CaSR Antagonist (Calcilytic) NPS 2143 Hinders the Release of Neuroinflammatory IL-6, Soluble ICAM-1, RANTES, and MCP-2 from A\u3b2-Exposed Human Cortical Astrocytes

Available evidence shows that human cortical neurons\u2019 and astrocytes\u2019 calcium-sensing receptors (CaSRs) bind Amyloid-beta (A\u3b2) oligomers triggering the overproduction/oversecretion of several Alzheimer\u2019s disease (AD) neurotoxinseffects calcilytics suppress. We asked whether A\u3b2CaSR signaling might also play a direct pro-neuroinflammatory role in AD. Cortical nontumorigenic adult human astrocytes (NAHAs) in vitro were untreated (controls) or treated with A\u3b225-35\uf020\ub1\uf020NPS 2143 (a calcilytic) and any proinflammatory agent in their protein lysates and growth media assayed via antibody arrays, enzyme-linked immunosorbent assays (ELISAs), and immunoblots. Results show A\u3b2\u2022CaSR signaling upregulated the synthesis and release/shedding of proinflammatory interleukin (IL)-6, intercellular adhesion molecule-1 (ICAM-1) (holoprotein and soluble [s] fragment), Regulated upon Activation, normal T cell Expressed and presumably Secreted (RANTES), and monocyte chemotactic protein (MCP)-2. Adding NPS 2143 (i) totally suppressed IL-6\u2032s oversecretion while remarkably reducing the other agents\u2019 over-release; and (ii) more effectively than A\u3b2 alone increased over controls the four agents\u2019 distinctive intracellular accumulation. Conversely, NPS 2143 did not alter A\u3b2-induced surges in IL-1\u3b2, IL-3, IL-8, and IL-16 secretion, consequently revealing their A\u3b2\u2022CaSR signaling-independence. Finally, A\u3b225-35\uf020\ub1\uf020NPS 2143 treatments left unchanged MCP-1\u2032s and TIMP-2\u2032s basal expression. Thus, NAHAs A\u3b2\u2022CaSR signaling drove four proinflammatory agents\u2019 over-release that NPS 2143 curtailed. Therefore, calcilytics would also abate NAHAs\u2019 A\u3b2\u2022CaSR signaling direct impact on AD\u2019s neuroinflammation

Calcium-sensing receptor antagonist NPS 2143 restores amyloid precursor protein physiological non-amyloidogenic processing in A\u3b2-exposed adult human astrocytes

Physiological non-amyloidogenic processing (NAP) of amyloid precursor holoprotein (hAPP) by \u3b1-secretases (e.g., ADAM10) extracellularly sheds neurotrophic/neuroprotective soluble (s)APP\u3b1 and precludes amyloid-\u3b2 peptides (A\u3b2s) production via \u3b2-secretase amyloidogenic processing (AP). Evidence exists that A\u3b2s interact with calcium-sensing receptors (CaSRs) in human astrocytes and neurons, driving the overrelease of toxic A\u3b242/A\u3b242-os (oligomers), which is completely blocked by CaSR antagonist (calcilytic) NPS 2143. Here, we investigated the mechanisms underlying NPS 2143 beneficial effects in human astrocytes. Moreover, because Alzheimer's disease (AD) involves neuroinflammation, we examined whether NPS 2143 remained beneficial when both fibrillary (f)A\u3b225-35 and a microglial cytokine mixture (CMT) were present. Thus, hAPP NAP prevailed over AP in untreated astrocytes, which extracellularly shed all synthesized sAPP\u3b1 while secreting basal A\u3b240/42 amounts. Conversely, fA\u3b225-35 alone dramatically reduced sAPP\u3b1 extracellular shedding while driving A\u3b242/A\u3b242-os oversecretion that CMT accelerated but not increased, despite a concurring hAPP overexpression. NPS 2143 promoted hAPP and ADAM10 translocation to the plasma membrane, thereby restoring sAPP\u3b1 extracellular shedding and fully suppressing any A\u3b242/A\u3b242-os oversecretion, but left hAPP expression unaffected. Therefore, as anti-AD therapeutics calcilytics support neuronal viability by safeguarding astrocytes neurotrophic/neuroprotective sAPP\u3b1 shedding, suppressing neurons and astrocytes A\u3b242/A\u3b242-os build-up/secretion, and remaining effective even under AD-typical neuroinflammatory conditions

Exosomes of adult human fibroblasts cultured on 3D silk fibroin nonwovens intensely stimulate neoangiogenesis

Bombyx mori silk fibroin (SF) is a biomacromolecule allowing to assemble scaffolds 7 for tissue engineering/regeneration purposes because of its cellular adhesiveness, high 8 biocompatibility, and low immunogenicity. Earlier work showed that two types of 3D-SF-based 9 nonwovens (3D-SFnws) implanted into mouse subcutaneous tissue were promptly vascularised via plates in exosome-depleted medium. DNA amounts and D-glucose consumption revealed HDFs 17 undefined molecular mechanisms. This study used nontumorigenic adult human dermal fibroblasts (HDFs) adhering to a third type of 3D-SFnws to test whether HDFs release exosomes whose contents promote neoangiogenesis

