Functionalized micro-capillary film for the rapid at-line analysis of IgG aggregates in a cell culture bioreactor.

A micro-capillary film has been developed that offers the potential for an at-line analytical tool for rapid aggregate analysis during biopharmaceutical antibody production. A non-porous walled micro-capillary film (NMCF) with cation exchange functionality was demonstrated to act as a chromatography medium that could be operated with high linear fluid velocities and was highly resistant to blockage by entrained particulates, including cells. The NMCF containing 19 parallel microcapillaries was prepared using a melt extrusion process from poly(ethylene-vinyl alcohol) copolymer (EVOH). The NMCF-EVOH was modified to have cation-exchange functionality (NMCF-EVOH-SP) and shown to differentially bind monomer and aggregated species of IgG antibody directly from a bioreactor. The use of NMCF-EVOH-SP to quantify aggregate concentrations in monoclonal antibody preparations in less than 20 minutes was demonstrated.The authors would like to thank the EPSRC for the provision of a CASE Award. This study was sponsored by MedImmune, the global biologics R&D arm of AstraZeneca.This is the final version of the article. It first appeared from Taylor & Francis via http://dx.doi.org/10.1080/19420862.2015.106536

Patients with schizophrenia show deficits of working memory maintenance components in circuit-specific tasks

Working memory (WM) deficits are a neuropsychological core finding in patients with schizophrenia and also supposed to be a potential endophenotype of schizophrenia. Yet, there is a large heterogeneity between different WM tasks which is partly due to the lack of process specificity of the tasks applied. Therefore, we investigated WM functioning in patients with schizophrenia using process- and circuit-specific tasks. Thirty-one patients with schizophrenia and 47 controls were tested with respect to different aspects of verbal and visuospatial working memory using modified Sternberg paradigms in a computer-based behavioural experiment. Total group analysis revealed significant impairment of patients with schizophrenia in each of the tested WM components. Furthermore, we were able to identify subgroups of patients showing different patterns of selective deficits. Patients with schizophrenia exhibit specific and, in part, selective WM deficits with indirect but conclusive evidence of dysfunctions of the underlying neural networks. These deficits are present in tasks requiring only maintenance of verbal or visuospatial information. In contrast to a seemingly global working memory deficit, individual analysis revealed differential patterns of working memory impairments in patients with schizophrenia

A systematic experimental neuropsychological investigation of the functional integrity of working memory circuits in major depression

Verbal and visuospatial working memory (WM) impairment is a well-documented finding in psychiatric patients suffering from major psychoses such as schizophrenia or bipolar affective disorder. However, in major depression (MDD) the literature on the presence and the extent of WM deficits is inconsistent. The use of a multitude of different WM tasks most of which lack process-specificity may have contributed to these inconsistencies. Eighteen MDD patients and 18 healthy controls matched with regard to age, gender and education were tested using process- and circuit-specific WM tasks for which clear brain-behaviour relationships had been established in prior functional neuroimaging studies. Patients suffering from acute MDD showed a selective impairment in articulatory rehearsal of verbal information in working memory. By contrast, visuospatial WM was unimpaired in this sample. There were no significant correlations between symptom severity and WM performance. These data indicate a dysfunction of a specific verbal WM system in acutely ill patients with MDD. As the observed functional deficit did not correlate with different symptom scores, further, longitudinal studies are required to clarify whether and how this deficit is related to illness acuity and clinical state of MDD patients

The fundamental cycle of concept construction underlying various theoretical frameworks

In this paper, the development of mathematical concepts over time is considered. Particular reference is given to the shifting of attention from step-by-step procedures that are performed in time, to symbolism that can be manipulated as mental entities on paper and in the mind. The development is analysed using different theoretical perspectives, including the SOLO model and various theories of concept construction to reveal a fundamental cycle underlying the building of concepts that features widely in different ways of thinking that occurs throughout mathematical learning

