Abstract Natriuretic peptide hormones secreted from the heart are important in maintaining the volume and electrolyte balance and in regulation of blood pressure. The secretion of natriuretic peptides is stimulated by myocyte stretch and paracrine factors. However, the intracellular actions of these stimuli and the cellular and molecular mechanisms involved in the processing and secretion of natriuretic peptides are still largely unknown. In this study, a new model for studies of the natriuretic peptide system was developed using a novel natriuretic peptide from salmon. Salmon (Salmo salar) maintains its water and salt homeostasis despite the volume gain in fresh water and electrolyte gain in sea water. Thus, salmon is an ideal model to study the mechanisms regulating the extracellular volume and salt balance, like natriuretic peptides. Furthermore, comparative studies revealing the common characteristics in phylogenetically distinct species suggest the importance of these factors in the regulation of the natriuretic peptide system. A novel natriuretic peptide, salmon cardiac peptide (sCP), was cloned from salmon heart. Distribution of sCP was studied in a variety of vertebrates and its physiological effects were examined in in vitro and in vivo experiments in salmon and rats. The storage and release of sCP was studied using a salmon ventricle perfusion system and by analysing the molecular forms of stored and secreted forms. Factors modulating the secretion and circulating concentration of sCP, and cardiac peptide and sCP mRNA level in salmon were examined as well. The biosynthesis of sCP is strictly restricted to the heart. sCP is stored in myocytes in the prohormone form, while the secreted form is a 29-amino acid peptide in salmon. Mechanical load on isolated salmon ventricle and volume overload in intact salmon induced a rapid release of sCP. Exposure to hyperosmotic environment decreased the plasma sCP level. sCP increased diuresis and natriuresis, as well as relaxed preconstricted arteries from salmon and rats. Thus the storage, processing and release of sCP resembles those of mammalian ANP. The circulating level of sCP in salmon was markedly upregulated at increased temperatures. Upregulation resulted from decreased elimination rather than increased secretion of sCP, providing the first direct evidence that elimination is used for the regulation of the natriuretic peptide system. In conclusion, sCP is a promising model for studying the general factors regulating the cardiac natriuretic peptides.

No description available.

The function of skeletal adaptation to mechanical load is to adjust the amount and distribution of bone tissue (geometry); such that stresses experienced within the bone are kept within certain physiological limits and fractures are prevented. Genetic, environmental or hormonal factors may cause heterogeneity in this adaptive response, altering geometry and consequently fragility. The purpose of this thesis was to explore the skeletal response to load in vivo, by evaluating stress at the hip under three different conditions: FRACTURE (Study 1), DIABETES (Study 2) and ESTROGEN deficiency (STUDY 3). We studied women 25 years of age or older who participated in the Canadian Multicentre Osteoporosis Study and had available Hip Structure Analysis (HSA) data from baseline dual energy x-ray absorptiometry (DXA) scans. Women were categorized according to fracture status (fracture or no fracture), diabetes status (diabetes or no diabetes) and estrogen use (current users or never users). We computed stress (megapascals=MPa) at the infero-medial margin of the femoral neck in a one-legged iii stance using a 2-D engineering beam analysis. We used linear regression to determine associations between femoral neck stress and each categorical variable. Study 1 (n=2168) demonstrated higher stresses in postmenopausal women with fractures compared to women without fractures (10.57 ± 2.19 vs. 10.30 ± 2.03 MPa; p=0.0031). Study 2 (n=3665) demonstrated higher stresses in women with Type 2 Diabetes Mellitus compared to non-diabetic women (10.98 ± 2.33 vs. 10.57 ± 2.20 MPa; p=0.0194). Study 3 (n=2447) demonstrated higher stresses in postmenopausal women not on estrogen than in premenopausal women (10.66 ± 2.14 vs. 10.09 ± 2.01 MPa; p<0.0001), but no differences in stresses between postmenopausal women on estrogen and premenopausal women (10.16 ± 2.00 vs. 10.09 ± 2.01 MPa; p=0.6102). Since stress is an indicator of underlying geometry, and geometry should be adapted to prevalent loads, higher stress indicates weaker geometry and suggests an impaired modeling response in these three conditions. Compromised modeling has important clinical implications in terms of treatment selection, as individuals with reduced load sensitivity may respond best to metabolic agents that would improve modeling responses to load stimuli.

