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Intra-abdominal infections are a common entity faced by general surgeons, which, when complicated by pneumonia lead to increased mortality. The susceptibility to a second infection is correlated with impaired polymorphonuclear neutrophil (PMN) migration to the remote site, perhaps due to inadequate PMN number. We tested the second front hypothesis that the elevated mortality arising from a second infection is due to insufficient circulating PMN, the majority being sequestered at the first site of infection. Further, augmentation of systemic PMN number by granulocyte colony stimulating factor (G-CSF) will decrease mortality. / We established a murine model for peritonitis and pneumonia. PMN counts in the blood, lung tissue, peritoneal cavity and bronchoalveolar system were measured. The results indicated significant PMN recruitment to the primary site of infection, whether lung or abdomen. Almost no PMN were found in the lung if it was the second site of infection. This was consistent with the second front hypothesis.

Cystic fibrosis (CF) patients have airway neutrophilic inflammation associated with high levels of interleukin-8 (IL-8) and often develop airway hyperresponsiveness (AHR). Aerosolized uridine triphosphate (UTP) has been used to improve mucosal clearance by inducing Cl- secretion. However, nucleotides such as UTP and ATP (adenine triphosphate) also evoke the release of intracellular Ca2+ ([Ca2+]i), which are enhanced by pro-inflammatory mediators, such as IL-1beta. Since most of the CF airway epithelium is lost during infection and inflammation, the underlying airway smooth muscle cells (ASMC) are exposed to IL-8 and extracellular nucleotides. Thus, we were particularly interested in evaluating the effects of IL-8, extracellular ATP and UTP on contractile responses of ASMC. For this study, cultured ASM from normal and CF lung specimens were used. We have found that human ASM cells (HASMC) express mRNA and protein for IL-8 receptors (CXCR1 and CXCR2) and IL-8 causes an increase in [Ca2+]; as well as cellular contraction and migration. Furthermore, changes in the [Ca2+] i were abrogated on blocking IL-8 receptors with specific neutralizing antibodies, and were also decreased following inhibition of phospholipase C (PLC), indicating that changes in [Ca2+]i were likely mediated by inositol trisphosphate. The effects of IL-8 on HASMC prompted us to investigate whether IL-8 induced ASMC contraction in CF was different from normal. IL-8 caused larger ASM contractions and greater phosphorylation of myosin light chain (MLC20) compared to normal cells. CF cells expressed more MLC20 compared to normal cells. However, the expression of CXCR1, CXCR2, IL-8 induced changes in [Ca2+]i and ASMC migration in CF cells were similar to normal cells. We also found that normal HASMC express receptors (subtypes of P2Y) for ATP and UTP, which stimulated the release of intracellular Ca2+. Brief exposure of normal HASMC with UTP enhanced Ca2+ transients in response t

Early growth response protein-1 (EGR-1) is a transcription factor induced by stress or injury, mitogens, and differentiation factors. It has been shown to be regulated by various cytokines, growth factors and by ischemic/hypoxic stress as well as shear stress and mechanical injury. These regulators have been linked to both the development as well the degeneration of the musculoskeletal system, namely articlar cartilage, intervertebral discs (IVDs) and bone. Furthermore, Egr-1 has been shown to regulate the expression of collagens and enzymes affecting the extracellular matrix. Polymorphisms of DNA binding sites for EGR-1 have shown to be associated with both disc degeneration and osteoporosis. The aim of this study was to determine the affects of EGR-1 deficiency on articular cartilage, IVD and bone phenotype. / Wild-type (+/+) C57Bl/6 or Egr-1-deficient (−/−) mice were sacrificed at the same age interval (8- to 9-months). Standard histological preparation and staining with Safranin-O/Fast Green were done. Also immuncohistochemistry was performed using anti-bodies to type X collagen, cleavage products of both type II collagen and aggrecan. Imaging of mice was with plain radiographs, bone mineral density measurements and microCT analysis. / Results revealed that these mice have differences including abnormal bone structure and density, structural and possibly compositional differences in articular cartilage and structural and biochemical changes in IVDs. This points to the importance of Egr-1 in the maintenance of normal bone, IVD and articular cartilage and makes it a possible target for initiating