Serum cortisol and immunoglobulin G (IgG) concentrations were measured to investigate the relationship between vitamin A status and immune function in lambs. Twenty-four crossbred ewe lambs were each fed 900 g·d-1 of a carotene—deficient diet composed of 95.5% whole oats, 3% molasses, .5% trace mineral salt and 1% limestone. All lambs were injected monthly with vitamins D and E and with selenium. The 12 control lambs also received a 100,000 IU oral dose of vitamin A palmitate in capsule form every 2 wk. All lambs were challenged by injecting them with 1 mg ovalbumin in 1 ml of Freund’s complete adjuvant. At the time of challenge, serum vitamin A levels for the control and A-deficient (A—def) lambs were 33.3 and 3.1 ug·dl-1 respectively. Blood was collected prior to and 6, 13, 20 and 34 d post—challenge. The lambs were then reschallenged using the same antigen and blood was obtained 1, 2, 6 and 22 d post—challenge. Lambs were sacrificed at the end of the second challenge period. Spleen weights were obtained and gross post—mortem observations were made at this time. / Master of Science

No / The formulation of poorly water-soluble drugs is a challenging problem within pharmaceutical development. Recently, formulation using nanoparticles was highlighted as showing great potential to improve the dissolution and solubility characteristics of poorly water soluble drugs.

Tableau d’honneur de la Faculté des études supérieures et postdoctorales, 2008-2009 / L'objectif de cette thèse de doctorat consiste à étudier l'étiologie génétique et environnementale de la sécrétion cortisolaire à la petite enfance, une période du développement caractérisée par une forte plasticité de certaines structures cérébrales. Une attention particulière est accordée à l'investigation des variations des contributions génétiques et environnementales en fonction de l'adversité familiale, indice d'une possible interaction entre les gènes et l'environnement. L'adversité familiale renvoie à une mesure composite de 7 facteurs périnataux et postnaux ayant été liés à la sécrétion cortisolaire (l'exposition prénatale à la cigarette, le faible poids à la naissance, le faible revenu familial, pas de diplôme d'études secondaires, mère monoparentale, le jeune âge de la mère et les comportements maternels hostiles ou réactifs envers les jumeaux). Considérant que les mesures cortisolaires ont été effectuées dans des contextes différents (prélèvements salivaires réalisés à deux temps au cours de la journée vs une réponse à un stress de nouveauté sociale) et à des âges distincts (6 vs 19 mois), la poursuite de cet objectif s'effectue séparément pour ces mesures. Dans un premier temps (article 1; mesure réactive à un contexte de nouveauté sociale à 19 mois), les résultats indiquent une variation des contributions génétiques et environnementales en fonction de l'adversité familiale, prenant la forme d'un effet organisationnel de l'adversité environnementale. Cet effet se traduit par une contribution réduite des facteurs génétiques combinée à une plus forte contribution de l'environnement partagé en contexte d'adversité familiale comparativement à une contribution génétique modérée du cortisol réactif en absence d'adversité familiale. Dans un deuxième temps (article 2; mesures diurnes à 6 mois), les résultats indiquent une variation des contributions génétiques et environnementales de la sécrétion cortisolaire matinale en fonction de l'adversité familiale. Cette variation prend cette fois la forme d'une plus forte héritabilité en présence d'adversité familiale alors que seuls les facteurs environnementaux uniques entrent en jeu en l'absence de ce contexte de vie adverse. Aucune variation des contributions génétiques et environnementales n'est notée sur la sécrétion cortisolaire mesurée au réveil en fonction de l'adversité familiale. Pour cette mesure, seuls les facteurs génétiques expliquent la ressemblance entre les jumeaux d'une même paire.