Histological Examination of Collagen and Proteoglycan Changes in Osteoarthritic Menisci

This study sought to examine collagen and proteoglycan changes in the menisci of patients with osteoarthritis (OA). Collagens were examined using picrosirius red, and hematoxylin and eosin. Proteoglycans were examined using safranin-O and alcian blue. Types I and II collagens and aggrecan were examined using immunochemistry. Severe loss of collagens was observed to occur in OA menisci, particularly in the middle and deep zones and collagen networks were less organized than those of normal menisci. In contrast, proteoglycan staining in the middle and deep zones of OA meniscus increased compared to normal control menisci. Immunohistochemistry indicated that types I and II collagens were co-localized and the loss of types I collagen in OA menisci appeared more severe in the middle and deep zones than that in the surface zones. The loss of type II collagen however was severe across all three zones. Immunohistochemistry also indicated elevated aggrecan staining in OA menisci. These findings together indicate that severe loss of collagens and intrameniscal degeneration are hallmarks of OA menisci and that extracellular matrix degeneration occurred in OA menisci follows a pathway different from that occurred in OA articular cartilage. These findings are not only important for a better understanding of the disease process but also important for the development of novel structure-modifying drugs for OA therapy

Three-dimensional culture of human meniscal cells: Extracellular matrix and proteoglycan production

<p>Abstract</p> <p>Background</p> <p>The meniscus is a complex tissue whose cell biology has only recently begun to be explored. Published models rely upon initial culture in the presence of added growth factors. The aim of this study was to test a three-dimensional (3D) collagen sponge microenvironment (without added growth factors) for its ability to provide a microenvironment supportive for meniscal cell extracellular matrix (ECM) production, and to test the responsiveness of cells cultured in this manner to transforming growth factor-β (TGF-β).</p> <p>Methods</p> <p>Experimental studies were approved prospectively by the authors' Human Subjects Institutional Review Board. Human meniscal cells were isolated from surgical specimens, established in monolayer culture, seeded into a 3D scaffold, and cell morphology and extracellular matrix components (ECM) evaluated either under control condition or with addition of TGF-β. Outcome variables were evaluation of cultured cell morphology, quantitative measurement of total sulfated proteoglycan production, and immunohistochemical study of the ECM components chondroitin sulfate, keratan sulfate, and types I and II collagen.</p> <p>Result and Conclusion</p> <p>Meniscal cells attached well within the 3D microenvironment and expanded with culture time. The 3D microenvironment was permissive for production of chondroitin sulfate, types I and II collagen, and to a lesser degree keratan sulfate. This microenvironment was also permissive for growth factor responsiveness, as indicated by a significant increase in proteoglycan production when cells were exposed to TGF-β (2.48 μg/ml ± 1.00, mean ± S.D., vs control levels of 1.58 ± 0.79, p < 0.0001). Knowledge of how culture microenvironments influence meniscal cell ECM production is important; the collagen sponge culture methodology provides a useful in vitro tool for study of meniscal cell biology.</p

The Role of the Immunological Synapse in Differential Effects of APC Subsets in Alloimmunization to Fresh, Non-stored RBCs

