A Universal Live Cell Barcoding-Platform for Multiplexed Human Single Cell Analysis.

Single-cell barcoding enables the combined processing and acquisition of multiple individual samples as one. This maximizes assay efficiency and eliminates technical variability in both sample preparation and analysis. Remaining challenges are the barcoding of live, unprocessed cells to increase downstream assay performance combined with the flexibility of the approach towards a broad range of cell types. To that end, we developed a novel antibody-based platform that allows the robust barcoding of live human cells for mass cytometry (CyTOF). By targeting both the MHC class I complex (beta-2-microglobulin) and a broadly expressed sodium-potassium ATPase-subunit (CD298) with platinum-conjugated antibodies, human immune cells, stem cells as well as tumor cells could be multiplexed in the same single-cell assay. In addition, we present a novel palladium-based covalent viability reagent compatible with this barcoding strategy. Altogether, this platform enables mass cytometry-based, live-cell barcoding across a multitude of human sample types and provides a scheme for multiplexed barcoding of human single-cell assays in general

Short guide to lunar sample information

Data pertaining to the collection, processing, and study of lunar samples, and the ways to gain access to that data are described. Data pertaining to the samples includes not only the normally published literature but also NASA and USGS interagency documents, sample processing information, and photography taken from lunar orbit, on the lunar surface, and of samples in the Lunar Receiving Laboratory

From the Outside Looking In: Employers’ Views of the Basic Course

This essay is designed to connect specific communication skills desired by employers in industry to basic course concepts. While communication is often identified as one of the most important skills for graduates seeking employment, this broad view makes it nearly impossible for basic course directors and instructors to design their pedagogy to satisfy students’ future needs. This manuscript examines a part of the 2014 Basic Course Conference where industry leaders were invited to present what they felt to be the most important communication skills and knowledge in employees and then engaged in a discussion with the attending basic course directors to clarify which specific communication skills they desired most. This study ties those communication skills and knowledge to concepts common to the basic course identified in the 2014 document produced by the National Communication Association Core Competencies for Introductory Communication Courses Task Force on the role of communication in general education, as well as the most recent Basic Communication Course Survey from 2010

The Invaluable Nature of Speech Evaluation Training for New Basic Course Instructors

Recent reforms in higher education recognize the centrality of communication in general education programs (e.g., Association of American Colleges and Universities, American Association of State Colleges and Universities, LEAP, Common Core State Standards). As oral communication knowledge and skills are becoming recognized as integral to general education programs across the country, many basic course directors are finding themselves in the position of offering multiple sections of the course taught by multiple instructors. Additionally, basic course directors find themselves with the responsibility of providing clear measures of what they do and how well they do it. Because oral communication assessment is key to remaining integral to general education (Allen, 2002), basic course directors must provide instructor training on how to fairly and consistently evaluate student performances. But before this training can take place, basic course directors need to have an evaluation system in place that is fair, consistent, and reflective of actual student performance. There are several challenges to speech evaluation that warrant such a process. This essay will address those challenges and propose a systematic evaluation process that can serve as an impetus to instructor training in this area

The parafibromin tumor suppressor protein interacts with actin-binding proteins actinin-2 and actinin-3

<p>Abstract</p> <p>Background</p> <p>Germline and somatic inactivating mutations in the <it>HRPT2 </it>gene occur in the inherited hyperparathyroidism-jaw tumor syndrome, in some cases of parathyroid cancer and in some cases of familial hyperparathyroidism. <it>HRPT2 </it>encodes parafibromin. To identify parafibromin interacting proteins we used the yeast two-hybrid system for screening a heart cDNA library with parafibromin as the bait.</p> <p>Results</p> <p>Fourteen parafibromin interaction positive preys representing 10 independent clones encoding actinin-2 were isolated. Parafibromin interacted with muscle alpha-actinins (actinin-2 and actinin-3), but not with non-muscle alpha-actinins (actinin-1 and actinin-4). The parafibromin-actinin interaction was verified by yeast two-hybrid, GST pull-down, and co-immunoprecipitation. Yeast two-hybrid analysis revealed that the N-terminal region of parafibromin interacted with actinins. In actin sedimentation assays parafibromin did not dissociate skeletal muscle actinins from actin filaments, but interestingly, parafibromin could also bundle/cross-link actin filaments. Parafibromin was predominantly nuclear in undifferentiated proliferating myoblasts (C2C12 cells), but in differentiated C2C12 myotubes parafibromin co-localized with actinins in the cytoplasmic compartment.</p> <p>Conclusion</p> <p>These data support a possible contribution of parafibromin outside the nucleus through its interaction with actinins and actin bundling/cross-linking. These data also suggest that actinins (and actin) participate in sequestering parafibromin in the cytoplasmic compartment.</p

Instructional Discussion: The Most Important Area of Training for New Basic Course Instructors

