Comparison of Magnetic Resonance Imaging and Serum Biomarkers for Detection of Human Pluripotent Stem Cell-Derived Teratomas.

The use of cells derived from pluripotent stem cells (PSCs) for regenerative therapies confers a considerable risk for neoplastic growth and teratoma formation. Preclinical and clinical assessment of such therapies will require suitable monitoring strategies to understand and mitigate these risks. Here we generated human-induced pluripotent stem cells (iPSCs), selected clones that continued to express reprogramming factors after differentiation into cardiomyocytes, and transplanted these cardiomyocytes into immunocompromised rat hearts post-myocardial infarction. We compared magnetic resonance imaging (MRI), cardiac ultrasound, and serum biomarkers for their ability to delineate teratoma formation and growth. MRI enabled the detection of teratomas with a volume >8 mm(3). A combination of three plasma biomarkers (CEA, AFP, and HCG) was able to detect teratomas with a volume >17 mm(3) and with a sensitivity of more than 87%. Based on our findings, a combination of serum biomarkers with MRI screening may offer the highest sensitivity for teratoma detection and tracking

Mitochondrial cyclophilin-D as a critical mediator of ischaemic preconditioning

It has been suggested that mitochondrial reactive oxygen species (ROS), Akt and Erk1/2 and more recently the mitochondrial permeability transition pore (mPTP) may act as mediators of ischaemic preconditioning (IPC), although the actual interplay between these mediators is unclear. The aim of the present study is to determine whether the cyclophilin-D (CYPD) component of the mPTP is required by IPC to generate mitochondrial ROS and subsequently activate Akt and Erk1/2.Mice lacking CYPD (CYPD-/-) and B6Sv129 wild-type (WT) mice were used throughout. We have demonstrated that under basal conditions, non-pathological mPTP opening occurs (indicated by the percent reduction in mitochondrial calcein fluorescence). This effect was greater in WT cardiomyocytes compared with CYPD-/- ones (53 +/- 2% WT vs. 17 +/- 3% CYPD-/-; P < 0.01) and was augmented by hypoxic preconditioning (HPC) (70 +/- 9% WT vs. 56 +/- 1% CYPD-/-; P < 0.01). HPC reduced cell death following simulated ischaemia-reperfusion injury in WT (23.2 +/- 3.5% HPC vs. 43.7 +/- 3.2% WT; P < 0.05) but not CYPD-/- cardiomyocytes (19.6 +/- 1.4% HPC vs. 24.4 +/- 2.6% control; P > 0.05). HPC generated mitochondrial ROS in WT (four-fold increase; P < 0.05) but not CYPD-/- cardiomyocytes. HPC induced significant Akt phosphorylation in WT cardiomyocytes (two-fold increase; P < 0.05), an effect which was abrogated by ciclosporin-A (a CYPD inhibitor) and N-2-mercaptopropionyl glycine (a ROS scavenger). Finally, in vivo IPC of adult murine hearts resulted in significant phosphorylation of Akt and Erk1/2 in WT but not CYPD-/- hearts.The CYPD component of the mPTP is required by IPC to generate mitochondrial ROS and phosphorylate Akt and Erk1/2, major steps in the IPC signalling pathway

PolyQ length-dependent metabolic alterations and DNA damage drive human astrocyte dysfunction in Huntington's disease

Huntington's Disease (HD) is a neurodegenerative disease caused by a polyglutamine (polyQ) expansion in the Huntingtin gene. Astrocyte dysfunction is known to contribute to HD pathology, however our understanding of the molecular pathways involved is limited. Transcriptomic analysis of patient-derived PSC (pluripotent stem cells) astrocyte lines revealed that astrocytes with similar polyQ lengths shared a large number of differentially expressed genes (DEGs). Notably, weighted correlation network analysis (WGCNA) modules from iPSC derived astrocytes showed significant overlap with WGCNA modules from two post-mortem HD cohorts. Further experiments revealed two key elements of astrocyte dysfunction. Firstly, expression of genes linked to astrocyte reactivity, as well as metabolic changes were polyQ length-dependent. Hypermetabolism was observed in shorter polyQ length astrocytes compared to controls, whereas metabolic activity and release of metabolites were significantly reduced in astrocytes with increasing polyQ lengths. Secondly, all HD astrocytes showed increased DNA damage, DNA damage response and upregulation of mismatch repair genes and proteins. Together our study shows for the first time polyQ-dependent phenotypes and functional changes in HD astrocytes providing evidence that increased DNA damage and DNA damage response could contribute to HD astrocyte dysfunction

