Adult Human Vascular Smooth Muscle Cells on 3D Silk Fibroin Nonwovens Release Exosomes Enriched in Angiogenic and Growth-Promoting Factors

Background. Our earlier works showed the quick vascularization of mouse skin grafted Bombyx mori 3D silk fibroin nonwoven scaffolds (3D-SFnws) and the release of exosomes enriched in angiogenic/growth factors (AGFs) from in vitro 3D-SFnws-stuck human dermal fibroblasts (HDFs). Here, we explored whether coronary artery adult human smooth muscle cells (AHSMCs) also release AGFs-enriched exosomes when cultured on 3D-SFnws in vitro. Methods. Media with exosome-depleted FBS served for AHSMCs and human endothelial cells (HECs) cultures on 3D-SFnws or polystyrene. Biochemical methods and double-antibody arrays assessed cell growth, metabolism, and intracellular TGF-β and NF-κB signalling pathways activation. AGFs conveyed by CD9+/CD81+ exosomes released from AHSMCs were double-antibody array analysed and their angiogenic power evaluated on HECs in vitro. Results. AHSMCs grew and consumed D-glucose more intensely and showed a stronger phosphorylation/activation of TAK-1, SMAD-1/-2/-4/-5, ATF-2, c-JUN, ATM, CREB, and an IκBα phosphorylation/inactivation on SFnws vs. polystyrene, consistent overall with a proliferative/secretory phenotype. SFnws-stuck AHSMCs also released exosomes richer in IL-1α/-2/-4/-6/-8; bFGF; GM-CSF; and GRO-α/-β/-γ, which strongly stimulated HECs’ growth, migration, and tubes/nodes assembly in vitro. Conclusions. Altogether, the intensified AGFs exosomal release from 3D-SFnws-attached AHSMCs and HDFs could advance grafts’ colonization, vascularization, and take in vivo—noteworthy assets for prospective clinical applications

Will silk fibroin nanofiber scaffolds ever hold a usefulplace in Translational Regenerative Medicine?

Presently, some view silk fibroin-based biomaterials as obsolete, being outperformed by a host of newly discovered biomaterials. But several lines of evidence support the notion that silk fibroin proteins, especially those from B. mori and spiders and their recombinant forms, particularly in the form of electrospun nanofiber scaffolds, still represent promising tools for human tissue engineering/regeneration. Inevitably, the allure of recently reported biomaterials turns away many scientists and resources from the aim of more deeply elucidating the biological interactions of the various kinds of silk fibroin nanofiber scaffolds in vivo. But, even the biological features of newly reported biomaterials are not investigated in adequate depth. Hence, collaborative efforts among biomaterialists, biomedical experts, and private firms must be undertaken on a much greater scale than hitherto done to assess the real usefulness of silk fibroin proteins, thereby allowing or denying their useful introduction into the fields of Translational Regenerative Medicine

Is Alzheimer\u2019s Disease at Least Partly a Ciliopathy?

The uninterrupted generation throughout life of the dentate gyrus [DGY] granule cells [GCs] (one site of adult neurogenesis), which initiate the encoding of novel memories, is driven by signals from the DGy GC precursors tiny, non-motile primary cilia. The hypothesis is surmised that the damage of such primary cilia be responsible of the crippling decline of memory formation in Alzheimer's Disease [AD]. Were human DGy CGs ciliated like their rodent counterparts, part of the AD cases might be indeed based upon a ciliopathy

Amyloid-\u3b225-35, an Amyloid-\u3b21-42 Surrogate, and Proinflammatory Cytokines Stimulate VEGF-A Secretion by Cultured, Early Passage, Normoxic Adult Human Cerebral Astrocytes

Cerebrovascular angiopathy affects late-onset Alzheimer's disease (LOAD) brains by possibly increasing vascular endothelial growth factor (VEGF). A expression, thereby stimulating endothelial cell proliferation and migration. Indeed, VEGF-A gene upregulation, with increased VEGF-A protein content of reactive astrocytes and microglia, occurs in LOAD brains, and neovascularization was observed one week after injecting amyloid-\u3b2 (A\u3b2)1-42 into rat hippocampus. We have now found, with cultured 'normoxic' normal adult human astrocytes (NAHAs), that fibrillar A\u3b225-35 (an active A\u3b21-42 fragment) or a cytokine mixture (the (CM)-trio (interleukin [IL]-1\u3b2+interferon [IFN]-\u3b3+tumor necrosis factor [TNF]-\u3b1), or pair (IFN-\u3b3+TNF-\u3b1) like those produced in LOAD brains) stimulates the nuclear translocation of stabilized hypoxia-inducible factor (HIF)-1\u3b1 protein and its binding to VEGF-A hypoxia-response elements; the mRNA synthesis for three VEGF-A splice variants (121, 165, 189); and the secretion of VEGF-A165. The CM-trio was the most powerful stimulus, IFN-\u3b3+TNF-\u3b1 was less potent, and other cytokine pairs or single cytokines or A\u3b235-25 were ineffective. While A\u3b225-35 did not change HIF-1\u3b2 protein levels, the CM-trio increased both HIF-1\u3b1 and HIF-1\u3b2 protein levels, thereby giving an earlier and stronger stimulus to VEGF-A secretion by NAHAs. Thus, increased VEGF-A secretion from astrocytes stimulated by A\u3b21-42 and by microglia-released cytokines might restore angiogenesis and A\u3b21-42 vascular clearance

