Masa pulsátil en el cuello: arteria subclavia sobre costilla cervical

A pulsatile mass in the posterior triangle of the neck may induce concern when it grows and starts to bother. We present the case of a young woman who consulted for this reason, in whom a verticalized subclavian artery was identified due to coexistence with cervical rib. Being, for the moment, asymptomatic, it only required a preventive attitude, in addition to an evolutionary follow-up for the possibility of the development of a compressive condition due to thoracic gorge syndrome and/or an arterial aneurysm.Una masa pulsátil en el triángulo posterior del cuello puede inducir preocupación cuando crece y comienza a molestar. Se presenta caso de mujer joven, que consulta por dicho motivo, en la que se identifica una arteria subclavia verticalizada por coexistencia con costilla cervical. Al ser, por el momento, asintomática únicamente requirió una actitud preventiva, además de un seguimiento evolutivo ante la posibilidad del desarrollo de un cuadro compresivo por el síndrome del desfiladero torácico y/o un aneurisma arterial

Matrix metalloproteinase 14 regulates HSV-1 infection in neuroblastoma cells

Growing evidence supports that chronic or latent infection of the central nervous system might be implicated in Alzheimer's disease (AD). Among them, Herpes simplex virus type 1 (HSV-1) has emerged as a major factor in the etiology of the disease. Our group is devoted to the study of the relationship among HSV-1, oxidative stress (OS) and neurodegeneration. We have found that HSV-1 induces the main neuropathological hallmarks of AD, including the accumulation of intracellular amyloid beta (Aβ), hyperphosphorylated tau protein and autophagic vesicles, that OS exacerbates these effects, and that matrix metalloproteinase 14 (MMP-14) participates in the alterations induced by OS. In this work, we focused on the role of MMP-14 in the degenerative markers raised by HSV-1 infection. Interestingly, we found that MMP-14 blockage is a potent inhibitor of HSV-1 infection efficiency, that also reduces the degeneration markers, accumulation of Aβ and hyperphosphorylated tau, induced by the virus. Our results point to MMP-14 as a potent antiviral target to control HSV-1 infection and its associated neurodegenerative effectsThis work was supported by the Ministerio de Ciencia, Innovación y Universidades (SAF2017-85747-R

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Matrix Metalloproteinase 14 Mediates APP Proteolysis and Lysosomal Alterations Induced by Oxidative Stress in Human Neuronal Cells

The alteration of amyloid precursor protein (APP) proteolysis is a hallmark of Alzheimer’s disease (AD). Recent studies have described noncanonical pathways of APP processing that seem partly executed by lysosomal enzymes. Our laboratory’s in vitro human SK-N-MC model has shown that oxidative stress (OS) alters the lysosomal degradation pathway and the processing/metabolism of APP. The present study identifies the lysosomal protein matrix metalloproteinase 14 (MMP14) as a protease involved in the APP noncanonical processing. Previous expression analyses of the above cells showed MMP14 to be overexpressed under OS. In the present work, its role in changes in OS-induced APP proteolysis and lysosomal load was examined. The results show that MMP14 mediates the accumulation of an ≈85 kDa N-terminal APP fragment and increases the lysosome load induced by OS. These results were validated in neurons and neural progenitor cells generated from the induced pluripotent stem cells of patients with sporadic AD, reinforcing the idea that MMP14 may offer a therapeutic target in this disease

Effects of oxidative stress on HSV-1 protein expression.

<p>The accumulation of viral proteins ICP4, VP16 and gC was analysed by immunoblotting in SK-N-MC cells simultaneously treated with X-XOD and infected with HSV-1 at a moi of 1 and 10 for 18 h (<b>A</b>) or at a moi of 0.1 for 42 h (<b>B</b>). The blots shown are representative of three independent experiments. A tubulin blot was performed as a loading control. In all blots, the ratio of viral proteins to tubulin is shown below the blots.</p

Oxidative stress reduces HSV-1 replication and increases cell viability of infected cells.

