Inhibition of CD26/DPP-IV enhances donor muscle cell engraftment and stimulates sustained donor cell proliferation

<p>Abstract</p> <p>Background</p> <p>Transplantation of myogenic stem cells possesses great potential for long-term repair of dystrophic muscle. In murine-to-murine transplantation experiments, CXCR4 expression marks a population of adult murine satellite cells with robust engraftment potential in <it>mdx </it>mice, and CXCR4-positive murine muscle-derived SP cells home more effectively to dystrophic muscle after intra-arterial delivery in <it>mdx^5cv</it>mice. Together, these data suggest that CXCR4 plays an important role in donor cell engraftment. Therefore, we sought to translate these results to a clinically relevant canine-to-canine allogeneic transplant model for Duchenne muscular dystrophy (DMD) and determine if CXCR4 is important for donor cell engraftment.</p> <p>Methods</p> <p>In this study, we used a canine-to-murine xenotransplantation model to quantitatively compare canine muscle cell engraftment, and test the most effective cell population and modulating factor in a canine model of DMD using allogeneic transplantation experiments.</p> <p>Results</p> <p>We show that CXCR4 expressing cells are important for donor muscle cell engraftment, yet FACS sorted CXCR4-positive cells display decreased engraftment efficiency. However, diprotin A, a positive modulator of CXCR4-SDF-1 binding, significantly enhanced engraftment and stimulated sustained proliferation of donor cells <it>in vivo</it>. Furthermore, the canine-to-murine xenotransplantation model accurately predicted results in canine-to-canine muscle cell transplantation.</p> <p>Conclusions</p> <p>Therefore, these results establish the efficacy of diprotin A in stimulating muscle cell engraftment, and highlight the pre-clinical utility of a xenotransplantation model in assessing the relative efficacy of muscle stem cell populations.</p

The genetic architecture of the human cerebral cortex

The cerebral cortex underlies our complex cognitive capabilities, yet little is known about the specific genetic loci that influence human cortical structure. To identify genetic variants that affect cortical structure, we conducted a genome-wide association meta-analysis of brain magnetic resonance imaging data from 51,665 individuals. We analyzed the surface area and average thickness of the whole cortex and 34 regions with known functional specializations. We identified 199 significant loci and found significant enrichment for loci influencing total surface area within regulatory elements that are active during prenatal cortical development, supporting the radial unit hypothesis. Loci that affect regional surface area cluster near genes in Wnt signaling pathways, which influence progenitor expansion and areal identity. Variation in cortical structure is genetically correlated with cognitive function, Parkinson's disease, insomnia, depression, neuroticism, and attention deficit hyperactivity disorder

Immunodetection of Human Double Homeobox 4

Double homeobox 4 (DUX4) is a candidate disease gene for facioscapulohumeral dystrophy (FSHD), one of the most common muscular dystrophies characterized by progressive skeletal muscle degeneration. Despite great strides in understanding precise genetics of FSHD, the molecular pathophysiology of the disease remains unclear. One of the major limitations has been the availability of appropriate molecular tools to study DUX4 protein. In the present study, we report the development of five new monoclonal antibodies targeted against the N- and C-termini of human DUX4, and characterize their reactivity using Western blot and immunofluorescence staining. Additionally, we show that expression of the canonical full coding DUX4 induces cell death in human primary muscle cells, whereas the expression of a shorter splice form of DUX4 results in no such toxicity. Immunostaining with these new antibodies reveals a differential effect of two DUX4 isoforms on human muscle cells. These antibodies will provide an excellent tool for investigating the role of DUX4 in FSHD pathogenesis

Fundamental differences in promoter CpG island DNA hypermethylation between human cancer and genetically engineered mouse models of cancer

Genetic and epigenetic alterations are essential for the initiation and progression of human cancer. We previously reported that primary human medulloblastomas showed extensive cancer-specific CpG island DNA hypermethylation in critical developmental pathways. To determine whether genetically engineered mouse models (GEMMs) of medulloblastoma have comparable epigenetic changes, we assessed genome-wide DNA methylation in three mouse models of medulloblastoma. In contrast to human samples, very few loci with cancer-specific DNA hypermethylation were detected, and in almost all cases the degree of methylation was relatively modest compared with the dense hypermethylation in the human cancers. To determine if this finding was common to other GEMMs, we examined a Burkitt lymphoma and breast cancer model and did not detect promoter CpG island DNA hypermethylation, suggesting that human cancers and at least some GEMMs are fundamentally different with respect to this epigenetic modification. These findings provide an opportunity to both better understand the mechanism of aberrant DNA methylation in human cancer and construct better GEMMs to serve as preclinical platforms for therapy development

