Syngeneic Cardiac and Bone Marrow Stromal Cells Display Tissue-Specific microRNA Signatures and microRNA Subsets Restricted to Diverse Differentiation Processes

MicroRNAs are key modulators at molecular level in different biological processes, including determination of cell fate and differentiation. Herein, microRNA expression profiling experiments were performed on syngeneic cardiac (CStC) and bone marrow (BMStC) mesenchymal stromal cells cultured in standard growth medium and then in vitro exposed to adipogenic, osteogenic, cardiomyogenic and endothelial differentiation media. Analysis identified a tissue-specific microRNA signature composed of 16 microRNAs that univocally discriminated cell type of origin and that were completely unaffected by in vitro differentiation media: 4 microRNAs were over-expressed in cardiac stromal cells, and 12 were overexpressed or present only in bone marrow stromal cells. Further, results revealed microRNA subsets specifically modulated by each differentiation medium, irrespective of the cell type of origin, and a subset of 7 microRNAs that were down-regulated by all media with respect to growth medium. Finally, we identified 16 microRNAs that were differentially modulated by the media when comparing the two tissues of origin. The existence of a tissue-specific microRNA signature surviving to any differentiation stimuli, strongly support the role if microRNAs determining cell identity related to tissue origin. Moreover, we identified microRNA subsets modulated by different culture conditions in a tissue-specific manner, pointing out their importance during differentiation processes.     

Comparative Analysis of rs12979860 SNP of the IFNL3 Gene in Children with Hepatitis C and Ethnic Matched Controls Using 1000 Genomes Project Data

The rs12979860 single nucleotide polymorphism located on chromosome 19q13.13 near the interferon L3 gene (formerly and commonly known as interleukin 28B gene) has been associated in adults with both spontaneous and treatment induced clearance of hepatitis C virus. Although the exact mechanism of these associations remains unclear, it suggests that variation in genes involved in the immune response against the virus favours viral clearance. Limited and preliminary data are available on this issue in children. The aim of the present study was to evaluate, in a representative cohort of children with perinatal infection, the potential association between rs12979860 single nucleotide polymorphism and the outcome of hepatitis C virus infection. Alleles and genotypes frequencies were evaluated in 30 children who spontaneously cleared the virus and in 147 children with persistent infection and were compared with a population sample of ethnically matched controls with unknown hepatitis C status obtained using the 1000 Genomes Project data. The C allele and the C/C genotype showed greater frequencies in the clearance group (76.7% and 56.7%, respectively) when compared with both children with viral persistence (C allele 56.5%, p = 0.004; C/C genotype 32.7%, p = 0.02) and with the ethnically matched individuals (C allele 59.7%, p = 0.02; C/C genotype 34.7%, p = 0.03). Children with the C/C genotype were 2 times more likely to clear hepatitis C virus relative to children with the C/T and T/T genotypes combined (odds ratio: 2.7; 90% confidence intervals: 1.3–5.8). The present study provides the evidence that the rs12979860 single nucleotide polymorphism influences the natural history of hepatitis C virus in children.     

Stigmergic algorithms for multiple minimalistic robots on an RFID floor

Stigmergy is a powerful principle in nature, which has been shown to have interesting applications to robotic systems. By leveraging the ability to store information in the environment, robots with minimal sensing, memory, and computational capabilities can solve complex problems like global path planning. In this paper, we discuss the use of stigmergy in minimalist multi-robot systems, in which robots do not need to use any internal model, long-range sensing, or position awareness. We illustrate our discussion with three case studies: building a globally optimal navigation map, building a gradient map of a sensed feature, and updating the above maps dynamically. All case studies have been implemented in a real environment with multiple ePuck robots, using a floor with 1,500 embedded radio frequency identification tags as the stigmergic medium. Results collected from tens of hours of real experiments and thousands of simulated runs demonstrate the effectiveness of our approach.     

Macrophage Cholesterol Depletion and Its Effect on the Phagocytosis of Cryptococcus neoformans

Cryptococcosis is a life-threatening infection caused by pathogenic fungi of the genus Cryptococcus. Infection occurs upon inhalation of spores, which are able to replicate in the deep lung. Phagocytosis of Cryptococcus by macrophages is one of the ways that the disease is able to spread into the central nervous system to cause lethal meningoencephalitis. Therefore, study of the association between Cryptococcus and macrophages is important to understanding the progression of the infection. The present study describes a step-by-step protocol to study macrophage infectivity by C. neoformansin vitro. Using this protocol, the role of host sterols on host-pathogen interactions is studied. Different concentrations of methyl--cyclodextrin (MCD) were used to deplete cholesterol from murine reticulum sarcoma macrophage-like cell line J774A.1. Cholesterol depletion was confirmed and quantified using both a commercially available cholesterol quantification kit and thin layer chromatography. Cholesterol depleted cells were activated using Lipopolysacharide (LPS) and Interferon gamma (IFNγ) and infected with antibody-opsonized Cryptococcus neoformans wild-type H99 cells at an effector-to-target ratio of 1:1. Infected cells were monitored after 2 hr of incubation with C. neoformans and their phagocytic index was calculated. Cholesterol depletion resulted in a significant reduction in the phagocytic index. The presented protocols offer a convenient method to mimic the initiation of the infection process in a laboratory environment and study the role of host lipid composition on infectivity.     