L’environnement maternel peut influencer le développement. Chez les poissons, ce phénomène serait modulé via le dépôt dans les œufs de facteurs maternels, tel le cortisol, une hormone de réponse au stress. L’augmentation de cortisol dans le plasma maternel et dans les œufs a été associée avec des altérations comportementales chez les jeunes. Pour séparer les effets du cortisol de ceux des autres facteurs dans cette reprogrammation comportementale due au stress maternel, nous avons pharmacologiquement et physiquement manipulé des femelles omble de fontaine, Salvelinus fontinalis, pendant l’oogénèse. Nous les avons soit laissées non dérangées (témoins), (1) nourries de cortisol additionné à leur nourriture ou (2) manipulées hebdomadairement. Nous avons aussi (3) exposé des œufs des femelles témoins à du cortisol avant la fécondation. Nous avons mesuré les capacités cognitives et l’audace chez les juvéniles. Nous n’avons détecté aucun effet des traitements sur leur comportement. / The maternal environment can influence development. In fish, this phenomenon could be modulated by egg deposition of maternal factors, like cortisol, a stress-response hormone. Increased maternal plasma and egg cortisol levels have been associated with behavioral alterations in offsrping. To discriminate between the effects of cortisol and those from other factors in the behavioral reprogramming due to maternal stress, we have pharmacologically and physically manipulated female brook trout, Salvelinus fontinalis, during oogenesis. We either left them undisturbed (controls), (1) fed them cortisol through their food or (2) handled them once a week. We also (3) exposed eggs from the control females to cortisol before fertilization. We measured the juveniles’ cognitive abilities and boldness. We found no effects of treatments on behavior.

Tableau d’honneur de la Faculté des études supérieures et postdoctorales, 2007-2008. / OBJECTIF : L’étomidate, un agent inducteur très utilisé pour l’intubation des traumatisés cranio-cérébraux (TCC), soulève des débats par rapport à son effet néfaste sur la fonction surrénalienne. Cette étude visait à évaluer le risque et la durée de l’insuffisance surrénalienne relative (ISR) induite par l’étomidate chez les TCC. Secondairement, elle visait à évaluer son effet sur la mortalité et la morbidité. MÉTHODES : Cette étude d’observation a adopté un devis longitudinal prospectif. Les sujets admissibles étaient tous des TCC modérés et sévères intubés, âgés de 16 ans ou plus, qui ont été admis à l'Hôpital de l'Enfant-Jésus entre août 2003 et novembre 2004. Un test de stimulation à l’ACTH (250 mcg) a été effectué 24, 48 et 168 heures après l’intubation. L’ISR a été définie par une augmentation de la cortisolémie une heure après l’ACTH (delta cortisol) de moins de 248,4 nmol/l. Des analyses de régression logistique et linéaire ont été effectuées afin de mesurer la force de l'association entre l’usage de l'étomidate et le risque d'ISR, la mortalité et les mesures de morbidité. RÉSULTATS : Parmi les 94 sujets admissibles à l’étude, 40 ont subi des tests à l’ACTH. Parmi ces 40 sujets, 15 ont reçu l’étomidate et 25 ont reçu un autre agent inducteur. À 24 heures, il n’y avait pas d’augmentation d’ISR associée à l’étomidate. Par contre, l’étomidate était associé à un delta cortisol plus faible (moyenne ajustée : 305,1 nmol/l, IC 95 % [214,7-384,8] versus 500,5 nmol/l, IC 95 % [441,8-565,7]; p=0,02). À 48 et à 168 heures, cette différence disparaissait. Pour tous les patients admissibles (n=94), l’étomidate était associé à un risque non statistiquement significatif de mortalité plus élevé (RC ajusté : 4,8, IC 95 % [0,6-35,9]). Par contre, il était associé à une hausse significative du risque de pneumonie (RC ajusté : 3,0, IC 95 % [1,0-8,7]; p=0,04). La durée moyenne ajustée de séjour aux soins intensifs n’était pas différente (10,2 jours pour l’étomidate versus 10,8 jours pour les autres agents). Au congé, le score moteur ajusté de la mesure d’indépendance fonctionnelle (MIF) était plus bas pour les sujets exposés à l’étomidate (32 versus 56, p=0,002), mais le score cognitif ajusté n’était pas différent entre les groupes (35 versus 46, p=0,15). CONCLUSION : L’étomidate inhibe la production de cortisol suite à une stimulation à l’ACTH jusqu’à 24 heures après une dose unique utilisée pour intuber des TCC modérés et sévères. Son impact sur la mortalité et sur la morbidité reste à préciser à l’aide d’un essai clinique randomisé et contrôlé. / INTRODUCTION: Etomidate is one of the most frequently used anesthetic induction agents for intubating patients with traumatic brain injury (TBI), although the clinical impacts of its adverse effects on adrenal function are debated. Therefore, it is important to assess the consequences of any adrenal suppression that could result from its use in patients with TBI. OBJECTIVE: The primary objective of this study was to determine the risk and the length of relative adrenal insufficiency (RAI) induced by etomidate in patients intubated for moderate and severe TBI. The secondary objective was to determine etomidate’s impact on mortality and morbidity. METHODS: This was a prospective cohort study. Eligible participants were moderate to severe TBI victims aged 16 years and over, intubated and admitted to a tertiary neurosurgical reference center between August 2003 and November 2004. ACTH stimulation tests (250 mcg) were performed on participants 24, 48 and 168 hours after intubation. RAI was defined as an increased of serum cortisol one hour post ACTH (delta cortisol) of less than 248.4 nmol/L (9 mcg/dl). Logistic and linear regression models assessed the association between the exposure to etomidate and the risk of RAI. RESULTS: Of the 94 subjects eligible to participate, 40 underwent ACTH testing. Fifteen subjects received etomidate and 25 received other induction agents. At 24 hours, etomidate did not change the risk of RAI. However, etomidate decreased the delta cortisol (adjusted mean: 305.1 nmol/l, 95% CI [214.7-384.8] vs. 500.5 nmol/l, 95% CI [441.8-565.7], p=0.02). At 48 and 168 hours, this difference disappeared. For all eligible subjects (n=94), there was a non significant trend for increased mortality in the etomidate group (adjusted OR: 4.8, 95% CI [0.6-35.9]). Etomidate was however associated with a significant increased risk of pneumonia (adjusted OR: 3.0, 95% CI [1.0-8.7]; p=0.04). The adjusted length of stay in the intensive care unit was not different (10.2 days for etomidate versus 10.8 days for the other agents). At discharge, the adjusted motor Functional Independence Measure score was significantly lower for subjects in the etomidate group (32 versus 56, p=0.002), but the adjusted cognitive score was not significantly different (35 versus 46, p=0.15). CONCLUSION: Etomidate decreases the adrenal response to an ACTH test up to 24 hours after a single dose used for the intubation of TBI victims. A large randomized controlled trial is needed to further assess its impact on morbidity and mortality.

