Impact of differential DNA methylation on transgene expression in cotton (Gossypium hirsutum L.) events generated by targeted sequence insertion

Targeted Genome Optimization (TGO) using site‐specific nucleases to introduce a DNA double‐strand break (DSB) at a specific target locus has broadened the options available to breeders for generation and combination of multiple traits. The use of targeted DNA cleavage in combination with homologous recombination (HR)‐mediated repair, enabled the precise targeted insertion of additional trait genes (2mepsps, hppd, axmi115) at a pre‐existing transgenic locus in cotton. Here we describe the expression and epigenome analyses of cotton Targeted Sequence Insertion (TSI) events over generations. In a subset of events, we observed variability in the level of transgene (hppd, axmi115) expression between independent but genetically identical TSI events. Transgene expression could also be differential within single events and variable over generations. This expression variability and silencing occurred independently of the transgene sequence and could be attributed to DNA methylation that was further linked to different DNA methylation mechanisms. The trigger(s) of transgene DNA methylation remains elusive but we hypothesize that targeted DSB induction and repair could be a potential trigger for DNA methylation.     

Non-rigid deformation to include subject-specific detail in musculoskeletal models of CP children with proximal femoral deformity and its effect on muscle and contact forces during gait

To account for proximal femoral deformities in children with cerebral palsy (CP), subject-specific musculoskeletal models are needed. Non-rigid deformation (NRD) deforms generic onto personalized bone geometry and thereby transforms the muscle points. The goal of this study was to determine to what extent the models and simulation outcomes in CP patients differ when including subject-specific detail using NRD or Magnetic Resonance Imaging (MRI)-based models. The NRD models slightly overestimated hip contact forces compared to MRI models and differences in muscle point positions and moment arm lengths (MALs) remained, although differences were smaller than for the generic model.     

Optimal fed batch experiment design for estimation of monod kinetics of Azospirillum brasilense: from theory to practice

In this paper the problem of reliable and accurate parameter estimation for unstructured models is considered. It is illustrated how a theoretically optimal design can be successfully translated into a practically feasible, robust, and informative experiment. The well-known parameter estimation problem of Monod kinetic parameters is used as a vehicle to illustrate our approach. As known for a long time, noisy batch measurements do not allow for unique and accurate estimation of the kinetic parameters of the Monod model. Techniques of optimal experiment design are, therefore, exploited to design informative experiments and to improve the parameter estimation accuracy. During the design process, practical feasibility has to be kept in mind. The designed experiments are easy to implement in practice and do not require additional monitoring equipment. Both design and experimental validation of informative fed batch experiments are illustrated with a case study, namely, the growth of the nitrogen-fixing bacteria Azospirillum brasilense.     

Hi-C deconvolution of a textile dye–related microbiome reveals novel taxonomic landscapes and links phenotypic potential to individual genomes

International Microbiology - Microbial biodiversity is represented by a variety of genomic landscapes adapted to dissimilar environments on Earth. These genomic landscapes contain functional...     

The Barnes Maze Task Reveals Specific Impairment of Spatial Learning Strategy in the Intrahippocampal Kainic Acid Model for Temporal Lobe Epilepsy

Neurochemical Research - Temporal lobe epilepsy (TLE) is an acquired form of focal epilepsy, in which patients not only suffer from unprovoked, devastating seizures, but also from severe...     

Untersuchungen über die Osteon- und Lamellenformen im Extremitätenskelet des Erwachsenen