Fundamental differences in promoter CpG island DNA hypermethylation between human cancer and genetically engineered mouse models of cancer

Genetic and epigenetic alterations are essential for the initiation and progression of human cancer. We previously reported that primary human medulloblastomas showed extensive cancer-specific CpG island DNA hypermethylation in critical developmental pathways. To determine whether genetically engineered mouse models (GEMMs) of medulloblastoma have comparable epigenetic changes, we assessed genome-wide DNA methylation in three mouse models of medulloblastoma. In contrast to human samples, very few loci with cancer-specific DNA hypermethylation were detected, and in almost all cases the degree of methylation was relatively modest compared with the dense hypermethylation in the human cancers. To determine if this finding was common to other GEMMs, we examined a Burkitt lymphoma and breast cancer model and did not detect promoter CpG island DNA hypermethylation, suggesting that human cancers and at least some GEMMs are fundamentally different with respect to this epigenetic modification. These findings provide an opportunity to both better understand the mechanism of aberrant DNA methylation in human cancer and construct better GEMMs to serve as preclinical platforms for therapy development

The Intensively Managed Landscape Critical Zone Observatory: A Scientific Testbed for Understanding Critical Zone Processes in Agroecosystems

In intensively managed landscapes, interactions between surface (tillage) and subsurface (tile drainage) management with prevailing climate/weather alter landscape characteristics, transport pathways, and transformation rates of surface/subsurface water, soil/sediment, and particulate/dissolved nutrients. To capture the high spatial and temporal variability of constituent transport and residence times in the critical zone (between the bedrock and canopy) of these altered landscapes, both storm event and continuous measurements are needed. The Intensively Managed Landscapes Critical Zone Observatory (IML-CZO) is comprised of three highly characterized, well instrumented, and representative watersheds (i.e., Clear Creek, Iowa; Upper Sangamon River, Illinois; and Minnesota River, Minnesota). It is organized to quantify the heterogeneity in structure and dynamic response of critical zone processes to human activities in the context of the glacial and management (anthropogenic) legacies. Observations of water, sediment, and nutrients are made at nested points of the landscape in the vertical and lateral directions during and between storm events (i.e., continuously). The measurements and corresponding observational strategy are organized as follows. First, reference measurements from surface soil and deep core extractions, geophysical surveys, lidar, and hyperspectral data, which are common across all Critical Zone Observatories, are available. The reference measurements include continuous quantification of energy, water, solutes, and sediment fluxes. The reference measurements are complemented with event-based measurements unique to IML-CZO. These measurements include water table fluctuations, enrichment ratios, and roughness as well as bank erosion, hysteresis, sediment sources, and lake/floodplain sedimentation. The coupling of reference and event-based measurements support testing of the central hypothesis (i.e., system shifts from transformer to transporter in IML-CZO due to the interplay between management and weather/climate). Data collected since 2014 are available through a data repository and through the Geodashboard interface, which can be used for process-based model simulations.This article is published as Wilson, Christopher G., Benjamin Abban, Laura L. Keefer, Kenneth Wacha, Dimitrios Dermisis, Christos Giannopoulos, Shengnan Zhou et al. "The intensively managed landscape critical zone observatory: a scientific testbed for understanding critical zone processes in agroecosystems." Vadose Zone Journal 17, no. 1 (2018): 1-21. doi:10.2136/vzj2018.04.0088. Posted with permission of INRC. Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted

The Intensively Managed Landscape Critical Zone Observatory: A Scientific Testbed for Understanding Critical Zone Processes in Agroecosystems

