The Role of EphA Receptor Signaling in Neuroblast Survival During In Vitro Differentiation

Mouse embryonic stem cells can be differentiated into multiple cell types and can serve as an excellent model for studying developmental processes in vitro. In particular, stem cells can be differentiated into forebrain-like neurons, allowing investigation of nervous system development at single cell resolution. Eph receptor tyrosine kinases and their ephrin ligands play a critical role during in vivo cortical development, particularly during axon guidance. Preliminary data has shown that EphRs and ephrins are expressed during in vitro differentiation. In addition, we see EphA7 localization at the face of neural rosettes, where it co-expresses with markers of neuroblast identity. However, the role of Eph receptors and ephrins in neurogenesis is not well understood. Previous literature had shown that ephrin-A5 and EphA7 can function to balance cortical apoptosis during embryogenesis. We hypothesized that EphR/ephrin signaling may be required to balance apoptosis during in vitro neural development. A monolayer differentiation protocol was used to generate cells of forebrain fate at high yield. RNAi knockdown of EphA3, EphA4, and EphA7, was used to identify which Eph receptor was contributing to the apoptotic effect individually. It was found that transient knockdown of EphA3 and EphA4 fail to cause changes in apoptosis levels. However, knockdown of EphA7 at differentiation days 4 through 8 lead to a reduction in apoptosis suggesting that EphA7 can positively regulate cell death during differentiation. These data confirm that in vivo developmental events such as apoptosis can be modeled in an in vitro system. The innovative methods developed throughout the process of this research project may eventually prove to be useful in the analysis of other neurodevelopmental processes

Cilengitide: an RGD pentapeptide ανβ3 and ανβ5 integrin inhibitor in development for glioblastoma and other malignancies

Cilengitide, a cyclicized arginine-glycine-aspartic acid-containing pentapeptide, potently blocks ανβ3 and ανβ5 integrin activation. Integrins are upregulated in many malignancies and mediate a wide variety of tumor-stroma interactions. Cilengitide and other integrin-targeting therapeutics have preclinical activity against many cancer subtypes including glioblastoma (GBM), the most common and deadliest CNS tumor. Cilengitide is active against orthotopic GBM xenografts and can augment radiotherapy and chemotherapy in these models. In Phase I and II GBM trials, cilengitide and the combination of cilengitide with standard temozolomide and radiation demonstrate consistent antitumor activity and a favorable safety profile. Cilengitide is currently under evaluation in a pivotal, randomized Phase III study (Cilengitide in Combination With Temozolomide and Radiotherapy in Newly Diagnosed Glioblastoma Phase III Randomized Clinical Trial [CENTRIC]) for newly diagnosed GBM. In addition, randomized controlled Phase II studies with cilengitide are ongoing for non-small-cell lung cancer and squamous cell carcinoma of the head and neck. Cilengitide is the first integrin inhibitor in clinical Phase III development for oncology

Shot Noise of Single-Electron Tunneling in 1D Arrays

We have used numerical modeling and a semi-analytical calculation method to find the low frequency value S_{I}(0) of the spectral density of fluctuations of current through 1D arrays of small tunnel junctions, using the ``orthodox theory'' of single-electron tunneling. In all three array types studied, at low temperature (kT << eV), increasing current induces a crossover from the Schottky value S_{I}(0)=2e to the ``reduced Schottky value'' S_{I}(0)=2e/N (where N is the array length) at some crossover current I_{c}. In uniform arrays over a ground plane, I_{c} is proportional to exp(-\lambda N), where 1/\lambda is the single-electron soliton length. In arrays without a ground plane, I_{c} decreases slowly with both N and \lambda. Finally, we have calculated the statistics of I_{c} for ensembles of arrays with random background charges. The standard deviation of I_{c} from the ensemble average is quite large, typically between 0.5 and 0.7 of , while the dependence of on N or \lambda is so weak that it is hidden within the random fluctuations of the crossover current.Comment: RevTex. 21 pages of text, 10 postscript figure

A trapped-ion local field probe

We introduce a measurement scheme that utilizes a single ion as a local field probe. The ion is confined in a segmented Paul trap and shuttled around to reach different probing sites. By the use of a single atom probe, it becomes possible characterizing fields with spatial resolution of a few nm within an extensive region of millimeters. We demonstrate the scheme by accurately investigating the electric fields providing the confinement for the ion. For this we present all theoretical and practical methods necessary to generate these potentials. We find sub-percent agreement between measured and calculated electric field values

MGMT Promoter Methylation Cutoff with Safety Margin for Selecting Glioblastoma Patients into Trials Omitting Temozolomide: A Pooled Analysis of Four Clinical Trials.

The methylation status of the O &lt;sup&gt;6&lt;/sup&gt; -methylguanine DNA methyltransferase (MGMT) gene promoter is predictive for benefit from temozolomide in glioblastoma (GBM). A clinically optimized cutoff was sought allowing patient selection for therapy without temozolomide, while avoiding to withhold it from patients who may potentially benefit.Experimental Design: Quantitative MGMT methylation-specific PCR data were obtained for newly diagnosed patients with GBM screened or treated with standard radiotherapy and temozolomide in four randomized trials. The pooled dataset was randomly split into a training and test dataset. The unsupervised cutoff was obtained at a 50% probability to be (un)methylated. ROC analysis identified an optimal cutoff supervised by overall survival (OS). For 4,041 patients valid MGMT results were obtained, whereof 1,725 were randomized. The unsupervised cutoff in the training dataset was 1.27 (log &lt;sub&gt;2&lt;/sub&gt; [1,000 × (MGMT+1)/ACTB]), separating unmethylated and methylated patients. The optimal supervised cutoff for unmethylated patients was -0.28 (AUC = 0.61), classifying "truly unmethylated" (≤-0.28) and "gray zone" patients (&gt;-0.28, ≤1.27), the latter comprising approximately 10% of cases. In contrast, for patients with MGMT methylation (&gt;1.27) more methylation was not related to better outcome. Both methylated and gray zone patients performed significantly better for OS than truly unmethylated patients [HR = 0.35, 95% confidence interval (CI), 0.27-0.45, P &lt; 0.0001; HR = 0.58, 95% CI, 0.43-0.78, P &lt; 0.001], validated in the test dataset. The MGMT assay was highly reproducible upon retesting of 218 paired samples (R &lt;sup&gt;2&lt;/sup&gt; = 0.94). Low MGMT methylation (gray zone) may confer some sensitivity to temozolomide treatment, hence the lower safety margin should be considered for selecting patients with unmethylated GBM into trials omitting temozolomide