Call number: LD2668 .T4 1985 P34 / Master of Science

Background: Trauma is a major cause of death and disability. Adverse events, such as prolonged prehospital time, hypoxia, hypotension and/or hyperventilation have been reported to correlate to poor outcome. Adequate cortisol levels are essential for survival after major trauma. In hypotensive critically ill patients, lack of sufficient amount of cortisol can be suspected, and a concept of critical illness related corticosteroid insufficiency has been proposed. Corticosteroid therapy has many adverse effects in critically ill patients and should only be given if life-saving. Correct measurement of serum cortisol levels is important but difficult in critically ill patients with capillary leakage. Estimation of the free and biologically active cortisol is preferable. In serum less than 10% of cortisol is free and biologically active and not possible to measure with routine laboratory methods. Salivary cortisol can be used as a surrogate for free cortisol, but salivary production is reduced in critically ill patients. Liver resection could reduce cortisol levels due to substrate deficiency. Aims: 1. Evaluate the occurrence of early adverse events in patients with traumatic brain injury and relate them to outcome. 2. Assess cortisol levels over time after trauma and correlate to severity of trauma, sedative/analgesic drugs and cardiovascular function. 3. Evaluate if saliva stimulation could be performed without interfering with salivary cortisol levels. 4. Assess cortisol levels over time after liver resection in comparison to other major surgery. Results: There was no significant correlation between prehospital time ³60 minutes, hypoxia (saturation <95%), hypotension (systolic blood pressure <90 mmHg), or hyperventilation (ETCO2 <4.5 kPa) and a poor outcome (Glasgow Outcome Scale 1-3) in patients with traumatic brain injury. Cortisol levels decreased significantly over time after trauma, but there was no correlation between low (<200 nmol/L) serum cortisol levels and severity of trauma. Infusion of sedative/analgesic drugs was the strongest predictor for a low (<200 nmol/L) serum cortisol. The odds ratio for low serum cortisol levels (<200 nmol/L) was 8.0 for patients receiving continuous infusion of sedative/analgesic drugs. There was no significant difference between unstimulated and stimulated salivary cortisol levels (p=0.06) in healthy volunteers. Liver resection was not associated with significantly lower cortisol levels compared to other major surgery. Conclusion: There was no significant correlation between early adverse events and outcome in patients with traumatic brain injury. Cortisol levels decreased significantly over time in trauma patients. Low cortisol levels (<200 nmol/L) were significantly correlated to continuous infusion of sedative/analgesic drugs. Saliva stimulation could be performed without interfering with salivary cortisol levels. Liver resection was not associated with low cortisol levels compared to other major surgery.

