Simplified Monopalmitoyl Toll-like Receptor 2 Ligand Mini-UPam for Self-Adjuvanting Neoantigen-Based Synthetic Cancer Vaccines

Synthetic vaccines, based on antigenic peptides that comprise MHC−I and MHC-II T-cell epitopes expressed by tumors, show great promise for the immunotherapy of cancer. For optimal immunogenicity, the synthetic peptides (SPs) should be adjuvanted with suitable immunostimulatory additives. Previously, we have shown that improved immunogenicity in vivo is obtained with vaccine modalities in which an SP is covalently connected to an adjuvanting moiety, typically a ligand to Toll-like receptor 2 (TLR2). SPs were covalently attached to UPam, which is a derivative of the classic TLR2 ligand Pam₃CysSK₄. A disadvantage of the triply palmitoylated UPam is its high lipophilicity, which precludes universal adoption of this adjuvant for covalent modification of various antigenic peptides as it renders the synthetic vaccine insoluble in several cases. Here, we report a novel conjugatable TLR2 ligand, mini-UPam, which contains only one palmitoyl chain, rather than three, and therefore has less impact on the solubility and other physicochemical properties of a synthetic peptide. In this study, we used SPs that contain the clinically relevant neoepitopes identified in a melanoma patient who completely recovered after T-cell therapy. Homogeneous mini-UPam-SP conjugates have been prepared in good yields by stepwise solid-phase synthesis that employed a mini-UPam building block pre-prepared in solution and the standard set of Fmoc-amino acids. The immunogenicity of the novel mini-UPam-SP conjugates was demonstrated by using the cancer patient's T-cells.     

Antisense Oligonucleotide-Based Rescue of Aberrant Splicing Defects Caused by 15 Pathogenic Variants in ABCA4

The discovery of novel intronic variants in the ABCA4 locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes ABCA4 a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1.     

Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach

Bacillus virus Bam35 is the model Betatectivirus and member of the family Tectiviridae, which is composed of tailless, icosahedral, and membrane-containing bacteriophages. Interest in these viruses has greatly increased in recent years as they are thought to be an evolutionary link between diverse groups of prokaryotic and eukaryotic viruses. Additionally, betatectiviruses infect bacteria of the Bacillus cereus group, which are known for their applications in industry and notorious since it contains many pathogens. Here, we present the first protein–protein interactions (PPIs) network for a tectivirus–host system by studying the Bam35–Bacillus thuringiensis model using a novel approach that integrates the traditional yeast two-hybrid system and high-throughput sequencing (Y2H-HTS). We generated and thoroughly analyzed a genomic library of Bam35′s host B. thuringiensis HER1410 and screened interactions with all the viral proteins using different combinations of bait–prey couples. Initial analysis of the raw data enabled the identification of over 4000 candidate interactions, which were sequentially filtered to produce 182 high-confidence interactions that were defined as part of the core virus–host interactome. Overall, host metabolism proteins and peptidases were particularly enriched within the detected interactions, distinguishing this host–phage system from the other reported host–phage PPIs. Our approach also suggested biological roles for several Bam35 proteins of unknown function, including the membrane structural protein P25, which may be a viral hub with a role in host membrane modification during viral particle morphogenesis. This work resulted in a better understanding of the Bam35–B. thuringiensis interaction at the molecular level and holds great potential for the generalization of the Y2H-HTS approach for other virus–host models.     

A practical evaluation of spectrum-based fault localization

Spectrum-based fault localization (SFL) shortens the test–diagnose–repair cycle by reducing the debugging effort. As a light-weight automated diagnosis technique it can easily be integrated with existing testing schemes. Since SFL is based on discovering statistical coincidences between system failures and the activity of the different parts of a system, its diagnostic accuracy is inherently limited. Using a common benchmark consisting of the Siemens set and the space program, we investigate this diagnostic accuracy as a function of several parameters (such as quality and quantity of the program spectra collected during the execution of the system), some of which directly relate to test design. Our results indicate that the superior performance of a particular similarity coefficient, used to analyze the program spectra, is largely independent of test design. Furthermore, near-optimal diagnostic accuracy (exonerating over 80% of the blocks of code on average) is already obtained for low-quality error observations and limited numbers of test cases. In addition to establishing these results in the controlled environment of our benchmark set, we show that SFL can effectively be applied in the context of embedded software development in an industrial environment.     

耳机放大器设计：品质VS效率

在最近几年中,D类放大器因其体积小,效率高,已经进入了消费类音频电子产品、汽车以及便携式音频产品市场.它的效率帮助减小了汽车放大器以及便携式媒体扩展插槽的尺寸,并以较小的波形因子提供了更大的功率,有助实现新式的消费类音频设备.     

A heuristic to manage perishable inventory with batch ordering,positive lead-times,and time-varying demand

So far the literature on inventory control for perishable products has mainly focused on (near-) optimal replenishment policies for a stylized environment, assuming no lead-time, no lot-sizing, stationary demand, a first in first out withdrawal policy and/or product life time equal to two periods. This literature has given fundamental insight in the behavior and the complexity of inventory systems for perishable products. In practice, many grocery retailers have recently automated the inventory replenishment for non-perishable products. They recognize they may need a different replenishment logic for perishable products, which takes into account e.g. the age of the inventory in the system. Due to new information technologies like RFID, it now also becomes more economically feasible to register this type of information. This paper suggests a replenishment policy for perishable products which takes into account the age of inventories and which requires only very simple calculations. It will be shown that in an environment, which contains important features of the real-life retail environment, this new policy leads to substantial cost reductions compared with a base policy that does not take into account the age of inventories.     

Application of high-throughput computing in bioinformatics

We describe the challenges faced when developing a Linux/PC-based cluster to apply bioinformatics algorithms to the rapidly increasing raw genomics data available. The calculations, which take around two months to complete, result in a powerful resource that can be used for data mining--most obviously for the human genome. Our current infrastructure consists of a 1314 node cluster with 1734 processors supporting both production and research. This paper highlights the problems in achieving high data throughput with such systems and shows that raw computer power is only one component of a complex problem.     

Perceived trustworthiness of knowledge sources: The moderating impact of relationship length.

Prior meta-analytic evidence has indicated no association between relationship length and perceived trustworthiness. Viewing trustors as information processors, the authors propose a model in which relationship length, although having no direct effect on perceived trustworthiness, moderates the association between perceived trustworthiness and the basis on which people decide to trust each other. Specifically, as trustors learn about others, they base their trust on different kinds of information (demographic similarity, trustworthy behavior, and shared perspective). Hierarchical multiple regression analyses of a field survey of supervisors and subordinates from 3 companies (N = 88) provide evidence consistent with this prediction: Perceived trustworthiness is associated with demographic similarity in newer relationships, with trustworthy behavior in relationships that are neither brand new nor old but in-between, and with shared perspective in older relationships. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Movie Making at Pixar: A Collaboration of Art and Technology

This talk takes you behind the scenes at Pixar Animation Studios for an in‐depth look at how its 3d computer graphics films are made. Making a computer animated film involves people with artistic talent and people with technical skills working together in close collaboration. The process starts with the development of the story and continues with modeling the geometry, adding articulation controls, using those controls to animate the characters, simulating things like water and cloth and hair, defining the look of the surfaces, putting lights in the scene, adding special effects, rendering, and post‐production. Special emphasis is given to the roles of technology and computer graphics research in supporting the filmmaker.