Impaired Reverse Cholesterol Transport is Associated with Changes in Fatty Acid Profile in Children and Adolescents with Abdominal Obesity.

BACKGROUND: Abdominal obesity is an important cardiovascular disease risk factor. Plasma fatty acids display a complex network of both pro and antiatherogenic effects. High density lipoproteins (HDL) carry out the antiatherogenic pathway called reverse cholesterol transport (RCT), which involves cellular cholesterol efflux (CCE), and lecithin:cholesterol acyltransferase (LCAT) and cholesteryl ester transfer protein (CETP) activities. OBJECTIVES: Our aim was to characterize RCT and its relation to fatty acids present in plasma in pediatric abdominal obesity. METHODS: Seventeen children and adolescents with abdominal obesity and 17 healthy controls were studied. Anthropometric parameters were registered. Glucose, insulin, lipid levels, CCE employing THP-1 cells, LCAT and CETP activities, plus fatty acids in apo B-depleted plasma were measured. RESULTS: The obese group showed a more atherogenic lipid profile, plus lower CCE (Mean±Standard Deviation) (6 ± 2 vs. 7 ± 2%; P < 0.05) and LCAT activity (11 ± 3 vs. 15 ±5 umol/dL.h; P < 0.05). With respect to fatty acids, the obese group showed higher myristic (1.1 ± 0.3 vs. 0.7 ± 0.3; P < 0.01) and palmitic acids (21.5 ± 2.8 vs. 19.6 ± 1.9; P < 0.05) in addition to lower linoleic acid (26.4 ± 3.3 vs. 29.9 ± 2.6; P < 0.01). Arachidonic acid correlated with CCE (r = 0.37; P < 0.05), myristic acid with LCAT (r = -0.37; P < 0.05), palmitioleic acid with CCE (r = -0.35; P < 0.05), linoleic acid with CCE (r = 0.37; P < 0.05), lauric acid with LCAT (r = 0.49; P < 0.05), myristic acid with LCAT (r = -0.37; P < 0.05) ecoisatrienoic acid with CCE (r = 0.40; P < 0.05) and lignoseric acid with LCAT (r = -0.5; P < 0.01). CONCLUSIONS: Children and adolescents with abdominal obesity presented impaired RCT, which was associated with modifications in proinflammatory fatty acids, such as palmitoleic and myristic, thus contributing to increased cardiovascular disease risk.

Changes in abscisic acid metabolism in relation to the maturation of grapevine (Vitis vinifera L., cv. Mencía) somatic embryos.

BACKGROUND: Somatic embryogenesis in grapevines is a complex process that depends on many physiological and genetic factors. One of its main limitations is the process of precocious germination of the somatic embryos in differentiation medium. This process lowers plant conversion rates from the somatic embryos, and it is probably caused by a low endogenous abscisic acid (ABA) content. RESULTS: Precocious germination of the somatic embryos was successfully avoided by culturing grapevine cv. Mencía embryogenic aggregates over a semipermeable membrane extended on top of the differentiation medium. The weekly analysis of the endogenous ABA and ABA-glucosyl ester (ABA-GE) contents in the aggregates showed their rapid accumulation. The expression profiles of 9-cis-epoxycarotenoid dioxygenase (VvNCED1), 8'-hydroxylase (VvHyd2), UDP-glucosyltransferase (VvUGT) and ß-glucosidase (VvBG2) genes in grapevine revealed that the occurrence of a first accumulation peak of endogenous ABA in the second week of culture over the semipermeable membrane was mainly dependent on the expression of the VvNCED1 gene. A second increase in the endogenous ABA content was observed in the fourth week of culture. At this point in the culture, our results suggest that of those genes involved in ABA accumulation, one (VvNCED1) was repressed, while another (VvBG2) was activated. Similarly, of those genes related to a reduction in ABA levels, one (VvUGT) was repressed while another (VvHyd2) was activated. The relative expression level of the VvNCED1 gene in embryogenic aggregates cultured under the same conditions and treated with exogenous ABA revealed the significant downregulation of this gene. CONCLUSIONS: Our results demonstrated the involvement of ABA metabolism in the control of the maturation of grapevine somatic embryos cultured over a semipermeable membrane and two important control points for their endogenous ABA levels. Thus, subtle differences in the expression of the antagonistic genes that control ABA synthesis and degradation could be responsible for the final level of ABA during the maturation of grapevine somatic embryos in vitro. In addition, the treatment of somatic embryos with exogenous ABA suggested the feedback-based control of the expression of the VvNCED1 gene by ABA during the maturation of grapevine somatic embryos.

In vitro and in vivo efficacy of SYL040012, a novel siRNA compound for treatment of glaucoma.

Glaucoma is a progressive ocular syndrome characterized by degeneration of the optic nerve and irreversible visual field loss. Elevated intraocular pressure (IOP) is the main risk factor for glaucoma. Increased IOP is the result of an imbalance between synthesis and outflow of aqueous humor (AH). Blocking ß2 adrenergic receptor (ADRB2) has shown to reduce IOP by decreasing production of AH at the ciliary body (CB). SYL040012 is a siRNA designed to specifically silence ADRB2 currently under development for glaucoma treatment. Here, we show that SYL040012 specifically reduces ADRB2 expression in cell cultures and eye tissues. The compound enters the eye shortly after administration in eye drops and is rapidly distributed among structures of the anterior segment of the eye. In addition, SYL040012 is actively taken up by cells of the CB but not by cells of systemic organs such as the lungs, where inhibition of ADRB2 could cause undesirable side effects. Moreover, SYL040012 reduces IOP in normotensive and hypertensive animal models and the effect appears to be long lasting and extremely well tolerated both locally and systemically.

