High Throughput Primer Walking of cDNA Clones

Primer walking of cloned DNA is a standard research tool. It has been used in the past to determine the sequence of individual clones of interest. With the expansion of DNA sequencing capacity the need to be able to walk larger numbers of clones has become necessary. Our laboratory is a mid-sized genomics facility. In conjunction with the Advanced Biomedical Computing Center (ABCC) we have developed methods for automating the primer selection, DNA sequencing, contig assembly and sequence analysis for clones arrayed in microtiter format. This approach has allowed us to walk 475 clones (five microtiter plates) selected from a cDNA library.     

Decision support aids with anthropomorphic characteristics influence trust and performance in younger and older adults

This study examined the use of deliberately anthropomorphic automation on younger and older adults' trust, dependence and performance on a diabetes decision-making task. Research with anthropomorphic interface agents has shown mixed effects in judgments of preferences but has rarely examined effects on performance. Meanwhile, research in automation has shown some forms of anthropomorphism (e.g. etiquette) have effects on trust and dependence on automation. Participants answered diabetes questions with no-aid, a non-anthropomorphic aid or an anthropomorphised aid. Trust and dependence in the aid was measured. A minimally anthropomorphic aide primarily affected younger adults' trust in the aid. Dependence, however, for both age groups was influenced by the anthropomorphic aid. Automation that deliberately embodies person-like characteristics can influence trust and dependence on reasonably reliable automation. However, further research is necessary to better understand the specific aspects of the aid that affect different age groups. Automation that embodies human-like characteristics may be useful in situations where there is under-utilisation of reasonably reliable aids by enhancing trust and dependence in that aid. Practitioner Summary: The design of decision-support aids on consumer devices (e.g. smartphones) may influence the level of trust that users place in that system and their amount of use. This study is the first step in articulating how the design of aids may influence user's trust and use of such systems.     

Preclinical Assessment of the Combination of PSMA-Targeting Radionuclide Therapy with PARP Inhibitors for Prostate Cancer Treatment

Prostate specific membrane antigen targeted radionuclide therapy (PSMA-TRT) is a promising novel treatment for prostate cancer (PCa) patients. However, PSMA-TRT cannot be used for curative intent yet, thus additional research on how to improve the therapeutic efficacy is warranted. A potential way of achieving this, is combining TRT with poly ADP-ribosylation inhibitors (PARPi), which has shown promising results for TRT of neuroendocrine tumor cells. Currently, several clinical trials have been initiated for this combination for PCa, however so far, no evidence of synergism is available for PCa. Therefore, we evaluated the combination of PSMA-TRT with three classes of PARPi in preclinical PCa models. In vitro viability and survival assays were performed using PSMA-expressing PCa cell lines PC3-PIP and LNCaP to assess the effect of increasing concentrations of PARPi veliparib, olaparib or talazoparib in combination with PSMA-TRT compared to single PARPi treatment. Next, DNA damage analyses were performed by quantifying the number of DNA breaks by immunofluorescent stainings. Lastly, the potential of the combination treatments was studied in vivo in mice bearing PC3-PIP xenografts. Our results show that combining PSMA-TRT with PARPi did not synergistically affect the in vitro clonogenic survival or cell viability. DNA-damage analysis revealed only a significant increase in DNA breaks when combining PSMA-TRT with veliparib and not in the other combination treatments. Moreover, PSMA-TRT with PARPi treatment did not improve tumor control compared to PSMA-TRT monotherapy. Overall, the data presented do not support the assumption that combining PSMA-TRT with PARPi leads to a synergistic antitumor effect in PCa. These results underline that extensive preclinical research using various PCa models is imperative to validate the applicability of the combination strategy for PCa, as it is for other cancer types.     

A Guide to Polysaccharide-Based Hydrogel Bioinks for 3D Bioprinting Applications

Three-dimensional (3D) bioprinting is an innovative technology in the biomedical field, allowing the fabrication of living constructs through an approach of layer-by-layer deposition of cell-laden inks, the so-called bioinks. An ideal bioink should possess proper mechanical, rheological, chemical, and biological characteristics to ensure high cell viability and the production of tissue constructs with dimensional stability and shape fidelity. Among the several types of bioinks, hydrogels are extremely appealing as they have many similarities with the extracellular matrix, providing a highly hydrated environment for cell proliferation and tunability in terms of mechanical and rheological properties. Hydrogels derived from natural polymers, and polysaccharides, in particular, are an excellent platform to mimic the extracellular matrix, given their low cytotoxicity, high hydrophilicity, and diversity of structures. In fact, polysaccharide-based hydrogels are trendy materials for 3D bioprinting since they are abundant and combine adequate physicochemical and biomimetic features for the development of novel bioinks. Thus, this review portrays the most relevant advances in polysaccharide-based hydrogel bioinks for 3D bioprinting, focusing on the last five years, with emphasis on their properties, advantages, and limitations, considering polysaccharide families classified according to their source, namely from seaweed, higher plants, microbial, and animal (particularly crustaceans) origin.     

