An important step for cholinergic transmission involves the vesicular storage of acetylcholine (ACh), a process mediated by the vesicular acetylcholine transporter (VAChT). In order to understand the physiological roles of the VAChT, we developed a genetically altered strain of mice with reduced expression of this transporter. Heterozygous and homozygous VAChT knockdown mice have a 45% and 65% decrease in VAChT protein expression, respectively. VAChT deficiency alters synaptic vesicle filling and affects ACh release. Whereas VAChT homozygous mutant mice demonstrate major neuromuscular deficits, VAChT heterozygous mice appear normal in that respect and could be used for analysis of central cholinergic function. Behavioral analyses revealed that aversive learning and memory are not altered in mutant mice; however, performance in cognitive tasks involving object and social recognition is severely impaired. These observations suggest a critical role of VAChT in the regulation of ACh release and physiological functions in the peripheral and central nervous system.

It is a widely accepted axiom that localized concentration of mechanical stress and large flexural deformation is closely related to the calcification and tissue degeneration in bioprosthetic heart valves (BHV). In order to investigate the complex BHV deformations and stress distributions throughout the cardiac cycle, it is necessary to perform an accurate dynamic analysis with a morphologically and physiologically realistic material specification for the leaflets. We have developed a stress resultant shell model for BHV leaflets incorporating a Fung-elastic constitutive model for in-plane and bending responses separately. Validation studies were performed by comparing the finite element predicted displacement and strain measures with the experimentally measured data under physiological pressure loads. Computed regions of stress concentration and large flexural deformation during the opening and closing phases of the cardiac cycle correlated with previously reported regions of calcification and/or mechanical damage on BHV leaflets. It is expected that the developed experimental and computational methodology will aid in the understanding of the complex dynamic behavior of native and bioprosthetic valves and in the development of tissue engineered valve substitutes.

Objective To develop a new bioactive gas delivery method using echogenic liposomes (ELIP) as the gas carrier. Background Nitric oxide (NO) is a bioactive gas with potent therapeutic effects. Bioavailability of NO by systemic delivery is low with potential systemic effects. Methods Liposomes containing phospholipids and cholesterol were prepared using a new freezing under pressure method. The encapsulation and release profile of NO from NO containing-ELIP (NO-ELIP) or a mixture of NO/Argon (NO/Ar-ELIP was studied. Uptake of NO from NO-ELIP by cultured vascular smooth muscle cells (VSMC) both in the absence and presence of hemoglobin was determined. The effect of NO-ELIP delivery to attenuate intimal hyperplasia in a balloon-injured artery was determined. Results Coencapsulation of NO with argon (Ar) enabled the adjustment the amount of encapsulated NO. A total of 10 µl of gas can be encapsulated into 1 mg liposomes. The release profile of NO from NO-ELIP demonstrated an initial rapid release followed by a slower release over 8 hours. Sixty-eight percent of cells remained viable when incubated with 80 µg/ml of NO/Ar-ELIP for 4 hours. NO delivery to VSMC using NO/Ar-ELIP was 7-fold higher than unencapsulated NO. NO/Ar-ELIP remained effective NO delivery to VSMC even in the presence of hemoglobin. Local NO-ELIP administration to balloon-injured carotid arteries attenuated the development of intimal hyperplasia and reduced arterial wall thickening by 41±9%. Conclusions Liposomes can protect and deliver a bioactive gas to target tissues with the potential for both visualization of gas delivery and controlled therapeutic gas release.

Most surgical procedures for patients with mitral regurgitation (MR) focus on optimization of annular dimension and shape utilizing ring annuloplasty to restore normal annular geometry, increase leaflet coaptation, and reduce regurgitation. Computational studies may provide insight on the effect of annular motion on mitral valve (MV) function through the incorporation of patient-specific MV apparatus geometry from clinical imaging modalities such as echocardiography. In the present study, we have developed a novel algorithm for modeling patient-specific annular motion across the cardiac cycle to further improve our virtual MV modeling and simulation strategy. The MV apparatus including the leaflets, annulus, and location of papillary muscle tips was identified using patient 3D echocardiography data at end diastole and peak systole and converted to virtual MV model. Dynamic annular motion was modeled by incorporating the ECG-gated time-varying scaled annular displacement across the cardiac cycle. We performed dynamic finite element (FE) simulation of two sets of patient data with respect to the presence of MR. Annular morphology, stress distribution across the leaflets and annulus, and contact stress distribution were determined to assess the effect of annular motion on MV function and leaflet coaptation. The effect of dynamic annular motion clearly demonstrated reduced regions with large stress values and provided an improved accuracy in determining the location of improper leaflet coaptation. This strategy has the potential to better quantitate the extent of pathologic MV and better evaluate functional restoration following MV repair.

