Global DNA hypomethylation prevents consolidation of differentiation programs and allows reversion to the embryonic stem cell state.

DNA methylation patterns change dynamically during mammalian development and lineage specification, yet scarce information is available about how DNA methylation affects gene expression profiles upon differentiation. Here we determine genome-wide transcription profiles during undirected differentiation of severely hypomethylated (Dnmt1⁻/⁻) embryonic stem cells (ESCs) as well as ESCs completely devoid of DNA methylation (Dnmt1⁻/⁻;Dnmt3a⁻/⁻;Dnmt3b⁻/⁻ or TKO) and assay their potential to transit in and out of the ESC state. We find that the expression of only few genes mainly associated with germ line function and the X chromosome is affected in undifferentiated TKO ESCs. Upon initial differentiation as embryoid bodies (EBs) wild type, Dnmt1⁻/⁻ and TKO cells downregulate pluripotency associated genes and upregulate lineage specific genes, but their transcription profiles progressively diverge upon prolonged EB culture. While Oct4 protein levels are completely and homogeneously suppressed, transcription of Oct4 and Nanog is not completely silenced even at late stages in both Dnmt1⁻/⁻ and TKO EBs. Despite late wild type and Dnmt1⁻/⁻ EBs showing a much higher degree of concordant expression, after EB dissociation and replating under pluripotency promoting conditions both Dnmt1⁻/⁻ and TKO cells, but not wild type cells rapidly revert to expression profiles typical of undifferentiated ESCs. Thus, while DNA methylation seems not to be critical for initial activation of differentiation programs, it is crucial for permanent restriction of developmental fate during differentiation

Visual and refractive outcome of one-site phacotrabeculectomy compared with temporal approach phacoemulsification

Background We aimed to compare visual and refractive outcome following phacoemulsification and intraocular lens implant (IOL) and combined one-site phacotrabeculectomy. Method We performed a retrospective study of case records of patients who had temporal incision phacoemulsification with IOL or one-site phacotrabeculectomy, between June 1997 and June 2001. The patients were matched for age group, operating list and IOL type. All patients were operated on under local anesthesia by the same surgeon. Each arm of the study had 90 patients, age range 60 to 75 years. We collected pre- and postoperative visual acuity, pre- and postoperative refraction within six months after surgery, and intended refraction. The intraocular pressure control was not recorded, as it was not the aim of our study. Results In the phacotrabeculectomy group, 76.6% of patients achieved aimed spherical equivalent, 15.5% of patients had against-the-rule (ATR) astigmatism induced by the surgery, and 90% of the patients had best corrected visual acuity (BCVA) more than 6/12. In the temporal incision phacoemulsification group, 81.1% of patients achieved aimed spherical equivalent, 10% of the patients had induced ATR by the surgery and 95.55% of patients achieved BCVA more than 6/12. Conclusion In this study the visual outcome of the phacotrabeculectomy group did not differ significantly from the visual outcome of temporal approach phacoemulsification

Effects of static and dynamic higher-order optical modes in balanced homodyne readout for future gravitational waves detectors

With the recent detection of Gravitational waves (GW), marking the start of the new field of GW astronomy, the push for building more sensitive laser-interferometric gravitational wave detectors (GWD) has never been stronger. Balanced homodyne detection (BHD) allows for a quantum noise (QN) limited readout of arbitrary light field quadratures, and has therefore been suggested as a vital building block for upgrades to Advanced LIGO and third generation observatories. In terms of the practical implementation of BHD, we develop a full framework for analyzing the static optical high order modes (HOMs) occurring in the BHD paths related to the misalignment or mode matching at the input and output ports of the laser interferometer. We find the effects of HOMs on the quantum noise limited sensitivity is independent of the actual interferometer configuration, e.g. Michelson and Sagnac interferometers are effected in the same way. We show that misalignment of the output ports of the interferometer (output misalignment) only effects the high frequency part of the quantum noise limited sensitivity (detection noise). However, at low frequencies, HOMs reduce the interferometer response and the radiation pressure noise (back action noise) by the same amount and hence the quantum noise limited sensitivity is not negatively effected in that frequency range. We show that the misalignment of laser into the interferometer (input misalignment) produces the same effect as output misalignment and additionally decreases the power inside the interferometer. We also analyze dynamic HOM effects, such as beam jitter created by the suspended mirrors of the BHD. Our analyses can be directly applied to any BHD implementation in a future GWD. Moreover, we apply our analytical techniques to the example of the speed meter proof of concept experiment under construction in Glasgow. We find that for our experimental parameters, the performance of our seismic isolation system in the BHD paths is compatible with the design sensitivity of the experiment

Regulation and function of the atypical cadherin FAT1 in hepatocellular carcinoma