pathological conditions of these tissues. / La protéine de croissance EGR-1 (Early Growth Response protein-1, en anglais) est un facteur de transcription qui est induit par la tension ou la blessure, les facteurs mitogènes, et les facteurs de différenciation. EGR-1 est ainsi régulé par divers cytokines, facteurs de croissance, par les conditions ischémique, ainsi que la tension et les blessures mécaniques. Ces régulateurs ont été reliés au développement ainsi que la dégradation du système squeletto-musculaire, particulièrement le cartilage articulaire, les disques intervertébraux (DIV) et l'os. De plus, il a été démontré que EGR-1 peut réguler l'expression des collagènes et d'enzymes contribuant à la matrice extracellulaire. Le polymorphisme de séquences d'ADN pour les des sites d'attachements d'EGR-1 a démontré être associé avec la dégradation de disques intervertébraux et l'ostéoporose. L'objectif de cette étude était de déterminer l'effet d'une expression réduite d'EGR-1 sur les phénotypes du cartilage articulaire, les DIV, et l'os. / Les souris C57Bl/6 de phénotype sauvage (+/+) ou ceux avec une expression réduite d'EGR-1 (−/−) ont été sacrifiées au même intervalle d'âge (8 à 9 mois). La préparation histologique standard et la coloration avec Safranin-O/Fast Green a été fait. Aussi l'immunohistochimie a été exécuté avec des anticorps pour le collagène de type X, et les produits de clivage du collagène de type II ainsi que les aggrécanes. L'imagerie de souris a été faite avec les radiographies simples, les mesures de densité minéraux de l'os, et avec l'analyse de micro-tomodensitomètre. / Les résultats ont révélé que ces souris ont des différences incluant la structure et densité d'os anormaux, les différences structurelles et possiblement compositionnelles dans le cartilage articulaire, et les changements structurels et biochimiques dans les DIV. Ceci indique à l'importance d'EGR-1 dans l'entretien d'os normal, des DIV et le cartilage articulaire, et le rend une cible possible pour initier les conditions pathologiques de ces tissus.

Preservation of female fertility is an important issue today. There are a few effective clinical options for preserving female fertility. Conventional IVF followed by embryo cryopreservation is the only established procedure but is not applicable to all women. Oocyte cryopreservation avoids the ethical and moral concerns related to cryopreservation of embryos but conventional slow freezing methods are associated with low survival rate of oocytes. The main objective of this translational research thesis was to develop an efficient and safe methodology for oocyte cryopreservation that is clinically applicable for female fertility preservation. Specific research objectives were to investigate cryobiology of oocytes in terms of: 1) cryoprotectant (CPA) toxicity effect on oocyte ultra-structures and embryonic developmental potential; 2) Vitrification versus conventional slow freezing of oocytes and their effects of oocyte structures and embryonic developmental potential; 3) Vitrification of embryos using the McGill Cryoleaf and its effect of embryonic development and DNA fragmentation; 4) Clinical efficacy of oocyte vitrification in prospective clinical trials; 5) Effects of oocyte vitrification in terms of the clinical obstetrical and perinatal outcomes; 6) Clinical applications of vitrification of oocytes for preservation of female fertility. The CPA mixture of ethylene glycol (EG) and 1,2-propanediol (PROH) was found to be the most suitable combination for oocyte vitrification, resulting in high embryo development and the least DNA fragmentation. Vitrification of oocyte using the CPA mixture of EG and PROH in combination with the McGill Cryoleaf system is superior to the conventional slow-cooling method, resulting in better preservation of egg ultra-structures and functions. The reduced embryonic development potential of cryopreserved oocyte is related to increased DNA fragmentation and activation of caspase enzymes. Vitrification of human oocytes using the McG / Préservation de la fécondité est un sujet important à ce jour. Il y a peu de traitements effectifs pour préserver la fécondité des femmes. La fécondation in vitro (FIV) conventionnelle suivie par la cryoconservation des embryons est la seule procédure bien établie. Cependant, celle-ci n'est pas possible pour certaines femmes. La cryopréservation des ovocytes évite les problèmes éthiques et moraux reliés à la cryoconservation des embryons. Cependant, les méthodes de congélation lente sont associées à des taux de survie des ovocytes faible. Les objectifs principaux de cette thèse de recherche translationnelle