Melanocytes in the developing and adult atrioventricular valves of the murine heart

The Neural Crest (NC) is a multipotential group of cells that arises from the dorsal aspect of the neural tube early in development. It is well established that a group of NC cells named Cardiac Neural Crest (CNC) migrates to the heart and plays a critical role in the remodeling of the aortic arch arteries and septation of the outflow tract. In this study, using the mouse mutant Pax3sp/sp that has CNC deficits I have identified a putative novel role for the CNC in regulating apoptosis in the atrioventricular (AV) endocardial cushion. The AV endocardial cushion undergoes remodeling to give rise to the cardiac AV valves. Using a transgenic mouse that carries the LacZ reporter gene under the control of the Dopachrome tautomerase promoter (Dct-LacZ), I found that another NC derived population, melanocyte precursors, also contribute to the AV endocardial cushion and developing AV valves. The analysis of Dct-LacZ embryos at different stages showed that NC cells already committed to the melanocytic fate migrate to the heart along the same initial pathway taken by those that will populate the skin. Hypopigmented mice carrying mutations in the Kit and Endothelin receptor b genes, that are critical for the proper development of skin melanocytes, do not have cardiac melanocytes indicating that cardiac and skin melanocyte precursors share the same initial signaling requirements. The analysis of murine adult hearts showed that melanocytes are mostly found in the atrial sides of the tricuspid and mitral valve leaflets. The distribution of melanocytes in the AV valves corresponds exactly to areas of high Versican B expression, a proteoglycan essential for the process of AV valve remodeling. To evaluate a potential role for melanocytes in the AV valves, a nanoindentation analysis of the tricuspid valves of wild type, hypopigmented and hyperpigmented mice was performed. The storage modulus, a measure of stiffness, for the leaflets obtained from hyperpigmented mice was considerably higher (10.5GPa) than that for the leaflets from wild type (7.5GPa) and hypopigmented animals (between 3.5 and 5.5 GPa) suggesting that melanocytes may contribute to the mechanical properties of the AV valves.

Durability Assessment of Polymer Trileaflet Heart Valves

The durability of a polymer trileaflet valve is dependent on leaflet stress concentrations, so valve designs that reduce stress can, hypothetically, increase durability. Design aspects that are believed to contribute to reduced leaflet stress include stent flexibility, parabolic coaptation curvature, and leaflet anisotropy. With this in mind, the purpose of this investigation was to elucidate what specific combinations of these parameters promote optimal acute and long-term valve function. A combination of four stent designs, seven leaflet reinforcement materials, and three coaptation geometries were evaluated through a combination of experimentation and modeling. Static tensile and Poisson’s ratio tests and dynamic tensile fatigue testing were used to evaluate the individual leaflet components; and hydrodynamic testing and accelerated valve fatigue was used to assess complete valve prototypes. The two most successful designs included a 0.40 mm thick knit-reinforced valve with a fatigue life of 10.35 years, and a 0.20 mm thick knit-reinforced valve with a 28.9 mmHg decrease in pressure drop over the former. A finite element model was incorporated to verify the impact of the above-mentioned parameters on leaflet stress concentrations. Leaflet anisotropy had a large impact on stress concentrations, and matching the circumferential modulus to that of the natural valve showed the greatest benefit. Varying the radial modulus had minimal impact. Varying coaptation geometry had no impact, but stent flexibility did have a marked effect on the stress at the top of the commissure, where a completely rigid stent resulted in a higher peak stress than a flexible stent (E = 385 MPa). In conclusion, stent flexibility and leaflet anisotropy do effect stress concentrations in the SIBS trileaflet valve, but coaptation geometry does not. Regions of high stress concentrations were linked to failure locations in vitro, so a fatigue prediction model was developed from the S/N curves generated during dynamic tensile testing of the 0.20 mm knit-reinforced leaflets. Failure was predicted at approximately 400 million cycles (10 years) at the top of the commissure. In vitro fatigue of this valve showed failure initiation after approximately 167 million cycles (4.18 years), but it was related to a design defect that is subsequently being changed.

Les valves sans suture dans le traitement de la sténose aortique sévère

Rapport présenté en vue de l’obtention du grade de M.Sc.A génie biomédical option génie clinique

Valves cardiaques par génie tissulaire - Simplification des essais et concept tubulaire

