Constitutive and Inducible Innate Responses in Cells Infected by HSV-1-Derived Amplicon Vectors

Amplicons are helper-dependent herpes simplex virus type 1 (HSV-1)-based vectors that can deliver very large foreign DNA sequences and, as such, are good candidates both for gene delivery and vaccine development. However, many studies have shown that innate constitutive or induced cellular responses, elicited or activated by the entry of HSV-1 particles, can play a significant role in the control of transgenic expression and in the induction of inflammatory responses. Moreover, transgene expression from helper-free amplicon stocks is often weak and transient, depending on the particular type of infected cells, suggesting that cellular responses could be also responsible for the silencing of amplicon-mediated transgene expression. This review summarizes the current experimental evidence underlying these latter concepts, focusing on the impact on transgene expression of very-early interactions between amplicon particles and the infected cells, and speculates on possible ways to counteract the cellular protective mechanisms, thus allowing stable transgene expression without enhancement of vector toxicity

Precise Determination of Minimum Achievable Temperature for Solid-State Optical Refrigeration

We measure the minimum achievable temperature (MAT) as a function of excitation wavelength in anti-Stokes fluorescence cooling of high purity Yb3+-doped LiYF4 (Yb:YLF) crystal. Such measurements were obtained by developing a sensitive noncontact thermometry that is based on a two-band differential luminescence spectroscopy using balanced photo-detectors. These measurements are in excellent agreement with the prediction of the laser cooling model and identify MAT of 110 K at 1020 nm, corresponding to E4-E5 Stark manifold transition in Yb:YLF crystal.Comment: 10 pages, 6 figure

Interaction of chemical patterns in coupled layers

We investigate the interaction between reaction-diffusion systems coupled by diffusion. The photosensitive CDIMA (chorine dioxide–iodine–malonic acid) reaction allows us to study experimentally the mutual influence of two layers of Turing patterns coupled via a diffusive interaction. By illuminating each of the layers with different intensities of homogeneous external light, the chemical conditions in each layer can be shifted, allowing us to study the result of diffusive interaction between Turing patterns with different spatial configurations. Our experiments suggest a complex scenario for the interaction between different patterns, strongly dependent on the spatial characteristics of the interacting patterns. Numerical simulations are also reported in full agreement with experimental observationsThis work has been supported by the DGI (Spain) under Project No. FIS2010-21023 and Xunta de Galicia (Spain) under Project Nos. PGIDIT05PXIC20607PN and INCITE07PXI206131ES and by the NSF (USA). D.G.M. acknowledges a Ramon y Cajal Fellowship from the Ministry of Science and Technology of Spain and a Marie Curie International Reintegration Grant from the EU248346-NMSSBLS, as well as financial support from the CSIC-SPAIN (JAE-DOC

Meeting public health objectives and supporting the resumption of tourist activity through COVID-19: a triangular perspective

This work was supported by FEDER/Junta de AndalucAa-ConsejerAa de TransformaciA3n EconA3mica, Industria, Conocimiento y Universidades [grant number Project CV20-09357].Non-pharmaceutical interventions (NPIs) implemented during the COVID- 19 pandemic (and previous health crises) have included measures to restrict interaction between people and minimize non-essential mobility. Therefore, tourism travel is one of the main areas affected by the restrictions. Even when the majority of the population is vaccinated, some risk of infection will remain, and governments are obliged to consider NPI measures that balance the health risk of outbreaks against the economic and social benefits of resuming tourist activity. This study analyzes the effect of each of four categories of NPIs (Social Distancing; Public Healthcare-System Improvements; Tourist Controls; and Capacity and Opening-Hours Regulation) on three major objectives (the resumption of tourism activity; tourist travel intention; and the minimization of public health risk), taking a triangular perspective (destination managers, domestic tourists, and public healthcare managers, respectively). While it is difficult to fulfil public healthcare objectives while simultaneously responding to the economic interests of tourism-industry stakeholders, the study finds that, under vaccinatedpopulation conditions, tourist controls (e.g. COVID Certificate) alongside improvements to the public healthcare system (e.g. adequate resourcing and an efficient epidemiological monitoring system) could constitute a viable combination of measures.FEDER/Junta de AndalucAa-ConsejerAa de TransformaciA3n EconA3mica, Industria, Conocimiento y Universidades CV20-0935

Universal Sequencing on an Unreliable Machine

We consider scheduling on an unreliable machine that may experience unexpected changes in processing speed or even full breakdowns. Our objective is to minimize ∑ wjf(Cj) for any nondecreasing, nonnegative, differentiable cost function f(Cj). We aim for a universal solution that performs well without adaptation for all cost functions for any possible machine behavior. We design a deterministic algorithm that finds a universal scheduling sequence with a solution value within 4 times the value of an optimal clairvoyant algorithm that knows the machine behavior in advance. A randomized version of this algorithm attains in expectation a ratio of e. We also show that both performance guarantees are best possible for any unbounded cost function. Our algorithms can be adapted to run in polynomial time with slightly increased cost. When jobs have individual release dates, the situation changes drastically. Even if all weights are equal, there are instances for which any universal solution is a factor of Ω(log n / log log n) worse than an optimal sequence for any unbounded cost function. Motivated by this hardness, we study the special case when the processing time of each job is proportional to its weight. We present a nontrivial algorithm with a small constant performance guarantee