The function of skeletal adaptation to mechanical load is to adjust the amount and distribution of bone tissue (geometry); such that stresses experienced within the bone are kept within certain physiological limits and fractures are prevented. Genetic, environmental or hormonal factors may cause heterogeneity in this adaptive response, altering geometry and consequently fragility. The purpose of this thesis was to explore the skeletal response to load in vivo, by evaluating stress at the hip under three different conditions: FRACTURE (Study 1), DIABETES (Study 2) and ESTROGEN deficiency (STUDY 3). We studied women 25 years of age or older who participated in the Canadian Multicentre Osteoporosis Study and had available Hip Structure Analysis (HSA) data from baseline dual energy x-ray absorptiometry (DXA) scans. Women were categorized according to fracture status (fracture or no fracture), diabetes status (diabetes or no diabetes) and estrogen use (current users or never users). We computed stress (megapascals=MPa) at the infero-medial margin of the femoral neck in a one-legged iii stance using a 2-D engineering beam analysis. We used linear regression to determine associations between femoral neck stress and each categorical variable. Study 1 (n=2168) demonstrated higher stresses in postmenopausal women with fractures compared to women without fractures (10.57 ± 2.19 vs. 10.30 ± 2.03 MPa; p=0.0031). Study 2 (n=3665) demonstrated higher stresses in women with Type 2 Diabetes Mellitus compared to non-diabetic women (10.98 ± 2.33 vs. 10.57 ± 2.20 MPa; p=0.0194). Study 3 (n=2447) demonstrated higher stresses in postmenopausal women not on estrogen than in premenopausal women (10.66 ± 2.14 vs. 10.09 ± 2.01 MPa; p<0.0001), but no differences in stresses between postmenopausal women on estrogen and premenopausal women (10.16 ± 2.00 vs. 10.09 ± 2.01 MPa; p=0.6102). Since stress is an indicator of underlying geometry, and geometry should be adapted to prevalent loads, higher stress indicates weaker geometry and suggests an impaired modeling response in these three conditions. Compromised modeling has important clinical implications in terms of treatment selection, as individuals with reduced load sensitivity may respond best to metabolic agents that would improve modeling responses to load stimuli.

Abstract The natriuretic peptides are a family of hormones secreted by the heart. They play a fundamental role in salt and water balance and blood pressure regulation. Atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) are the known members of the mammalian natriuretic peptide family. A major stimulus for the secretion of cardiac natriuretic peptides is myocyte stretch. Therefore, the secretion of natriuretic peptides is increased in response to elevated blood volume. Natriuretic peptide production and release is also affected by several other factors, such as endothelin-1 (ET-1), acting in paracrine fashion. The aim of this study was to elucidate factors regulating the novel cardiac peptide hormone, salmon cardiac peptide (sCP), belonging to the family of natriuretic peptides. The role of mechanical load, temperature and ET-1 in sCP secretion and production was studied using in vitro (isolated perfused ventricle preparation) and in vivo methods. Comparisons between the natriuretic peptide systems in fish and mammals were done to clarify functional evolution of this hormone family. Salmon (Salmo salar) was selected as a model, since it has an outstanding adaptability to wide variations in environmental salinity and has developed defence mechanisms against volume or salt load. The results showed that salmon ventricle stores large amounts of the prohormone of sCP, whereas the secreted form is the mature 29-amino acid form. The N-terminal fragment of pro-sCP is co-secreted with sCP in equimolar amounts. sCP is released rapidly in response to appropriate stimulus, whereas induction of its gene expression is slower. Mechanical load is an important regulator of sCP secretion. Temperature also plays a major role in regulating sCP plasma concentration by affecting its elimination from circulation. Additionally, ET-1 is a potent secretagogue of the sCP system and an inotropic agent in salmon heart. Furthermore, the present results reveal remarkable synergism between the cardiac effects of ET-1 and β-adrenergic stimulation. In conclusion, the sCP system in salmon ventricle largely resembles the ANP system in mammalian atrium, while also having specific characteristics, such as a regulated ventricular natriuretic peptide secretion pathway. Therefore, the sCP system offers a unique model for studying mechanisms of natriuretic peptide biology.

