1,562 research outputs found
Muscle-directed gene therapy corrects Pompe disease and uncovers species-specific GAA immunogenicity
Pompe disease is a severe disorder caused by loss of acid Ī±-glucosidase (GAA), leading to glycogen accumulation in tissues and neuromuscular and cardiac dysfunction. Enzyme replacement therapy is the only available treatment. AT845 is an adeno-associated viral vector designed to express human GAA specifically in skeletal muscle and heart. Systemic administration of AT845 in Gaaā/ā mice led to a dose-dependent increase in GAA activity, glycogen clearance in muscles and heart, and functional improvement. AT845 was tolerated in cynomolgus macaques at low doses, while high doses caused anti-GAA immune response, inflammation, and cardiac abnormalities resulting in unscheduled euthanasia of two animals. Conversely, a vector expressing the macaque GAA caused no detectable pathology, indicating that the toxicity observed with AT845 was an anti-GAA xenogeneic immune response. Western blot analysis showed abnormal processing of human GAA in cynomolgus muscle, adding to the species-specific effects of enzyme expression. Overall, these studies show that AAV-mediated GAA delivery to muscle is efficacious in Gaaā/ā mice and highlight limitations in predicting the toxicity of AAV vectors encoding human proteins in non-human species
Spectral properties of zero temperature dynamics in a model of a compacting granular column
The compacting of a column of grains has been studied using a one-dimensional
Ising model with long range directed interactions in which down and up spins
represent orientations of the grain having or not having an associated void.
When the column is not shaken (zero 'temperature') the motion becomes highly
constrained and under most circumstances we find that the generator of the
stochastic dynamics assumes an unusual form: many eigenvalues become
degenerate, but the associated multi-dimensional invariant spaces have but a
single eigenvector. There is no spectral expansion and a Jordan form must be
used. Many properties of the dynamics are established here analytically; some
are not. General issues associated with the Jordan form are also taken up.Comment: 34 pages, 4 figures, 3 table
IRX-2, a Novel Immunotherapeutic, Enhances Functions of Human Dendritic Cells
Background: In a recent phase II clinical trial for HNSCC patients, IRX-2, a cell-derived biologic, promoted T-cell infiltration into the tumor and prolonged overall survival. Mechanisms responsible for these IRX-2-mediated effects are unknown. We hypothesized that IRX-2 enhanced tumor antigen-(TA)-specific immunity by up-regulating functions of dendritic cells (DC). Methodology/Principal Findings: Monocyte-derived DC obtained from 18 HNSCC patients and 12 healthy donors were matured using IRX-2 or a mix of TNF-Ī±, IL-1Ī² and IL-6 ("conv. mix"). Multicolor flow cytometry was used to study the DC phenotype and antigen processing machinery (APM) component expression. ELISPOT and cytotoxicity assays were used to evaluate tumor-reactive cytotoxic T lymphocytes (CTL). IL-12p70 and IL-10 production by DC was measured by LuminexĀ® and DC migration toward CCL21 was tested in transwell migration assays. IRX-2-matured DC functions were compared with those of conv. mix-matured DC. IRX-2-matured DC expressed higher levels (p<0.05) of CD11c, CD40, CCR7 as well as LMP2, TAP1, TAP2 and tapasin than conv. mix-matured DC. IRX-2-matured DC migrated significantly better towards CCL21, produced more IL-12p70 and had a higher IL12p70/IL-10 ratio than conv. mix-matured DC (p<0.05 for all). IRX-2-matured DC carried a higher density of tumor antigen-derived peptides, and CTL primed with these DC mediated higher cytotoxicity against tumor targets (p<0.05) compared to the conv. mix-matured DC. Conclusion: Excellent ability of IRX-2 to induce ex vivo DC maturation in HNSCC patients explains, in part, its clinical benefits and emphasizes its utility in ex vivo maturation of DC generated for therapy. Ā© 2013 Schilling et al
Controlled assembly of SNAP-PNA-fluorophore systems on DNA templates to produce fluorescence resonance energy transfer
The SNAP protein is a widely used self-labeling tag that can be used for tracking protein localization and trafficking in living systems. A model system providing controlled alignment of SNAP-tag units can provide a new way to study clustering of fusion proteins. In this work, fluorescent SNAP-PNA conjugates were controllably assembled on DNA frameworks forming dimers, trimers, and tetramers. Modification of peptide nucleic acid (PNA) with the O6-benzyl guanine (BG) group allowed the generation of site-selective covalent links between PNA and the SNAP protein. The modified BG-PNAs were labeled with fluorescent Atto dyes and subsequently chemo-selectively conjugated to SNAP protein. Efficient assembly into dimer and oligomer forms was verified via size exclusion chromatography (SEC), electrophoresis (SDS-PAGE), and fluorescence spectroscopy. DNA directed assembly of homo- and hetero-dimers of SNAP-PNA constructs induced homo- and hetero-FRET, respectively. Longer DNA scaffolds controllably aligned similar fluorescent SNAP-PNA constructs into higher oligomers exhibiting homo-FRET. The combined SEC and homo-FRET studies indicated the 1:1 and saturated assemblies of SNAP-PNA-fluorophore:DNA formed preferentially in this system. This suggested a kinetic/stoichiometric model of assembly rather than binomially distributed products. These BG-PNA-fluorophore building blocks allow facile introduction of fluorophores and/or assembly directing moieties onto any protein containing SNAP. Template directed assembly of PNA modified SNAP proteins may be used to investigate clustering behavior both with and without fluorescent labels which may find use in the study of assembly processes in cells
Global Production of Marine Bivalves. Trends and Challenges
The global production of marine bivalves for human consumption is
more than 15 million tonnes per year (average period 2010ā2015), which is about
14% of the total marine production in the world. Most of the marine bivalve production
(89%) comes from aquaculture and only 11% comes from the wild fishery.