Quaking RNA bindings proteins as mediator of oncolytic HSV vectors in huma hepatoma cells

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Development and assessment of herpes simplex virus type 1 (HSV-1) amplicon vectors with sensory neuron-selective promoters

Background: Neurogenic detrusor overactivity (NDO) is a severe pathological condition characterized by involuntary detrusor contractions leading to urine leakage. This condition is frequent after spinal cord injury (SCI). Gene therapy for NDO requires the development of vectors that express therapeutic transgenes driven by sensory neuron-specific promoters. The aim of this study was to develop and assess tools for the characterization of sensory neuron-specific promoters in dorsal root ganglia (DRG) neurons after transduction with herpes simplex virus type 1 (HSV-1)-based amplicon defective vectors. Methods: The HSV-1 vector genome encoded two independent transcription cassettes: one expressed firefly luciferase (FLuc) driven by different promoters’ candidates (rTRPV1, rASIC3, rCGRP, or hCGRP), and the other expressed a reporter gene driven by an invariable promoter. The strength and selectivity of promoters was assessed in organotypic cultures of explanted adult DRG, or sympathetic and parasympathetic ganglia from control and SCI rats. Results: The rCGRP promoter induced selective expression in the DRG of normal rats. The rTRPV-1 promoter, which did not display selective activity in control rats, induced selective expression in DRG explanted from SCI rats. Conclusions: This study provides a methodology to assess sensory neuron-specific promoters, opening new perspectives for future gene therapy for ND

Induction of humoral responses to BHV-1 glycoprotein D expressed by HSV-1 amplicon vectors

Herpes simplex virus type-1 (HSV-1) amplicon vectors are versatile and useful tools for transferring genes into cells that are capable of stimulating a specific immune response to their expressed antigens. In this work, two HSV-1-derived amplicon vectors were generated. One of these expressed the full-length glycoprotein D (gD) of bovine herpesvirus 1 while the second expressed the truncated form of gD (gDtr) which lacked the trans-membrane region. After evaluating gD expression in the infected cells, the ability of both vectors to induce a specific gD immune response was tested in BALB/c mice that were intramuscularly immunized. Specific serum antibody responses were detected in mice inoculated with both vectors, and the response against truncated gD was higher than the response against full-length gD. These results reinforce previous findings that HSV-1 amplicon vectors can potentially deliver antigens to animals and highlight the prospective use of these vectors for treating infectious bovine rhinotracheitis disease

Quantitative immunocytochemical study of islet cell populations in diabetic calmodulin-transgenic mice

The present study describes the changes in the endocrine pancreas of severely diabetic calmodulin-transgenic mice using light microscopic immunocyto-chemical and morphometric techniques. A marked reduction in the number and volume of islets, together with distortion of their normal architecture, was found in diabetic mice. In addition, the volume density of both endocrine tissue and B-cells was decreased. An irregular distribution of non-B-cells was also observed in diabetic animals. The volume density and the percentage of A-cells appeared increased. However, when quantified per area unit, the number of all the islet cell types diminished, although only the decrease in B-cell number was statistically significant. The decrease in B-cell mass might account for the diabetic state developed in this animal model.Facultad de Ciencias Médica

Reduced expression of hippocampal GluN2A-NMDAR increases seizure susceptibility and causes deficits in contextual memory

N-methyl-D-aspartate receptors are heterotetramers composed of two GluN1 obligatory subunits and two regulatory subunits. In cognitive-related brain structures, GluN2A and GluN2B are the most abundant regulatory subunits, and their expression is subjected to tight regulation. During development, GluN2B expression is characteristic of immature synapses, whereas GluN2A is present in mature ones. This change in expression induces a shift in GluN2A/GluN2B ratio known as developmental switch. Moreover, modifications in this relationship have been associated with learning and memory, as well as different pathologies. In this work, we used a specific shRNA to induce a reduction in GluN2A expression after the developmental switch, both in vitro in primary cultured hippocampal neurons and in vivo in adult male Wistar rats. After in vitro characterization, we performed a cognitive profile and evaluated seizure susceptibility in vivo. Our in vitro results showed that the decrease in the expression of GluN2A changes GluN2A/GluN2B ratio without altering the expression of other regulatory subunits. Moreover, rats expressing the anti-GluN2A shRNA in vivo displayed an impaired contextual fear-conditioning memory. In addition, these animals showed increased seizure susceptibility, in terms of both time and intensity, which led us to conclude that deregulation in GluN2A expression at the hippocampus is associated with seizure susceptibility and learning–memory mechanisms.Fil: Acutain, Maria Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; ArgentinaFil: Griebler Luft, Jordana. Universidade Federal do Rio Grande do Sul; BrasilFil: Vázquez, Cecilia Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; ArgentinaFil: Popik, Bruno. Universidade Federal do Rio Grande do Sul; BrasilFil: Cercato, Magalí Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; ArgentinaFil: Epstein, Alberto. Universite Lyon 2; FranciaFil: Salvetti, Anna. Inserm; FranciaFil: Jerusalinsky, Diana Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; ArgentinaFil: de Oliveira Alvares, Lucas. Universidade Federal do Rio Grande do Sul; BrasilFil: Baez, Maria Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Biología Celular y Neurociencia "Prof. Eduardo de Robertis". Universidad de Buenos Aires. Facultad de Medicina. Instituto de Biología Celular y Neurociencia; Argentin