men in Europe, yet no current imaging technique is capable of detecting with precision tumours in the prostate. The histology slices are the gold standard for the diagnosis. Therefore, in order to evaluate each imaging technique, the histology slices must be precisely registered to the imaged data. As it cannot be assumed that the histology slices are cut along the same plane as the imaged data is acquired, the registration must be considered as a 3D problem. An apparatus has been developed that enables internal fiducial markers to be created in the histology slices in a rapid and standardised manner. An algorithm has been developed that automatically detects and identifies these markers, enabling the alignment of the histology slices. The method has been tested on 10 prostate specimens, with 19.2 slices on average per specimen. The accuracy of the alignment at the fiducial markers was on average 0.18±0.13 mm. A second algorithm was developed to 3D register the aligned histology slices with the MR images. The registration is designed to be guided by the ejaculatory ducts, an anatomical landmark present in every prostate and visible in both histology and MR images acquired at standard clinical resolution. The algorithm was first tested by using the fiducial needles to guide the registration. The average registration accuracy was 0.45 ± 0.25 mm at the fiducial needles and 1.04±0.21 mm at the ejaculatory ducts. The algorithm was then tested by using the ejaculatory ducts to guide the registration. The average registration accuracy was 0.16±0.05 mm at the ejaculatory ducts and 2.82 ± 0.41 mm at the fiducial needles. The results suggest that the histology shrinkage factor is of the order 1.07±0.03 and the tilt of the histology slicing plane is 13.6◦ ±9.61◦, with both parameters showing significant variance

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No. of bitstreams: 2 Dissertação - Rodrigo Fernandes de Lima - 2015.pdf: 2077886 bytes, checksum: 41cc0e061b83ff79f5098eb054d1fcda (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-03-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The neonatal development is stimulated by various hormonal interactions. Environmental pollutants that mimic steroid hormones, such as bisphenol A (BPA) can cause changes in the pattern of development of the prostate, leading to lifelong lesions. The objective of this study was to determine whether neonatal exposure to BPA affects the development of prostate gerbils males and females. For this, the newborns were exposed to environmental levels (LBPA - 40 μ / kg / day) and high dose (HBPA - 4 mg / kg / day) of BPA, from 1st to the 7th day of life. On the eighth day, the prostatic complexes were collected and processed for morphological analysis, stereological and immunohistochemistry. It was observed through the three-dimensional reconstruction that in males the prostatic buds have elongated in a "V" form towards the ventral condensed mesenchyme (VMP). The stereological data of males showed a significant increase in the relative frequency of the mesenchymal compartment in the LBPA group compared to other groups (p ≤ 0.05). The AR-positive cells became significantly more frequent in the periurethral mesenchyme (PEM), ventral epithelial buds (VB) and muscle layer (SM) in the BPA group. There was a reduction in the frequency of PCNA-positive cells in the periurethral buds (PeB) and in the SM of the LBPA group. In females, the three-dimensional reconstruction showed that PeBs can emerge from one or both sides of the urethra and it was also visible a significant paraurethral mesenchyme (MAP) reduction in the BPA group. Only females of the HBPA group apparently presented paraurethral buds (PAB) more developed. In the LBPA group it was observed an increase in the AR-positive cells frequency in the PeM and a decrease of these cells in the PeB and mesenchyme paraurethral (PaM). In the HBPA group the AR-positive cells were more frequent in the PeM and PaB and reduced in PeB and PaM. PCNA-positive cells became significantly less frequent in the PaB and PaM of LBPA and HBPA groups. Regarding the ER-α positive cells in the group LBPA females showed a reduction in the immunoblots of PeM and MS, whereas in the group HBPA this reduction occurred only in the PaM. This study revealed that the postnatal development of the prostate of female gerbils occurs earlier and are morphologically distinct from what occurs in males of the same species. Furthermore, it can be seen that the BPA exerted a proliferative effect on the prostate gland of males and females with eight days of postnatal life, though females being more susceptible to this environmental chemicals. / O desenvolvimento neonatal é estimulado por diversas interações hormonais. Os poluentes ambientais que mimetizam hormônios esteróides, como o bisfenol-A (BPA), podem causar alterações no padrão de desenvolvimento da próstata, levando a lesões ao longo da vida. O objetivo deste estudo foi verificar se a exposição neonatal ao BPA afeta o desenvolvimento da próstata de gerbilos machos e fêmeas. Para isso, os recém-nascidos foram expostos a níveis ambientais (LBPA – 40 μ/kg/dia) e doses elevadas (HBPA – 4 mg/kg/dia) de BPA, do 1° ao 7º dia de vida. No oitavo dia, os complexos prostáticos foram coletados e processados para as análises morfológica, estereológica e imunohistoquímica. Com a reconstrução tridimensional observamos que nos machos os brotos prostáticos se alongaram em forma de “V” em direção ao mesênquima condensado ventral (VMP). Os dados estereológicos dos machos demonstraram um aumento significativo na frequência relativa do compartimento mesenquimal do grupo LBPA em relação aos demais grupos (p ≤ 0,05). As células AR-positivas tornaram-se significativamente mais frequentes no mesênquima periuretral (PeM), brotos epiteliais ventrais (VB) e camada muscular (SM) dos grupos tratados com BPA. Ocorreu uma redução na frequência de células PCNA-positivas nos brotos periuretrais (PeB) e na SM do grupo LBPA. Em fêmeas, a reconstrução tridimensional demonstrou que os PeBs podem emergir de um ou ambos os lados da uretra e também foi visível uma significativa redução do mesênquima parauretral (PaM) nos grupos tratados com BPA. Apenas as fêmeas do grupo HBPA apresentaram brotos parauretrais (PaB) aparentemente mais desenvolvidos. No grupo LBPA observou-se um aumento na frequência de células AR-positivas no PeM e uma diminuição destas células no PeB e mesênquima parauretral (PaM). No grupo HBPA as células AR-positivas tornaram-se mais frequentes no PeM e PaB, contudo sofreram redução no PeB e PaM. Células PCNA-positivas tornaram-se significativamente menos frequentes no PaB e PaM dos grupos LBPA e HBPA. Em relação às células ER-α positivas, nas fêmeas do grupo LBPA observou-se uma redução das imunomarcações no PeM e SM, enquanto que no grupo HBPA essa redução ocorreu apenas no PaM. Este estudo revelou que o desenvolvimento pós-natal da próstata de fêmeas de gerbilos é mais precoce e morfologicamente distinto do que ocorre nos machos da mesma espécie. Além disso, pode-se observar que o BPA exerceu um efeito antiproliferativo sobre a glândula prostática de machos e fêmeas com oito dias de vida pós-natal, sendo as fêmeas mais susceptíveis a este químico ambiental.