Utilisation de la spectroscopie de fluorescence pour la vérification du nettoyage d'un ingrédient pharmaceutique actif sur les surfaces des équipements de production

La vérification de nettoyage des équipements pharmaceutiques permet de déterminer si la concentration du résidu sur la surface est inférieure à une limite acceptable afin de limiter la contamination croisée. La façon de procéder est de prélever un échantillon sur la surface par écouvillon et de l’analyser par chromatographie liquide à haute performance. Le problème avec cette approche est qu’il peut s’écouler jusqu’à 2 jours avant que le résultat soit obtenu et, durant ce temps, l’équipement nettoyé ne peut être libéré pour produire un autre lot de médicaments. Il existe une opportunité pour le développement d’une nouvelle méthode analytique permettant la quantification en temps réel et sans prise d’échantillon. La nouvelle méthode évaluée utilise la fluorescence ciblée pour la quantification directe de l’ibuprofène. Dans cette preuve de concept, les différents paramètres pouvant influencer le signal sont évalués. La façon de préparer les standards pour l’étalonnage de l’appareil (TraC) est déterminée ainsi que la façon de les analyser. Une première courbe d’étalonnage pour des surfaces en acier inoxydable est validée lors d’une collecte de données in situ et il est déterminé qu’il est impossible d’évaluer la propreté de certains équipements dû à la limite de quantification du TraC et que le TraC possède un biais positif par rapport à la méthode traditionnelle. Suite à cette étude, l’appareil a subi des modifications, une nouvelle courbe d’étalonnage est réalisée et la limite de quantification est réduite. Il est déterminé que la méthode est exacte (pourcentage de récupération situé entre 85 et 115%), précise (écart-type relatif inférieur à 5%) et que les excipients testés n’ont aucun impact sur le signal. L’étalonnage sur l’aluminium, le bronze, le polypropylène et le laiton est réalisé. La preuve de concept pour l’utilisation du TraC pour la quantification de l’ibuprofène est démontrée et il est recommandé d’implanter la méthode dans une première usine.Cleaning verification is used to assess the cleanliness of pharmaceutical process equipment. Cleanliness is established when the concentration of the residue of a pharmaceutical ingredient is lower than an acceptance limit. The method used to perform this verification consists of collecting a sample using a swab, and to analyze it using high performance liquid chromatography. The issue with this method is that it can take up to 2 days before the results are obtained, and before the equipment can be released for the production of another batch of drugs. Therefore, there is an opportunity to develop a new analytical method to quantify cleanliness in real-time without sampling (contactless). The new method investigated in this thesis uses fluorescence for direct quantification of ibuprofen residue. In this proof of concept, the impact of various parameters on the fluorescence signal was assessed. The best way to prepare the standards and how to analyze them with the instrument (the TraC) were determined. Following the verification of a first calibration curve using in situ data collection, the equipment was modified to allow the evaluation of equipment with an acceptance limit lower than the quantification limit of the TraC. The plant tests also revealed that the TraC always gives a higher concentration than the traditional method. An upgraded unit was calibrated, and the limit of quantification was lowered. It was established that the method is accurate (recovery is between 85 and 115%) and precise (relative standard deviation is lower than 5%), and that the excipients studied have no impact on the signal. Calibration curves were also built for aluminum, bronze, polyethylene and brass. Finally, the proof of concept for the use of the TraC for the cleaning verification of ibuprofen was demonstrated, and it is recommended to implement the method at a first plant

Specific Humoral Immunity versus Polyclonal B Cell Activation in Trypanosoma cruzi Infection of Susceptible and Resistant Mice