Tonic inhibition in the brain is mediated through an activation of extrasynaptic GABA receptors by the tonically released GABA, resulting in a persistent GABAergic inhibitory action. It is one of the key regulators for neuronal excitability, exerting a powerful action on excitation/inhibition balance. We have previously reported that astrocytic GABA, synthesized by monoamine oxidase B (MAOB), mediates tonic inhibition via GABA-permeable bestrophin 1 (Best1) channel in the cerebellum. However, the role of astrocytic GABA in regulating neuronal excitability, synaptic transmission, and cerebellar brain function has remained elusive. Here, we report that a reduction of tonic GABA release by genetic removal or pharmacological inhibition of Best1 or MAOB caused an enhanced neuronal excitability in cerebellar granule cells (GCs), synaptic transmission at the parallel fiber-Purkinje cell (PF-PC) synapses, and motor performance on the rotarod test, whereas an augmentation of tonic GABA release by astrocyte-specific overexpression of MAOB resulted in a reduced neuronal excitability, synaptic transmission, and motor performance. The bidirectional modulation of astrocytic GABA by genetic alteration of Best1 or MAOB was confirmed by immunostaining and in vivo microdialysis. These findings indicate that astrocytes are the key player in motor coordination through tonic GABA release by modulating neuronal excitability and could be a good therapeutic target for various movement and psychiatric disorders, which show a disturbed excitation/inhibition balance.

Background-Ischemia-related neurological injury is a primary cause of stroke disability. Studies have demonstrated that xenon (Xe) may have potential as an effective and nontoxic neuroprotectant. Xe delivery is, however, hampered by lack of suitable administration methods. We have developed a pressurization-freeze method to encapsulate Xe into echogenic liposomes (Xe-ELIP) and have modulated local gas release with transvascular ultrasound exposure. Methods and Results-Fifteen microliters of Xe were encapsulated into each 1 mg of liposomes (70% Xe and 30% argon).Xe delivery from Xe-ELIP into cells and consequent neuroprotective effects were evaluated with oxygen/glucosedeprived and control neuronal cells in vitro. Xe-ELIP were administered into Sprague-Dawley rats intravenously or intra-arterially after right middle cerebral artery occlusion. One-megahertz low-amplitude (0.18 MPa) continuous wave ultrasound directed onto the internal carotid artery triggered Xe release from circulating Xe-ELIP. Effects of Xe delivery on ischemia-induced neurological injury and disability were evaluated. Xe-ELIP delivery to oxygen/glucose-deprived neuronal cells improved cell viability in vitro and resulted in a 48% infarct volume decrease in vivo. Intravenous Xe-ELIP administration in combination with the ultrasound directed onto the carotid artery enhanced local Xe release from circulating Xe-ELIP and demonstrated 75% infarct volume reduction. This was comparable to the effect after intra-arterial administration. Behavioral tests on limb placement and grid and beam walking correlated with infarct reduction. Conclusions-This novel methodology may provide a noninvasive strategy for ultrasound-enhanced local therapeutic gas delivery for cerebral ischemia-related injury while minimizing systemic side effects. (Circulation. 2010;122:1578-1587.)

Technological advances of mankind, through the development of electrical and communication technologies, have resulted in the exposure to artificial electromagnetic fields (EMF). Technological growth is expected to continue; as such, the amount of EMF exposure will continue to increase steadily. In particular, the use-time of smart phones, that have become a necessity for modern people, is steadily increasing. Social concerns and interest in the impact on the cranial nervous system are increased when considering the area where the mobile phone is used. However, before discussing possible effects of radiofrequency-electromagnetic field (RF-EMF) on the human body, several factors must be investigated about the influence of EMFs at the level of research using in vitro or animal models. Scientific studies on the mechanism of biological effects are also required. It has been found that RF-EMF can induce changes in central nervous system nerve cells, including neuronal cell apoptosis, changes in the function of the nerve myelin and ion channels; furthermore, RF-EMF act as a stress source in living creatures. The possible biological effects of RF-EMF exposure have not yet been proven, and there are insufficient data on biological hazards to provide a clear answer to possible health risks. Therefore, it is necessary to study the biological response to RF-EMF in consideration of the comprehensive exposure with regard to the use of various devices by individuals. In this review, we summarize the possible biological effects of RF-EMF exposure.

In order to achieve a more realistic and accurate computational simulation of native and bioprosthetic heart valve dynamics, a finite shell element model was developed. Experimentally derived and uncoupled in-plane and bending behaviors were implemented into a fully nonlinear stress resultant shell element. Validation studies compared the planar biaxial extension and three-point bending simulations to the experimental data and demonstrated excellent fidelity. Dynamic simulations of a pericardial bioprosthetic heart valve with the developed shell element model showed significant differences in the deformation characteristics compared to the simulation with an assumed isotropic bending model. The new finite shell element model developed in the present study can also incorporate various types of constitutive models and is expected to help us to understand the complex dynamics of native and bioprosthetic heart valve function in physiological and pathological conditions.