Design of the fill/transfer station cryostat for the OMEGA cryogenic target system

General Atomics is designing, testing and fabricating a system for supplying cryogenic targets for the University of Rochester`s OMEGA laser system. A prototype system has demonstrated the filling of 1 mm diameter, 3 {micro}m wall plastic spheres to 111 MPa (1,100 atm) with deuterium and then cooling to 18 K to condense the fuel. The production design must be capable of routinely filling and cooling targets with a 50/50 mix of deuterium and tritium and transferring them to a device which places the targets into the focus of 60 laser beams. This paper discusses the design and analysis of the production Fill/Transfer Station cryostat. The cryostat has two major components, a fixed base and a removable dome. The joint between the base and the dome is similar to a bayonet fitting and is sealed by a room temperature elastomeric o-ring. Since the cryostat must be housed in a glovebox, its design is driven strongly by maintenance requirements. To reach the equipment inside the cryostat, the dome is simply unbolted and lifted. The inside of the cryostat is maintained at 16 K by a closed loop helium flow system. Gaseous helium at about 1.4 MPa (200 psi) flows through tubes which are brazed to the inner walls. Cooling is provided by several cryocoolers which are located external to the cryostat. Liquid nitrogen is used as a heat intercept and to precool the helium gas

Tumor-initiating cell frequency is relevant for glioblastoma aggressiveness

Glioblastoma (GBM) is maintained by a small subpopulation of tumor-initiating cells (TICs). The arduous assessment of TIC frequencies challenges the prognostic role of TICs in predicting the clinical outcome in GBM patients. We estimated the TIC frequency in human GBM injecting intracerebrally in mice dissociated cells without any passage in culture.All GBMs contained rare TICsand were tumorigenic in vivo but only 54% of them grew in vitro as neurospheres. We demonstrated that neurosphere formation in vitro did not foretell tumorigenic ability in vivo and frequencies calculated in vitro overestimated the TIC content.Our findings assert the pathological significance of GBM TICs. TIC number correlated positively with tumor incidence and inversely with survival of tumor-bearing mice. Stratification of GBM patients according to TIC content revealed that patients with low TIC frequency experienced a trend towards a longer progression free survival. The expression of either putative stem-cell markers or markers associated with different GBM molecular subtypes did not associate with either TIC content or neurosphere formation underlying the limitations of TIC identification based on the expression of some putative stem cell-markers

Regulatory T cells with multiple suppressive and potentially pro-tumor activities accumulate in human colorectal cancer

Tregs can contribute to tumor progression by suppressing antitumor immunity. Exceptionally, in human colorectal cancer (CRC), Tregs are thought to exert beneficial roles in controlling pro-tumor chronic inflammation. The goal of our study was to characterize CRC-infiltrating Tregs at multiple levels, by phenotypical, molecular and functional evaluation of Tregs from the tumor site, compared to non-tumoral mucosa and peripheral blood of CRC patients. The frequency of Tregs was higher in mucosa than in blood, and further significantly increased in tumor. Ex vivo, those Tregs suppressed the proliferation of tumor-infiltrating CD8(+) and CD4(+) T cells. A differential compartmentalization was detected between Helioshigh and Helios(low) Treg subsets (thymus-derived versus peripherally induced): while Helios(low) Tregs were enriched in both sites, only Helios(high) Tregs accumulated significantly and specifically in tumors, displayed a highly demethylated TSDR region and contained high proportions of cells expressing CD39 and OX40, markers of activation and suppression. Besides the suppression of T cells, Tregs may contribute to CRC progression also through releasing IL-17, or differentiating into Tfr cells that potentially antagonize a protective Tfh response, events that were both detected in tumor-associated Tregs. Overall, our data indicate that Treg accumulation may contribute through multiple mechanisms to CRC establishment and progression

Preoperative Magnetic Resonance and Intraoperative Ultrasound Fusion Imaging for Real-Time Neuronavigation in Brain Tumor Surgery = Präoperative MRI- und intraoperative Ultraschallfusion für die Echtzeit-Neuronavigation in der Neurochirurgie von Hirntumoren