Mesenchymal stromal cells-exosomes: a promising cell-free therapeutic tool for wound healing and cutaneous regeneration

Cutaneous regeneration at the wound site involves several intricate and dynamic processes which require a series of coordinated interactions implicating various cell types, growth factors, extracellular matrix (ECM), nerves, and blood vessels. Mesenchymal stromal cells (MSCs) take part in all the skin wound healing stages playing active and beneficial roles in animal models and humans. Exosomes, which are among the key products MSCs release, mimic the effects of parental MSCs. They can shuttle various effector proteins, messenger RNA (mRNA) and microRNAs (miRNAs) to modulate the activity of recipient cells, playing important roles in wound healing. Moreover, using exosomes avoids many risks associated with cell transplantation. Therefore, as a novel type of cell-free therapy, MSC-exosome -mediated administration may be safer and more efficient than whole cell. In this review, we provide a comprehensive understanding of the latest studies and observations on the role of MSC-exosome therapy in wound healing and cutaneous regeneration. In addition, we address the hypothesis of MSCs microenvironment extracellular vesicles (MSCs-MEVs) or MSCs microenvironment exosomes (MSCs-MExos) that need to take stock of and solved urgently in the related research about MSC-exosomes therapeutic applications. This review can inspire investigators to explore new research directions of MSC-exosome therapy in cutaneous repair and regeneration

Three-Layered Silk Fibroin Tubular Scaffold for the Repair and Regeneration of Small Caliber Blood Vessels: From Design to in vivo Pilot Tests

Silk fibroin (SF) is an eligible biomaterial for the development of small caliber vascular grafts for substitution, repair, and regeneration of blood vessels. This study presents the properties of a newly designed multi-layered SF tubular scaffold for vascular grafting (SilkGraf). The wall architecture consists of two electrospun layers (inner and outer) and an intermediate textile layer. The latter was designed to confer high mechanical performance and resistance on the device, while electrospun layers allow enhancing its biomimicry properties and host\u2019s tissues integration. In vitro cell interaction studies performed with adult Human Coronary Artery Endothelial Cells (HCAECs), Human Aortic Smooth Muscle Cells (HASMCs), and Human Aortic Adventitial Fibroblasts (HAAFs) demonstrated that the electrospun layers favor cell adhesion, survival, and growth. Once cultured in vitro on the SF scaffold the three cell types showed an active metabolism (consumption of glucose and glutamine, release of lactate), and proliferation for up to 20 days. HAAF cells grown on SF showed a significantly lower synthesis of type I procollagen than on polystyrene, meaning a lower fibrotic effect of the SF substrate. The cytokine and chemokine expression patterns were investigated to evaluate the cells\u2019 proliferative and pro-inflammatory attitude. Interestingly, no significant amounts of truly pro-inflammatory cytokines were secreted by any of the three cell types which exhibited a clearly proliferative profile. Good hemocompatibility was observed by complement activation, hemolysis, and hematology assays. Finally, the results of an in vivo preliminary pilot trial on minipig and sheep to assess the functional behavior of implanted SF-based vascular graft identified the sheep as the more apt animal model for next medium-to-long term preclinical trials

Istologia per le professioni sanitarie

Istologia per le professioni sanitarie con 314 figure a colori e in b/n e 10 tabell

Family C G-Protein-coupled receptors in Alzheimer\u2019s disease and therapeutic implications

Alzheimer\u2019s disease (AD), particularly its sporadic or late-onset form (SAD/LOAD), is the most prevalent (96\u201398% of cases) neurodegenerative dementia in aged people. AD\u2019s neuropathology hallmarks are intrabrain accumulation of amyloid-\u3b2 peptides (A\u3b2s) and of hyperphosphorylated Tau (p-Tau) proteins, diffuse neuroinflammation, and progressive death of neurons and oligodendrocytes. Mounting evidences suggest that family C G-protein-coupled receptors (GPCRs), which include \u3b3-aminobutyric acid B receptors (GABABRs), metabotropic glutamate receptors (mGluR1-8), and the calcium-sensing receptor (CaSR), are involved in many neurotransmitter systems that dysfunction in AD. This review updates the available knowledge about the roles of GPCRs, particularly but not exclusively those expressed by brain astrocytes, in SAD/ LOAD onset and progression, taking stock of their respective mechanisms of action and of their potential as anti-AD therapeutic targets. In particular, GABABRs prevent A\u3b2s synthesis and neuronal hyperexcitability and group I mGluRs play important pathogenetic roles in transgenic AD-model animals. Moreover, the specific binding of A\u3b2s to the CaSRs of human cortical astrocytes and neurons cultured in vitro engenders a pathological signaling that crucially promotes the surplus synthesis and release of A\u3b2s and hyperphosphorylated Tau proteins, and also of nitric oxide, vascular endothelial growth factor-A, and proinflammatory agents. Concurrently, A\u3b2s\u2022CaSR signaling hinders the release of soluble (s)APP-\u3b1 peptide, a neurotrophic agent and GABABR1a agonist. Altogether these effects progressively kill human cortical neurons in vitro and likely also in vivo. Several CaSR\u2019s negative allosteric modulators suppress all the noxious effects elicited by A\u3b2s\u2022CaSR signaling in human cortical astrocytes and neurons thus safeguarding neurons\u2019 viability in vitro and raising hopes about their potential therapeutic benefits in AD patients. Further basic and clinical investigations on these hot topics are needed taking always heed that activation of the several brain family C GPCRs may elicit divergent upshots according to the models studied