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, affects 10–12 million people in Latin America. Patent parasitemia develops during acute disease. During this phase, polyclonal B cell activation has been reported to generate high levels of serum antibody with low parasite specificity, and delayed protective humoral immunity, which is necessary to prevent the host from succumbing to infection. In this manuscript, data show that relatively resistant mice have improved parasite-specific humoral immunity and decreased polyclonal B cell activation compared to susceptible mice. Parasite-specific humoral immunity was associated with differential expansion of B cell subsets and T cells in the spleen, as well as with increased Th1 and decreased Th2 cytokine production. These data suggest that host susceptibility/genetic biases impact the development of humoral responses to infection. Th2 cytokines are generally associated with improved antibody responses. In the context of T. cruzi infection of susceptible mice, Th2 cytokines were associated with increased total antibody production concomitant with delayed pathogen-specific humoral immunity. This study highlights the need to consider the effect of host biases when investigating humoral immunity to any pathogen that has reported polyclonal B cell activation during infection

Estrogens, age, and, neonatal stress: panic disorders and novel views on the contribution of non-medullary structures to respiratory control and CO2 responses

CO2 is a fundamental component of living matter. This chemical signal requires close monitoring to ensure proper match between metabolic production and elimination by lung ventilation. Besides ventilatory adjustments, CO2 can also trigger innate behavioral and physiological responses associated with fear and escape but the changes in brain CO2/pH required to induce ventilatory adjustments are generally lower than those evoking fear and escape. However, for patients suffering from panic disorder (PD), the thresholds for CO2-evoked hyperventilation, fear and escape are reduced and the magnitude of those reactions are excessive. To explain these clinical observations, Klein proposed the false suffocation alarm hypothesis which states that many spontaneous panics occur when the brain’s suffocation monitor erroneously signals a lack of useful air, thereby maladaptively triggering an evolved suffocation alarm system. After 30 years of basic and clinical research, it is now well established that anomalies in respiratory control (including the CO2 sensing system) are key to PD. Here, we explore how a stress-related affective disorder such as PD can disrupt respiratory control. We discuss rodent models of PD as the concepts emerging from this research has influenced our comprehension of the CO2 chemosensitivity network, especially structure that are not located in the medulla, and how factors such as stress and biological sex modulate its functionality. Thus, elucidating why hormonal fluctuations can lead to excessive responsiveness to CO2 offers a unique opportunity to gain insights into the neuroendocrine mechanisms regulating this key aspect of respiratory control and the pathophysiology of respiratory manifestations of PD

Patient and work flow and costs associated with staff time and facility usage at a comprehensive cancer centre in Quebec, Canada – a time and motion study

Abstract Background Mapping patient and work flow and cost analysis studies can help determine the most efficient and cost effective way of providing health services while still maintaining the best standards of care. This study used both time and motion methodology and hospital data to assess the contribution of staff time and facility usage to the overall cost of cancer care during patient visits to a comprehensive cancer centre in Quebec, using metastatic colorectal cancer as a model. Methods A workflow diagram was created mapping direct and indirect steps involved during a patient’s physician or treatment (FOLFOX/bevacizumab or XELOX/bevacizumab) visit. Staff were timed as they performed each task and this data together with compensation amounts were used to calculate personnel costs. Mean work times and 95% confidence intervals (CI) were calculated. Operation and maintenance (O&M) costs for the Centre were calculated using information from hospital databases. All costs were presented in constant Canadian dollars for the 2010–2011 fiscal year period. Results For physician visits, direct and indirect personnel costs were $9.25 (95$7.00-11.51) and O&M costs were 60.21, for a total of $69.46 (95$67.21-71.72). For treatment visits, personnel and O&M costs were 71.91 (95%CI:$45.53-$98.29) and $62.00 respectively for a total of$133.91 (95%CI:$107.53-$160.29). When calculated for treatment alone, the total cost was $136.06 (95$109.16-$162.95) for FOLFOX/bevacizumab and$119.94 (95%CI:$96.89-$142.99) for XELOX/bevacizumab. The highest cumulative personnel costs were for the pharmacists and nurses ($38.87 and$34.82 respectively). Regarding patient flow, total time in between steps was 77.6 and 49.5 minutes for a physician or treatment visit respectively. Conclusions This study from a health care provider’s perspective, demonstrated that in the context of increasingly expensive therapies, costs associated with staff time and facility usage do not contribute greatly to the overall cost of treating cancer at this cancer centre. It also illustrated the need for improvements in patient and work flow to reduce wait times in the clinic.</p

