Characterization of surrogate parameters for blood pressure regulation in neurally-mediated syncope.

Neurally Mediated Syncope (NMS) is often cited as the most common cause of syncope. It can lead to severe consequences such as injuries, high rates of hospitalization and reduced quality of life, especially in elderly populations. Therefore, information about the syncope triggers and reflex mechanisms would be of a great value in the development of a cost-effective p-health system for the prediction of syncope episodes, by enhancing patients' quality of life and reducing the incidence of syncope related disorders/conditions. In the present paper we study the characterization of syncope reflex mechanisms and blood pressure changes from the analysis of several non-invasive modalities (ECG, ICG and PPG). Several parameters were extracted in order to characterize the chronotropic, inotropic and vascular tone changes. Thus, we evaluate the ability of parameters such as Heart Rate (HR), Pre-Ejection Period (PEP) and Left Ventricular Ejection Time (LVET) to characterize the physiological mechanisms behind the development of reflex syncope and their potential syncope prediction capability. The significant parameter changes (e.g. HR from 12.9% to -12.4%, PEP from 14.9% to -3.8% and LVET from -14.4% to 12.3%) found in the present work suggest the feasibility of these surrogates to characterize the blood pressure regulation mechanisms during impending syncope.

Detection of hemodynamic adaptations during impending syncope: implementation of a robust algorithm based on pulse arrival time measurements only.

Syncopes are a major public health concern since they can cause severe injuries e.g. by associated falls. We previously demonstrated the feasibility of syncope prediction based on the pulse arrival time (PAT) analysis. Importantly, algorithms for early detection of impending syncope need to be robust against measurement noise, in particular photoplethysmography (PPG) artifacts, causing false detection. We introduce in this work an algorithm concept to deal with artifacts as well as to detect the onset of syncope based on tracking of relative PAT changes only. Our method has been shown useful to improve detection performance for measurements during impending syncope in patients undergoing head-up tilt table testing which might improve syncope prediction.

Improved methods for reprogramming human dermal fibroblasts using fluorescence activated cell sorting.

Current methods to derive induced pluripotent stem cell (iPSC) lines from human dermal fibroblasts by viral infection rely on expensive and lengthy protocols. One major factor contributing to the time required to derive lines is the ability of researchers to identify fully reprogrammed unique candidate clones from a mixed cell population containing transformed or partially reprogrammed cells and fibroblasts at an early time point post infection. Failure to select high quality colonies early in the derivation process results in cell lines that require increased maintenance and unreliable experimental outcomes. Here, we describe an improved method for the derivation of iPSC lines using fluorescence activated cell sorting (FACS) to isolate single cells expressing the cell surface marker signature CD13(NEG)SSEA4(POS)Tra-1-60(POS) on day 7-10 after infection. This technique prospectively isolates fully reprogrammed iPSCs, and depletes both parental and "contaminating" partially reprogrammed fibroblasts, thereby substantially reducing the time and reagents required to generate iPSC lines without the use of defined small molecule cocktails. FACS derived iPSC lines express common markers of pluripotency, and possess spontaneous differentiation potential in vitro and in vivo. To demonstrate the suitability of FACS for high-throughput iPSC generation, we derived 228 individual iPSC lines using either integrating (retroviral) or non- integrating (Sendai virus) reprogramming vectors and performed extensive characterization on a subset of those lines. The iPSC lines used in this study were derived from 76 unique samples from a variety of tissue sources, including fresh or frozen fibroblasts generated from biopsies harvested from healthy or disease patients.

Pulse Arrival Time as surrogate for systolic blood pressure changes during impending neurally mediated syncope.

Blood pressure regulation failures cause neurally mediated syncope often resulting in a fall. A warning device might help to make patients aware of an impending critical event or even trigger the patient to perform countermeasures such as lying down or isometric exercises. We previously demonstrated that the Pulse Arrival Time (PAT) methodology is a potential approach to enable early detection of impending faints. The aim of the present study was to evaluate whether PAT can be used as an easy to measure beat-to-beat surrogate for systolic blood pressure (SBP) changes during a passive standing exercise (head-up tilt table testing (HUTT)). A significant PAT increase of more than 10 % was accompanied with a critical SBP decrease in syncope patients. Although PAT is in general not considered as a good measure of absolute blood pressure we found strong correlations (R>0.89, P<0.01) of SBP and PAT after PAT began to increase. Therefore, our data suggest that the pulse arrival time is useful to monitor blood pressure changes in patients with neurally mediated syncope. This might open up new avenues to prevent falls in these patients.

NOA1 is an essential GTPase required for mitochondrial protein synthesis.

Nitric oxide associated-1 (NOA1) is an evolutionarily conserved guanosine triphosphate (GTP) binding protein that localizes predominantly to mitochondria in mammalian cells. On the basis of bioinformatic analysis, we predicted its possible involvement in ribosomal biogenesis, although this had not been supported by any experimental evidence. Here we determine NOA1 function through generation of knockout mice and in vitro assays. NOA1-deficient mice exhibit midgestation lethality associated with a severe developmental defect of the embryo and trophoblast. Primary embryonic fibroblasts isolated from NOA1 knockout embryos show deficient mitochondrial protein synthesis and a global defect of oxidative phosphorylation (OXPHOS). Additionally, Noa1⁻/⁻ cells are impaired in staurosporine-induced apoptosis. The analysis of mitochondrial ribosomal subunits from Noa1⁻/⁻ cells by sucrose gradient centrifugation and Western blotting showed anomalous sedimentation, consistent with a defect in mitochondrial ribosome assembly. Furthermore, in vitro experiments revealed that intrinsic NOA1 GTPase activity was stimulated by bacterial ribosomal constituents. Taken together, our data show that NOA1 is required for mitochondrial protein synthesis, likely due to its yet unidentified role in mitoribosomal biogenesis. Thus, NOA1 is required for such basal mitochondrial functions as adenosine triphosphate (ATP) synthesis and apoptosis.