Zusammenfassung Auf Grund der Untersuchung von Knochenquerschnitten eines gesunden 43jährigen Mannes, bestätigt durch noch zu veröffentlichende Untersuchungen an Tibien verschiedener Individuen, werden die Strukturformen des Knochens nach ihrem Querschnittsbild beschrieben.Es wird zwischen Osteonen und Tangentiallamellen unterschieden. Zu den Tangentiallamellen mit dem mehr oder minder parallelen Verlauf zur Knochenoberfläche gehören die bisher als Generallamellen und Schaltlamellen beschriebenen Systeme.Soweit eine Lamellengliederung vorhanden ist, zeichnen sich die Tangentiallamellen durch den strengen Wechsel zwischen flach und steil gewickelten aus.Auf Grund des Querschnittsbildes werden verschiedene Osteonformen unterschieden. Die Größe des einzelnen Osteonquerschnittes wird mit Hilfe der Lamellenzahl bestimmt. Gleichzeitig wird die Steigungsfolge beachtet, d. h. der Wechsel des Kollagenfaserverlaufs von Lamelle zu Lamelle.Es ergibt sich, daß die kleineren Osteone überwiegend in der peripheren Schnitthälfte, die größeren dagegen in der zentralen liegen. Der regelmäßige Wechsel der Steigungsfolge nimmt von den kleineren zu den größeren Osteonen hin ab, die mehr steile Verlaufsweise dagegen zu. Die kleineren Größenklassen lassen häufiger die lamelläre Gliederung vermissen als die großen.Abschließend wird erörtert, daß sowohl das Osteon wie die Lamelle nur als eine besondere Lagerungsform der Kollagenfasern im Knochen angesehen werden können. Der Begriff Osteon wurde in Anlehnung an die Begriffe der überwiegend zellulären Einheiten Neuron und Chondron bzw. der sog. Entwicklungs- und Funktionseinheit Nephron gebildet. Die zirkuläre Lagerung der Kollagenfasern hat aller Voraussicht nach eine besondere festigkeitstheoretische Bedeutung. Sie ist aber abhängig vom Gefäßbaum, an dessen Verzweigungen Doppelbildungen auftreten. Diese Doppelbildungen teilen sich und begleiten die Gefäßäste. Sie werden damit zum Osteon, das sich nach Querschnittsgröße und Wicklung in benachbarten Querschnitten verschieden verhalten kann. Die zirkuläre Wicklung führt nicht zu individuellen Gebilden, die den Knochen wie Bausteine aufbauen. Sie stellt ein System dar, das den Gefäßbaum im Knochen in mehr oder minder kontinuierlichem Zusammenhang begleitet. Die zirkulären Wicklungen gehen ohne Abgrenzung in die übrigen Lamellensysteme, die Tangentiallamellen, über. Osteone und Tangentiallamellen erscheinen damit als eine übergeordnete Lagerungsform der Kollagenfasern. Die nächstniedere Stufe der Lagerung ist die Lamelle.Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Untreated silicone breast implant rupture

Implant rupture is a well-known complication of breast implant surgery that can pass unnoticed by both patient and physician. To date, no prospective study has addressed the possible health implications of silicone breast implant rupture. The aim of the present study was to evaluate whether untreated ruptures are associated with changes over time in magnetic resonance imaging findings, serologic markers, or self-reported breast symptoms. A baseline magnetic resonance imaging examination was performed in 1999 on 271 women who were randomly chosen from a larger cohort of women having cosmetic breast implants for a median period of 12 years (range, 3 to 25 years). A follow-up magnetic resonance imaging examination was carried out in 2001, excluding women who underwent explantation in the period between the two magnetic resonance imaging examinations (n = 44). On the basis of these examinations, the authors identified 64 women who had at least one ruptured implant at the first magnetic resonance imaging examination and, for comparison, all women who had intact implants at both examinations (n = 98). Magnetic resonance images from the two examinations were compared and changes in rupture configuration were evaluated. Comparisons were also made for self-reported breast symptoms occurring during the study period and for changes in serum values of antinuclear antibodies, rheumatoid factor, and cardiolipin antibodies immunoglobulin G and immunoglobulin M. The majority of the women with implant rupture had no visible magnetic resonance imaging changes of their ruptured implants. For 11 implants (11 percent) in 10 women, the authors observed progression of silicone seepage, either as a conversion from intracapsular into extracapsular rupture (n = 7), as progression of extra-capsular silicone (n = 3), or as increasing herniation of the silicone within the fibrous capsule (n = 1); however, in most cases, these changes were minor. Some changes could be ascribed to trauma, but others seemed spontaneous. There was no increase in levels of autoantibodies during the study period in either study group. Women with untreated implant ruptures reported a significant increase in nonspecific breast changes (odds ratio, 2.1; 95 percent confidence interval, 1.2 to 3.8) compared with women without ruptures. On the basis of this first study of women with untreated silicone breast implant rupture, the authors conclude that implant rupture is a relatively harmless condition, which only rarely progresses and gives rise to notable symptoms. Even so, because of a small risk of silicone spread, the authors suggest that women with implant ruptures be followed clinically, if not operated on. Because implant ruptures often occur asymptomatically, any woman with silicone implants, regardless of rupture status, should be evaluated at regular intervals.     

Glomulin is predominantly expressed in vascular smooth muscle cells in the embryonic and adult mouse

Mutations in the glomulin gene result in dominantly inherited vascular lesions of the skin known as glomuvenous malformations (GVMs). These lesions are histologically distinguished by their distended vein-like channels containing characteristic 'glomus cells', which appear to be incompletely or improperly differentiated vascular smooth muscle cells (VSMCs). The function of glomulin is currently unknown. We studied glomulin expression during murine development (E9.5 days post-coitum until adulthood) by non-radioactive in situ hybridization. Glomulin was first detected at E10.5 dpc in cardiac outflow tracts. Later, it showed strong expression in VSMCs as well as a limited expression in the perichondrium. At E11.5-14.5 dpc glomulin RNA was most abundant in the walls of the large vessels. At E16.5 dpc expression was also detectable in smaller arteries and veins. The high expression of glomulin in murine vasculature suggests an important role for glomulin in blood vessel development and/or maintenance, which is supported by the vascular phenotype seen in GVM patients with mutations in this gene.