Connaissances et compétences des pharmaciens communautaires et qualité de la pharmacothérapie des patients atteints d’insuffisance rénale chronique : Résultats provisoires du programme ProFiL

Résumé Objectif : Dans le cadre d’une analyse provisoire comprenant l’Hôpital de la Cité-de-la Santé de Laval et l’Hôpital Maisonneuve- Rosemont, évaluer les retombées d’un programme de formation et de liaison en néphrologie (ProFiL) sur les connaissances et compétences des pharmaciens, leurs pratiques cliniques et la qualité de la pharmacothérapie de leurs patients. Méthodologie : Des patients de cliniques de prédialyse et leur pharmacie ont été répartis aléatoirement dans les groupes ProFiL et Soins habituels. Les connaissances et compétences des pharmaciens ont été évaluées au moment de l’inclusion des patients dans l’étude et après une année. La qualité de la pharmacothérapie et les pratiques cliniques des pharmaciens ont été évaluées au cours des années précédant et suivant le recrutement. Résultats : Après un an de suivi, les pharmaciens ProFiL (n = 55) ont obtenu une amélioration plus importante de leurs scores de connaissances (différence avant-après entre les groupes : 5,6 % [IC 95 % : 0,08 % à 11,1 %]) et de compétences (10,8 % [5,5 % à 16,1 %]) que les pharmaciens du groupe Soins habituels (n = 27). Le nombre de problèmes liés à la pharmacothérapie des patients ProFiL (n = 117) est passé de 3,2 à 1,7 problèmes par patient, comparativement au groupe Soins habituels, qui n’est passé que de 2,6 à 2,1 par patient (n = 51), soit une diminution incrémentale de -1,1 problème par patient (-1,8 à -0,4), notamment pour les médicaments nécessitant un ajustement en insuffisance rénale chronique (-0,3 problème lié à la pharmacothérapie par patient [-0,5 à -0,02]) et l’adhésion au traitement (-0,5 problème lié à la pharmacothérapie par patient [-0,9 à -0,2]). Un nombre supérieur de recommandations aux patients et aux médecins, nommées opinions pharmaceutiques, et de refus d’exécuter une ordonnance ont été émis pour les patients ProFiL (0,4 opinion par patient versus 0,1 opinion par patient). Conclusion : Après une année, ProFiL améliore les connaissances et compétences des pharmaciens communautaires, leurs pratiques cliniques et la qualité de la pharmacothérapie de leurs patients. Abstract Objective: To evaluate the impact of a training and liaison program in nephrology (ProFil) on the knowledge and competencies of pharmacists, their clinical practice, and the quality of their patient pharmacotherapy. Methods: Patients in predialysis clinics in two academic medical centers and their community pharmacy were randomly assigned either to the ProFiL group or the Usual Care group. The knowledge and competencies of pharmacists were evaluated at the time of patient inclusion in the study and after one year. The quality of pharmacotherapy and the clinical practice of pharmacists were evaluated prior to and following patient recruitment. Results: After one year of follow-up, pharmacists from the ProFiL group (n=55) had significant improvement in their scores for knowledge (before-after difference between groups: 5.6% [CI 95 %: 0.08 % to 11.1 %]) and competency (10.8 % [5.5 % to 16.1 %]) than pharmacists in the Usual Care group (n=27). The number of drug-related problems in the ProFiL group patients (n=117) decreased from 3.2 to 1.7 problems per patient, compared to the patients in the Usual Care group where drug-related problems decreased from 2.6 to 2.1 per patient (n=51), an incremental decrease of -1.1 problems per patient (-1.8 to -0.4), primarily for medications requiring dosage adjustment in chronic renal failure (-0.3 drug-related problems per patient [-0.5 to -0.02]) and treatment adherence (-0.5 drug-related problems [-0.9 to -0.2]). A greater number of recommendations to patients and physicians, either as pharmaceutical opinions or refusal to fill a prescription, were made for patients in the ProFiL group (0.4 opinions per patient versus 0.1 opinions per patient). Conclusion: After one year, the ProFil program improved the knowledge and competencies of community pharmacists, their clinical practice, and the quality of patient pharmacotherapy. Key words: Chronic renal failure, community pharmacy, continuing education, drug-related problems, knowledge and competencies, pre-dialysis clinic