Danger\u2010Sensing/Pattern Recognition Receptors and Neuroinflammation in Alzheimer\u2019s Disease

Fibrillar aggregates and soluble oligomers of both Amyloid\u2010\u3b2 peptides (A\u3b2s) and hyperphosphorylated Tau proteins (p\u2010Tau\u2010es), as well as a chronic neuroinflammation are the main drivers causing progressive neuronal losses and dementia in Alzheimer\u2019s disease (AD). However, the underlying pathogenetic mechanisms are still much disputed. Several endogenous neurotoxic ligands, including A\u3b2s, and/or p\u2010Tau\u2010es activate innate immunity\u2010related danger\u2010sensing/pattern recognition receptors (PPRs) thereby advancing AD\u2019s neuroinflammation and progression. The major PRR families involved include scavenger, Toll\u2010like, NOD\u2010like, AIM2\u2010like, RIG\u2010like, and CLEC\u2010 2 receptors, plus the calcium\u2010sensing receptor (CaSR). This quite intricate picture stresses the need to identify the pathogenetically topmost A\u3b2\u2010activated PRR, whose signaling would trigger AD\u2019s three main drivers and their intra\u2010brain spread. In theory, the candidate might belong to any PRR family. However, results of preclinical studies using in vitro nontumorigenic human cortical neurons and astrocytes and in vivo AD\u2010model animals have started converging on the CaSR as the pathogenetically upmost PRR candidate. In fact, the CaSR binds both Ca2+ and A\u3b2s and promotes the spread of both Ca2+ dyshomeostasis and AD\u2019s three main drivers, causing a progressive neurons\u2019 death. Since CaSR\u2019s negative allosteric modulators block all these effects, CaSR\u2019s candidacy for topmost pathogenetic PRR has assumed a growing therapeutic potential worth clinical testing

Maladaptive remodeling of pulmonary artery root autografts after Ross procedure: A proteomic study

OBJECTIVE: Pulmonary autograft root dilatation is the major long-term complication after Ross procedure and the leading cause for reoperation. However, the mechanisms underlying dilatation remain to be elucidated. This study analyzed the proteomic changes seen in the dilated pulmonary autograft compared with normal pulmonary artery and aorta tissues. METHODS: Pulmonary autograft surgical samples were taken from 9 consecutive patients (mean age 37 \ub1 14; 15-51 years) with mean diameters of 5.2 \ub1 0.5 cm (4.6-5.8 cm) reoperated 8 to 16 years after Ross procedure. Control pulmonary artery and aorta samples were from 7 age- and sex-matched cardiac donors. Tunicae mediae from all samples were processed for proteomic analysis via 2-dimensional electrophoresis, matrix-assisted-laser-desorption-ionization-time of flight/mass spectrometry, and bioinformatics. The thus-identified putatively relevant proteins were validated via Western immunoblotting. RESULTS: Pulmonary autograft proteome features differed markedly from control pulmonary arteries, since proteins related to focal adhesions (eg, paxillin), cytoskeleton (eg, vimentin), and metalloprotease-regulating proteoglycans (eg, testican-2) were significantly up-regulated, whereas significant decreases occurred in microfibril-associated glycoprotein1, which controls elastic fiber buildup. Profound changes also occurred in cell-signaling proteins, ie, increases in soluble Jagged-1 fragment and ectodysplasin-2 receptor, and decreases in Notch-1 intracellular domain fragment. Moreover, pulmonary autograft expression levels of Paxillin, Vimentin, Jagged-1 fragment, and Notch1 intracellular domain fragment also differed from those of control aorta. CONCLUSIONS: This study provides the first description of the specific proteomic features of dilated pulmonary autograft tunica media, which separate them sharply not only from those of control pulmonary artery and aorta but also of aortic aneurysms. These findings suggest that dilated pulmonary autografts undergo a unique maladaptive remodeling process deserving further investigation

A paradigm-changing surprise from dentate gyrus granule cells- cilium-localized p75NTR may drive their progenitor cell proliferation

Peer reviewed: YesNRC publication: Ye