Leptin, Sonic Hedgehogs, and Neurogenesis--A Primary Cilium's Tale

Leptin-induced signals from from Leptin receptor (R)-b stationed in the cell membrane could stimulate the primary cilium-dependent proliferation of transit amplifying cells [TANs] generated by radial glial neuronal stem cells [RG-NSCs] in the dentate gyrus of the adult hippocampal formation

A stable panel comprising 18 urinary proteins in the human healthy population.

Just as biomarkers specific for diseases, biomarkers indicative of healthy conditions are valuable for the early diagnosis, monitoring, and prognosis of diseases. Our study focused on discovering via proteomics a stable panel of urinary proteins in the human healthy population. Urine samples were collected three times during 4 months from 100 male and 100 female healthy donors and analyzed through four different fractionation techniques (i.e. in-gel, 2D-LC, OFFGEL, and mRP) coupled with HPLC-Chip-MS/MS. Thus, 1641 urinary proteins were identified with a high confidence, among which 70 exhibiting an intergender/day variation <0.25 were selected and matched with the previously published five largest urinary proteomes to get 56 candidate proteins. Next, a panel comprising 18 intact urinary proteins was constructed by comparing the urinary proteomes via SDS-PAGE and 2DE. Finally, such 18 urinary proteins were validated via enzyme-linked immunosorbent assay in eight healthy individuals. Most of these proteins had been related to multiple rather than to single diseases. Therefore, we surmise that this protein set could be used as a biomarker to assess the human health status. Further determinations of the normal fluctuations of the single urinary proteins in this series using samples from large numbers of healthy individuals are required prior to any application in clinical settings

Digital chest radiography: an update on modern technology, dose containment and control of image quality

The introduction of digital radiography not only has revolutionized communication between radiologists and clinicians, but also has improved image quality and allowed for further reduction of patient exposure. However, digital radiography also poses risks, such as unnoticed increases in patient dose and suboptimum image processing that may lead to suppression of diagnostic information. Advanced processing techniques, such as temporal subtraction, dual-energy subtraction and computer-aided detection (CAD) will play an increasing role in the future and are all targeted to decrease the influence of distracting anatomic background structures and to ease the detection of focal and subtle lesions. This review summarizes the most recent technical developments with regard to new detector techniques, options for dose reduction and optimized image processing. It explains the meaning of the exposure indicator or the dose reference level as tools for the radiologist to control the dose. It also provides an overview over the multitude of studies conducted in recent years to evaluate the options of these new developments to realize the principle of ALARA. The focus of the review is hereby on adult applications, the relationship between dose and image quality and the differences between the various detector systems

Search for Eccentric Black Hole Coalescences during the Third Observing Run of LIGO and Virgo

Despite the growing number of confident binary black hole coalescences observed through gravitational waves so far, the astrophysical origin of these binaries remains uncertain. Orbital eccentricity is one of the clearest tracers of binary formation channels. Identifying binary eccentricity, however, remains challenging due to the limited availability of gravitational waveforms that include effects of eccentricity. Here, we present observational results for a waveform-independent search sensitive to eccentric black hole coalescences, covering the third observing run (O3) of the LIGO and Virgo detectors. We identified no new high-significance candidates beyond those that were already identified with searches focusing on quasi-circular binaries. We determine the sensitivity of our search to high-mass (total mass $M>70$ $M_\odot$) binaries covering eccentricities up to 0.3 at 15 Hz orbital frequency, and use this to compare model predictions to search results. Assuming all detections are indeed quasi-circular, for our fiducial population model, we place an upper limit for the merger rate density of high-mass binaries with eccentricities $0 < e \leq 0.3$ at $0.33$ Gpc$^{-3}$ yr$^{-1}$ at 90\% confidence level.Comment: 24 pages, 5 figure

Il Midollo Spinale

Nuova trattazione completamente rifatta e aggiornata dell'Anatomia macro- e microscopica, della neurocitologia e della neurofisiologia del midollo spinale dell'Uom