était de développer une méthode efficace et sécuritaire pour la cryopréservation des ovocytes qui est cliniquement applicable pour la préservation de la fécondité des femmes. Les objectifs spécifiques de cette recherche étaient d'étudier la cryobiologie des ovocytes à propos de : 1) l'effet toxique du cryoprotectant sur l'ultra-structures des ovocytes et le potentiel de développement des embryons; 2) la vitrification versus la congélation lente des ovocytes et leur effets sur la structure des ovocytes et le potentiel de développement embryonnaire; 3) la vitrification des embryons en utilisant le McGill Cryoleaf et son effet sur le développement embryonnaire et la fragmentation de l'ADN; 4) l'efficacité clinique de vitrification des ovocytes dans des études prospectives cliniques; 5) les effets de long terme de vitrification des ovocytes au niveau des résultats obstétriques et périnatals; 6) l'application clinique de vitrification des ovocytes pour la préservation de la fécondité des femmes. Le mélange de CPA de l'éthylène glycol (EG) et de 1-2 propanediol (PROH) a été trouvé d'être la combinaison la plus convenable pour la vitrification des ovocytes en donnant des résultats de développement embryonnaire supérieur et à un taux de fragmentation de l'ADN diminué. La vitrification des ovocytes en utilisant l

Background: Renal transplant patients who are highly sensitized linger on the transplant list due to HLA specific antibodies. Current desensitization protocols are unreliable. Interleukin-21 is a type I cytokine that signals through a receptor shared by the all T-cell growth factors (TCGFs). Secreted by T-cells, IL-21 acts as a regulator of B-cell anti-body production and differentiation into plasma cells as well as immunoglobulin class switching. It co-stimulates proliferation and memory response in T-cells. We hypothesized that IL-21 could be a significant contributing factor in induction of sensitization and antibody production by B-lymphocytes. We evaluated the effect of IL-21 on the anti-body production capacity of B-cells and compared the expression IL-21 Receptor (IL-21R) on the peripheral B-cells in highly sensitized and non-sensitized patients on dialysis. / Methods: In-vitro studies: B-cells isolated from peripheral blood of healthy individuals, were stimulated with mouse anti-Human CD40 monoclonal antibody (R&D), recombinant IL-21 or combination of both. Produced IgG and IgM levels were measured by classic Eliza method. Ex-vivo studies: Patients selected from the transplant waiting list and assigned into two groups: non-sensitized with (PRA<5%), sensitized (PRA>50%). After obtaining consent, peripheral blood was used for evaluation of IL-21R Expression on PBMNCs by Flowcytometry. Results: (1) In-vitro Co-stimulation of B-cells with IL-21 and anti-CD40 results in higher antibody production compared with isolated anti-CD-40 or IL-21 stimulation. (2) In-vitro stimulation of human B-cells with anti-CD40 Antibody increases expression of IL-21R. (3) The expression of IL-21R is significantly higher on the peripheral B-cells in highly sensitized patients compared to non-sensitized patients. (4) A significant correlation exists between PRA levels and the percentage of peripheral cells expressing IL21R. / Conclusion: IL-21 and IL-21R may be an important cytokine interaction in B-cells and antibody production in highly sensitized renal transplant recipients. This may offer a novel target for desensitization protocols. / Antécédent: les patients transplantés rénaux hautement sensibilisés sont sur la liste de transplantation en raison d'anticorps anti-HLA. Les protocoles actuels de désensibilisation sont non fiables. Interleukin-21 est une cytokine de type I qui signale par intermédiaire d'un récepteur commun entre les facteurs de croissance des cellules T (TCGFs). Sécrétée par les cellules T, l'IL-21 agit comme un régulateur de la production d'anticorps anti cellule B, de la différenciation des plasmocytes, ainsi de la commutation de classe d'immunoglobuline. Il co-stimule la prolifération et la réponse de la mémoire dans les cellules T. Nous avons évoqué l'hypothèse que l'IL-21 pourrait être un facteur contributif important pour l'induction de la sensibilisation et pour la production d'anticorps par les lymphocytes B. Nous avons évalué l'effet de l'IL-21 sur la capacité de production d'anticorps par les cellules B et avons comparé l'expression des récepteurs IL-21 (IL-21R) sur les cellules B périphériques chez les patients hautement sensibilisés et nonsensibilisés étant sous dialyse. / Méthodes: Les patients sélectionnés dans la liste d'attente de transplantation et sont enrôlé en deux groupes: les non sensibilisés avec (PRA <5%), sensibilisés (PRA> 50%). Après l'obtention du consentement, du sang périphérique a été utilisée pour l'évaluation de l'IL-21R expression sur PBMNCs par la Cytométrie de Flux. Résultats: (1) La co-stimulation in vitro des cellules B avec l'IL-21 et anti-CD40 résulte à la production d'anticorps plus élevée comparativement à la production liée à la stimulation isolée d'anticorps anti-CD40 ou à celle d'IL-21. (2) La stimulation in vitro des cellules B humaines par des anticorps anti-CD40 augmente l'expression de l'IL-21R. (3) L'expression de l'IL-21R est significativement plus élevée dans les cellules B périphériques chez les patients hautement sensibilisés par rapport aux patients non sensibilisés. (4) Il existe une corrélation significative entre les niveaux de PRA et le pourcentage de cellules périphériques exprimant IL21R. / Conclusion: L'IL-21 et l'IL-21R peuvent être une cytokine d'interaction importante entre les cellules B et la production d'anticorps chez les receveurs de greffe rénale hautement sensibilisés. Il se peut que ceci offre une nouvelle cible pour les protocoles de désensibilisation.

The purpose of this thesis was to evaluate microinvasive technology for the management of non-palpable mammographic findings with high risk for breast cancer. The specific objectives were, for the first part, to evaluate the Advanced Breast Biopsy Instrumentation System (ABBI) as a diagnostic and therapeutic tool for breast disease. For the second part, the objective was to evaluate lymphoscintigraphy with a malignancy specific radioactive agent (99m Technetium Sestamibi) as a non-surgical staging tool for breast cancer. / Design. The thesis consists of two observational studies. The first study was partially retrospective and partially prospective. The second study was prospective. The research was conducted between September 1997 and December 2001. / Methods. For the first study 262 women with non-palpable mammographic abnormalities were enrolled. Of these, 260 women underwent an ABBI. The ABBI procedure was performed by 3 surgeons in two hospitals in Montreal. The files of 531 women with similar findings who underwent a wire localization excisional biopsy in the same hospitals by the same surgeons were retrospectively reviewed. Data obtained included technical success, macroscopic and microscopic clarity of margins, need for supplemental surgery and residual cancer, volume of breast tissue excised, patient satisfaction, surgeon volume-performance association and health care costs. / For the second study, 110 women with confirmed operable primary breast cancer who required axillary lymph node dissection were enrolled. All the patients underwent two lymphoscintigraphies. The first (preliminary) lymphoscintigraphy took place at least 2 weeks before surgery and the second (preoperative) took place 2-24 hours before surgery. For the preliminary lymphoscintigraphy, 47 (42.7%) of the 110 women were tested with 99mTechnetium Sestamibi and 63 (57.3%) were tested with 99mTechnetium Sulphur Colloid. For the preoperative lymphoscintigraphy all patients were tested with 99mTechnetium Sulphur Colloid. All patients underwent axillary lymph nodes dissection. The data reviewed was the presence of metastasis in the axillary lymph nodes excised and the lymphoscintigraphy gamma camera imaging and gamma probe digital outcomes. The ROC curve was used to define the most probable cut-off points for the distinction between gamma counts representing positive and negative for metastasis axillary lymph nodes. / Results. For the first study, diagnostic effectiveness of the ABBI was 99.6% and the mean volume of excised breast tissue was 40cc. No serious complications were reported. For wire localization, the diagnostic effectiveness was 98.0% and the mean volume of excised breast tissue was 102cc. severity of complications was similar between the two techniques. The therapeutic effectiveness of the ABBI was 76.3%, supplementary surgery was required in 27.6% of the patients, and residual cancer was detected in 23.8%. The therapeutic effectiveness of wire localization biopsy was 47.0%, 53% of the patients underwent supplementary surgery and residual tumor was found in 48.0%. Of the 260 patients who underwent an ABBI, 51% reported a good level of satisfaction and 23% reported excellent. A significant association was shown between volume and therapeutic effectiveness for all 3 surgeons. On the average, the ABBI resulted in a cost reduction of $5,352 CAD (2000). / For the second study, the sensitivity of 99mTechnetium Sestamibi lymphoscintigraphy imaging was 50%, the negative predictive value was 