In intensively managed landscapes, interactions between surface (tillage) and subsurface (tile drainage) management with prevailing climate/weather alter landscape characteristics, transport pathways, and transformation rates of surface/subsurface water, soil/sediment, and particulate/dissolved nutrients. To capture the high spatial and temporal variability of constituent transport and residence times in the critical zone (between the bedrock and canopy) of these altered landscapes, both storm event and continuous measurements are needed. The Intensively Managed Landscapes Critical Zone Observatory (IML-CZO) is comprised of three highly characterized, well instrumented, and representative watersheds (i.e., Clear Creek, Iowa; Upper Sangamon River, Illinois; and Minnesota River, Minnesota). It is organized to quantify the heterogeneity in structure and dynamic response of critical zone processes to human activities in the context of the glacial and management (anthropogenic) legacies. Observations of water, sediment, and nutrients are made at nested points of the landscape in the vertical and lateral directions during and between storm events (i.e., continuously). The measurements and corresponding observational strategy are organized as follows. First, reference measurements from surface soil and deep core extractions, geophysical surveys, lidar, and hyperspectral data, which are common across all Critical Zone Observatories, are available. The reference measurements include continuous quantification of energy, water, solutes, and sediment fluxes. The reference measurements are complemented with event-based measurements unique to IML-CZO. These measurements include water table fluctuations, enrichment ratios, and roughness as well as bank erosion, hysteresis, sediment sources, and lake/floodplain sedimentation. The coupling of reference and event-based measurements support testing of the central hypothesis (i.e., system shifts from transformer to transporter in IML-CZO due to the interplay between management and weather/climate). Data collected since 2014 are available through a data repository and through the Geodashboard interface, which can be used for process-based model simulations

RNA transcripts, miRNA-sized fragments and proteins produced from D4Z4 units: new candidates for the pathophysiology of facioscapulohumeral dystrophy

Deletion of a subset of the D4Z4 macrosatellite repeats in the subtelomeric region of chromosome 4q causes facioscapulohumeral muscular dystrophy (FSHD) when occurring on a specific haplotype of 4qter (4qA161). Several genes have been examined as candidates for causing FSHD, including the DUX4 homeobox gene in the D4Z4 repeat, but none have been definitively shown to cause the disease, nor has the full extent of transcripts from the D4Z4 region been carefully characterized. Using strand-specific RT–PCR, we have identified several sense and antisense transcripts originating from the 4q D4Z4 units in wild-type and FSHD muscle cells. Consistent with prior reports, we find that the DUX4 transcript from the last (most telomeric) D4Z4 unit is polyadenylated and has two introns in its 3-prime untranslated region. In addition, we show that this transcript generates (i) small si/miRNA-sized fragments, (ii) uncapped, polyadenylated 3-prime fragments that encode the conserved C-terminal portion of DUX4 and (iii) capped and polyadenylated mRNAs that contain the double-homeobox domain of DUX4 but splice-out the C-terminal portion. Transfection studies demonstrate that translation initiation at an internal methionine can produce the C-terminal polypeptide and developmental studies show that this peptide inhibits myogenesis at a step between MyoD transcription and the activation of MyoD target genes. Together, we have identified new sense and anti-sense RNA transcripts, novel mRNAs and mi/siRNA-sized RNA fragments generated from the D4Z4 units that are new candidates for the pathophysiology of FSHD

Nonlinear model-based control of thin-film drying for continuous pharmaceutical manufacturing

This paper considers the model-based control of composition and thickness for a thin-film drying process used in the continuous manufacturing of pharmaceutical tablets. In this nonlinear distributed dynamical system, a drug formulation solution is coated onto a moving surface and then dried to form thin films of approximately 250 μm in thickness. A dynamic optimizer is designed that employs a first-principles process model to simulate the spatial distribution of solvent concentration in the film and the thin film shrinkage during drying. A critical parameter to describe the highly nonlinear dynamics of the thin-film drying is the mutual polymer-solvent diffusion coefficient, which strongly depends on solvent concentration and film temperature. Two optimal control problems are studied for set point tracking of solvent concentration and minimization of energy consumption in the dryer while satisfying various operational and product quality constraints. An unscented Kalman filter is designed to facilitate the output feedback implementation of the dynamic optimizer and to estimate unmeasured thin-film quality attributes such as the film thickness. The performance of the model-based controller is compared to that of a proportional integral controller in two simulation case studies. The nonlinear model-based control strategy has improved versatility and the potential to reduce production of off spec material. © 2013 American Chemical Society