Ischemic Preconditioning and Postconditioning Protect the Heart by Preserving the Mitochondrial Network

BACKGROUND: Mitochondria fuse to form elongated networks which are more tolerable to stress and injury. Ischemic pre- and postconditioning (IPC and IPost, respectively) are established cardioprotective strategies in the preclinical setting. Whether IPC and IPost modulates mitochondrial morphology is unknown. We hypothesize that the protective effects of IPC and IPost may be conferred via preservation of mitochondrial network. METHODS: IPC and IPost were applied to the H9c2 rat myoblast cells, isolated adult primary murine cardiomyocytes, and the Langendorff-isolated perfused rat hearts. The effects of IPC and IPost on cardiac cell death following ischemia-reperfusion injury (IRI), mitochondrial morphology, and gene expression of mitochondrial-shaping proteins were investigated. RESULTS: IPC and IPost successfully reduced cardiac cell death and myocardial infarct size. IPC and IPost maintained the mitochondrial network in both H9c2 and isolated adult primary murine cardiomyocytes. 2D-length measurement of the 3 mitochondrial subpopulations showed that IPC and IPost significantly increased the length of interfibrillar mitochondria (IFM). Gene expression of the pro-fusion protein, Mfn1, was significantly increased by IPC, while the pro-fission protein, Drp1, was significantly reduced by IPost in the H9c2 cells. In the primary cardiomyocytes, gene expression of both Mfn1 and Mfn2 were significantly upregulated by IPC and IPost, while Drp1 was significantly downregulated by IPost. In the Langendorff-isolated perfused heart, gene expression of Drp1 was significantly downregulated by both IPC and IPost. CONCLUSION: IPC and IPost-mediated upregulation of pro-fusion proteins (Mfn1 and Mfn2) and downregulation of pro-fission (Drp1) promote maintenance of the interconnected mitochondrial network, ultimately conferring cardioprotection against IRI

Experimental demonstration of the relationship between the second- and third-order polarizabilities of conjugated donor-acceptor molecules

The dependence of the second- and third-order polarizabilities ((beta) and (gamma) ) on ground-state polarization was measured for a series of donor-acceptor polyenes using electric field induced second harmonic generation and third harmonic generation, respectively. The changes in ground-state polarization, associated with the donor/acceptor strength or solvent polarity, were probed by x-ray crystallography, 1H-NMR, electronic absorption, and Raman spectroscopies. The observed behavior of (beta) and (gamma) as a function of ground- state polarization agrees well with theoretical predictions. In particular, positive and negative peaks, as well as sign changes, were observed for both (beta) and (gamma) . The dependences for (beta) and (gamma) are consistent with a derivative relationship between them. In addition, the third-order polarizability of a series of molecules possessing zero bond length alternation was found to be negative, in agreement with predictions based on the relationship between the polarizabilities and ground-state geometry

International Space Station (ISS) Plasma Contactor Unit (PCU) Utilization Plan Assessment Update