Nitric oxide and cytochrome oxidase: reaction mechanisms from the enzyme to the cell

The aim of this work is to review the information available on the molecular mechanisms by which the NO radical reversibly downregulates the function of cytochrome c oxidase (CcOX). The mechanisms of the reactions with NO elucidated over the past few years are described and discussed in the context of the inhibitory effects on the enzyme activity. Two alternative reaction pathways are presented whereby NO reacts with the catalytic intermediates of CcOX populated during turnover. The central idea is that at "cellular" concentrations of NO (相似文献   

A multi‐data ensemble approach for predicting woodland type distribution: Oak woodland in Britain

Interactions between soil, topography, and climatic site factors can exacerbate and/or alleviate the vulnerability of oak woodland to climate change. Reducing climate‐related impacts on oak woodland habitats and ecosystems through adaptation management requires knowledge of different site interactions in relation to species tolerance. In Britain, the required thematic detail of woodland type is unavailable from digital maps. A species distribution model (SDM) ensemble, using biomod2 algorithms, was used to predict oak woodland. The model was cross‐validated (50%:50% ‐ training:testing) 30 times, with each of 15 random sets of absence data, matching the size of presence data, to maximize environmental variation while maintaining data prevalence. Four biomod2 algorithms provided stable and consistent TSS‐weighted ensemble mean results predicting oak woodland as a probability raster. Biophysical data from the Ecological Site Classification (forest site classification) for Britain were used to characterize oak woodland sites. Several forest datasets were used, each with merits and weaknesses: public forest estate subcompartment database map (PFE map) for oak‐stand locations as a training dataset; the national forest inventory (NFI) “published regional reports” of oak woodland area; and an “NFI map” of indicative forest type broad habitat. Broadleaved woodland polygons of the NFI map were filled with the biomod2 oak woodland probability raster. Ranked pixels were selected up to the published NFI regional area estimate of oak woodland and matched to the elevation distribution of oak woodland stands, from “NFI survey” sample squares. Validation using separate oak woodland data showed that the elevation filter significantly improved the accuracy of predictions from 55% (p = .53) to 83% coincidence success rate (p < .0001). The biomod2 ensemble, with masking and filtering, produced a predicted oak woodland map, from which site characteristics will be used in climate change interaction studies, supporting adaptation management recommendations for forest policy and practice.     

Recognition mechanism of p63 by the E3 ligase Itch: Novel strategy in the study and inhibition of this interaction

The HECT-containing E3 ubiquitin ligase Itch mediates the degradation of several proteins, including p63 and p73, involved in cell specification and fate. Itch contains four WW domains, which are essential for recognition on the target substrate, which contains a short proline-rich sequence. Several signaling complexes containing these domains have been associated with human diseases such as muscular dystrophy, Alzheimer’s or Huntington’s diseases. To gain further insight into the structural determinants of the Itch-WW2 domain, we investigated its interaction with p63. We assigned, by 3D heteronuclear NMR experiments, the backbone and side chains of the uniformly ¹³C-¹⁵N-labeled Itch-WW2. In vitro interaction of Itch-WW2 domain with p63 was studied using its interactive p63 peptide, pep63. Pep63 is an 18-mer peptide corresponding to the region from 534–551 residue of p63, encompassing the PPxY motif that interacts with the Itch-WW domains, and we identified the residues involved in this molecular recognition. Moreover, here, a strategy of stabilization of the conformation of the PPxY peptide has been adopted, increasing the WW-ligand binding. We demonstrated that cyclization of pep63 leads to an increase of both the biological stability of the peptide and of the WW-ligand complex. Stable metal-binding complexes of the pep63 have been also obtained, and localized oxidative damage on Itch-WW2 domain has been induced, demonstrating the possibility of use of metal-pep63 complexes as models for the design of metal drugs to inhibit the Itch-WW-p63 recognition in vivo. Thus, our data suggest a novel strategy to study and inhibit the recognition mechanism of Itch E3-ligase.     