Purpose: Brain shift and tissue deformation during surgery for intracranial lesions are the main actual limitations of neuro-navigation (NN), which currently relies mainly on preoperative imaging. Ultrasound (US), being a real-time imaging modality, is becoming progressively more widespread during neurosurgical procedures, but most neurosurgeons, trained on axial computed tomography (CT) and magnetic resonance imaging (MRI) slices, lack specific US training and have difficulties recognizing anatomic structures with the same confidence as in preoperative imaging. Therefore real-time intraoperative fusion imaging (FI) between preoperative imaging and intraoperative ultrasound (ioUS) for virtual navigation (VN) is highly desirable. We describe our procedure for real-time navigation during surgery for different cerebral lesions. Materials and Methods: We performed fusion imaging with virtual navigation for patients undergoing surgery for brain lesion removal using an ultrasound-based real-time neuro-navigation system that fuses intraoperative cerebral ultrasound with preoperative MRI and simultaneously displays an MRI slice coplanar to an ioUS image. Results: 58 patients underwent surgery at our institution for intracranial lesion removal with image guidance using a US system equipped with fusion imaging for neuro-navigation. In all cases the initial (external) registration error obtained by the corresponding anatomical landmark procedure was below 2mm and the craniotomy was correctly placed. The transdural window gave satisfactory US image quality and the lesion was always detectable and measurable on both axes. Brain shift/deformation correction has been successfully employed in 42 cases to restore the co-registration during surgery. The accuracy of ioUS/MRI fusion/overlapping was confirmed intraoperatively under direct visualization of anatomic landmarks and the error was < \u30083mm in all cases (100%). Conclusion: Neuro-navigation using intraoperative US integrated with preoperative MRI is reliable, accurate and user-friendly. Moreover, the adjustments are very helpful in correcting brain shift and tissue distortion. This integrated system allows true real-time feedback during surgery and is less expensive and time-consuming than other intraoperative imaging techniques, offering high precision and orientation.Brain Shift und Gewebeverschiebung w\ue4hrend der chirurgischen Entfernung intrakranialer Raumforderungen sind die limitierenden Faktoren bei der Neuronavigation (NN), welche aktuell haupts\ue4chlich pr\ue4operative Bilder einsetzt. Ultraschall (US) als Echtzeit-Bildgebung wird bei neurochirurgischen Prozeduren zunehmend angewandt. Vielen Neurochirurgen fehlt aber die US Expertise, da schon in der Ausbildung standarisierte (typisch axiale) CT und MRT Schnittbilder f\ufcr die Navigation bevorzugt eingesetzt werden und somit die Sicherheit bei der sonografischen Identifikation anatomischer Strukturen fehlt. Daher ist eine intraoperative Echtzeitfusion zwischen pr\ue4operativen CT bzw. MRT Bildern und intraoperativem Ultraschall (ioUS) im Rahmen der virtuellen Navigation (VN) au ferordentlich w\ufcnschenswert. Wir pr\ue4sentieren hier die bei uns angewandte Methode f\ufcr dieEchtzeitnavigation bei der Entfernung verschiedener Hirntumoren. Material und Methoden: Wir wandten die Bildfusion mit virtueller Navigation bei der chirurgischen Entfernung von Hirntumoren an. Zum Einsatz kam ein Neuronavigationssystem, welches intraoperative Ultraschallbilder mit pr\ue4operativen MRT Bildern in Echtzeit \ufcberlagert und zu jedem US Bild simultan die dazu passende ko-planare MRTSchnittebene anzeigt. Ergebnisse: Die US-basierte Neuronavigation wurde bei der Operation von 58 Patienten mit Hirntumoren eingesetzt. In allen F\ue4llen war der Fehler der initialen (externen) Registrierung, welche anhand von anatomischen Landmarken erfolgte, unterhalb von 2mm und die Kraniotomie konnte korrekt angesetzt werden. Die Bildqualit\ue4t des transduralen Ultraschalls war gut und die L\ue4sion konnte bei allen Patienten detektiert und in allen Achsen vermessen werden. Die Korrektur von Brain Shift sowie Gewebeverschiebung gelang erfolgreich in 42 F\ue4llen zur Wiederherstel lung der intraoperativen Co-Registrierung. Die Genauigkeit der cberlagerung von ioUS und MRT wurde intraoperativ anhand der Visualisierung anatomischerLandmarken \ufcberpr\ufcft und der Fehler lag in allen F\ue4llen (100 %) unterhalb von 3mm. Schlussfolgerung: Neuronavigation mit Hilfe von in pr\ue4operative MRT Bilder integrierten intraoperativen US Bildern ist eine zuverl\ue4ssige, genaue und anwenderfreundliche neue Technologie. Brain Shift und Gewebeverlagerungen k\uf6nnen anhand verschiedener Einstellungsm\uf6glichkeiten am System erfolgreich intraoperativ korrigiert werden. Das integrierte System erm\uf6glicht eine intraoperative cberpr\ufcfung der Navigation in Echtzeit und ist dabei kosteng\ufcnstiger und weniger Zeit aufw\ue4ndig als andere intraoperative Bild-gebende Verfahren, trotzdem aber hoch pr\ue4zise

Peripheral blood biomarkers as prognostic factors for immunotherapy in advanced non-small cell lung cancer (aNSCLC) patients

n/

Base-editing-mediated dissection of a γ-globin cis-regulatory element for the therapeutic reactivation of fetal hemoglobin expression

: Sickle cell disease and β-thalassemia affect the production of the adult β-hemoglobin chain. The clinical severity is lessened by mutations that cause fetal γ-globin expression in adult life (i.e., the hereditary persistence of fetal hemoglobin). Mutations clustering ~200 nucleotides upstream of the HBG transcriptional start sites either reduce binding of the LRF repressor or recruit the KLF1 activator. Here, we use base editing to generate a variety of mutations in the -200 region of the HBG promoters, including potent combinations of four to eight γ-globin-inducing mutations. Editing of patient hematopoietic stem/progenitor cells is safe, leads to fetal hemoglobin reactivation and rescues the pathological phenotype. Creation of a KLF1 activator binding site is the most potent strategy - even in long-term repopulating hematopoietic stem/progenitor cells. Compared with a Cas9-nuclease approach, base editing avoids the generation of insertions, deletions and large genomic rearrangements and results in higher γ-globin levels. Our results demonstrate that base editing of HBG promoters is a safe, universal strategy for treating β-hemoglobinopathies