Calcium-activated potassium channels:Implications for aging and age-related neurodegeneration

Population aging, as well as the handling of age-associated diseases, is a worldwide increasing concern. Among them, Alzheimer's disease stands out as the major cause of dementia culminating in full dependence on other people for basic functions. However, despite numerous efforts, in the last decades, there was no new approved therapeutic drug for the treatment of the disease. Calcium-activated potassium channels have emerged as a potential tool for neuronal protection by modulating intracellular calcium signaling. Their subcellular localization is determinant of their functional effects. When located on the plasma membrane of neuronal cells, they can modulate synaptic function, while their activation at the inner mitochondrial membrane has a neuroprotective potential via the attenuation of mitochondrial reactive oxygen species in conditions of oxidative stress. Here we review the dual role of these channels in the aging phenotype and Alzheimer's disease pathology and discuss their potential use as a therapeutic tool

Mitochondrial dysfunction in neurodegenerative diseases:A focus on iPSC-derived neuronal models

Progressive neuronal loss is a hallmark of many neurodegenerative diseases, including Alzheimer's and Parkinson's disease. These pathologies exhibit clear signs of inflammation, mitochondrial dysfunction, calcium deregulation, and accumulation of aggregated or misfolded proteins. Over the last decades, a tremendous research effort has contributed to define some of the pathological mechanisms underlying neurodegenerative processes in these complex brain neurodegenerative disorders. To better understand molecular mechanisms responsible for neurodegenerative processes and find potential interventions and pharmacological treatments, it is important to have robust in vitro and pre-clinical animal models that can recapitulate both the early biological events undermining the maintenance of the nervous system and early pathological events. In this regard, it would be informative to determine how different inherited pathogenic mutations can compromise mitochondrial function, calcium signaling, and neuronal survival. Since post-mortem analyses cannot provide relevant information about the disease progression, it is crucial to develop model systems that enable the investigation of early molecular changes, which may be relevant as targets for novel therapeutic options. Thus, the use of human induced pluripotent stem cells (iPSCs) represents an exceptional complementary tool for the investigation of degenerative processes. In this review, we will focus on two neurodegenerative diseases, Alzheimer's and Parkinson's disease. We will provide examples of iPSC-derived neuronal models and how they have been used to study calcium and mitochondrial alterations during neurodegeneration

Stem cells in embryonic skin development

The skin is a complex stratified organ which acts not only as a permeability barrier and defense against external agents, but also has essential thermoregulatory, sensory and metabolic functions. Due to its high versatility and activity, the skin undergoes continuous self-renewal to repair damaged tissue and replace old cells. Consequently, the skin is a reservoir for adult stem cells of different embryonic origins. Skin stem cell populations reside in the adult hair follicle, sebaceous gland, dermis and epidermis. However, the origin of most of the stem cell populations found in the adult epidermis is still unknown. Far more unknown is the embryonic origin of other stem cells that populate the other layers of this tissue. In this review we attempt to clarify the emergence, structure, markers and embryonic development of diverse populations of stem cells from the epidermis, dermis and related appendages such as the sebaceous gland and hair follicle

Post-Transcriptional Control of RNA Expression in Cancer

Approximately 80% of the human genome contains functional DNA, including protein coding genes, non-protein coding regulatory DNA elements and non-coding RNAs (ncRNAs). An altered transcriptional signature is not only a cause, but also a consequence of the characteristics known as the hallmarks of cancer, such as sustained proliferation, replicative immortality, evasion of growth suppression and apoptotic signals, angiogenesis, invasion, metastasis, evasion of immune destruction and metabolic re-wiring. Post-transcriptional events play a major role in determining this signature, which is evidenced by the fact that alternative RNA splicing takes place in more than half of the human genes, and, among protein coding genes, more than 60% contain at least one conserved miRNA-binding site. In this chapter, we will discuss the involvement of post-transcriptional events, such as RNA processing, the action of non-coding RNAs and RNA decay in cancer development, and how their machinery may be used in cancer diagnosis and treatment

The Unfolded Protein Response Sensor PERK Mediates Stiffness-Dependent Adaptation in Glioblastoma Cells