In order to determine the most important concept to teach new basic course instructors, it is important to know what we want students to be able to do as a result of the basic course and what teaching method will best reach that outcome. One main goal of the basic course is to teach students to communicate orally and give them practice doing so. This can be accomplished through what Muller (2014) defines as instructional discussion, or “an instructional interaction where teachers and students engage together in an exploration of problems, ideas, and questions in ways that incorporate the knowledge of all participants to generate a collective wisdom or understanding that would not have emerged without the interaction” (p. 326). This definition illustrates the importance of engagement and interaction, both important goals within the basic communication course. Additionally, instructional discussion highlights the central role of communication in the teaching and learning process. Thus, it is imperative that training programs for basic course instructors address how to plan, facilitate, and assess an instructional discussion as well as teach students how to engage in the process

Preventing Perinatal Transmission of HIV Costs and Effectiveness of a Recommended Intervention

In the United States, nearly all new human immunodeficiency virus (HIV) infections in children are acquired through perinatal (mother-to-infant) transmission. Each year, approximately 7000 infants are born to HIV-infected women in the United States.1 Without intervention, an estimated 15-30% of these infants would become infected.2 In 1994, results of the AIDS Clinical Trial Group (ACTG) Protocol 076 showed that treatment of infected pregnant women and their infants with zidovudine (ZDV) reduced the rate of perinatal HIV transmission from 25% to 8%.3,4 Following these findings, the Public Health Service (PHS) issued recommendations for ZDV therapy to prevent perinatal HIV transmission5 and for HIV counseling and voluntary testing of pregnant women.

GateFinder: projection-based gating strategy optimization for flow and mass cytometry

Motivation: High-parameter single-cell technologies can reveal novel cell populations of interest, but studying or validating these populations using lower-parameter methods remains challenging.Results: Here, we present GateFinder, an algorithm that enriches high-dimensional cell types with simple, stepwise polygon gates requiring only two markers at a time. A series of case studies of complex cell types illustrates how simplified enrichment strategies can enable more efficient assays, reveal novel biomarkers and clarify underlying biology

The EIF4EBP3 translational repressor is a marker of CDC73 tumor suppressor haploinsufficiency in a parathyroid cancer syndrome

Germline mutation of the tumor suppressor gene CDC73 confers susceptibility to the hyperparathyroidism-jaw tumor syndrome associated with a high risk of parathyroid malignancy. Inactivating CDC73 mutations have also been implicated in sporadic parathyroid cancer, but are rare in sporadic benign parathyroid tumors. The molecular pathways that distinguish malignant from benign parathyroid transformation remain elusive. We previously showed that a hypomorphic allele of hyrax (hyx), the Drosophila homolog of CDC73, rescues the loss-of-ventral-eye phenotype of lobe, encoding the fly homolog of Akt1s1/ PRAS40. We report now an interaction between hyx and Tor, a central regulator of cell growth and autophagy, and show that eukaryotic translation initiation factor 4E-binding protein (EIF4EBP), a translational repressor and effector of mammalian target of rapamycin (mTOR), is a conserved target of hyx/CDC73. Flies heterozygous for Tor and hyx, but not Mnn1, the homolog of the multiple endocrine neoplasia type 1 (MEN1) tumor suppressor associated with benign parathyroid tumors, are starvation resistant with reduced basal levels of Thor/4E-BP. Human peripheral blood cell levels of EIF4EBP3 were reduced in patients with CDC73, but not MEN1, heterozygosity. Chromatin immunoprecipitation demonstrated occupancy of EIF4EBP3 by endogenous parafibromin. These results show that EIF4EBP3 is a peripheral marker of CDC73 function distinct from MEN1-regulated pathways, and suggest a model whereby starvation resistance and/or translational de-repression contributes to parathyroid malignant transformation

Inhibition of G-protein signalling in cardiac dysfunction of intellectual developmental disorder with cardiac arrhythmia (IDDCA) syndrome

BACKGROUND: Pathogenic variants of GNB5 encoding the β5 subunit of the guanine nucleotide-binding protein cause IDDCA syndrome, an autosomal recessive neurodevelopmental disorder associated with cognitive disability and cardiac arrhythmia, particularly severe bradycardia. METHODS: We used echocardiography and telemetric ECG recordings to investigate consequences of Gnb5 loss in mouse. RESULTS: We delineated a key role of Gnb5 in heart sinus conduction and showed that Gnb5-inhibitory signalling is essential for parasympathetic control of heart rate (HR) and maintenance of the sympathovagal balance. Gnb5-/- mice were smaller and had a smaller heart than Gnb5+/+ and Gnb5+/- , but exhibited better cardiac function. Lower autonomic nervous system modulation through diminished parasympathetic control and greater sympathetic regulation resulted in a higher baseline HR in Gnb5-/- mice. In contrast, Gnb5-/- mice exhibited profound bradycardia on treatment with carbachol, while sympathetic modulation of the cardiac stimulation was not altered. Concordantly, transcriptome study pinpointed altered expression of genes involved in cardiac muscle contractility in atria and ventricles of knocked-out mice. Homozygous Gnb5 loss resulted in significantly higher frequencies of sinus arrhythmias. Moreover, we described 13 affected individuals, increasing the IDDCA cohort to 44 patients. CONCLUSIONS: Our data demonstrate that loss of negative regulation of the inhibitory G-protein signalling causes HR perturbations in Gnb5-/- mice, an effect mainly driven by impaired parasympathetic activity. We anticipate that unravelling the mechanism of Gnb5 signalling in the autonomic control of the heart will pave the way for future drug screening