Tendon injuries are ubiquitous in the sporting and occupational environment. Clinically they present a challenge to Orthopaedic surgeons as they account for up to half of all sports injuries and almost half of reported work related ailments. The capacity for tendons to heal subsequent to injury is restricted due to their poor blood supply. Moreover, healed tendon tissue may be inferior to the intact tendon, having diminished biochemical and biomechanical properties and this brings about an ever increasing need for optimized treatment methods for tendon repair. Mechanobiology is concerned with how mechanical forces influence physiological and pathological aspects of the living tissue. However, the complex and poorly controlled loading environment in living organisms prevent the establishment of direct relationships between mechanical stimuli and tissue response. By developing a novel in vitro loading system (IVLS), the work in this thesis investigates the potential of a new and exciting biophysical loading intervention, High Frequency Low Magnitude (HFLM) mechanical loading, for stimulation of tendon repair and remodelling. Following a pre-defined stimulation period, healthy rat tail tendon fascicles (RTTFs) were evaluated for tissue viability and metabolism, Glycosaminoglycan (GAG) content, collagen arrangement and tangent modulus, using staining and biochemical assays, together with microscopy techniques, and mechanical testing. HFLM mechanically loaded tendons showed a trend for a higher tangent modulus than fresh tissue, and significantly higher modulus than unloaded. Further, when varying mechanical loading parameters of frequencies and dosages over clinically relevant ranges, a frequency dependent response was observed with increased tangent modulus and GAG content with increasing frequency. An association between high tendon crimp pattern and elevated tendon modulus as a result of HFLM mechanical loading was also demonstrated. Concomitantly, an injury model was developed to evaluate the effects of in vitro static, low frequency cyclic and HFLM mechanical loading conditions on the biochemical and biomechanical properties of in vitro damaged tendons. HFLM mechanically loaded damaged tendons again demonstrated significantly higher modulus and metabolism than unloaded tissue, although these were reduced below those of fresh damaged tissue. The findings in this thesis together with the newly developed IVLS reveal the potential for an exciting and unique biophysical therapeutic loading intervention for treatment of tendon injuries, and provide a scientific platform for further investigation of the effects of HFLM mechanical loads, potentially leading to an application within the clinic for enhanced connective tissue repair and remodelling.

[Truncated abstract] The extent to which an airway narrows is strongly influenced by mechanical loads on airway smooth muscle (ASM). This thesis considers both dynamic and static mechanical loads. Dynamic load describes the time varying load on airways produced by oscillatory breathing movements. Static load is that present at a fixed lung volume ie. without breathing. In the intact lung static load principally comprises the pressure across the airway wall, that is transmural pressure (Ptm), and elastic after-load arising from distortion of airway and lung tissue by the narrowing airway. The experiments performed in this thesis were designed to answer several outstanding questions relating to how dynamic and static loads regulate airway narrowing. Dynamic load from breathing movements cyclically stretches ASM, which produces a number of physiological and cellular effects. For example in ASM strips a period of cyclical stretch reduces subsequent ASM contraction. However the response of the whole airway to dynamic load may differ from isolated ASM where non-muscle tissue also contributes. The first aim of this thesis was to characterise the response of the whole airway to dynamic load and determine whether the airway wall modifies the effects produced by ASM length cycling. Static after-loads restrict ASM shortening providing a limit to airway narrowing. Two primary sources of airway wall load include cartilage and the mucosal membrane which contribute to airway compliance. The relative importance of cartilage and mucosa to airway wall compliance and airway narrowing is unclear. ... Results demonstrate that airway narrowing is restricted by Ptm but not by parenchymal elastic after-load. The major findings of this thesis are: (1) dynamic loads produced by breathing movements regulate airway responsiveness through cyclical airway expansion and elongation; (2) the reported effects of cyclical stretch on ASM contraction differs in situ 8 possibly due to modification by one or more biomechanical or physiological properties of the airway wall; (3) parenchymal elastic after-loads, previously thought to be important during bronchoconstriction, do not restrict airway narrowing. Given the absence of an effect of parenchymal elastic after-load on airway narrowing, the static mechanical load on ASM therefore comprises Ptm and airway wall stiffness, with important contributions from cartilage and mucosa depending on lung volume.