Targeting CD10 on B-Cell Leukemia Using the Universal CAR T-Cell Platform (UniCAR)

Chimeric antigen receptor (CAR)-expressing T-cells are without a doubt a breakthrough therapy for hematological malignancies. Despite their success, clinical experience has revealed several challenges, which include relapse after targeting single antigens such as CD19 in the case of B-cell acute lymphoblastic leukemia (B-ALL), and the occurrence of side effects that could be severe in some cases. Therefore, it became clear that improved safety approaches, and targeting multiple antigens, should be considered to further improve CAR T-cell therapy for B-ALL. In this paper, we address both issues by investigating the use of CD10 as a therapeutic target for B-ALL with our switchable UniCAR system. The UniCAR platform is a modular platform that depends on the presence of two elements to function. These include UniCAR T-cells and the target modules (TMs), which cross-link the T-cells to their respective targets on tumor cells. The TMs function as keys that control the switchability of UniCAR T-cells. Here, we demonstrate that UniCAR T-cells, armed with anti-CD10 TM, can efficiently kill B-ALL cell lines, as well as patient-derived B-ALL blasts, thereby highlighting the exciting possibility for using CD10 as an emerging therapeutic target for B-cell malignancies.     

Ethical challenges for psychologists conducting humanitarian work.

This article describes the ethical challenges that psychologists face when doing international humanitarian aid work. It is based on personal experience and uses the framework of the Canadian Code of Ethics for Psychologists to reflect on the unique challenges when doing such work. The article is organized around themes of ethical challenges under the following headings: limitation of resources, local political pressure, local cultural and social realities, challenges to confidentiality, problems in obtaining consent, sequelae of colonial history, individual versus population interests, and tolerance of differences in standards. Examples illustrate these points. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

A Content Analysis of Print News Coverage of Media Violence and Aggression Research

We conducted a content analysis of news articles (N = 540) to examine whether news coverage of media violence accurately reflects scientific knowledge about exposure to media violence and its effects on viewer aggression. The analysis revealed that over the past 30 years, news articles generally suggested that a link between media violence and aggression exists. However, the tone shifted sharply back toward a neutral conclusion since 2000. This shift may be attributable to the type of medium discussed (e.g., television vs. video games), the number of unaffiliated sources that are cited in the news article, and the sex of the journalist. Implications for how this news coverage may influence news readers are discussed.     

Measurement of Onset of Structural Relaxation in Melt-Quenched Phase Change Materials

Chalcogenide phase change materials enable non-volatile, low-latency storage-class memory. They are also being explored for new forms of computing such as neuromorphic and in-memory computing. A key challenge, however, is the temporal drift in the electrical resistance of the amorphous states that encode data. Drift, caused by the spontaneous structural relaxation of the newly recreated melt-quenched amorphous phase, has consistently been observed to have a logarithmic dependence in time. Here, it is shown that this observation is valid only in a certain observable timescale. Using threshold-switching voltage as the measured variable, based on temperature-dependent and short timescale electrical characterization, the onset of drift is experimentally measured. This additional feature of the structural relaxation dynamics serves as a new benchmark to appraise the different classical models to explain drift.     

How Modifying Third‐Party Information Affects Interpersonal Impressions and the Evaluation of Collaborative Online Media

Previous research has drawn upon warranting theory to help explain how viewers evaluate people and entities online. Extending previous research, this study assesses how the ability of a target to modify third‐party information affects perceptions of warranting value, and in turn, interpersonal impressions and the perceived legitimacy of online media that host evaluations. Additionally, this work explores how the perceived objectivity of a third‐party evaluator affects impressions in online settings. The results provide support for warranting theory and help clarify how impressions are formed in online environments when people have the ability to generate and modify content collectively. The theoretical implications this study has for warranting theory and future research directions are discussed.