Optimizing The Melanoma Tropism Of Mouse Parvovirus 1a For Use As A Viral Immunotherapy Vector

Melanoma is a prevalent, deadly disease with poor outcomes following metastasis. It is an immunogenic cancer with many unique tumor antigens, but the tumor microenvironment inhibits the immune system. Current monoclonal antibody treatment has been shown to be beneficial in metastatic melanoma by blocking inhibitory immune signals such as PD-1 or CTLA-4. The expression of immunostimulatory proteins such as B7.1 by tumor cells could provide additional immune targeting. Viral vectors based on the parvovirus genome could efficiently and selectively express B7.1 in melanoma cells, such that they become competent to directly activate cognate T cells. Once activated, T cells will become armed as cytotoxic effectors against cells expressing melanoma tumor antigens. To generate a viral vector, we first needed a parvovirus that infected melanoma. Parvoviruses are uniquely adapted to infect tumor cells as viral replication is dependent on the host cell advancing through S phase. However, most parvoviruses tested were unable to establish infection in the murine melanoma cell line B16F10. The most infectious parvovirus in B16F10 was mouse parvovirus 1a (MPV1a). B16F10 cells were infected with MPV1a, and the progeny virus was harvested and used to re-infect more B16F10 cells. After five serial passages of virus, the mutant polyclonal stock named MPV P5 was analyzed. At 24 hours following infection at a multiplicity of 5000 virions per cell, MPV1a infected 16% of cells, whereas MPV P5 infected 74%. Construction of a molecular clone from the polyclonal MPV P5 mixture yielded a construct consisting of the non-structural and virion polypeptide (NS and VP) genes that was able to infect 45% of cells under the same conditions. The mutations that were most responsible for the increase in B16F10 infectivity were isolated to the VP2 region, which encodes the major capsid protein. By incorporating the viral proteins from the clonal version of MPV P5 into a parvoviral vector system, we have generated a vector capable of transducing B16F10 cells and simultaneously expressing the Green Fluorescent Protein marker and B7.1. Further experiments are needed to determine if the B7.1 expression is sufficient at its current rate of infectivity to modulate an immune response, and to examine whether viral products within the tumor cells generate additional immunogenic signaling. Other aspects of the MPV P5 genome may be engineered into the vector to increase its efficiency

Letter From the Editor

Welcome to the third issue of The Richmond Journal of Law and Technology\u27s seventh publication term! The 2000-2001 academic year has proved to be one of the most productive and exciting in the Journal\u27s decorated history. Our Editorial Board and staff have done a phenomenal job on the Journal\u27s seventh volume and we are very proud of the issue we have worked to prepare for you today

Linear Tetraphenylmethane-Based Thioether Oligomers Stabilising an Entire Gold Nanoparticle by Enwrapping

The design and synthesis of a novel linear thioether-based ligand subunit with a tetraphenylmethane core used in the stabilisation of gold nanoparticles (AuNPs) are presented. Mono-, tri, penta- and heptamers of the ligand have been synthesised and used to stabilise AuNPs by enwrapping. With the exception of the monomer, all ligands provide reliable long-term stability and redispersibility for the coated nanoparticles in common organic solvents. Despite variation of the oligomer length, all stable particles were of the same size within error tolerance (1.16±0.32 nm for the trimer, 1.15±0.30 nm for the pentamer, 1.17±0.34 nm for the heptamer), as investigated by transmission electron microscopy (TEM). These findings suggest that not only the number of sulfur atoms in the ligand, but also its bulkiness play a crucial role in stabilising the AuNPs. These findings are supported by thermogravimetric analysis (TGA), showing that AuNPs stabilised by the penta- or heptamer are passivated by a single ligand. Thermal stability measurements suggest a correlation between ligand coverage and thermal stability, further supporting these findings

Emotional arousal modulates oscillatory correlates of targeted memory reactivation during NREM, but not REM sleep

Rapid eye movement (REM) sleep is considered to preferentially reprocess emotionally arousing memories. We tested this hypothesis by cueing emotional vs. neutral memories during REM and NREM sleep and wakefulness by presenting associated verbal memory cues after learning. Here we show that cueing during NREM sleep significantly improved memory for emotional pictures, while no cueing benefit was observed during REM sleep. On the oscillatory level, successful memory cueing during NREM sleep resulted in significant increases in theta and spindle oscillations with stronger responses for emotional than neutral memories. In contrast during REM sleep, solely cueing of neutral (but not emotional) memories was associated with increases in theta activity. Our results do not support a preferential role of REM sleep for emotional memories, but rather suggest that emotional arousal modulates memory replay and consolidation processes and their oscillatory correlates during NREM sleep

Zur Geschichte von Rechentechnik und Datenverarbeitung in der DDR 1946 - 1968

„Die DDR-Führung suchte in den sechziger Jahren durch gezielten Ressourceneinsatz die Technologielücke zu verkleinern, die die DDR auf dem Gebiet der Rechentechnik zu den westlichen Industrienationen hatte ... Die wissenschaftlichen Institutionen und die Wirtschaft der DDR agierten beim Einführen von Rechentechnik und Datenverarbeitung in dem ihnen vorgegebenen Rahmen durchaus nicht erfolglos ...” [... aus der Einleitung