Η παρούσα διπλωματική εργασία πραγματεύεται την μελέτη, τον σχεδιασμό και την κατασκευή ενός ηλεκτρονικά ελεγχόμενου μηχανικού φορτίου. Η εργασία αυτή εκπονήθηκε στο Εργαστήριο Ηλεκτρομηχανικής Μετατροπής Ενέργειας του Τμήματος Ηλεκτρολόγων Μηχανικών και Τεχνολογίας Υπολογιστών της Πολυτεχνικής Σχολής του Πανεπιστημίου Πατρών. Σκοπός είναι η δημιουργία ενός μηχανικού φορτίου του οποίου το προφίλ μπορεί να ελέγχεται με την χρήση ψηφιακού μικροελεγκτή. Αρχικά, εξετάζεται η θέση που κατέχει το μηχανικό φορτίο σε ένα ηλεκτρικό κινητήριο σύστημα και παρουσιάζονται οι χαρακτηριστικές καμπύλες των διάφορων τύπων μηχανικών φορτίων. Στη συνέχεια, μελετώνται η βασική αρχή λειτουργίας, οι μαθηματικές εξισώσεις καθώς και τα κατασκευαστικά χαρακτηριστικά των μηχανών συνεχούς ρεύματος αλλά και της μηχανής που χρησιμοποιήθηκε στην κατασκευή. Στο επόμενο βήμα, γίνεται μια θεωρητική ανάλυση του κυκλώματος της πλήρους γέφυρας που κατασκευάστηκε καθώς και όλων των υπόλοιπων κυκλωμάτων και στοιχείων που είναι αναγκαία για την λειτουργία της. Επιπροσθέτως, αναλύεται η μέθοδος παλμοδότησης των διακοπτικών στοιχείων του μετατροπέα, που είναι η Διαμόρφωση Εύρους Παλμών (PWM) καθώς και τα ιδιαίτερα χαρακτηριστικά του μικροελεγτή που χρησιμοποιήθηκε. Για το συνολικό σύστημα, αποτελούμενο από την μηχανή συνεχούς ρεύματος και τον μετατροπέα τύπου πλήρους γέφυρας, δημιουργήθηκε ένα μαθηματικό μοντέλο στο περιβάλλον Simulink για να ελεγχθεί η συμπεριφορά του. Τέλος, αναλύονται τα τεχνικά χαρακτηριστικά όλων των κυκλωμάτων που κατασκευάστηκαν και παρατίθενται τα παλμογραφήματα που προέκυψαν από τα πειράματα που διενεργήθηκαν μετά την ολοκλήρωση της κατασκευής. / This thesis deals with the study, design and construction of an electronically controlled mechanical load. The work was conducted in the Laboratory of Electromechanical Energy Conversion, Department of Electrical and Computer Engineering School of Engineering, University of Patras. The aim is to create a mechanical load whose profile can be controlled using digital microcontroller. First, was considered the position of the mechanical load in an electric driving system and shows the characteristic curves of various types of mechanical loads. Then was studied the basic principle of operation, mathematical equations and the construction of DC machines and the machine used in construction. The next step is a theoretical analysis of the full bridge circuit that is constructed and all other circuits and components necessary for its operation. In addition, explains the method of interrupted pulses data converter, which is shaping pulse width (PWM) and the characteristics of icroconroler which is used. For the total system, consisting of the DC machine and type full bridge converter, a mathematical model in Simulink environment to verify its behavior. Finally, analyzing the technical characteristics of all circuits built out and the curves obtained from experiments carried out after the completion of construction.

Abstract Clarification of the mechanisms of cardiac-specific gene expression provides not only basic knowledge about how the gene expression is regulated in the heart, but also about the changes in the gene expression during the development of cardiovascular diseases. The purpose of this study was to analyze the mechanisms of cardiac chamber-specific gene expression and cardiac gene activation induced by mechanical load. In the present study, the experiments were carried out by using two cardiac genes, salmon cardiac peptide (sCP) and rat B-type natriuretic peptide (BNP) genes as models. sCP was discovered previously in our laboratory and turned out to be extremely cardiac-specific, representing A-type natriuretic peptide characters in an exaggerated way. In neonatal rat cardiomyocytes, the sCP promoter activity was shown to be strictly restricted to atrial cells and the promoter to be inert to cardiac hypertrophy-inducing factors. In order to find out the mechanisms of earlier proved BNP gene activation by mechanical load, BNP promoter activity was studied in vivo in adult rat hearts. The tandem GATA transcription factor binding site at position -80/-91 was shown to be essential for the BNP gene induction by angiotensin II. To clarify the possiblity to transfer the characters of the BNP gene into the sCP gene, short BNP fragments were inserted to the sCP gene promoter. The otherwise atrial-restricted sCP promoter was shown to be switched on in rat ventricular cardiomyocytes by adding a short BNP proximal promoter element to the sCP promoter, preferably near to the transcription start site. This activity was partly dependent on the -80/-91 GATA sites in the BNP promoter. Thus, A-type natriuretic peptide regulation can be switched to B-type regulation by a short proximal BNP promoter element. In conclusion, these studies reveal certain basic differences in cardiac atrial and ventricular gene expression.

One of the serious problems encountered in the use of various materials in technology is the occurrence of fatigue phenomena as an undesirable material damage under cyclic mechanical load. For aluminium alloys this issue is of extremely high importance in case of their utilisation for aircraft purposes, e.g., where a very wide spectrum of frequencies occur. Moreover, the cyclic loading may be joined by the presence of specific electrolyte media. Therefore, the material candidates must be thoroughly examined in view of their sensitivity to fatigue as well as to corrosion fatigue. Usually, the Cu-containing alloy EN-AW 2024 T3 is applied besides 7075 T6 in Airbus aircrafts, but the weldable alloy 6013 T6 is considered to be a potential alternative. Referring to former investigations on the environmental sensitivity (ES) in the fatigue behaviour /1-6/ this paper brings up experimental findings as well as expanded considerations about damaging mechanisms and modelling. The situation with the alloy 6013 T6 is emphasized. The propagation of cracks under cyclic load in different environments, such as vacuum, air or aqueous media, is described by means of fracture mechanics. This enables discrimination in view of the influence of environmental factors and, hence, the participation of corrosion processes.