No abstract

Glucocorticoids are vital for the structural and functional maturation of foetal organs, yet excessive foetal exposure is detrimental to adult cardiovascular health. To elucidate the role of glucocorticoid signalling in late-gestation cardiovascular maturation, we have generated mice with conditional disruption of glucocorticoid receptor (GR) in cardiomyocytes and vascular smooth muscle cells using smooth muscle protein 22-driven Cre recombinase (SMGRKO mice) and compared them with mice with global deficiency in GR (GR(-/-)). Echocardiography shows impaired heart function in both SMGRKO and GR(-/-) mice at embryonic day (E)17.5, associated with generalized oedema. Cardiac ultrastructure is markedly disrupted in both SMGRKO and GR(-/-) mice at E17.5, with short, disorganized myofibrils and cardiomyocytes that fail to align in the compact myocardium. Failure to induce critical genes involved in contractile function, calcium handling and energy metabolism underpins this common phenotype. However, although hearts of GR(-/-) mice are smaller, with 22% reduced ventricular volume at E17.5, SMGRKO hearts are normally sized. Moreover, while levels of mRNA encoding atrial natriuretic peptide are reduced in E17.5 GR(-/-) hearts, they are normal in foetal SMGRKO hearts. These data demonstrate that structural, functional and biochemical maturation of the foetal heart is dependent on glucocorticoid signalling within cardiomyocytes and vascular smooth muscle, though some aspects of heart maturation (size, ANP expression) are independent of GR at these key sites.

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Myocardial motion was quantified in normal cats (n = 25) and cats with hypertrophic cardiomyopathy (HCM) (n = 23) using the pulsed tissue Doppler imaging (TDI) technique. A physiologic nonuniformity was documented in the myocardial motion of normal cats, which was detected as higher early diastolic velocities, acceleration, and deceleration in the interventricular septum compared with the left ventricular free wall (LVFW). HCM cats exhibited lower early diastolic velocities, acceleration, and deceleration and also prolonged isovolumic relaxation time compared with normal cats. These differences were detected mainly along the longitudinal axis of the heart. A cutoff value of E' in the LVFW along the longitudinal axis >7.2 cm/s discriminated normal from HCM cats with a sensitivity of 92% and a specificity of 87%. The physiologic nonuniformity of myocardial motion during diastole was lost in affected cats. Systolic impairment (decreased late-systolic velocities in most segments along the longitudinal axis and decreased early systolic acceleration in both mitral annular sites) was evident in HCM cats irrespective of the presence of left ventricular outflow tract obstruction and congestive heart failure. Postsystolic thickening was recorded in the LVFW along the longitudinal axis only in affected cats (n = 6) and was another finding indicative of systolic impairment in the HCM of this species. This study identified both diastolic and systolic impairment in cats with HCM compared with normal cats. The study also documents the normal physiologic nonhomogeneity in myocardial motion in cats and the subsequent loss of this feature in the HCM diseased state.

The objective of this study was to determine the intraoperator, intraobserver, and interobserver repeatability in a series of conventional echocardiographic parameters and in some of the newer measurements of diastolic function, including color M-mode flow propagation velocity, isovolumic relaxation time and pulsed-wave Doppler tissue imaging velocities. Four healthy cats were each scanned five times over a 3-day period. The repeatability of these echocardiographic analyses was compared using Bland-Altman analysis (intraoperator repeatability). After a minimum of 5 weeks, one scan was randomly selected from each cat, and was remeasured by the original observer and the results compared using a standard paired Student's t-test (intraobserver repeatability). One scan from each cat was then randomly selected and two observers, with similar levels of experience, measured each of these scans. The repeatability of these echocardiographic analyses was compared using Bland-Altman analysis (interobserver repeatability). The conventional two-dimensional (2D), M-mode and spectral Doppler measurements were repeatable in both their acquisition and measurement by a single investigator; there was a greater degree of variation between the two observers. The predominant (S', E', and A') pulsed-wave Doppler tissue imaging velocities from the left apical four-chambered view, generally had a coefficient of variation of approximately 20% (range 9.62-34.08%). However, with pulsed-wave Doppler tissue imaging, velocities recorded during the isovolumic phases, the velocity of the tricuspid annulus, and the radial fiber velocity within the interventricular septum, frequently had coefficients of variation in excess of 20% and should therefore be interpreted with caution.