Analysis of nerve supply pattern in thoracic duct in young and elderly men

BACKGROUND: Analysis of the innervation pattern of the thoracic duct in young and elderly human subjects has been performed. The subdivision of the vessels in cervical and lumbar region were taken in consideration. MeTHODS AND RESULTS: Immunostaining for general nerve fibers with a PGP 9.5 marker disclosed a diffuse innervation of the thoracic duct in young subjects, which was strongly reduced in elderly subjects. In young subjects, tyrosine hydroxylase (TH) and neuropeptide Y (NPY) immunoreactive fibers, markers of noradrenergic postganglionic sympathetic fibers, were frequent; choline acetyltransferase (ChAT) immunoreactive fibers, marker of cholinergic parasympathetic nerve fibers, were also well represented. Therefore, the influence of sympathetic and parasympathetic nerve systems on the thoracic duct can be confirmed. The immunoreactivity of vasoactive intestinal peptide (VIP), a neuropeptide frequently present in cholinergic parasympathetic nerve fibers, was scarcely present. Dopamine-positive fibers were observed in few short nerve fibers. Substance P (SP)-positive fibers were widely distributed in the medial and intimal smooth muscle layers, suggesting their involvement as contractile modulating fibers and sensitive fibers. In elderly subjects, an evident reduction of all specific nerve fibers analyzed was detected, the ChAT-positive fibers being the most affected. CONCLUSIONS: The lymphatic vessel thoracic duct is able to regulate hydrodynamic lymph flow by intrinsic contraction of its smooth muscle layer. Therefore, analysis of the thoracic duct innervation pattern may be important in assessing the regulation of vessel contraction. These findings called attention to the reduction of lymphatic drainage functionality affecting fluid balance in the elderly.     

Increasing Strawberry Fruit Sensorial and Nutritional Quality Using Wild and Cultivated Germplasm

Background

Increasing antioxidant levels in fruit through breeding is an important option to support higher antioxidant intake particularly when fruit consumption is low. Indeed, if nutritional components are also combined with a high standard of sensorial fruit quality, the perspective for consumer health can be further improved by encouraging more fruit consumption. Wild species are valued by strawberry breeders as sources of novel traits, especially for pest resistance and abiotic stress tolerance. Furthermore, previous investigations have shown improvements in fruit nutritional quality in breeding material that originated from Fragaria virginiana ssp. glauca (FVG) inter-species crosses. Recently, commercial varieties of strawberries have also shown interesting variability in fruit nutritional quality.

Results

Strawberry fruit sensorial and nutritional qualities generated by Fragaria inter-species and intra-species crosses were evaluated on 78 offspring derived from 8 families: two that originated from F. × ananassa intra-species crossing; three from back-crossing of F1– FVG × F. × ananassa; and three from back-crossing of BC1– FVG × F. × ananassa. The genetic variability from the three types of cross combinations was analyzed by calculation of the correlations among the fruit sensorial and nutritional parameters. The results obtained show that two subsequent back-crossing generations from an inter-species crossing combination with F. virginiana ssp. glauca provides useful improvement of the fruit nutritional and sensorial qualities that is combined with agronomic standards that are close to those requested at the commercial level. Improvements of these traits can also be achieved by programming F. × ananassa intra-species crosses and producing progeny with productivity traits more similar to those of the commercial cultivars.

Conclusions

The two types of combination programs (inter-species back-crosses, and intra-species crosses) can be used to improve strawberry nutritional quality.     

Lipoprotein-Associated Phospholipase A2 Activity Predicts Cardiovascular Events in High Risk Coronary Artery Disease Patients

Objective

Lipoprotein-associated phospholipase A2 (Lp-PLA2) is deemed to play a role in atherosclerosis and plaque destabilization as demonstrated in animal models and in prospective clinical studies. However, most of the literature is either focused on high-risk, apparently healthy patients, or is based on cross sectional studies. Therefore, we tested the hypothesis that serum Lp-PLA2 mass and activity are useful for predicting cardiovascular (CV) events over the coronary atherosclerotic burden and conventional risk factors in high-risk coronary artery disease patients.

Methods and Results

In a prospective cohort study of 712 Caucasian patients, who underwent coronary angiography and measurement of both Lp-PLA2 mass and activity at baseline, we determined incident CV events at follow-up after splitting the patients into a high and a low Lp-PLA2 mass and activity groups based on ROC analysis and Youden index. Kaplan-Meier and propensity score matching analysis were used to compare CV event-free survival between groups. Follow-up data were obtained in 75% of the cohort after a median of 7.2 years (range 1–12.7 years) during which 129 (25.5%) CV events were observed. The high Lp-PLA2 activity patients showed worse CV event-free survival (66.7% vs. 79.5%, p = 0.023) and acute coronary syndrome-free survival (75.4% vs. 85.6%, p = 0.04) than those in low Lp-PLA2 group.

Conclusions

A high Lp-PLA2 activity implies a worse CV prognosis at long term follow up in high-risk Caucasian patients referred for coronary angiography.