Arterial Glycosaminoglycans (GAGs) have gained importance in artherogenesis due to their ability to trap lipid inside the vessel wall. Atherosclerotic lesions have displayed an altered GAG content and distribution. Diabetes is a recognized risk factor for atherosclerosis, but no information is available on the arterial GAGs in human diabetes. To improve our understanding of the atherogenic proccss we examined GAGs in normal and atherosclerotic intima of nondiabetic and type-II diabetic humans. Intima was stripped from the autopsy samples of thoracic aortas, normal and plaque areas were separated. GAGs were isolated by delipidatlon, proteolytic digestion, and precipitation. They were assayed biochemically and their distribution evaluated by electrophoresis and densitometry. Results indicate a significant decrease in total GAGs and a change in GAG distribution in plaques of nondiabetics. Similar changes of lesser magnitude were found in normal intima of diabetics, while changes in plaque areas were more pronounced. This indicates that changes in arterial GAGs precede the development of lesions in diabetes and may be important in atherogenesis.

Estrogen and progesterone receptors were localized in fresh frozen sections of human endometrial tissues, in both health and disease, using the ER-ICA kit and a mouse monoclonal antiprogesterone receptor antibody ($ alpha$PR6), respectively. Estrogen and progesterone receptors were detected exclusively in the nuclei of epithelial and stromal cells of the endometrium. Their highest levels in both components were found during the late proliferative phase of the normal menstrual cycle. Estrogen receptors decreased faster in the stroma than in the epithelium throughout the post ovulatory phase, whereas progesterone receptors decreased more rapidly in the epithelium during the mid and late secretory phases. Estrogen and progesterone receptor levels were high in the epithelium of hyperplasia without cytologic atypia. They were low in the epithelium of endometrial intraepithelial neoplasia (hyperplasia with cytologic atypia) and the majority of invasive carcinomas. The stroma contained relatively high estrogen and progesterone receptors levels, irrespective of whether the epithelium was hyperplastic or neoplastic.

No description available.

Objective: Bone invasion represent significant problem in managing head and neck cancers, however the mechanisms of interactions between oral squamous cell carcinoma (OSCC) and bone cells are poorly understood. We hypothesized that tumor cells can directly stimulate osteoclastogenesis. Methods: OSCC cell lines, bone-invasive BHY and metastatic but not bone-invasive HN were cultured and conditioned medium (CM) was collected. Osteoclast formation from RAW 264.7 mouse monocytic cell-line was assessed. Results: When RAW 264.7 were primed with receptor activator of nuclear factor κB ligand (RANKL) and then treated with BHY-CM, marked 2-6 fold induction of osteoclastogenesis was observed. In contrast, HN-CM did not significantly affect osteoclastogenesis. In addition, BHY-CM, but not HN-CM promoted survival of mature osteoclasts. Using pharmacological inhibitors, we found that Protein kinase C (PKC)/Extracellular signal-regulated kinase (ERK)1/2/Mitogen activated protein kinase (MAPK) p38 as well as Phosphatidyl-inositol 3-kinases (PI3K)/Serine/threonine protein kinase Akt/Mammalian target of rapamycin (mTOR) pathways mediate BHY-CM induced osteoclastogenesis. Conclusion: OSCC-cells produce soluble factors that stimulate osteoclastogenesis from RANKL-primed precursors. Tumor-derived factors act by stimulating ERK1/2 and p38 MAPK pathways in osteoclast precursors. / Objectif: L'invasion du tissu osseux est un problème majeur dans le traitement des cancers de la tête et du cou, cependant les mécanismes d'interactions entre le carcinome de cellules de squamous oral (OSCC) et les cellules du tissu osseux sont mal compris. Nous avons posé comme hypothèse que les cellules tumorales peuvent stimuler directement le phénomène d'ostéoclastogenèse. Méthodes: Deux différentes populations cellulaires de la lignée OSCC furent utilisées: les cellules BHY ayant un potentiel de colonisation du tissu osseux et les cellules HN ayant un potentiel métastatique mais non colonisant. Ces deux lignées cellulaires ont été cultivées et le milieu de culture conditionné (CM) a été collecté. La formation de cellules ostéoclastiques à partir de cellules de la lignée monocytaire de souris RAW 264.7 a été évaluée. Résultats: Une augmentation significative du phénomène d'ostéoclastogenèse d'un facteur 2 à 6 fut observée lors d'une activation des cellules RAW 264.7 avec RANKL suivit d'un traitement avec BHY-CM. De plus, la survie des cellules ostéoclastiques matures était favorisée en présence de BHY-CM uniquement. L'utilisation d'inhibiteurs pharmacologiques nous a permis de mettre en évidence que la stimulation du phénomène d'ostéoclastogenèse induite par BHY-CM est médiée par les voies de signalisations PKC/ERK/p38 et PI3K/AKT/mTOR. Conclusion : Les cellules OSCC produisent des facteurs solubles stimulant la formation d'ostéoclastes à partir de précurseurs activées par RANKL. Les facteurs dérivés de tumeurs agissent en stimulant les voies de signalisation ERK1/2 et p38 dans les précurseurs ostéoclastiques.