<p><b>A)</b> Quantification of viral DNA by real-time quantitative PCR in SK-N-MC cells simultaneously treated with X-XOD and infected with HSV-1 at a moi of 1 and 10 for 18 h or at a moi of 0.1 for 42 h. <b>B)</b> Intracellular (intra) and extracellular (extra) viral titres were determined by plaque assays in SK-N-MC cells infected under the same conditions as in (<b>A</b>). In (<b>A</b>) and (<b>B</b>), the data represent the mean ± SEM of five experiments performed in triplicate and are expressed as a percentage with respect to untreated cells (- X-XOD) (*p<0.05; **p<0.01; ***p<0.001). <b>C)</b> The cell viability of mock and HSV-1-infected SK-N-MC cells exposed to X-XOD was monitored using the MTT reduction assay. Cells were infected with HSV-1 at a moi of 1 and 10 for 18 h or at a moi of 0.1 for 42 h. Values are expressed relative to the optical density of untreated mock-infected cells. The data shown represent the mean ± SEM for four independent experiments performed in triplicate (*p<0.05; **p<0.01).</p

Cholesterol Modulation Attenuates the AD-like Phenotype Induced by Herpes Simplex Virus Type 1 Infection

Cholesterol, a crucial component of cell membranes, influences various biological processes, including membrane trafficking, signal transduction, and host-pathogen interactions. Disruptions in cholesterol homeostasis have been linked to congenital and acquired conditions, including neurodegenerative disorders such as Alzheimer&rsquo;s disease (AD). Previous research from our group has demonstrated that herpes simplex virus type I (HSV-1) induces an AD-like phenotype in several cell models of infection. This study explores the interplay between cholesterol and HSV-1-induced neurodegeneration. The impact of cholesterol was determined by modulating its levels with methyl-beta-cyclodextrin (M&beta;CD) using the neuroblastoma cell lines SK-N-MC and N2a. We have found that HSV-1 infection triggers the intracellular accumulation of cholesterol in structures resembling endolysosomal/autophagic compartments, a process reversible upon M&beta;CD treatment. Moreover, M&beta;CD exhibits inhibitory effects at various stages of HSV-1 infection, underscoring the importance of cellular cholesterol levels, not only in the viral entry process but also in subsequent post-entry stages. M&beta;CD also alleviated several features of AD-like neurodegeneration induced by viral infection, including lysosomal impairment and intracellular accumulation of amyloid-beta peptide (A&beta;) and phosphorylated tau. In conclusion, these findings highlight the connection between cholesterol, neurodegeneration, and HSV-1 infection, providing valuable insights into the underlying mechanisms of AD

Effects of oxidative stress on HSV-1 entry.

<p><b>A)</b> Immunofluorescence analysis of HSV-1-infected SK-N-MC cells at a moi of 10 in the presence and absence of X-XOD. The immunoreactivity of ICP4 protein is shown at 3 and 5 hours post-infection (h.p.i.). Nuclei are stained with DAPI. Scale bar: 20 µm. <b>B)</b> Quantification of infected cells by ICP4 staining. The graph shows the percentage of ICP4-positive cells. At least 400 nuclei were counted for each condition. <b>C)</b> Analysis of ICP4 levels by Western blotting in SK-N-MC cells infected with HSV-1 at a moi of 1 and 10 for 3 h and 5 h, in the presence and absence of X-XOD.</p

Oxidative stress does not affect the autophagosomal localization of Aβ in HSV-1-infected cells.

<p>Confocal images obtained with anti-Aβ and anti-LC3 antibodies showing endogenous LC3 and Aβ patterns in HSV-1-infected SK-APP cells at a moi of 10 for 18 h in the presence and absence of oxidative stress (X-XOD). Colocalization is shown by yellow fluorescence signals in the merged panels. Scale bar: 10 µm.</p