Autoantibody-associated kappa light chain variable region gene expressed in chronic lymphocytic leukemia with little or no somatic mutation. Implications for etiology and immunotherapy.

Recently the minor B cell subpopulation that expresses the CD5 (Leu-1) antigen has been implicated as a source of IgM autoantibodies. Chronic lymphocytic leukemia (CLL), the most common leukemia in humans, represents a malignancy of small B lymphocytes that also express the CD5 antigen. However, little is known concerning the antibody variable region genes (V genes) that are used by these malignant CD5 B cells. We have found that a relatively high frequency of CLL patients have leukemic B cells with surface immunoglobulin (sIg) recognized by 17.109, a murine mAb specific for a kappa light chain associated crossreactive idiotype (CRI) associated with rheumatoid factor and other IgM autoantibodies. Flow cytometric analyses revealed that the relative expression of the 17.109-CRI by circulating leukemic B cells was directly proportional to the levels of sIg kappa light chain, indicating that there exists stable idiotype expression in the leukemic population. To examine this at the molecular level, the nucleic acid sequences encoding the Ig kappa light chains of two unrelated patients with CLL bearing sIg with the 17.109-CRI were determined. Analyses of multiple independent kappa light chain cDNA clones did not reveal any evidence for sequence heterogeneity in the CLL cell population. Furthermore, the nucleic acid sequences expressed by the leukemic cells of these two patients were identical or very homologous to a germline V kappa gene isolated from placental DNA, designated Humkv 325, or "V kappa RF" because of its association with IgM autoantibodies. This study suggests; (a) that the malignant CD5+ B lymphocytes in CLL use the same V kappa gene that has been highly associated with IgM autoantibodies and (b) that the expression of V genes is stable in CLL, in contrast to other B cell malignancies examined to date. We propose that many CLL cases represent malignancies of autoreactive CD5 B cells that use a restricted set of conserved V genes. This property may render CLL particularly amenable to immunotherapy with antiidiotypic antibodies

An analysis of the State

The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file.Title from title screen of research.pdf file (viewed on July 31, 2009)Thesis (Ph. D.) University of Missouri-Columbia 2008.What the state is remains far from clear in political philosophy. However, the state is also a key concept at work in many discussions in political philosophy. For example, there is a debate about anarchism, the question of whether or not the state is legitimate in some way. However, if we are unclear about what the state is, then we cannot be clear about what the position of anarchism amounts to. To this end, I have attempted to find some necessary features of statehood. I have done this by considering two debates in political philosophy that concern the state. The first is the already mentioned issue of anarchism. The second is the issue of state sovereignty in international relations. The worry in this second debate is about the interference by some states with the affairs of other states. The guiding question for the dissertation has been: What features are required for the debates in question to make sense? If a debate requires a feature, then that feature is necessary for statehood. The central feature for statehood which has emerged concerns control. Basically, a state is an organization that controls the lives of its citizens in some way. Usually, this minimally means that a state says when citizens can and cannot use force and that this control is ensured through the use of forcible coercion. Any other features that we may be able to point to as necessary will be necessary only to ensure that states do control the lives if their citizens in some way.Includes bibliographical reference

Assessing Transit Access to Ramsey County Service Facilities

Report, presentation and poster completed by students enrolled in GIS 5578: GIS Programming, taught by David Haynes in Fall 2018.This project was completed as part of the 2018-2019 Resilient Communities Project (rcp.umn.edu) partnership with Ramsey County. Ramsey County wanted to assess the accessibility for visitors and clients of County-owned service facilities to make informed decisions about future capital investment in or relocation of these facilities to better meet the needs of the public. Ramsey County project lead Max Holdhusen worked with a graduate student in David Haynes' GIS 5578: GIS Programming, who used GIS analysis to determine public transit access to three Ramsey County Corrections Facilities. Based on this analysis, the student concluded that midday transit access is inadequate for residents of northwestern Ramsey County, and that if the central corrections facility were moved slightly to the northwest, it would be located in a region of higher client density and transit accessibility. The student's final report, presentation, and a poster summarizing the project are available.This project was supported by the Resilient Communities Project (RCP), a program at the University of Minnesota whose mission is to connect communities in Minnesota with U of MN faculty and students to advance community resilience through collaborative, course-based projects. RCP is a program of the Center for Urban and Regional Affairs (CURA). More information at http://www.rcp.umn.edu