Les substituts valvulaires disponibles actuellement comportent encore plusieurs lacunes. La disponibilité restreinte des allogreffes, les risques de coagulation associés aux valves mécaniques et la durabilité limitée des bioprothèses en tissu animal sont toutes des problématiques que le génie tissulaire a le potentiel de surmonter. Avec la méthode d’auto-assemblage, le seul support des cellules consiste en leur propre matrice extracellulaire, permettant la fabrication d’un tissu entièrement libre de matériau exogène. Ce projet a été précédé par ceux des doctorantes Catherine Tremblay et Véronique Laterreur, ayant respectivement développé une méthode de fabrication de valves moulées par auto-assemblage et une nouvelle version de bioréacteur. Au cours de cette maîtrise, le nouveau bioréacteur a été adapté à une utilisation stérile avec des tissus vivants et la méthode de fabrication de valves moulées a été modifiée puis éprouvée avec la production de 4 prototypes. Ces derniers n’ont pas permis d’obtenir des performances satisfaisantes en bioréacteur, motivant la conception d’une nouvelle méthode. Plutôt que de tenter de répliquer la forme native des valves cardiaques, des études récentes ont suggéré une géométrie tubulaire. Cela permettrait une fabrication simplifiée, une implantation rapide, et un encombrement minimal en vue d’opérations percutanées. Cette approche minimaliste s’accorde bien avec la méthode d’auto-assemblage, qui a déjà été utilisée pour la production de vaisseaux de petits diamètres. Un total de 11 tubes ont été produits par l’enroulement de feuillets fibroblastiques auto-assemblés, puis ont été transférés sur des mandrins de diamètre inférieur, leur permettant de se contracter librement. La caractérisation de deux tubes contrôles a démontré que cette phase de précontraction était bénéfique pour les propriétés du tissu en plus de prévenir la contraction en bioréacteur. Les prototypes finaux pouvaient supporter un écoulement physiologique pulmonaire. Cette nouvelle méthode montre que le procédé d’auto-assemblage a le potentiel d’être utilisé pour fabriquer des valves cardiaques tubulaires.

Experimental and analytical investigation of refrigerant and lubricant migration

The off-cycle refrigerant mass migration has a direct influence on the on-cycle performance since compressor energy is necessary to redistribute the refrigerant mass. No studies, as of today, are available in the open literature which experimentally measured the lubricant migration within a refrigeration system during cycling or stop/start transients. Therefore, experimental procedures measuring the refrigerant and lubricant migration through the major components of a refrigeration system during stop/start transients were developed and implemented. Results identifying the underlying physics are presented. The refrigerant and lubricant migration of an R134a automotive A/C system-utilizing a fixed orifice tube, minichannel condenser, plate and fin evaporator, U-tube type accumulator and fixed displacement compressor-was measured across five sections divided by ball valves. Using the Quick-Closing Valve Technique (QCVT) combined with the Remove and Weigh Technique (RWT) using liquid nitrogen as the condensing agent resulted in a measurement uncertainty of 0.4 percent regarding the total refrigerant mass in the system. The determination of the lubricant mass distribution was achieved by employing three different techniques-Remove and Weigh, Mix and Sample, and Flushing. To employ the Mix and Sample Technique a device-called the Mix and Sample Device-was built. A method to separate the refrigerant and lubricant was developed with an accuracy-after separation-of 0.04 grams of refrigerant left in the lubricant. When applying the three techniques, the total amount of lubricant mass in the system was determined to within two percent. The combination of measurement results-infrared photography and high speed and real time videography-provide unprecedented insight into the mechanisms of refrigerant and lubricant migration during stop-start operation. During the compressor stop period, the primary refrigerant mass migration is caused by, and follows, the diminishing pressure difference across the expansion device. The secondary refrigerant migration is caused by a pressure gradient as a result of thermal nonequilibrium within the system and causes only vapor phase refrigerant migration. Lubricant migration is proportional to the refrigerant mass during the primary refrigerant mass migration. During the secondary refrigerant mass migration lubricant is not migrating. The start-up refrigerant mass migration is caused by an imbalance of the refrigerant mass flow rates across the compressor and expansion device. The higher compressor refrigerant mass flow rate was a result of the entrainment of foam into the U-tube of the accumulator. The lubricant mass migration during the start-up was not proportional to the refrigerant mass migration. The presence of water condensate on the evaporator affected the refrigerant mass migration during the compressor stop period. Caused by an evaporative cooling effect the evaporator held 56 percent of the total refrigerant mass in the system after three minutes of compressor stop time-compared to 25 percent when no water condensate was present on the evaporator coil. Foam entrainment led to a faster lubricant and refrigerant mass migration out of the accumulator than liquid entrainment through the hole at the bottom of the U-tube. The latter was observed for when water condensate was present on the evaporator coil because-as a result of the higher amount of refrigerant mass in the evaporator before start-up-the entrainment of foam into the U-tube of the accumulator ceased before the steady state refrigerant mass distribution was reached.

Analysis of the moisture effect on the mechanical behaviour of an adhesively bonded joint under proportional multi-axial loads

The objective of the study is to identify the 3D behaviour of an adhesive in an assembly, and to take into account the effect of ageing in a marine environment. To that end, three different tests were employed. Gravimetric analyses were used to determine the water diffusion kinetics in the adhesive. Bulk tensile tests were performed to highlight the effects of humid ageing on the adhesive behaviour. Modified Arcan tests were performed for several ageing times to obtain the experimental database which was necessary to identify constitutive models. A Mahnken-Schlimmer type model was determined for the unaged state according to a procedure developed in a previous study. This identification used inverse techniques. It was based on the unaged modified Arcan results and on a coupling between an optimisation routine and finite-element analysis. Then, a global inverse identification procedure was developed. Its aim was to relate the unaged parameters to the moisture concentration and overcome the difficulties usually associated with ageing of bonded assemblies in a humid environment: a non-uniformity of the stress state and a gradient of mechanical properties in the adhesive. This procedure was similar to the one used in the first part but needed modified Arcan results for several ageing times. It also required an initial assumption for the evolution of the Mahnken-Schlimmer parameters with the moisture concentration.

