Precise temporal regulation of alternative splicing during neural development

Alternative splicing (AS) is one crucial step of gene expression that must be tightly regulated during neurodevelopment. However, the precise timing of developmental splicing switches and the underlying regulatory mechanisms are poorly understood. Here we systematically analyze the temporal regulation of AS in a large number of transcriptome profiles of developing mouse cortices, in vivo purified neuronal subtypes, and neurons differentiated in vitro. Our analysis reveals early-switch and late-switch exons in genes with distinct functions, and these switches accurately define neuronal maturation stages. Integrative modeling suggests that these switches are under direct and combinatorial regulation by distinct sets of neuronal RNA-binding proteins including Nova, Rbfox, Mbnl, and Ptbp. Surprisingly, various neuronal subtypes in the sensory systems lack Nova and/or Rbfox expression. These neurons retain the “immature” splicing program in early-switch exons, affecting numerous synaptic genes. These results provide new insights into the organization and regulation of the neurodevelopmental transcriptome

Randomized controlled phase 2 trial of hydroxychloroquine in childhood interstitial lung disease

Background No results of controlled trials are available for any of the few treatments offered to children with interstitial lung diseases (chILD). We evaluated hydroxychloroquine (HCQ) in a phase 2, prospective, multicentre, 1:1-randomized, double-blind, placebo-controlled, parallel-group/crossover trial. HCQ (START arm) or placebo were given for 4 weeks. Then all subjects received HCQ for another 4 weeks. In the STOP arm subjects already taking HCQ were randomized to 12 weeks of HCQ or placebo (= withdrawal of HCQ). Then all subjects stopped treatment and were observed for another 12 weeks. Results 26 subjects were included in the START arm, 9 in the STOP arm, of these four subjects participated in both arms. The primary endpoint, presence or absence of a response to treatment, assessed as oxygenation (calculated from a change in transcutaneous O 2 -saturation of ≥ 5%, respiratory rate ≥ 20% or level of respiratory support), did not differ between placebo and HCQ groups. Secondary endpoints including change of O 2 -saturation ≥ 3%, health related quality of life, pulmonary function and 6-min-walk-test distance, were not different between groups. Finally combining all placebo and all HCQ treatment periods did not identify significant treatment effects. Overall effect sizes were small. HCQ was well tolerated, adverse events were not different between placebo and HCQ. Conclusions Acknowledging important shortcomings of the study, including a small study population, the treatment duration, lack of outcomes like lung function testing below age of 6 years, the small effect size of HCQ treatment observed requires careful reassessments of prescriptions in everyday practice (EudraCT-Nr.: 2013-003714-40, www.clinicaltrialsregister.eu , registered 02.07.2013)

Influence of pH, Temperature and Protease Inhibitors on Kinetics and Mechanism of Thermally Induced Aggregation of Potato Proteins

Understanding aggregation in food protein systems is essential to control processes ranging from the stabilization of colloidal dispersions to the formation of macroscopic gels. Patatin rich potato protein isolates (PPI) have promising techno-functionality as alternatives to established proteins from egg white or milk. In this work, the influence of pH and temperature on the kinetics of PPI denaturation and aggregation was investigated as an option for targeted functionalization. At a slightly acidic pH, rates of denaturation and aggregation of the globular patatin in PPI were fast. These aggregates were shown to possess a low amount of disulfide bonds and a high amount of exposed hydrophobic amino acids (S0). Gradually increasing the pH slowed down the rate of denaturation and aggregation and alkaline pH levels led to an increased formation of disulfide bonds within these aggregates, whereas S0 was reduced. Aggregation below denaturation temperature (Td) favored aggregation driven by disulfide bridge formation. Aggregation above Td led to fast unfolding, and initial aggregation was less determined by disulfide bridge formation. Inter-molecular disulfide formation occurred during extended heating times. Blocking different protein interactions revealed that the formation of disulfide bond linked aggregation is preceded by the formation of non-covalent bonds. Overall, the results help to control the kinetics, morphology, and interactions of potato protein aggregation for potential applications in food systems