Glioblastoma multiforme (GBM) is the most aggressive brain tumor in adults. In addition to genetic causes, the tumor microenvironment (TME), including stiffening of the extracellular matrix (ECM), is a main driver of GBM progression. Mechano-transduction and the unfolded protein response (UPR) are essential for tumor-cell adaptation to harsh TME conditions. Here, we studied the effect of a variable stiff ECM on the morphology and malignant properties of GBM stem cells (GSCs) and, moreover, examined the possible involvement of the UPR sensor PERK herein. For this, stiffness-tunable human blood plasma (HBP)/alginate hydrogels were generated to mimic ECM stiffening. GSCs showed stiffness-dependent adaptation characterized by elongated morphology, increased proliferation, and motility which was accompanied by F-Actin cytoskeletal remodeling. Interestingly, in PERK-deficient GSCs, stiffness adaptation was severely impaired, which was evidenced by low F-Actin levels, the absence of F-Actin remodeling, and decreased cell proliferation and migration. This impairment could be linked with Filamin-A (FLN-A) expression, a known interactor of PERK, which was strongly reduced in PERK-deficient GSCs. In conclusion, we identified a novel PERK/FLNA/F-Actin mechano-adaptive mechanism and found a new function for PERK in the cellular adaptation to ECM stiffening

A-Kinase Anchoring Proteins Diminish TGF-β1/Cigarette Smoke-Induced Epithelial-To-Mesenchymal Transition

Epithelial-to-mesenchymal transition (EMT) plays a role in chronic obstructive pulmonary diseases (COPD). Cyclic adenosine monophosphate (cAMP) can inhibit transforming growth factor-β1 (TGF-β1) mediated EMT. Although compartmentalization via A-kinase anchoring proteins (AKAPs) is central to cAMP signaling, functional studies regarding their therapeutic value in the lung EMT process are lacking. The human bronchial epithelial cell line (BEAS-2B) and primary human airway epithelial (pHAE) cells were exposed to TGF-β1. Epithelial (E-cadherin, ZO-1) and mesenchymal markers (collagen Ӏ, α-SMA, fibronectin) were analyzed (mRNA, protein). ELISA measured TGF-β1 release. TGF-β1-sensitive AKAPs Ezrin, AKAP95 and Yotiao were silenced while using siRNA. Cell migration was analyzed by wound healing assay, xCELLigence, Incucyte. Prior to TGF-β1, dibutyryl-cAMP (dbcAMP), fenoterol, rolipram, cilostamide, and forskolin were used to elevate intracellular cAMP. TGF-β1 induced morphological changes, decreased E-cadherin, but increased collagen Ӏ and cell migration, a process that was reversed by the inhibitor of δ/epsilon casein kinase I, PF-670462. TGF-β1 altered (mRNA, protein) expression of Ezrin, AKAP95, and Yotiao. St-Ht31, the AKAP antagonist, decreased E-cadherin (mRNA, protein), but counteracted TGF-β1-induced collagen Ӏ upregulation. Cigarette smoke (CS) increased TGF-β1 release, activated TGF signaling, augmented cell migration, and reduced E-cadherin expression, a process that was blocked by TGF-β1 neutralizing antibody. The silencing of Ezrin, AKAP95, and Yotiao diminished TGF-β1-induced collagen Ӏ expression, as well as TGF-β1-induced cell migration. Fenoterol, rolipram, and cilostamide, in AKAP silenced cells, pointed to distinct cAMP compartments. We conclude that Ezrin, AKAP95, and Yotiao promote TGF-β1-mediated EMT, linked to a TGF-β1 release by CS. AKAP members might define the ability of fenoterol, rolipram, and cilostamide to modulate the EMT process, and they might represent potential relevant targets in the treatment of COPD

Chemical structure and biological activity of the (1 → 3)-linked β-D-glucan isolated from marine diatom Conticribra weissflogii

Several polysaccharides are considered to be "biological response modifiers" (BRM) - these refer to biomolecules that augment immune responses and can be derived from a variety of sources. Microalgae produce a diverse range of polysaccharides and could be an excellent source of BRM. Here, we describe the chemical structure and biological activity of water-soluble polysaccharide isolated from the marine diatom Conticribra weissflogii. Using chemical and NMR spectroscopic methods, the polysaccharide was identified as a (1 → 3)-linked β-D-glucan with a low proportion of C-6 substitution by single β-glucose units. The biological activity of this low molecular weight β-glucan (11.7 kDa) was investigated with respect to glioblastoma cell lines (U87 MG and U251) and macrophages (RAW 264.7). We observed that this β-D-glucan did not exhibit cytotoxic activity against glioblastoma cells, but did enhance the phagocytic activity of macrophages, suggesting that it possesses immunomodulatory properties.</p