Pre-operative radiotherapy for patients with locally advanced rectal carcinoma has been shown to improve survival rates and local tumour control. The ability to identify tumours most likely to undergo a complete or partial response would improve the selection of patients for radiotherapy and potentially modify post-treatment planning. The aim of this study was to develop a multi-marker model of tumour response to pre-operative high-dose rate endorectal brachytherapy (HDREB). Immunohistochemistry (IHC) for p53, Bcl-2, VEGF, APAF-1 and EGFR was carried out on 104 pre-treatment rectal tumour biopsies from patients undergoing a pre-operative HDREB protocol. Immunoreactivity was scored by at least three pathologists using a semi-quantitative scoring method. The reproducibility of the scoring system was evaluated. Receiver operating characteristic curve (ROC) analysis was performed for each protein to determine clinically relevant cutoff scores for defining tumour positivity. Multivariate logistic regression analysis was carried out to identify independent predictive factors of tumour response. Both the semi-quantitative scoring system and ROC curve analysis were found to be reproducible. In addition, the combined analysis of VEGF and EGFR was highly predictive of complete pathologic response to radiotherapy. EGFR was found to independently predict complete or partial tumour regression but only with low sensitivity and specificity. A large-scale prospective study is necessary to confirm these findings. Moreover, the novel methodology proposed and validated in this study to assess immunoreactivity could significantly enhance the value of IHC findings in colorectal cancer as well as other tumour types.