Giant cadherin FAT1 is increased in HCC and is a protumorigenic factors in HCC cell

Demonstration of a switchable damping system to allow low-noise operation of high-Q low-mass suspension systems

Low mass suspension systems with high-Q pendulum stages are used to enable quantum radiation pressure noise limited experiments. Utilising multiple pendulum stages with vertical blade springs and materials with high quality factors provides attenuation of seismic and thermal noise, however damping of these high-Q pendulum systems in multiple degrees of freedom is essential for practical implementation. Viscous damping such as eddy-current damping can be employed but introduces displacement noise from force noise due to thermal fluctuations in the damping system. In this paper we demonstrate a passive damping system with adjustable damping strength as a solution for this problem that can be used for low mass suspension systems without adding additional displacement noise in science mode. We show a reduction of the damping factor by a factor of 8 on a test suspension and provide a general optimisation for this system.Comment: 5 pages, 5 figure

Mutations in GATA2 cause primary lymphedema associated with a predisposition to acute myeloid leukemia (Emberger syndrome).

We report an allelic series of eight mutations in GATA2 underlying Emberger syndrome, an autosomal dominant primary lymphedema associated with a predisposition to acute myeloid leukemia. GATA2 is a transcription factor that plays an essential role in gene regulation during vascular development and hematopoietic differentiation. Our findings indicate that haploinsufficiency of GATA2 underlies primary lymphedema and predisposes to acute myeloid leukemia in this syndrome

Model Checking Classes of Metric LTL Properties of Object-Oriented Real-Time Maude Specifications

This paper presents a transformational approach for model checking two important classes of metric temporal logic (MTL) properties, namely, bounded response and minimum separation, for nonhierarchical object-oriented Real-Time Maude specifications. We prove the correctness of our model checking algorithms, which terminate under reasonable non-Zeno-ness assumptions when the reachable state space is finite. These new model checking features have been integrated into Real-Time Maude, and are used to analyze a network of medical devices and a 4-way traffic intersection system.Comment: In Proceedings RTRTS 2010, arXiv:1009.398

Stress-induced TRAILR2 expression overcomes TRAIL resistance in cancer cell spheroids

The influence of 3D microenvironments on apoptosis susceptibility remains poorly understood. Here, we studied the susceptibility of cancer cell spheroids, grown to the size of micrometastases, to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Interestingly, pronounced, spatially coordinated response heterogeneities manifest within spheroidal microenvironments: In spheroids grown from genetically identical cells, TRAIL-resistant subpopulations enclose, and protect TRAIL-hypersensitive cells, thereby increasing overall treatment resistance. TRAIL-resistant layers form at the interface of proliferating and quiescent cells and lack both TRAILR1 and TRAILR2 protein expression. In contrast, oxygen, and nutrient deprivation promote high amounts of TRAILR2 expression in TRAIL-hypersensitive cells in inner spheroid layers. COX-II inhibitor celecoxib further enhanced TRAILR2 expression in spheroids, likely resulting from increased ER stress, and thereby re-sensitized TRAIL-resistant cell layers to treatment. Our analyses explain how TRAIL response heterogeneities manifest within well-defined multicellular environments, and how spatial barriers of TRAIL resistance can be minimized and eliminated

Limited Value of Bladder Wash Cytology During Follow-Up of Patients With Non-muscle Invasive Bladder Cancer

Aims We aimed to assess the performance of bladder wash cytology (BWC) in daily clinical practice in a pure follow-up cohort of patients previously diagnosed with non-muscle invasive bladder cancer (NMIBC). Materials and methods We analyzed 2064 BWCs derived from 314 patients followed for NMIBC (2003-2016). Follow-up investigations were performed using cystoscopy (CS) in combination with BWC. Patients with suspicious CS and/or positive BWC underwent bladder biopsy or transurethral resection. BWC was considered positive if malignant or suspicious cells were reported. Sensitivity (Sn) and specificity (Sp) were calculated for the entire cohort and separately for low-grade (LG) and high-grade (HG) tumors, and carcinoma in situ (CIS) subgroups. Results A total of 95 recurrences were detected, of which only three were detected by BWC alone. Overall, Sn and Sp of BWC were 17.9% and 99.5%, respectively. For LG disease, these numbers were 14.0% and 100%, and for HG disease, these were 22.2% and 99.1%, respectively. For patients with CIS at initial diagnosis, Sn and Sp were 11.0% and 71.4%, respectively. For isolated primary CIS, Sn was 50.0%, and Sp was 98.2%. Conclusion Routine use of BWC in the follow-up for NMIBC is of limited value even in HG tumors. In the presence of isolated primary CIS, adjunct BWC might be justified