THEORETICAL APPROACHES IN OCCUPATIONAL HEALTH OF NURSING WORKERS: AN INTEGRATIVE REVIEW

This integrative review aims to identify the theoretical approaches mentioned by authors in studies about the health of nursing workers. The search for publications between 2008 and 2012 occurred in three databases, using the descriptors of ‘occupational health’ and ‘nursing’. The sample was made up of 18 ar ticles which dealt specifically with the occupational health of nursing workers. Of the studies selected, the vast majority did not make clear the theoretical framework adopted, and among those which did, two approaches predominated: work and subjectivity, and health policies. The different theoretical approaches enriched the discussion showing the strong relation between organization, work conditions, and subjectivity and situations of pleasure and suffering in the nursing work.Revisão integrativa com o objetivo de identificar as abordagens teóricas referidas por autores de estudos atuais sobre a saúde do trabalhador de enfermagem. A busca das publicações entre 2008 e 2012 ocorreu em três bases de dados, utilizando-se os descritores saúde do trabalhador e enfermagem. A amostra constituiu-se de 18 artigos. Dentre os artigos excluídos, por não conterem a abordagem teórica explicitada, encontraramse estudos quantitativos, estudos transversais e de coorte. Os estudos selecionados possuem características semelhantes, relacionadas à subjetividade do trabalhador e as formas de organização do trabalho. As abordagens teóricas enriquecem a discussão sobre prazer e sofrimento no trabalho de enfermagem, ofertando um leque de possibilidades de interpretação para as situações vivenciadas pelos trabalhadores, e que independentemente da teoria adotada, os estudos corroboram entre si, no sentido de enriquecer o debate acerca da temática, alguns levantando situações emergentes, outros fornecendo subsídios para a melhoria das condições de trabalho.Revisión integrativa con el objetivo de identificar los abordajes teóricos referidos por autores de estudios actuales sobre la salud del trabajador de enfermería. La búsqueda de las publicaciones entre 2008 y 2012 ocurrió en tres bases de datos, utilizándose los descriptores salud del trabajador y enfermería. La muestra fue constituida de 18 artículos. Entre los artículos excluidos, por no contener el abordaje teórico explicitado, están estudios cuantitativos, estudios transversales y de cohorte. Los estudios seleccionados poseen características semejantes, relacionadas a la subjetividad del trabajador y las formas de organización del trabajo. Los abordajes teóricos enriquecen la discusión sobre placer y sufrimiento en el trabajo de enfermería, ofreciendo una serie de posibilidades de interpretación para las situaciones vivenciadas por los trabajadores, y que independientemente de la teoría adoptada, los estudios corroboran entre sí, en el sentido de enriquecer el debate acerca de la temática, algunos verificando situaciones, otros proveyendo subsídios para la mejoría de las condiciones de trabajo

Extracellular Matrix Proteome Remodeling in Human Glioblastoma and Medulloblastoma

Medulloblastomas (MBs) and glioblastomas (GBMs) are high-incidence central nervous system tumors. Different origin sites and changes in the tissue microenvironment have been associated with the onset and progression. Here, we describe differences between the extracellular matrix (ECM) signatures of these tumors. We compared the proteomic profiles of MB and GBM decellularized tumor samples between each other and their normal decellularized brain site counterparts. Our analysis revealed that 19, 28, and 11 ECM proteins were differentially expressed in MBs, GBMs, and in both MBs and GBMs, respectively. Next, we validated key findings by using a protein tissue array with 53 MB and 55 GBM cases and evaluated the clinical relevance of the identified differentially expressed proteins through their analysis on publicly available datasets, 763 MB samples from the GSE50161 and GSE85217 studies, and 115 GBM samples from RNAseq-TCGA. We report a shift toward a denser fibrillary ECM as well as a clear alteration in the glycoprotein signature, which influences the tumor pathophysiology. MS data have been submitted to the PRIDE repository, project accession: PXD023350

Pharmacological Inhibition of Epac1 Averts Ferroptosis Cell Death by Preserving Mitochondrial Integrity

Exchange proteins directly activated by cAMP (Epac) proteins are implicated in a wide range of cellular functions including oxidative stress and cell survival. Mitochondrial-dependent oxidative stress has been associated with progressive neuronal death underlying the pathology of many neurodegenerative diseases. The role of Epac modulation in neuronal cells in relation to cell survival and death, as well as its potential effect on mitochondrial function, is not well established. In immortalized hippocampal (HT-22) neuronal cells, we examined mitochondria function in the presence of various Epac pharmacological modulators in response to oxidative stress due to ferroptosis. Our study revealed that selective pharmacological modulation of Epac1 or Epac2 isoforms, exerted differential effects in erastin-induced ferroptosis conditions in HT-22 cells. Epac1 inhibition prevented cell death and loss of mitochondrial integrity induced by ferroptosis, while Epac2 inhibition had limited effects. Our data suggest Epac1 as a plausible therapeutic target for preventing ferroptosis cell death associated with neurodegenerative diseases