Purpose and objectives: Molecular mechanisms underlying prostate cancer progression from hormone -responsive to -refractory remain poorly understood. Studies in the host lab suggest that the up-regulated Fer kinase contributes to prostate cancer progression by its ability to phosphorylate and then bind pSTAT3Y705, pARY223 and pActinY53. As pSTAT3 and AR are known progression markers, our goal was to assess Fer, pSTAT3 and pActin expression in the human prostate along with the cell AR status. Methods: Prostate sections (425 cases) include benign hyperplasia, localized cancer of low, intermediate and high risks (untreated and neoadjuvant hormone treatment) and advanced cases under androgen deprivation and also bone metastases. Cells were counted (500-1500 cells/case) and their nuclear intensity assessed on a 0-3+ scale to obtain H Scores that were then analyzed for significance (ANOVA and Spearman Rho). Results: Fer and pActin were absent in the epithelium of pure BPH while sporadic weak pSTAT3 staining was observed. AR was at moderate levels in almost all luminal cells and weakly expressed in 40% of basal cells. Benign cells in prostate of localized cancer behave similarly to tumor cells of localized cases, with weak nuclear Fer and pActin and a patchy pSTAT3 pattern. Among predominant features was the increased intensity of Fer, pSTAT3, pActin, and AR in augmenting percentages of stained tumor cells with adjuvant hormonal therapy and disease progression, whereas weakly stained cells decreased in parallel. Intriguingly the percentage of AR negative (-) tumor cells, interspersed among AR positive (3+) cells, increased concomitantly with progression. Significant correlations were obtained for Fer/pSTAT3, pSTAT3/AR and Fer/pActin. Conclusions: Altogether the increased expression of nuclear Fer along with pSTAT3, pActin and AR in tumor cells of the human prostate with progression, along with established correlations between Fer and each of these proteins support its role in pathways and crosstalks helping cells to survive, grow, and resist therapy. Fer, pSTAT3, pActin and AR may thus be considered as interrelated markers of progression and targets for new therapies aimed at alleviating all tumor cell subtypes, both AR(+) and AR(-). / Préambule et objectifs: Les mécanismes moléculaires sous-jacents à la progression du cancer de la prostate, d'un état sensible à réfractaire aux androgènes, demeurent méconnus. Les études du laboratoire d'accueil permettent de proposer que la kinase Fer, qui est surexprimée dans ce cancer, contribue à la progression par sa capacité de phosphoryler et ensuite lier pSTAT3Y705, pARY223 et pActineY53. Puisque pSTAT3 and AR sont des marqueurs de progression connus, le but était d'étudier l'expression de Fer, pSTAT3 et pActine dans la prostate humaine en parallèle au statut du AR dans les cellules. Méthodes: Les lames de tissu prostatique (425 cas) comprennent de l'hyperplasie bénigne, du cancer localisé de faible, intermédiaire, et haut risque (non-traité, et avec thérapie hormonale néo-adjuvante) et du cancer avancé soient des cas dépourvus d'androgènes et des métastases osseuses. Les cellules ont été comptées (500-1500 par cas) et leur intensité de marquage nucléaire établie de 0 à 3+ afin d'obtenir des « scores » H, ensuite analysés statistiquement (ANOVA et Spearman Rho). Résultats: Fer et la pActine sont absents de l'épithelium bénin alors que pSTAT3 est sporadiquement observée à de faibles niveaux. Le AR est exprimé modérément dans les cellules luminales et faiblement dans 40% des cellules basales. Les cellules bénignes des cas de cancer localisé ressemblent aux cellules tumorales avec des niveaux faibles de Fer, pActine et un patron mixte de pSTAT3. Parmi nos observations, notons les niveaux de Fer, pSTAT3, pActine, et AR par noyau augmentant de pair avec plus de cellules tumorales marquées, sous thérapie néo-adjuvante et avec la progression, alors que les cellules faiblement réactives diminuent. Paradoxalement, le pourcentage de cellules AR (-) clairsemées entre les cellules AR (3+), augmente avec la progression. Des corrélations significatives ont été obtenues pour Fer/pSTAT3, pSTAT3/AR et Fer/pActine.Conclusions: L'expression accrue de Fer dans le noyau des cellules tumorales de la prostate humaine ainsi que celle de pSTAT3, pActine et AR avec la progression, de même que les corrélations entre Fer et ses partenaire supportent un rôle majeur dans des voies de signalisation facilitant la survie et croissance des cellules, et leur résistance aux thérapies. Ainsi Fer, pSTAT3, pActine et AR peuvent être considérés comme des marqueurs de progression inter-reliés, pouvant aussi servir de cibles thérapeutiques afin d'éliminer toutes les cellules tumorales, AR(+) et AR(-).