83.8% and the proportion of false negative results was 17.1%. Logistic regression showed a statistically significant (p&lt;0.001) association between the gamma probe counts and axillary lymph node metastasis. Using 3,080 counts as cut-off point on the ROC curve the sensitivity and false negative rate were 100%, the specificity was 94.3%, and the positive predictive value was 83.3%. / Conclusion. The results of the first study show that the ABBI offers a safe and well tolerated microinvasive surgical technique, with superior therapeutic effectiveness, less invasiveness, and better cost effectiveness than wire localization excisional biopsy. The strong volume-performance association shows that the therapeutic effectiveness of the ABBI can improve with proper training and experience. / The results of the second study show that 99mTechnetium Sestamibi lymphoscintigraphy imaging was 50% the negative predictive value was 83.8% and the proportion of false negative results was 17.1%. A significant association was shown between the intensity of radioactivity in the hot node as measured by the gamma-probe counts and the presence of axillary lymph node metastasis. / In conclusion, this thesis demonstrates that both the ABBI and lymphoscintigraphy with 99mTechnetium Sestamibi are superior to conventional techniques used for the management of mammographic abnormalities and specifically breast cancer. Proper training of surgeons in highly qualified training centers with a high volume of cases can help further improve the effectiveness and increase the practise of these techniques in Canadian hospitals. This may in turn increase the benefits for the patients and reduce the healthcare costs for the management of breast disease and specifically breast cancer in Canada.

Intravenous lidocaine infusion for colorectal surgery has been shown to provide superior analgesia compared with systemic opioids and facilitate hospital discharge. While epidural analgesia has definite advantages over systemic opioids in term of return of bowel function and quality of postoperative pain control, there is no study comparing lidocaine infusion with epidural technique in the setting of enhanced recovery program (ERP) for laparoscopic colorectal surgery. In addition, functional recovery and quality of life have not been assessed and compared with other analgesic techniques. This project is designed to evaluate the impact of lidocaine on surgical and functional outcomes. In these randomized studies, patients scheduled for elective laparoscopic colorectal surgery were prospectively randomized to receive thoracic epidural analgesia (TEA group), intravenous lidocaine infusion (IL group) or patient-controlled analgesia with morphine (PCA group). All patients received similar surgical care in the context of ERP. The average time to return of bowel function and median duration of hospital stay were similar in IL and TEA groups. TEA provided better postoperative analgesia than intravenous lidocaine in patients undergoing rectal surgery; otherwise there was no difference for colon resection. IL, TEA and PCA facilitated the return of postoperative functional walking capacity to baseline, and this was independent of the analgesic techniques use. However physical functioning and fatigue levels were impaired at 3 weeks after surgery with no difference between the 3 groups. The present study demonstrated that the restoration of bowel function and diet intake were similar in both groups receiving either lidocaine infusion or epidural. Functional walking capacity at 3 weeks after surgery returned to baseline in all the groups and this was independent of the analgesic technique used. However, in all groups physical function decreased and fatigue increased and this was also independent of the type of analgesia used. / Lors de chirurgie colorectale, il a été démontré que la Lidocaine intra-veineuse provoque un niveau d'analgésie comparable aux opiacés mais facilite la récupération postopératoire. L'épidurale est nettement supérieure aux opiacés systémiques en terme de fonction intestinale et d'analgésie. Il n'existe hélas pas d'étude comparant la lidocaine versus l'épidurale en termes de réhabilitation fonctionnelle et qualité de vie dans le cadre d'un programme de réhabilitation accélérée après chirurgie colorectale. L'objectif du présent protocole est d'évaluer l'utilité de la lidocaine en termes de récupération fonctionnelle et chirurgicale. Cet essai randomisé inclut des patients requérant une chirurgie colorectale par laparoscopie. Les patients sont prospectivement randomisés en 3 groupes: Epidurale (Groupe TEA), lidocaine intraveineuse (Groupe IL) ou