In einer prospektiven, randomisierten, doppelblinden, Placebo-kontrollierten Cross-over-Studie wurden hämodynamische und immunologische Effekte einer dreitägigen adjunktiven Therapie mit niedrig dosiertem Hydrocortison (HC) (100 mg Bolus + 10 mg/Stunde) bei 40 Patienten im septischen Schock untersucht. Die Therapie mit HC führte zum Anstieg des mittleren arteriellen Drucks und des systemischen Gefäßwiderstands sowie zur Reduktion des Herzzeitvolumens und der Herzfrequenz, die pulmonalvaskulären Widerstände blieben unverändert. Die Nitrit/Nitrat-Plasmaspiegel (Stickstoffmonoxid-Synthese) und der Katecholaminverbrauch nahmen ab. Die Immunreaktionen waren komplex: Abnahme proinflammatorischer (Interleukin-(IL)-6, 8) und antiinflammatorischer (IL-10, lösliche Tumor-Nekrosefaktor-Rezeptoren) Mediatoren, Anstieg proinflammatorischer Zytokine (IL-12 und Interferon-?), Reduktion der Endothel- (E-Selektin) und Granulozytenaktivierung (CD11b, CD64), Reduktion der T-Helfer- und Suppressorzellzahl und der eosinophilen und basophilen Granulozyen, die Monozytenzahl stieg an und die neutrophilen Granulozyten sowie die Gesamtleukozytenzahl blieben unverändert. Parameter der unspezifischen (Respiratory Burst, Phagozytose) und der spezifischen Immunreaktion (HLA-DR auf Monozyten, Antigenpräsentation) wurden nicht oder nicht wesentlich supprimiert, die Phagozytosefähigkeit von Monozyten nahm zu. Eine Beendigung der HC-Therapie führte zu ausgeprägten hämodynamischen und immunologischen Rebound-Phänomenen. Die Wirkung von niedrig dosiertem HC im septischen Schock kann daher als kreislaufstabilisierend und immunmodulatorisch charakterisiert werden, Zeichen einer ausgeprägten Immunsuppression fanden sich nicht. / In a prospective, double-blind, randomised, placebo-controlled cross-over study, hemodynamic and immune effects of a three-day adjunctive treatment with low doses of hydrocortisone (HC) (100 mg bolus followed by 10 mg per hour) were investigated in forty patients with septic shock. HC-therapy induced a rise of mean arterial pressure and systemic vascular resistance and a decline of cardiac index and heart rate without altering pulmonary vascular resistance. Both, nitrite/nitrate levels (nitric oxide formation) and cathecholamine requirement were reduced. Immune responses were complex and included: reduction of proinflammatory (interleukin-(IL)-6, 8) and antiinflammatory (IL-10, soluble tumor necrosis factor receptors) mediators, an increase of proinflammatory cytokines (IL-12 and interferon-?), a reduction of endothelial (E-selectin) and granulocyte activation (CD11b, CD64), and a decrease of T-helper and suppressor cells as well as eosinophil and basophil granulocytes; monocytes increased and total granulocyte and leukocyte counts remained unaltered. Parameters of innate (respiratory burst, phagocytosis) and adaptive immune responses (HLA-DR-expression on monocytes, antigen presentation) were not essentially affected, monocyte phagocytosis rather increased. HC-withdrawal induced marked hemodynamic and immunologic rebound effects. In conclusion, effects of low dose HC-therapy in septic shock is characterised by hemodynamic stabilisation and immunomodulation, without inducing severe immunosuppression.