Background: Each year, over 5 million red blood cell (RBC) transfusions are administered to patients in the USA. Despite the therapeutic benefits of RBC transfusions, there are associated risks. RBC-specific alloantibodies may form in response to antigenic differences between RBC donors and recipients; these alloantibodies can be a problem as they may mediate hemolysis or pose barriers to future transfusion support. While there is currently no reliable way to predict which RBC recipients will make an alloantibody response, risk factors such as inflammation have been shown to correlate with increased rates of RBC alloimmunization. The underlying mechanisms behind how inflammation mediates alloantibody production are incompletely defined.Methods: To assess erythrophagocytosis, mice were treated with PBS or inflammatory stimuli followed by a transfusion of allogeneic RBCs labeled with a lipophilic dye. At multiple time points, RBC consumption and expression of activation makers by leukocytes was evaluated. To determine which antigen presenting cell (APC) subset(s) were capable of promoting allogeneic T cell activation, sorted leukocyte populations (which had participated in erythrophagocytosis) were co-cultured in vitro with allogeneic CD4+ T cells; T cell proliferation and ability to form immunological synapses with APCs were determined.Results: Upon transfusion of fresh allogeneic RBCs, multiple APCs consumed transfused RBCs. However, only CD8+ and CD11b+ dendritic cells formed productive immunological synapses with allogeneic T cells and stimulated proliferation. Importantly, allogeneic T cell activation and RBC alloantibody production occurred in response to RBC transfusion alone, and transfusion in the context of inflammation enhanced RBC consumption, the number of immune synapses, allogeneic T cell proliferation, and the rate and magnitude of alloantibody production.Conclusions: These data demonstrate that regardless of the ability to participate in RBC consumption, only a subset of APCs are capable of forming an immune synapse with T cells thereby initiating an alloantibody response. Additionally, these data provide mechanistic insight into RBC alloantibody generation

Billions of impoverished people deserve to be better served: A call to action for the service research community

Purpose – The purpose of this paper is to create a movement within the service research community that aspires to help the billions of impoverished people across the world achieve better service from each other, from their communities, from corporations, from their governments, and from nongovernmental organizations. The authors believe every human being is worthy of being served properly. To achieve this purpose, understanding and learning from this huge low-income segment of society known as the base of the pyramid (BoP) is essential. There are myths about the BoP that need to be dispelled and there is a fundamental lack of service research on this important problem. Design/methodology/approach – The existence of an extensive BoP literature combined with service research priorities has called attention to drafting research agendas. Human service systems are explored historically and systems theory provides a perspective for understanding and reducing poverty. Transformative service research, service design research, and community action research are presented to illustrate three research approaches that can contribute to understanding and then better serving the needs of the neglected billions of humanity. Findings – First, the authors present a practical and meaningful call to action by making the case for the service research community to contribute to poverty alleviation with the creation of fresh ideas and research agendas. Second, the authors describe the ample opportunity for conducting service research in and with the BoP and thereby expanding service knowledge about the BoP. Third, the authors suggest a number of approaches for service researchers to join this new movement and help improve the well-being of billions of impoverished people. Social implications – Most existing service research comes from highly developed Anglo-Saxon countries and concerns the service problems of customers in affluent societies. Therefore, there is a fundamental lack of service research at the BoP. The social implications are truly global. Poverty is a global service system problem that can be reduced. Effective poverty alleviation solutions in one part of the world can be adapted to other parts of the world. Originality/value – This paper is a new and very original call to action to the service research community. First, with the exception of a few previous manuscripts calling for research on the BoP, this is the first time a collaborative effort has been made to start systematically changing this knowledge gap. Second, the service research community has never worked on a project of this magnitude. The authors hope to offer a role model to other academic communities as to how to marshal their resources to have a collective, positive impact on the well-being of the world’s impoverished

The Fermi GBM Gamma-Ray Burst Spectral Catalog: Four Years Of Data

In this catalog we present the updated set of spectral analyses of GRBs detected by the Fermi Gamma-Ray Burst Monitor (GBM) during its first four years of operation. It contains two types of spectra, time-integrated spectral fits and spectral fits at the brightest time bin, from 943 triggered GRBs. Four different spectral models were fitted to the data, resulting in a compendium of more than 7500 spectra. The analysis was performed similarly, but not identically to Goldstein et al. 2012. All 487 GRBs from the first two years have been re-fitted using the same methodology as that of the 456 GRBs in years three and four. We describe, in detail, our procedure and criteria for the analysis, and present the results in the form of parameter distributions both for the observer-frame and rest-frame quantities. The data files containing the complete results are available from the High-Energy Astrophysics Science Archive Research Center (HEASARC).Comment: Accepted for publication in ApJ