Les valves sans suture dans le traitement de la sténose aortique sévère

Rapport présenté en vue de l’obtention du grade de M.Sc.A génie biomédical option génie clinique

The Value Of The 2005 International Society Of Urological Pathology (isup) Modified Gleason Grading System As A Predictor Of Biochemical Recurrence After Radical Prostatectomy.

To compare time and risk to biochemical recurrence (BR) after radical prostatectomy of two chronologically different groups of patients using the standard and the modified Gleason system (MGS). Cohort 1 comprised biopsies of 197 patients graded according to the standard Gleason system (SGS) in the period 1997/2004, and cohort 2, 176 biopsies graded according to the modified system in the period 2005/2011. Time to BR was analyzed with the Kaplan-Meier product-limit analysis and prediction of shorter time to recurrence using univariate and multivariate Cox proportional hazards model. Patients in cohort 2 reflected time-related changes: striking increase in clinical stage T1c, systematic use of extended biopsies, and lower percentage of total length of cancer in millimeter in all cores. The MGS used in cohort 2 showed fewer biopsies with Gleason score ≤ 6 and more biopsies of the intermediate Gleason score 7. Time to BR using the Kaplan-Meier curves showed statistical significance using the MGS in cohort 2, but not the SGS in cohort 1. Only the MGS predicted shorter time to BR on univariate analysis and on multivariate analysis was an independent predictor. The results favor that the 2005 International Society of Urological Pathology modified system is a refinement of the Gleason grading and valuable for contemporary clinical practice.

Cd8+ Tumour-infiltrating Lymphocytes And Cox2 Expression May Predict Relapse In Differentiated Thyroid Cancer.

There is an increasing rate of papillary thyroid carcinomas that may never progress to cause symptoms or death. Predicting outcome and determining tumour aggressiveness could help diminish the number of patients submitted to aggressive treatments. We aimed to evaluate whether markers of the immune system response and of tumour-associated inflammation could predict outcome of differentiated thyroid cancer (DTC) patients. Retrospective cohort study. We studied 399 consecutive patients, including 325 papillary and 74 follicular thyroid carcinomas. Immune cell markers were evaluated using immunohistochemistry, including tumour-associated macrophages (CD68) and subsets of tumour-infiltrating lymphocytes (TIL), such as CD3, CD4, CD8, CD16, CD20, CD45RO, GRANZYME B, CD69 and CD25. We also investigated the expression of cyclooxygenase 2 (COX2) in tumour cells and the presence of concurrent lymphocytic infiltration characterizing chronic thyroiditis. Concurrent lymphocytic infiltration characterizing chronic thyroiditis was observed in 29% of the cases. Among all the immunological parameters evaluated, only the enrichment of CD8+ lymphocytes (P = 0·001) and expression of COX2 (P =0·01) were associated with recurrence. A multivariate model analysis identified CD8+ TIL/COX2 as independent risk factor for recurrence. A multivariate analysis using Cox's proportional-hazards model adjusted for the presence of concurrent chronic thyroiditis demonstrated that the presence of concurrent chronic thyroiditis had no effect on prognostic prediction mediated by CD8+ TIL and COX2. In conclusion, we suggest the use of a relatively simple pathology tool to help select cases that may benefit of a more aggressive approach sparing the majority of patients from unnecessary procedures.

Obesity Versus Osteoarthritis: Beyond The Mechanical Overload.

Obesity is currently considered a major public health problem in the world, already reaching epidemic characteristics, according to the World Health Organization. Excess weight is the major risk factor associated with various diseases, such as type 2 diabetes mellitus, hypertension, dyslipidemia and osteometabolic diseases, including osteoporosis and osteoarthritis. Osteoarthritis is the most prevalent rheumatic disease and the leading cause of physical disability and reduced quality of life of the population over 65 years. It mainly involves the joints that bear weight - knees and hips. However, along with the cases of obesity, its prevalence is increasing, and even in other joints, such as hands. Thus, it is assumed that the influence of obesity on the development of OA is beyond mechanical overload. The purpose of this review was to correlate the possible mechanisms underlying the genesis and development of these two diseases. Increased fat mass is directly proportional to excessive consumption of saturated fatty acids, responsible for systemic low-grade inflammation condition and insulin and leptin resistance. At high levels, leptin assumes inflammatory characteristics and acts in the articular cartilage, triggering the inflammatory process and changing homeostasis this tissue with consequent degeneration. We conclude that obesity is a risk factor for osteoarthritis and that physical activity and changes in diet composition can reverse the inflammatory and leptin resistance, reducing progression or preventing the onset of osteoarthritis.