Generation of Amyloid-β Is Reduced by the Interaction of Calreticulin with Amyloid Precursor Protein,Presenilin and Nicastrin

Dysregulation of the proteolytic processing of amyloid precursor protein by γ-secretase and the ensuing generation of amyloid-β is associated with the pathogenesis of Alzheimer''s disease. Thus, the identification of amyloid precursor protein binding proteins involved in regulating processing of amyloid precursor protein by the γ-secretase complex is essential for understanding the mechanisms underlying the molecular pathology of the disease. We identified calreticulin as novel amyloid precursor protein interaction partner that binds to the γ-secretase cleavage site within amyloid precursor protein and showed that this Ca²⁺- and N-glycan-independent interaction is mediated by amino acids 330–344 in the C-terminal C-domain of calreticulin. Co-immunoprecipitation confirmed that calreticulin is not only associated with amyloid precursor protein but also with the γ-secretase complex members presenilin and nicastrin. Calreticulin was detected at the cell surface by surface biotinylation of cells overexpressing amyloid precursor protein and was co-localized by immunostaining with amyloid precursor protein and presenilin at the cell surface of hippocampal neurons. The P-domain of calreticulin located between the N-terminal N-domain and the C-domain interacts with presenilin, the catalytic subunit of the γ-secretase complex. The P- and C-domains also interact with nicastrin, another functionally important subunit of this complex. Transfection of amyloid precursor protein overexpressing cells with full-length calreticulin leads to a decrease in amyloid-β₄₂ levels in culture supernatants, while transfection with the P-domain increases amyloid-β₄₀ levels. Similarly, application of the recombinant P- or C-domains and of a synthetic calreticulin peptide comprising amino acid 330–344 to amyloid precursor protein overexpressing cells result in elevated amyloid-β₄₀ and amyloid-β₄₂ levels, respectively. These findings indicate that the interaction of calreticulin with amyloid precursor protein and the γ-secretase complex regulates the proteolytic processing of amyloid precursor protein by the γ-secretase complex, pointing to calreticulin as a potential target for therapy in Alzheimer''s disease.     

Can habitat characteristics shape vertebral morphology in dolphins? An example of two phylogenetically related species from southern South America

Fast swimming pelagic cetacean species have osteological characteristics that promote a more stable spine in comparison to that of coastal species. The Peale's dolphin (Lagenorhynchus australis) and the hourglass dolphin (Lagenorhynchus cruciger) have a close phylogenetic relationship and are found in coastal and pelagic waters in the Southern Hemisphere, respectively. The aim of this work was to study the relationship between the vertebral column's morphology and its flexibility, across these species of contrasting habitats. Vertebral counts and multiple measurements of each vertebra were used to infer intervertebral flexibility. Bivariate plots and discriminant multivariate analyses were employed to compare each functional region along the vertebral column. Both species displayed a regionalization of the column into three stable regions and two flexible areas, which statistically differ in the proportion of the skeleton occupied in each species. While the Peale's dolphin has rounder vertebrae, associated with higher flexibility, the hourglass dolphin has disk‐shaped vertebrae and strongly inclined processes related to high stability. Although the species are closely related phylogenetically, vertebral morphology is influenced by a diverse set of ecological and behavioral factors, reflecting a high degree of vertebral plasticity within the genus.     

Developmental trends of theta–beta interelectrode power correlation during resting state in normal children

The possibility that power-to-power (theta–beta) frequency coupling increases during development was analyzed. Three minutes of spontaneous EEG in an open eyes condition were recorded in a sample of 160 subjects ranging from 6 to 26 years old. Theta (4–7 Hz) and beta band (15–20 Hz) power was calculated in a trial-by-trial basis. Inter-electrode power correlations (IPC) were computed in each subject as the correlation between the power of two frequency bands recorded in two electrodes. An increase in theta–beta IPC with age was obtained. IPCs were higher when theta was seeded in posterior regions than in anterior or central regions. Moreover, the significant correlations between each individual IPC and age were calculated, making it possible to draw IPC versus age correlation maps in order to capture the IPC development topography. An increase was found in significant correlations in the left hemisphere compared to the right hemisphere. There were no differences in the inter-hemispheric versus intra-hemispheric IPC maturation spatial patterns. An increase in power-to-power–frequency coupling in theta–beta occurs during development, suggesting an increase in functional connectivity with age. Frequency coupling between theta and beta rhythms would be one of the mechanisms facilitating integration of long distance functional networks during development.     

Solution NMR assignment of the C-terminal domain of human chTOG

The microtubule regulatory protein colonic and hepatic tumor overexpressed gene (chTOG), also known as cytoskeleleton associated protein 5 (CKAP5) plays an important role in organizing the cytoskeleton and in particular in the assembly of k-fibres in mitosis. Recently, we dissected the hitherto poorly understood C-terminus of this protein by discovering two new domains—a cryptic TOG domain (TOG6) and a smaller, helical domain at the very C-terminus. It was shown that the C-terminal domain is important for the interaction with the TACC domain in TACC3 during the assembly of k-fibres in a ternary complex that also includes clathrin. Here we now present the solution NMR assignment of the chTOG C-terminal domain which confirms our earlier prediction that it is mainly made of α-helices. However, the appearance of the ¹H–¹⁵N HSQC spectrum is indicative of the presence of a considerable amount of unstructured and possibly flexible portions of protein in the domain.     

Novel materials based on DNA‐CTMA and lanthanide (Ce3+, Pr3+)

New, deoxyribonucleic acid (DNA) based compounds, functionalized with hexadecyltrimethylammonium chloride (CTMA) and lanthanide hydroxide nanoparticles were synthesized. The spectral measurements suggest that between the DNA‐CTMA complex and the lanthanide (III) ions a chemical interaction takes place. The obtained materials exhibit an improved fluorescence efficiency, showing a potential interest for application in photonics, and more particularly, in light emitting devices. © 2016 Wiley Periodicals, Inc. Biopolymers 105: 613–617, 2016.     