Thrombin Primes Responsiveness of Selective Chemoattractant Receptors at a Site Distal to G Protein Activation

To define the molecular basis of human chemoattractant receptor regulation, rat basophilic leukemia RBL-2H3 cells, which are thrombin- responsive, were transfected to stably express epitope-tagged receptors for C5a, interleukin-8 (IL-8), formylpeptides (e.g. N-formylmethionyl-leucyl- phenylalanine (fMLP)), and platelet-activating factor (PAF). Here we demonstrate that both thrombin and a synthetic peptide ligand for the thrombin receptor (sequence SFLLRN) caused phosphorylation and heterologous desensitization of the receptors for C5a, IL-8, and PAF but not that for formylpeptides as measured by agonist-stimulated [35S]guanosine 5\u27-3-O- (thio)triphosphate binding to membranes. Consistent with the PAF receptor phosphorylation, both thrombin and thrombin receptor peptide inhibited phosphoinositide hydrolysis, Ca2+ mobilization, and degranulation stimulated by PAF. Unexpectedly, despite heterologous desensitization at the level of receptor/G protein activation, there was enhancement (\u27priming\u27) by thrombin of subsequent activities stimulated by C5a and IL-8 as well as fMLP. The priming effect of thrombin was blocked by its inhibitor, hirudin. However, two other activators of the thrombin receptor, the peptide SFLLRN and trypsin, stimulated Ca2+ mobilization in RBL-2H3 cells but did not cause priming. In addition, SFLLRN and the thrombin receptor antagonist peptide FLLRN both inhibited thrombin-induced Ca2+ mobilization but not priming. Furthermore, the proteolytically active γ-thrombin, which does not stimulate the tethered ligand thrombin receptor and caused little or no Ca2+ mobilization in RBL-2H3 cells, effectively primed the response to fMLP. These data demonstrate that heterologous receptor phosphorylation and attenuation of G protein activation are not, by themselves, sufficient for the inhibition of biological responses mediated by C5a and IL-8. Moreover, thrombin appears to utilize mechanism(s) independent of its tethered ligand receptor to selectively prime phospholipase C-mediated biological responses of the C5a, IL-8, and formylpeptide receptors but not PAF. Because C5a, IL- 8, and formylpeptide activate phospholipase Cβ2, whereas PAF stimulates a different phospholipase C, the striking selectivity of thrombin\u27s priming may be mediated via its ability to enhance receptor-mediated activation of phospholipase Cβ2

Cross-Desensitization of Chemoattractant Receptors Occurs at Multiple Levels: Evidence for a Role for Inhibition of Phospholipase C Activity

To define the molecular mechanisms of cross-regulation among chemoattractant receptors, we stably coexpressed, in a rat basophilic leukemia (RBL-2H3) cell line, epitope-tagged receptors for the chemoattractants formylmethionylleucylphenylalanine (fMLP), a peptide of the fifth component of the complement system (C5a), and interleukin-8 (IL-8). All the expressed receptors underwent homologous phosphorylation and desensitization upon agonist stimulation. When co-expressed, epitope-tagged C5a receptor (ET-C5aR) and epitope-tagged IL-8 receptor (ET-IL-8RA) were cross-phosphorylated by activation of the other. Activation of epitope- tagged fMLP receptor (ET-FR) also cross-phosphorylated ET-C5aR and ET-IL- 8RA, but ET-FR was totally resistant to cross-phosphorylation. Similarly, C5a and IL-8 stimulation of [35S]guanosine 5\u27-3-P-(thio) triphosphate (GTPγS) binding and Ca2+ mobilization were cross-desensitized by each other and by fMLP. Stimulation of [35S]GTPγS binding by fMLP was also not cross- desensitized by C5a or IL-8, however, Ca2+ mobilization was, suggesting a site of inhibition distal to G protein activation. Consistent with this desensitization of Ca2+ mobilization, inositol 1,4,5-trisphosphate release in RBL-2H3 cells expressing both ET-C5aR and ET-FR revealed that fMLP and C5a cross-desensitized each other\u27s ability to stimulate phosphoinositide hydrolysis. Taken together, these results indicate that receptor cross- phosphorylation correlates directly with desensitization at the level of G protein activation. The ET-FR was resistant to this process. Of note, cross- desensitization of ET-FR at the level of phosphoinositide hydrolysis and Ca2+ mobilization was demonstrated in the absence of receptor phosphorylation. This suggests a new form of chemoattractant cross-regulation at a site distal to receptor/G protein coupling, involving the activity of phospholipase C

How do you say watermelon?