The nanomechanical properties of biocompatible thin-shell hollow polymer microspheres with approximately constant ratio of shell thickness to microsphere diameter were measured by nanocompression tests in aqueous conditions. These microspheres encapsulate an inert gas and are used as ultrasound contrast agents by releasing free microbubbles in the presence of an ultrasound field as a result of free gas leakage from the shell. The tests were performed using an atomic force microscope (AFM) employing the force-distance curve technique. An optical microscope, on which the AFM was mounted, was used to guide the positioning of tipless cantilevers on top of individual microspheres. We performed a systematic study using several cantilevers with spring constants varying from 0.08 to 2.3 N/m on a population of microspheres with diameters from about 2 to 6 microm. The use of several cantilevers with various spring constants allowed a systematic study of the mechanical properties of the microsphere thin shell at different regimes of force and deformation. Using thin-shell mechanics theory for small deformations, the Young's modulus of the thin wall material was estimated and was shown to exhibit a strong size effect: it increased as the shell became thinner. The Young's modulus of thicker microsphere shells converged to the expected value for the macroscopic bulk material. For high applied forces, the force-deformation profiles showed a reversible and/or irreversible nonlinear behavior including "steps" and "jumps" which were attributed to mechanical instabilities such as buckling events.

Hypoxic pulmonary vasoconstriction (HPV), which aids ventilation-perfusion matching in the lungs, is triggered by mechanisms intrinsic to pulmonary arterial smooth muscles. The unique sensitivity of these muscles to hypoxia is conferred by mitochondrial cytochrome c oxidase subunit 4 isoform 2, the inhibition of which has been proposed to trigger HPV through increased generation of mitochondrial reactive oxygen species. Contrary to this model, we have shown that the LKB1–AMPK-α1 signaling pathway is critical to HPV. Spectral Doppler ultrasound revealed that deletion of the AMPK-α1 catalytic subunit blocked HPV in mice during mild (8% O2) and severe (5% O2) hypoxia, whereas AMPK-α2 deletion attenuated HPV only during severe hypoxia. By contrast, neither of these genetic manipulations affected serotonin-induced reductions in pulmonary vascular flow. HPV was also attenuated by reduced expression of LKB1, a kinase that activates AMPK during energy stress, but not after deletion of CaMKK2, a kinase that activates AMPK in response to increases in cytoplasmic Ca2+. Fluorescence imaging of acutely isolated pulmonary arterial myocytes revealed that AMPK-α1 or AMPK-α2 deletion did not affect mitochondrial membrane potential during normoxia or hypoxia. However, deletion of AMPK-α1, but not of AMPK-α2, blocked hypoxia from inhibiting KV1.5, the classical “oxygen-sensing” K+ channel in pulmonary arterial myocytes. We conclude that LKB1–AMPK-α1 signaling pathways downstream of mitochondria are critical for the induction of HPV, in a manner also supported by AMPK-α2 during severe hypoxia.

Nine ultrasound transducers from six ultrasound scanners were assessed for their utility for preclinical ultrasound imaging. The transducers were: L8-16, L10-22 (Diasus; Dynamic Imaging Ltd., Livingston, UK); L17-5, L15-7io (iU22; Philips, Seattle, WA, USA), HFL38/13-6 (MicroMaxx; Sonosite Inc., Bothell, WA, USA); il3Lv (Vivid 5; GE, Fairfield, CT, USA), RMV 704 (Vevo 770; Visualsonics Inc., Toronto, Canada) and MS550S, MS550D (Vevo 2100; Visualsonics Inc.). A quantitative analysis of the ultrasound images from all nine transducers employed measurements of the resolution integral as an indication of the versatility and technology of the ultrasound scanners. Two other parameters derived from the resolution integral, the characteristic resolution and depth of field, were used to characterise imaging performance. Six of these transducers were also assessed qualitatively by ultrasonically scanning 59 female common marmosets (Callithrix jacchus) yielding a total of 215 scans. The quantitative measurements for each of the transducers were consistent with the results obtained in the qualitative in vivo assessment. Over a 0–10 mm imaging depth, the values of the resolution integral, characteristic resolution and depth of field, measured using the Edinburgh Pipe Phantom, ranged in magnitude from 7–72, 93–930 μm and 3.3–9.2 mm respectively. The largest resolution integrals were obtained using the Vevo 770 and Vevo 2100 scanners. The Edinburgh Pipe Phantom provides a quantitative method of characterising the imaging performance of preclinical imaging scanners. (E-mail: Carmel.Moran@ed.ac.uk)