Association of pre-radiotherapy tumour burden and overall survival in newly diagnosed glioblastoma adjusted for <i>MGMT </i>promoter methylation status

PURPOSE: We retrospectively evaluated the association between postoperative pre-radiotherapy tumour burden and overall survival (OS) adjusted for the prognostic value of O$^{6}$-methylguanine DNA methyltransferase (MGMT) promoter methylation in patients with newly diagnosed glioblastoma treated with radio-/chemotherapy with temozolomide. MATERIALS AND METHODS: Patients were included from the CENTRIC (EORTC 26071-22072) and CORE trials if postoperative magnetic resonance imaging scans were available within a timeframe of up to 4weeks before radiotherapy, including both pre- and post-contrast T1w images and at least one T2w sequence (T2w or T2w-FLAIR). Postoperative (residual) pre-radiotherapy contrast-enhanced tumour (CET) volumes and non-enhanced T2w abnormalities (NT2A) tissue volumes were obtained by three-dimensional segmentation. Cox proportional hazard models and Kaplan Meier estimates were used to assess the association of pre-radiotherapy CET/NT2A volume with OS adjusted for known prognostic factors (age, performance status, MGMT status). RESULTS: 408 tumour (of which 270 MGMT methylated) segmentations were included. Median OS in patients with MGMT methylated tumours was 117 weeks versus 61weeks in MGMT unmethylated tumours (p < 0.001). When stratified for MGMT methylation status, higher CET volume (HR 1.020; 95% confidence interval CI [1.013-1.027]; p < 0.001) and older age (HR 1.664; 95% CI [1.214-2.281]; p = 0.002) were significantly associated with shorter OS while NT2A volume and performance status were not. CONCLUSION: Pre-radiotherapy CET volume was strongly associated with OS in patients receiving radio-/chemotherapy for newly diagnosed glioblastoma stratified by MGMT promoter methylation status

Transport of charged particles by adjusting rf voltage amplitudes

We propose a planar architecture for scalable quantum information processing (QIP) that includes X-junctions through which particles can move without micromotion. This is achieved by adjusting radio frequency (rf) amplitudes to move an rf null along the legs of the junction. We provide a proof-of-principle by transporting dust particles in three dimensions via adjustable rf potentials in a 3D trap. For the proposed planar architecture, we use regularization techniques to obtain amplitude settings that guarantee smooth transport through the X-junction.Comment: 16 pages, 10 figure

Modeling Single Electron Transfer in Si:P Double Quantum Dots

Solid-state systems such as P donors in Si have considerable potential for realization of scalable quantum computation. Recent experimental work in this area has focused on implanted Si:P double quantum dots (DQDs) that represent a preliminary step towards the realization of single donor charge-based qubits. This paper focuses on the techniques involved in analyzing the charge transfer within such DQD devices and understanding the impact of fabrication parameters on this process. We show that misalignment between the buried dots and surface gates affects the charge transfer behavior and identify some of the challenges posed by reducing the size of the metallic dot to the few donor regime.Comment: 11 pages, 7 figures, submitted to Nanotechnolog

Association of pre-radiotherapy tumour burden and overall survival in newly diagnosed glioblastoma adjusted for MGMT promoter methylation status

PURPOSE: We retrospectively evaluated the association between postoperative pre-radiotherapy tumour burden and overall survival (OS) adjusted for the prognostic value of O$^{6}$-methylguanine DNA methyltransferase (MGMT) promoter methylation in patients with newly diagnosed glioblastoma treated with radio-/chemotherapy with temozolomide. MATERIALS AND METHODS: Patients were included from the CENTRIC (EORTC 26071-22072) and CORE trials if postoperative magnetic resonance imaging scans were available within a timeframe of up to 4weeks before radiotherapy, including both pre- and post-contrast T1w images and at least one T2w sequence (T2w or T2w-FLAIR). Postoperative (residual) pre-radiotherapy contrast-enhanced tumour (CET) volumes and non-enhanced T2w abnormalities (NT2A) tissue volumes were obtained by three-dimensional segmentation. Cox proportional hazard models and Kaplan Meier estimates were used to assess the association of pre-radiotherapy CET/NT2A volume with OS adjusted for known prognostic factors (age, performance status, MGMT status). RESULTS: 408 tumour (of which 270 MGMT methylated) segmentations were included. Median OS in patients with MGMT methylated tumours was 117 weeks versus 61weeks in MGMT unmethylated tumours (p < 0.001). When stratified for MGMT methylation status, higher CET volume (HR 1.020; 95% confidence interval CI [1.013-1.027]; p < 0.001) and older age (HR 1.664; 95% CI [1.214-2.281]; p = 0.002) were significantly associated with shorter OS while NT2A volume and performance status were not. CONCLUSION: Pre-radiotherapy CET volume was strongly associated with OS in patients receiving radio-/chemotherapy for newly diagnosed glioblastoma stratified by MGMT promoter methylation status