Cerebral vasospasm is a common cause of ischemia and death in patients suffering from cerebral hemorrhage, most notably subarachnoid hemorrhage (SAH), and it usually occurs 4-14 days after the initial episode of bleeding. Many mediators of cerebral vasospasm (CVS) have been proposed, but at this time the pathogenesis of CVS is poorly understood, and oxyhemoglobin is thought to be the principal pathogenic agent. / This project was designed to examine the effects of oxyhemoglobin on the vascular tone of the middle and basilar cerebral arteries in vitro and to evaluate the validity of using bovine vessels as an animal model of the human cerebral vasculature in which to study cerebral vasospasm. Human vessels were studied in parallel with the bovine arteries to test the validity of the animal model. To further evaluate the model, some experiments were also carried out on canine vessels, because the dog has frequently been used for cerebrovascular studies in the past. / The cow is a better model of the human cerebral vasculature than the dog. Arteries respond to a variety of vasoactive stimuli, in a manner closely resembling human vessels, and can be induced into a prolonged vasospastic state by exposure to oxyhemoglobin. Vasospasm in all three species was independent of endothelial status, functional innervation, or morphological evidence of atherosclerosis. Vasospasm could not be prevented by diltiazem, nicardipine or ascorbic acid, but was partially reversed in some samples by verapamil. Bovine vessels would appear to be exceptionally useful for studies designed to test pharmacological agents for the prevention or therapy of post-hemorrhagic vasospasm and also to determine the pathophysiological mechanisms involved.

In humans, it has been demonstrated that Th-2 type cytokines (interleukin (IL)-4 and IL-5) are highly expressed in bronchial biopsies from asthmatics. Our hypothesis is that a distinct cytokine response comprising Th-2 cytokines occurs during the late airway response (LR) after antigen challenge and that a different Th-1 response (interferon (IFN)-$ gamma$) is related to the absence of a physiological response after antigen challenge. We have tested this hypothesis in different strains of rats, Brown Norway (BN) rats are high IgE producers that develop LR and increased airway responsiveness after antigen challenge, whilst Sprague Dawley (SD) rats are low IgE producers that do not develop physiological changes following antigen challenge. We assessed the expression of cytokines in these two strains of rats using semi-quantitative polymerase chain reaction (SQPCR). Fourteen days after sensitisation to ovalbumin (OA), the rats underwent general anaesthesia and were killed either immediately or eight hours after OA challenge. The lungs were frozen in liquid nitrogen for total RNA preparation. cDNA was prepared from total RNA, and mRNA expression for IL-4, IL-5 and IFN-$ gamma$ was assessed before and after challenge in BN and SD rats using Cyclophilin as a housekeeping gene. (Abstract shortened by UMI.)

Nitric oxide (NO) and endothelin-1 (ET-1) are ubiquitous molecules whose actions have been implicated in several airway and vascular pathologies. NO is a free radical produced by the action of inducible and constitutive nitric oxide syntheses (NOS II and NOS III, respectively). NO possesses a wide range of actions including cellular damage which is mediated in part by peroxynitrite (ONOO-), an oxidant formed by the rapid reaction of NO with superoxide. ET-1 is a vasoconstrictor with well known mitogenic properties. In view of the physiologic properties of NO and ET-1, as well as the implication of these molecules in several pulmonary pathologies, we hypothesized that fibroproliferative lung disorders are associated with the cellular induction of NOS, ONOO-, and ET-1. Localization of these agents were qualitatively and quantitatively analyzed using immunohistochemistry and in situ hybridization in several proliferative lung disorders. / These findings are essential for our understanding of the pathogenesis of lung disease and the biology of vasoactive substances. They also suggest a possible role for both the NO and ET pathways in the functional and morphological abnormalities in proliferative disorders of the lung. (Abstract shortened by UMI.)

Airway eosinophilia is a prominent feature of asthma that may be mediated in part through the expression of eotaxin, a potent eosinophil-active chemokine that is highly expressed by epithelial cells and inflammatory cells in asthmatic airways. / The major aim of this study was to determine whether human airway smooth muscle (ASM) cells may be a source of eotaxin in asthma. The evidence presented in this thesis shows constitutive eotaxin gene expression in ASM in vitro that is markedly increased following stimulation with the proinflammatory cytokines TNF-alpha and IL-1beta. Release of eotaxin was confirmed in ASM culture supernatants which contained significant chemoattractant activity for eosinophils that was partly inhibited with antibodies directed against eotaxin or RANTES, and maximally inhibited by a combined blockade of both chemokines. Strong signals for eotaxin immunoreactivity were also observed in vivo in smooth muscle in asthmatic airways. / In conclusion, the results of this study suggest that ASM may contribute to airway inflammation in asthma through the production and release of eotaxin.