Asia, especially China, is by far the largest producer of marine bivalves, accounting
for 85% of the world production and responsible for the production growth. In
other continents, the production is stabilizing or decreasing (Europe) the last
decades. In order to stimulate growth, sustainability (Planet, Profit, People) of the
aquaculture activities is a key issue. Environmental (Planet) aspects for sustainable
aquaculture include the fishery on seed resources, carrying capacity, invasive species
and organic loading. Food safety issues due to environmental contaminants
and biotoxines should be minimized to increase the reliability of marine bivalves
as a healthy food source and to stimulate market demands. Properly designed monitoring
programs are important tools to accomplish sustainable growth of marine
bivalve production
A case report of erythroderma in a patient with borderline leprosy on reversal reaction: a result of the exacerbated reaction?
Economic Benefits of Intensive Insulin Therapy in Critically Ill Patients: The Targeted Insulin Therapy to Improve Hospital Outcomes (TRIUMPH) Project
OBJECTIVEāThe purpose of this study was to analyze the economic outcomes of a clinical program implemented to achieve strict glycemic control with intensive insulin therapy in patients admitted to the intensive care unit (ICU)
Apical Sparing Pattern of Longitudinal Strain and Positive Bone Scintigraphy in Metastatic Myocardial Calcification
An apical sparing pattern of longitudinal strain and positive radionuclide bone scintigraphy are believed to be specific for the diagnosis of transthyretin cardiac amyloidosis. We report on a young woman with apical sparing of longitudinal strain and positive bone scintigraphy who was found to have metastatic myocardial calcification at autopsy
Effects of acutely inhibiting PI3K isoforms and mTOR on regulation of glucose metabolism in vivo
In in vitro studies class-I PI3Ks (phosphoinositide 3-kinases), class-II PI3Ks and mTOR (mammalian target of rapamycin) have all been described as having roles in the regulation of glucose metabolism. The relative role each plays in the normal signalling processes regulating glucose metabolism in vivo is less clear. Knockout and knockin mouse models have provided some evidence that the class-I PI3K isoforms p110Ī±, p110Ī², and to a lesser extent p110Ī³, are necessary for processes regulating glucose metabolism and appetite. However, in these models the PI3K activity is chronically reduced. Therefore we analysed the effects of acutely inhibiting PI3K isoforms alone, or PI3K and mTOR, on glucose metabolism and food intake. In the present study impairments in glucose tolerance, insulin tolerance and increased hepatic glucose output were observed in mice treated with the pan-PI3K/mTOR inhibitors PI-103 and NVP-BEZ235. The finding that ZSTK474 has similar effects indicates that these effects are due to inhibition of PI3K rather than mTOR. The p110Ī±-selective inhibitors PIK75 and A66 also induced these phenotypes, but inhibitors of p110Ī², p110Ī“ or p110Ī³ induced only minor effects. These drugs caused no significant effects on BMR (basal metabolic rate), O2 consumption or water intake, but BEZ235, PI-103 and PIK75 did cause a small reduction in food consumption. Surprisingly, pan-PI3K inhibitors or p110Ī± inhibitors caused reductions in animal movement, although the cause of this is not clear. Taken together these studies provide pharmacological evidence to support a pre-eminent role for the p110Ī± isoform of PI3K in pathways acutely regulating glucose metabolism
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