The International Space Station (ISS) vehicle undergoes spacecraft charging as it interacts with Earth's ionosphere and magnetic field. The interaction can result in a large potential difference developing between the ISS metal chassis and the local ionosphere plasma environment. If an astronaut conducting extravehicular activities (EVA) is exposed to the potential difference, then a possible electrical shock hazard arises. The control of this hazard was addressed by a number of documents within the ISS Program (ISSP) including Catastrophic Safety Hazard for Astronauts on EVA (ISS-EVA-312-4A_revE). The safety hazard identified the risk for an astronaut to experience an electrical shock in the event an arc was generated on an extravehicular mobility unit (EMU) surface. A catastrophic safety hazard, by the ISS requirements, necessitates mitigation by a two-fault tolerant system of hazard controls. Traditionally, the plasma contactor units (PCUs) on the ISS have been used to limit the charging and serve as a "ground strap" between the ISS structure and the surrounding ionospheric plasma. In 2009, a previous NASA Engineering and Safety Center (NESC) team evaluated the PCU utilization plan (NESC Request #07-054-E) with the objective to assess whether leaving PCUs off during non-EVA time periods presented risk to the ISS through assembly completion. For this study, in situ measurements of ISS charging, covering the installation of three of the four photovoltaic arrays, and laboratory testing results provided key data to underpin the assessment. The conclusion stated, "there appears to be no significant risk of damage to critical equipment nor excessive ISS thermal coating damage as a result of eliminating PCU operations during non- EVA times." In 2013, the ISSP was presented with recommendations from Boeing Space Environments for the "Conditional" Marginalization of Plasma Hazard. These recommendations include a plan that would keep the PCUs off during EVAs when the space environment forecast input to the ISS charging model indicates floating potentials (FP) within specified limits. These recommendations were based on the persistence of conditions in the space environment due to the current low solar cycle and belief in the accuracy and completeness of the ISS charging model. Subsequently, a Noncompliance Report (NCR), ISS-NCR-232G, Lack of Two-fault Tolerance to EVA Crew Shock in the Low Earth Orbit Plasma Environment, was signed in September 2013 specifying new guidelines for the use of shock hazard controls based on a forecast of the space environment from ISS plasma measurements taken prior to the EVA [ISS-EVA-312-AC, 2012]. This NESC assessment re-evaluates EVA charging hazards through a process that is based on over 14 years of ISS operations, charging measurements, laboratory tests, EMU studies and modifications, and safety reports. The assessment seeks an objective review of the plasma charging hazards associated with EVA operations to determine if any of the present hazard controls can safely change the PCU utilization plan to allow more flexibility in ISS operations during EVA preparation and execution

TIA1 Mutations in Amyotrophic Lateral Sclerosis and Frontotemporal Dementia Promote Phase Separation and Alter Stress Granule Dynamics.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are age-related neurodegenerative disorders with shared genetic etiologies and overlapping clinical and pathological features. Here we studied a novel ALS/FTD family and identified the P362L mutation in the low-complexity domain (LCD) of T cell-restricted intracellular antigen-1 (TIA1). Subsequent genetic association analyses showed an increased burden of TIA1 LCD mutations in ALS patients compared to controls (p = 8.7 × 1

International Space Station (ISS) Plasma Contactor Unit (PCU) Utilization Plan Assessment Update

The NASA Engineering and Safety Center (NESC) received a request to support the Assessment of the International Space Station (ISS) Plasma Contactor Unit (PCU) Utilization Update. The NESC conducted an earlier assessment of the use of the PCU in 2009. This document contains the outcome of the assessment update

Comparison of Pittsburgh compound B and florbetapir in cross-sectional and longitudinal studies.

IntroductionQuantitative in vivo measurement of brain amyloid burden is important for both research and clinical purposes. However, the existence of multiple imaging tracers presents challenges to the interpretation of such measurements. This study presents a direct comparison of Pittsburgh compound B-based and florbetapir-based amyloid imaging in the same participants from two independent cohorts using a crossover design.MethodsPittsburgh compound B and florbetapir amyloid PET imaging data from three different cohorts were analyzed using previously established pipelines to obtain global amyloid burden measurements. These measurements were converted to the Centiloid scale to allow fair comparison between the two tracers. The mean and inter-individual variability of the two tracers were compared using multivariate linear models both cross-sectionally and longitudinally.ResultsGlobal amyloid burden measured using the two tracers were strongly correlated in both cohorts. However, higher variability was observed when florbetapir was used as the imaging tracer. The variability may be partially caused by white matter signal as partial volume correction reduces the variability and improves the correlations between the two tracers. Amyloid burden measured using both tracers was found to be in association with clinical and psychometric measurements. Longitudinal comparison of the two tracers was also performed in similar but separate cohorts whose baseline amyloid load was considered elevated (i.e., amyloid positive). No significant difference was detected in the average annualized rate of change measurements made with these two tracers.DiscussionAlthough the amyloid burden measurements were quite similar using these two tracers as expected, difference was observable even after conversion into the Centiloid scale. Further investigation is warranted to identify optimal strategies to harmonize amyloid imaging data acquired using different tracers