opiacés intraveineux (Groupe PCA). Les 3 groupes de patients reçoivent des soins chirurgicaux et anesthésique identiques dans le cadre d'un programme de réhabilitation accélérée. La récupération fonctionnelle intestinale et la durée d'hospitalisation est similaire entre les groupes lidocaine et épidurale. L'épidurale apporte une meilleure analgésie que la lidocaine chez les patients ayant des chirurgies rectales mais l'analgésie est similaire chez les patients subissant une colectomie. Les trois stratégies furent similaires en termes de récupération fonctionnelle. Néanmoins, a 3 semaines postopératoire l'état fonctionnel physique et la fatigue ne sont toujours pas retournés a leurs valeurs pré-opératoires dans aucun des groupes. La présente étude montre que la récupération fonctionnelle intestinale et la prise alimentaire est comparable entre les 3 groupes. A 3 semaines postopératoires, la capacité à la marche est retournée aux valeurs pré-opératoires dans les 3 groupes, indépendamment de la technique d'analgésie. Néanmoins a 3 semaines l'évaluation fonctionnelle physique restait diminuée et le niveau de fatigue accru par rapport aux évaluations pré-opératoires, indépendamment de la technique d'analgésie.

Context and objective: The Information Assessment Method (IAM) is a unique tool for continuing education and knowledge translation research. IAM allows health professionals to report search objectives, cognitive impact, use and patient health benefit associated with objects of clinical information retrieved from electronic knowledge resources. While IAM has been previously validated in the information delivery context (PUSH), this thesis examines the content validity, relevance and representativeness, of IAM items in the context of information retrieval (PULL). Methods: The study was conducted in three steps. In step 1, the relevance and representativeness of IAM items were assessed. In this step, data from a mixed methods triangulation study combining a prospective observational study with a qualitative multiple case study involving 40 family physicians were analyzed. Step 2 consisted of analysis and modification of every IAM item based on a set of guiding principles. Step 3 consisted of a multi-disciplinary expert panel discussion on all modified items and the development of a new version of IAM. Results: The content validity of 16 IAM items was supported, and these items were not changed. Nine other items were modified. Three new items were added; two were extensions based on one existing item, and one was obtained from data analysis and literature review. Conclusion: The final result of this thesis is a content validated version of IAM in the PULL context (IAM 2011). / Contexte et objectif: La Méthode d'Évaluation des Informations (MEI) est un outil unique pour les domaines de la formation continue et de l'application des connaissances. La MEI permet aux professionnels de la santé d'évaluer les objectifs des recherches d'information, les impacts cognitifs, les utilisations et les bienfaits sur la santé des patients associés à des objets d'information clinique, trouvées dans des ressources électroniques. Alors que la MEI a été validée pour évaluer plusieurs types d'informations envoyées par courriel (PUSH), ce mémoire examine la validité de contenu, la pertinence et la représentativité de chaque élément de la MEI pour évaluer les informations trouvées dans des ressources électroniques (PULL). Méthodes: L'étude a été menée en trois étapes. Durant la première, la pertinence et la représentativité des éléments de la MEI ont été évaluées. Au cours de cette étape, les données d'une recherche mixte ont été analysées. Cette étude combinait une étude quantitative prospective longitudinale et une étude qualitative de cas multiple, et elle a été menée auprès de 40 médecins de famille. La deuxième étape consistait à analyser et modifier chaque élément de la MEI, en se basant sur un ensemble de principes directeurs. Enfin, la troisième étape consistait à discuter tous les éléments modifiés avec un panel d'experts provenant de plusieurs disciplines, puis à élaborer une nouvelle version de la MEI. Résultats: La validité de contenu de 16 éléments de la MEI a été soutenue, et ces éléments n'ont pas été changés. Neuf autres éléments ont été modifiés. Trois nouveaux éléments ont été ajoutés : deux ont été construits à partir d'un élément existant, et le troisième a été suggéré à la fois par la revue de la littérature et l'analyse des données. Conclusion: Le résultat final de ce mémoire est une nouvelle version de la MEI pour évaluer les informations trouvées dans des ressources électroniques 'PULL' (MEI 2011).