Daily nurse requirements planning based on simulation of patient flows

Nurses account for approximately 50 % of total hospital budgets and their allocation to medical units and shifts can significantly affect the quality of care provided to patients. The adoption of flexible shift schedules and the assessment of actual nursing time can enable sensible resource planning, balancing the quality of care with efficiency in resource use. Starting from the concept that nurse requirements are triggered by patient needs, which are stochastic in nature both for clinical activities and their duration, this paper proposes an innovative Nurse Requirement Planning model grounded on the concept of the clinical pathway (the “standard” sequence of diagnostic, therapeutic and care activities a patient with certain pathology should undertake over time) with its inner routing probability and patient dependence on nurses, which can be correlated to the time needed to perform nursing tasks. In merging and modelling these two aspects, the method summarizes the best features of acuity-quality and timed-task/activity techniques, well known although not usually applied for reasons of demands on clinicians’ time. Instead, in this paper, for each shift of the day, hospital management is enabled to choose the optimal number of nurses to meet actual requirements according to a desired service level and personnel saturation by means of a tool that simulates the patient flow in a medical unit based on automatic data retrieval from hospital databases. The validation and verification of the proposal were undertaken in a stroke unit.     

Rotifer Nervous System Visualized by FMRFamide and 5-HT Immunocytochemistry and Confocal Laser Scanning Microscopy

The characteristic ecology of floodplain lakes is in part due to their relatively strong water-level fluctuations. We analyzed the factors determining water-level fluctuations in 100 floodplain lakes (during non-flooded conditions) in the active floodplains of the Lower Rhine in the Netherlands. Furthermore, we explored the relationship between water-level fluctuations and macrophyte species richness, and analyzed the suitability of artificially created lakes for macrophyte vegetation. During non-flooded conditions along the Rhine, lake water-level fluctuations are largely driven by groundwater connection to the river. Hence, water-level fluctuations are largest in lakes close to the main channel in strongly fluctuating sectors of the river and smallest in isolated lakes. Additionally, water-level fluctuations are usually small in old lakes, mainly due to reduced groundwater hydraulic conductivity resulting from accumulated clay and silt on the bottom. Species richness of floating-leaved and emergent macrophytes was reduced at both small and large water-level fluctuations, whereas species richness of submerged macrophytes was reduced at small water-level fluctuations only. In addition, species richness of submerged macrophytes was higher in lakes that experienced drawdown, whereas no similar pattern was detected for floating-leaved and emergent macrophytes. The decline in amplitude of lake water-level with lake age implies that the number of hydrologically dynamic lakes will decrease over time. Therefore, we suggest that excavation of new lakes is essential to conserve the successional sequence of floodplain water bodies including conditions of high biodiversity. Shallow, moderately isolated, lakes with occasional bottom exposure have the highest potential for creating macrophyte-rich floodplain lakes along large lowland rivers. The water-level regime of such lakes can in part be designed, through choice of the location along the river, the distance away from the river and the depth profile of the lake.     

Mitochondrial complex III ROS regulate adipocyte differentiation

Adipocyte differentiation is characterized by an increase in mitochondrial metabolism. However, it is not known whether the increase in mitochondrial metabolism is essential for differentiation or a byproduct of the differentiation process. Here, we report that primary human mesenchymal stem cells undergoing differentiation into adipocytes display an early increase in mitochondrial metabolism, biogenesis, and reactive oxygen species (ROS) generation. This early increase in mitochondrial metabolism and ROS generation was dependent on mTORC1 signaling. Mitochondrial-targeted antioxidants inhibited adipocyte differentiation, which was rescued by the addition of exogenous hydrogen peroxide. Genetic manipulation of mitochondrial complex III revealed that ROS generated from this complex is required to initiate adipocyte differentiation. These results indicate that mitochondrial metabolism and ROS generation are not simply a consequence of differentiation but are a causal factor in promoting adipocyte differentiation.     

Plasma and salivary steroid hormone responses of men to high-intensity cycling and resistance exercise

Hormonal responses to exercise could be used as a marker of overreaching. A short exercise protocol that induces robust hormonal elevations in a normal trained state should be able to highlight hormonal changes during overreaching. This study compared plasma and salivary cortisol and testosterone responses to 4 exercise trials; these were (a) continuous cycle to fatigue at 75% peak power output (Wmax) (FAT); (b) 30-minute cycle alternating 1-minute 60% and 1 minute 90% Wmax (60/90); (c) 30-minute cycle alternating 1-minute 55% and 4-minute 80% Wmax (55/80); and (d) Squatting 8 sets of 10 repetitions at 10 repetition maximum (RESIST). Blood and saliva samples were collected pre-exercise and at 0, 10, 20, 30, 40, 50, and 60 minute postexercise. Pre- to postexercise plasma cortisol increased in all exercise trials, except 60/90. Increases in 55/80 remained above pre-exercise levels for the entire postexercise period. Salivary cortisol increased from pre- to postexercise in FAT and 55/80 trials only. Once elevated after 55/80, it remained so for the postexercise period. Plasma testosterone increased from pre- to postexercise in all trials except 55/80. Saliva testosterone increased from pre- to postexercise in all trials with the longest elevation occurring after 55/80. Area under the curve analysis indicated that the exercise response of salivary hormones was greater in all cycle trials (cortisol) and in the 60/90 and 55/80 trials (testosterone) compared with the other trials. This study indicates that the 55/80 cycle protocol induces a prolonged salivary and plasma cortisol and salivary testosterone response compared with the other trials and so may be a useful diagnostic tool of overreaching.