A exodontia de terceiros molares pode afetar significativamente a qualidade de vida dos pacientes no período pós-operatório. A resposta inflamatória gerada pelo dano tecidual inerente ao procedimento pode apresentar edema e dor de duração e intensidade variáveis. No presente estudo foi avaliada a eficácia do corticosteroide hidrocortisona, como solução de irrigação durante exodontias de terceiros molares no controle do edema e da dor pós-operatória. Vinte e oito pacientes foram incluídos no estudo e após randomização foram submetidos a exodontia de ambos os terceiros molares inferiores em tempos cirúrgicos distintos, pelo mesmo cirurgião. Foram distribuídos aleatoriamente para receber 250ml de solução contendo 500mg de hidrocortisona (grupo H) ou 250ml de solução salina a 0,9% (grupo C). Para avaliação da eficácia da hidrocortisona no controle da dor pós-operatória três mensurações foram utilizadas: a quantidade de medicação analgésica de resgate consumida, o tempo entre o término da cirurgia e o uso do primeiro comprimido de resgate e a EVA em seis momentos. Para aferição da eficácia da hidrocortisona no controle do edema foram criadas 3 medidas faciais e o edema foi medido por meio da diferença em milímetros entre o segundo dia de pós-operatório e o pré-operatório. A comparação entre os grupos mostrou que a hidrocortisona foi eficaz no controle pós-operatório do edema em duas das medidas aferidas. A dor acompanhou a diminuição do edema, sendo que na média a EVA foi menor no grupo H, no entanto não houve diferença estatisticamente significativa nas outras duas aferições de dor. Conclui-se que a utilização local da hidrocortisona, como solução de irrigação trans-operatória em exodontias de terceiros molares é efetiva na redução do edema e da percepção de dor pós-operatória. / The third molar removal can significantly affect the quality of life of patients in the postoperative period. The inflammatory response caused by tissue damage inherent to the procedure may include edema and pain with variable duration and intensity. In the present study, we evaluated the efficacy of hydrocortisone as irrigation solution during third molars removal for postoperative pain and edema control. Twenty-eight patients were included in the study after randomization and were subjected to removal of both mandibular third molars in different surgical times by the same surgeon. They were randomly assigned to receive 250ml solution containing 500 mg of hydrocortisone (group H) or 250ml 0.9% saline solution (group C). To evaluate the efficacy of hydrocortisone in controlling postoperative pain three measurements were applied: the amount of analgesic medication taken, the time between the end of surgery and the use of the first analgesic tablet and VAS in six moments. To measure the effectiveness of hydrocortisone in edema control three facial measurements were created and swelling was measured by the difference in millimeters between the second postoperative day and preoperative. The comparison between the groups showed that hydrocortisone was effective in postoperative edema control in two of the measurements taken. Pain accompanied the edema reduction and the average VAS was lower in H group, however there was no statistically significant difference in the other two measurements of pain. We concluded that the local use of hydrocortisone as intraoperative irrigation solution for third molar removal is effective in reducing postoperative edema and pain perception.

Chronic inflammation is intimately linked with cancer development and progression and therefore reducing or eliminating inflammation represents a logical treatment and prevention strategy. Studies have shown that anti-inflammatory agents have anti-tumour effects in cancers, with reduced metastases and mortality. Current use of anti-inflammatory agents in the treatment and prevention of cancer is limited by their toxicity and side effects. The emerging field of nanotechnology allows the fundamental properties of a drug to be altered, creating a product with improved reactivity and bioavailability, leading to more targeted treatments and reduced dosage. In the present study, the genotoxic effects of three commonly used anti-inflammatory drugs; aspirin, ibuprofen and hydrocortisone, in their bulk and nano forms were evaluated on peripheral blood lymphocytes of healthy donors using the comet assay and the micronucleus assay. In order to determine any anti-cancer effects, these agents were also tested in peripheral blood lymphocytes in patients with haematological cancers. The glucocorticoid hydrocortisone was also evaluated for anti-oxidant capacity. Our results demonstrate that the nano versions of each drug produced a different response than the bulk counterpart, indicating that a reduction in particle size had an impact on the reactivity of the drug. Our results also indicate that the nano versions of each drug were less genotoxic than the bulk formulation, further emphasising the potential of nanoparticles as an improvement to current treatment options. We also found an anti-oxidant effect with hydrocortisone, with a more profound effect seen with the nano formulation.