Laser-speckle-based detection of fluid pulsation in the presence of motion artifacts: in vitro and in vivo study.

We have performed an in vitro and in vivo study, based on laser speckle contrast analysis, to detect fluid pulsation in the presence of artifacts caused by the relative motion between the sample and the illumination source. We observe that the pulsation signal is clearly detectable for a range of motion amplitudes and oscillation frequencies; however, for higher amplitudes and oscillation frequencies of motion, the signal, due to pulsation, becomes increasingly difficult to detect.

Direct effects of doxorubicin on skeletal muscle contribute to fatigue.

Chemotherapy-induced fatigue is a multidimensional symptom. Oxidative stress has been proposed as a working mechanism for anthracycline-induced cardiotoxicity. In this study, doxorubicin (DOX) was tested on skeletal muscle function. Doxorubicin induced impaired ex vivo skeletal muscle relaxation followed in time by contraction impediment, which could be explained by DOX-induced changes in Ca(2+) responses of myotubes in vitro. The Ca(2+) responses in skeletal muscle, however, could not be explained by oxidative stress.

Design of scaffolds for blood vessel tissue engineering using a multi-layering electrospinning technique.

Aiming to develop a scaffold architecture mimicking morphological and mechanically that of a blood vessel, a sequential multi-layering electrospinning (ME) was performed on a rotating mandrel-type collector. A bi-layered tubular scaffold composed of a stiff and oriented PLA outside fibrous layer and a pliable and randomly oriented PCL fibrous inner layer (PLA/PCL) was fabricated. Control over the level of fibre orientation of the different layers was achieved through the rotation speed of the collector. The structural and mechanical properties of the scaffolds were examined using scanning electron microscopy (SEM) and tensile testing. To assess their capability to support cell attachment, proliferation and migration, 3T3 mouse fibroblasts and later human venous myofibroblasts (HVS) were cultured, expanded and seeded on the scaffolds. In both cases, the cell-polymer constructs were cultured under static conditions for up to 4 weeks. Environmental-scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), histological examination and biochemical assays for cell proliferation (DNA) and extracellular matrix production (collagen and glycosaminoglycans) were performed. The findings suggest the feasibility of ME to design scaffolds with a hierarchical organization through a layer-by-layer process and control over fibre orientation. The resulting scaffolds achieved the desirable levels of pliability (elastic up to 10% strain) and proved to be capable to promote cell growth and proliferation. The electrospun PLA/PCL bi-layered tube presents appropriate characteristics to be considered a candidate scaffold for blood vessel tissue engineering.

Management of pediatric delirium in critical illness: a practical update.

Pediatric delirium (PD) is an acute state of brain dysfunction and is often seen in the pediatric intensive care unit (PICU). There is a growing awareness of its clinical interdisciplinary importance. The aim of this article was to describe the three clinical presentations, to evaluate the differential diagnosis and to give a concise and practical update for the pharmacological and non-pharmacological treatment of PD at the PICU, based on recent literature and expert opinions. We discuss an interdisciplinary flow chart which helps the reader dealing with the diagnosis and management of any acute emotional and or behavioral disturbance, of which PD is a special case.

A novel passive left heart platform for device testing and research.

Integration of biological samples into in vitro mock loops is fundamental to simulate real device's operating conditions. We developed an in vitro platform capable of simulating the pumping function of the heart through the external pressurization of the ventricle. The system consists of a fluid-filled chamber, in which the ventricles are housed and sealed to exclude the atria from external loads. The chamber is connected to a pump that drives the motion of the ventricular walls. The aorta is connected to a systemic impedance simulator, and the left atrium to an adjustable preload. The platform reproduced physiologic hemodynamics, i.e. aortic pressures of 120/80 mmHg with 5 L/min of cardiac output, and allowed for intracardiac endoscopy. A pilot study with a left ventricular assist device (LVAD) was also performed. The LVAD was connected to the heart to investigate aortic valve functioning at different levels of support. Results were consistent with the literature, and high speed video recordings of the aortic valve allowed for the visualization of the transition between a fully opening valve and a permanently closed configuration. In conclusion, the system showed to be an effective tool for the hemodynamic assessment of devices, the simulation of surgical or transcatheter procedures and for visualization studies.

Transfer of primitive stem/progenitor bone marrow cells from LT alpha-/- donors to wild-type hosts: implications for the generation of architectural events in lymphoid B cell domains.

To analyze whether the phenotypic abnormalities observed in lymphotoxin-alpha(-/-) (LT alpha-/-) mice are intrinsic to the hemolymphoid system itself or dependent on stromal elements, wild-type (WT) mice were reconstituted with bone marrow (BM) cells enriched for hemopoietic stem cells from LT alpha-/- animals. WT mice reconstituted with LT alpha-/- c-kit+ Lin- Sca-1+ BM cells do not maintain follicular dendritic cell (FDC) networks and do not form primary follicles, while clear segregation of B and T cells could be observed. Furthermore, IgM+ IgD- B cells, MOMA-1 (anti-metallophilic macrophages), ERTR-9 (anti-marginal zone macrophages), and MECA-367 (anti-MAdCAM-1) were all absent from the splenic marginal zone. Surprisingly, however, the expression of MOMA-1, ERTR-9, and MAdCAM-1 was normal in the lymph nodes of mice reconstituted with LT alpha-/- cells. In addition, peanut agglutinin-positive germinal centers were observed in both the spleen and mesenteric lymph nodes, although in the absence of detectable FDC. Furthermore, in animals reconstituted with a mixture of LT alpha-/- and WT c-kit+ Lin- Sca-1+, GC contained either predominantly LT alpha-/- B cells or WT B cells. These results suggest that although the formation of primary follicles, FDC networks, and the splenic marginal zone are all dependent on hemopoietically derived LT alpha, germinal center formation and the expression of MAdCAM-1, MOMA-1, and ERTR-9 in lymph nodes are not. Our results also suggest that the disturbed B-T cell separation in LT alpha-/- mice is unrelated to defects in the marginal zone.

Effects of tumor necrosis factor and lymphotoxin on peripheral lymphoid tissue development.

Previously, we have reported that neutralization of surface lymphotoxin (LT-alphabeta) in mice which expressed an LT-beta receptor-Fc fusion protein, driven by the cytomegalovirus promoter, resulted in an array of anatomic abnormalities. We now report that mice which express a tumor necrosis factor (TNF) receptor p60-Fc fusion protein (which neutralizes TNF and soluble LT-alpha3 activity) develop unique lymphoid abnormalities. Our data demonstrate that some aspects of peripheral lymphoid organ development require both surface LT-alphabeta and TNF interacting with their specific receptors. However, these related cytokines are also capable of signaling distinct developmental events. Splenic MAdCAM-1 expression, follicular dendritic cell localization and normal Peyer's patch development all require both surface LT-alphabeta and TNF activity. Marginal zone formation and splenic B cell localization primarily require surface LT-alphabeta-LT-beta receptor interactions. Primary follicle formation was dependent upon TNF receptor(s) engagement. Interestingly spleen, lymph nodes and Peyer's patches from TNF receptor p60-Fc-expressing mice all develop different abnormalities, suggesting distinct pathways of development in these lymphoid organs. Thymus development appears to be independent of these signaling pathways. These results demonstrate that TNF and LT are crucial for normal peripheral, but not central lymphoid organ development.