Aggravation de l’insuffisance rénale chronique et de ses facteurs de risque : Résultats provisoires du programme ProFiL

Résumé Objectif : Un essai clinique evalue presentement les retombees d’un Programme de formation et de liaison en nephrologie (ProFiL) sur la qualite de la pharmacotherapie de patients souffrant d’insuffisance renale chronique. L’objectif de cette analyse provisoire vise a evaluer la pertinence d’augmenter le nombre de patients a l’etude de maniere a pouvoir determiner les retombees du Programme sur l’aggravation de la fonction renale et de ses facteurs de risque. Méthodologie : Des patients de deux cliniques de predialyse (Hopital de la Cite-de-la-Sante de Laval et Hopital Maisonneuve- Rosemont) et leur pharmacie communautaire ont ete repartis aleatoirement dans les groupes ProFiL et Soins habituels et suivis pendant une annee. Les variables cliniques ont ete documentees au moment de l’inclusion des patients dans l’etude et un an plus tard. La comparaison de l’evolution des deux groupes a eu lieu a l’aide de modeles lineaires multivaries a effet mixte. Résultats : L’evolution du debit de filtration glomerulaire etait similaire dans les deux groupes. Une baisse additionnelle de -9,8 mmHg (IC 95 % : -15,8 a -3,7) de la tension arterielle systolique a ete observee dans le groupe ProFiL (n = 117) par rapport aux patients Soins habituels (n = 51). Parmi les patients diabetiques (n = 100), une reduction incrementale de l’hemoglobine glyquee de 0,4 % (IC 95 % : -0,9 a -0,1) a ete observee dans le groupe ProFiL alors que parmi les patients dyslipidemiques (n = 96), l’evolution du cholesterol-LDL etait similaire dans les deux groupes. Conclusion : Apres une annee, le Programme ProFiL a un effet positif sur le controle de la tension arterielle et de l’hemoglobine glyquee mais n’aurait pas d’effets significatifs sur le cholesterol-LDL et l’aggravation de l’insuffisance renale. Il n’est donc pas necessaire d’augmenter le nombre de patients recrutes. Abstract Objective: : A clinical trial is presently assessing the impact of a nephrology training and liaison program “ProFiL” on the quality of pharmacotherapy in patients with chronic renal failure. The objective of this interim analysis is to determine the usefulness of increasing the number of study patients to be able to assess the program’s impact on the progression of impaired renal function and of its risk factors. Methods: Patients from two predialysis clinics (Hopital de la Cite-de-la-Sante de Laval and Hopital Maisonneuve- Rosemont) and their community pharmacies were randomized to the ProFiL group or the usual care group and followed for one year. The clinical variables were recorded at study inclusion and one year later, and the changes in the two groups were compared using linear, mixed-effects multivariate models. Results: The changes in the glomerular filtration rate were similar in both groups. There was an additional decrease of -9.8 mm Hg (95% CI: -15.8 to -3.7) in systolic blood pressure in the ProFiL group (n = 117) compared to the patients receiving usual care (n = 51). In the diabetic patients (n = 100), an incremental reduction in the glycosylated hemoglobin concentration of 0.4% (95% CI: -0.9 to -0.1) was observed in the ProFiL group, while in the dyslipidemic patients (n = 96), the changes in the LDL cholesterol levels were similar in both groups. Conclusion: After one year, the ProFil program had a positive effect on blood pressure control and glycosylated hemoglobin control but did not appear to have a significant impact on LDL cholesterol or the progression of renal failure. Therefore, it is not necessary to increase the number of patients recruited. Key words: Chronic renal failure, community pharmacy, diabetes, dyslipidemia, predialysis, renal failure risk facto