Sleep benefits emotional and neutral associative memories equally

Background: Emotions modulate memory. It has been suggested that sleep contributes to improved memory of emotional events by preferentially consolidating emotional memories, presumably because of a selective off-line reactivation of information relevant to future behavior. Objectives: We aimed to validate sleep-dependent memory consolidation in a new associative emotional memory paradigm suitable for inducing memory reactivations during sleep. We hypothesized that sleep preferentially might benefit the consolidation of emotional associations independently of their negative vs positive emotional valence. Methods: Seventy-two healthy young participants performed an associative emotional memory task in either the evening or the morning. During the task, they were asked to associate neutrally spoken words to neutral, negative or positive pictures. Cued recall was tested after a 12-h retention interval filled with either night-time sleep or daytime wakefulness. Results: Generally, emotional associations were better remembered than neutral ones. However, we were not able to replicate a selective benefit of sleep on emotional memory. Sleep robustly improved the cued recall performance of all picture types compared with wakefulness, without any modulating influence of emotional arousal or valence. Conclusions: We conclude that the consolidation of explicitly learned associations benefits from sleep, independent of emotional arousal or valence. Selective emotional memory consolidation during sleep may be restricted to non-associative item memory or incidentally learned emotional associations

Effects of Psychotherapy on Glutamatergic Neurotransmission

Introduction: Psychodynamic psychotherapy is an effective and widely used treatment for major depressive disorder (MDD); however, little is known about neurobiological changes associated with induced symptom improvement. Methods: Proton magnetic resonance spectroscopy with a two-dimensional J-resolved sequence served to test the relationship between glutamate (Glu) and glutamine (Gln) levels, measured separately in pregenual anterior cingulate cortex (pgACC) and the anterior midcingulate cortex (aMCC) as a control region, with change in depression symptoms after 6 months of weekly psychodynamic psychotherapy sessions in MDD patients. Depressed (N = 45) and healthy (N = 30) subjects participated in a baseline proton magnetic resonance spectroscopy measurement and a subgroup of MDD subjects (N = 21) then received once-a-week psychodynamic psychotherapy and participated in a second proton magnetic resonance spectroscopy measurement after 6 months. Change in depression symptoms was assessed using the Hamilton Depression Rating Scale (HAMD). Results: Higher pretreatment pgACC Gln concentrations in MDD patients compared to healthy controls were associated with symptom severity. Patients and controls did not differ regarding Gln levels in aMCC nor regarding Glu levels in both regions. The association of pgACC Gln concentration and severity of depressive symptoms was reversed after 6 months of psychotherapy in MDD subjects. Regarding Gln in aMCC as well as Glu in both regions, there were no significant associations with improvement of depressive symptoms in the course of psychotherapy. Discussion: Findings indicate specific regional effects of psychodynamic psychotherapy on glutamatergic neurotransmission and thereby highlight the key role of the pgACC in both depression pathophysiology and recovery

Rising Sound Intensity: An Intrinsic Warning Cue Activating the Amygdala

Human subjects overestimate the change of rising intensity sounds compared with falling intensity sounds. Rising sound intensity has therefore been proposed to be an intrinsic warning cue. In order to test this hypothesis, we presented rising, falling, and constant intensity sounds to healthy humans and gathered psychophysiological and behavioral responses. Brain activity was measured using event-related functional magnetic resonance imaging. We found that rising compared with falling sound intensity facilitates autonomic orienting reflex and phasic alertness to auditory targets. Rising intensity sounds produced neural activity in the amygdala, which was accompanied by activity in intraparietal sulcus, superior temporal sulcus, and temporal plane. Our results indicate that rising sound intensity is an elementary warning cue eliciting adaptive responses by recruiting attentional and physiological resources. Regions involved in cross-modal integration were activated by rising sound intensity, while the right-hemisphere phasic alertness network could not be supported by this stud

ESTRO-ACROP guideline on surface guided radiation therapy

Surface guidance systems enable patient positioning and motion monitoring without using ionising radiation. Surface Guided Radiation Therapy (SGRT) has therefore been widely adopted in radiation therapy in recent years, but guidelines on workflows and specific quality assurance (QA) are lacking. This ESTRO-ACROP guideline aims to give recommendations concerning SGRT roles and responsibilities and highlights common challenges and potential errors. Comprehensive guidelines for procurement, acceptance, commissioning, and QA of SGRT systems installed on computed tomography (CT) simulators, C-arm linacs, closed-bore linacs, and particle therapy treatment systems are presented that will help move to a consensus among SGRT users and facilitate a safe and efficient implementation and clinical application of SGRT. Keywords: ACROP; ESTRO; Guideline; SGRT; Surface guided radiation therapy