This paper discusses the relationships between contemporary indigenous games and those played historically on Turtle Island. With Sla’hal as an example, we look for ancestral philosophies informing old games that might be used today in development of new indigenous games of survivance and survivance games. Using indigenous pedagogies of Anishinaabe, Choctaw and Lushootseed speaking peoples, in addition to some of Vizenor’s theories, we modeled the content of this paper with playful formats to encourage readers to think about their own gaming practices. Beginning with story, we offer a bit of history, philosophy, visuals, a podcast transcript, and our system of Indigenous Game Tags to assist your creative understandings

Detection of invasive species in Wetlands: Practical dl with heavily imbalanced data

Deep Learning (DL) has become popular due to its ease of use and accuracy, with Transfer Learning (TL) effectively reducing the number of images needed to solve environmental problems. However, this approach has some limitations which we set out to explore: Our goal is to detect the presence of an invasive blueberry species in aerial images of wetlands. This is a key problem in ecosystem protection which is also challenging in terms of DL due to the severe imbalance present in the data. Results for the ResNet50 network show a high classification accuracy while largely ignoring the blueberry class, rendering these results of limited practical interest to detect that specific class. Moreover, by using loss function weighting and data augmentation results more akin to our practical application, our goals can be obtained. Our experiments regarding TL show that ImageNet weights do not produce satisfactory results when only the final layer of the network is trained. Furthermore, only minor gains are obtained compared with random weights when the whole network is retrained. Finally, in a study of state-of-the-art DL architectures best results were obtained by the ResNeXt architecture with 93.75 True Positive Rate and 98.11 accuracy for the Blueberry class with ResNet50, Densenet, and wideResNet obtaining close results. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Analysis of UAV-acquired wetland orthomosaics using GIS, computer vision, computational topology and deep learning

Invasive blueberry species endanger the sensitive environment of wetlands and protection laws call for management measures. Therefore, methods are needed to identify blueberry bushes, locate them, and characterise their distribution and properties with a minimum of disturbance. UAVs (Unmanned Aerial Vehicles) and image analysis have become important tools for classification and detection approaches. In this study, techniques, such as GIS (Geographical Information Systems) and deep learning, were combined in order to detect invasive blueberry species in wetland environments. Images that were collected by UAV were used to produce orthomosaics, which were analysed to produce maps of blueberry location, distribution, and spread in each study site, as well as bush height and area information. Deep learning networks were used with transfer learning and unfrozen weights in order to automatically detect blueberry bushes reaching True Positive Values (TPV) of 93.83% and an Overall Accuracy (OA) of 98.83%. A refinement of the result masks reached a Dice of 0.624. This study provides an efficient and effective methodology to study wetlands while using different techniques. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This

Concepts of GPCR-controlled navigation in the immune system

G-protein-coupled receptor (GPCR) signaling is essential for the spatiotemporal control of leukocyte dynamics during immune responses. For efficient navigation through mammalian tissues, most leukocyte types express more than one GPCR on their surface and sense a wide range of chemokines and chemoattractants, leading to basic forms of leukocyte movement (chemokinesis, haptokinesis, chemotaxis, haptotaxis, and chemorepulsion). How leukocytes integrate multiple GPCR signals and make directional decisions in lymphoid and inflamed tissues is still subject of intense research. Many of our concepts on GPCR-controlled leukocyte navigation in the presence of multiple GPCR signals derive from in vitro chemotaxis studies and lower vertebrates. In this review, we refer to these concepts and critically contemplate their relevance for the directional movement of several leukocyte subsets (neutrophils, T cells, and dendritic cells) in the complexity of mouse tissues. We discuss how leukocyte navigation can be regulated at the level of only a single GPCR (surface expression, competitive antagonism, oligomerization, homologous desensitization, and receptor internalization) or multiple GPCRs (synergy, hierarchical and non-hierarchical competition, sequential signaling, heterologous desensitization, and agonist scavenging). In particular, we will highlight recent advances in understanding GPCR-controlled leukocyte navigation by intravital microscopy of immune cells in mice