Regulation of brain proteolytic activity is necessary for the in vivo function of NMDA receptors

Serine proteases are considered to be involved in plasticity-related events in the nervous system, but their in vivo targets and the importance of their control by endogenous inhibitors are still not clarified. Here, we demonstrate the crucial role of a potent serine protease inhibitor, protease nexin-1 (PN-1), in the regulation of activity-dependent brain proteolytic activity and the functioning of sensory pathways. Neuronal activity regulates the expression of PN-1, which in turn controls brain proteolytic activity. In PN-1-/- mice, absence of PN-1 leads to increased brain proteolytic activity, which is correlated with an activity-dependent decrease in the NR1 subunit of the NMDA receptor. Correspondingly, reduced NMDA receptor signaling is detected in their barrel cortex. This is coupled to decreased sensory evoked potentials in the barrel cortex and impaired whisker-dependent sensory motor function. Thus, a tight control of serine protease activity is critical for the in vivo function of the NMDA receptors and the proper function of sensory pathways

An investigation on the implementation of product development process models in construction companies

Process models have been developed by academia and industry to enhance design and construction activities. However, effective and widespread adoption and use of such models in practice has been limited. This research investigated the dynamics of product development process (PDP) models implementation in construction companies. Four case studies were undertaken, and key findings emphasize the need to consider the design and implementation of PDP models in an integrated fashion within the organizational context in which it takes place; and the need for a shift in the role of PDP models from a rational ‘planning and control’ perspective to a softer ‘learning’ approach

Long-term performance of oxidized zirconium on conventional and highly cross-linked polyethylene in total hip arthroplasty

Introduction: Polyethylene wear and subsequent osteolysis remain obstacles to the long-term survivorship of total hip arthroplasty (THA). Highly cross-linked polyethylene (XLPE) with radical quenching represents a massive leap forward with dramatically improved wear rates compared to ultra-high molecular weight polyethylene (UHMWPE). In this study we evaluate the wear of UHMWPE and XLPE coupled with oxidized zirconium (OxZr) femoral heads. Methods: A longitudinal, retrospective analysis was performed identifying consecutive patients who received a 28-mm OxZr-on-polyethylene primary THA from 2003 to 2004 by a single, high-volume arthroplasty surgeon. Patients were divided into two groups: those that received (1) UHMWPE liner and (2) a highly XLPE liner. Patients were included if clinical follow-up was complete to 2014 or later. Radiographic analysis was performed by two blinded observers. Measures included cup position, annual linear wear rate, and presence of osteolysis. Pairwise comparisons, correlations, and inter-rater reliability were calculated. Results: Eighty patients were in the UHMWPE group with an average follow-up of 10 ± 1.23 years and 88 patients in the XLPE group with an average of 10 ± 1.03-year follow-up. Average age (68) was similar between groups (p = 0.288). Observer reliability was excellent for cup abduction (ICC = 0.940), anteversion (ICC = 0.942), and detection of osteolysis (ICC = 0.811). Annual linear wear rates were significantly higher (p = 1 × 10−19) with UHMWPE (0.21 ± 0.12 mm/year) compared to XLPE (0.05 ± 0.03 mm/year). Linear wear rate was significantly correlated to decreasing acetabular abduction (p = 0.035). Osteolysis was noted only in the UHMWPE group, with 17 patients (21.2%) exhibiting acetabular osteolysis and 37 (46.3%) patients exhibiting femoral osteolysis. Conclusions: OxZr coupled with XLPE showed minimal wear and no osteolysis at 10-year follow up. The yearly linear penetration rate is similar to that seen in other studies of XLPE THA. A careful longitudinal follow-up will be required to determine if advanced bearings such as OxZr or ceramic can show improved performance in the second decade of implantation

Bulletin der deutschen Slavistik 16.2010

Bulletin der deutschen Slawistik 16, 201