Phase I clinical trial of SYL040012, a small interfering RNA targeting ß-adrenergic receptor 2, for lowering intraocular pressure.

The objective of this study was to evaluate ocular tolerance, safety, and effect on intraocular pressure (IOP) of a topically administered small interfering RNA; SYL040012, on healthy volunteers. The study was an open-label, controlled, single-center study comprised of two intervals that enrolled 30 healthy subjects having IOP below 21 mmHg. SYL040012 was administered to one eye as a single dose to six subjects during interval 1. During interval 2 two different doses of SYL040012 were administered to one eye on a daily basis to two separate groups of 12 subjects each, over a period of 7 days. The contralateral eye was evaluated but not administered and served as control for the tolerance study. SYL040012 was well tolerated locally. No local or systemic adverse events related to the product developed in response to any of the doses studied. SYL040012 was not detected in plasma at any time point. Administration of SYL040012 over a period of 7 days reduced IOP values in 15 out of 24 healthy subjects regardless of the dose used. IOP decrease was statistically significant in response to one of the doses tested and responsiveness to SYL040012 seemed to be greater in individuals with higher baseline IOP.

Prognostic and diagnostic value of epithelial to mesenchymal transition markers in pulmonary neuroendocrine tumors.

BACKGROUND: Pulmonary neuroendocrine tumors (Pulmonary NETs) include a wide spectrum of tumors, from the low-grade typical carcinoid (TC) and the intermediate-grade atypical carcinoid (AC), to the high-grade large-cell neuroendocrine carcinoma (LCNEC) and the small-cell carcinoma (SCLC). Epithelial Mesenchymal Transition (EMT) is a process initially recognised during several critical stages of embryonic development, which has more recently been implicated in promoting carcinoma invasion and metastasis. The initial stage of the EMT process begins with the deregulation of adhesion molecules, such as E-cadherin, due to transcriptional repression carried out by factors such as Snail family members, Twist and Foxc2. METHODS: Immunohistochemistry for EMT markers and E-cadherin/ ß-catenin complex in 134 patients with pulmonary NETs between 1990 - 2009. Analysis of potential associations with clinicopathological variables and survival. RESULTS: Pulmonary NETs of high malignant potential (LCNEC and SCLC) had reduced expression of the adhesion molecules and high level expression of transcriptional repressors (Snail1, Snail2, Twist and Foxc2). Snail high expression levels and the loss of E-cadherin/ß-catenin complex integrity had the strongest negative effect on the five-year survival rates. E-cadherin/ß-catenin complex integrity loss independently predicted lymph node involvement and helped in Atypical Carcinoid (AC) vs Typical Carcinoid (TC) differential diagnosis. Importantly, among the TC group, the loss of E-cadherin/ß-catenin complex integrity identified patients with an adverse clinical course despite favourable clinicopathological features. CONCLUSION: The immunohistochemical determination of E-cadherin/ß-catenin complex integrity loss and EMT markers in the clinical setting might be a potential useful diagnostic and prognostic tool especially among the TC patients.

EMS mutagenesis and qPCR-HRM prescreening for point mutations in an embryogenic cell suspension of grapevine.

KEY MESSAGE: Embryogenic suspension cultures are suitable for EMS mutagenesis in grapevine, and HRM prescreening of EMS-treated somatic embryo clusters allows rapid detection of point mutations before plant regeneration. ABSTRACT: Somatic embryogenesis is an excellent system for induced mutagenesis and clonal propagation in woody plants. Our work was focused on establishing a procedure for inducing ethyl methanesulfonate (EMS) mutagenesis in grapevine. Embryogenic cell aggregates (ECAs) growing in liquid medium were treated with increasing concentrations of EMS. We found that EMS dramatically affects the viability of ECAs at concentrations above 20 mM (25.5 ± 2.9 % survival), whereas concentrations above 10 mM affect embryogenic potential (22.1 ± 1.7 % of ECAs gave rise to embryos). Embryo masses generated from EMS-treated embryogenic cell aggregates were prescreened by quantitative PCR-High Resolution Melting (qPCR-HRM) to detect single nucleotide polymorphisms (SNPs) in a 1,000-bp VvNCED1-encoding DNA fragment, which served as the target gene. Detected mutations were verified in regenerated plants by PCR and sequencing. qPCR-HRM analysis of the difference plots for the fluorescence signals allowed detection of a mutation in a sample from an embryogenic aggregate treated with 10 mM EMS. To confirm the nature of the mutation, embryos from this aggregate were recovered and germinated, and leaves were collected for PCR and sequencing analysis. The alignment of sequences from regenerated plants with the wild-type sequence revealed a transitional mutation (G/C to A/T) in the 1,000-bp VvNCED1-encoding region. To our knowledge, this is the first time that EMS mutagenesis has been performed using an embryogenic cell suspension of grapevine.

[Dengue encephalitis in Argentina]. / Encefalitis por dengue en Argentina.

Dengue virus is an endemic virus in Argentina that, although it was initially considered to be non-neurotropic, it is currently recognized to be neuroinvasive; thus conditioning a prevalence of neurological manifestations of up to 15% among patients. Even being considered severe symptoms, there is underdiagnoses of dengue encephalitis due to its varied clinical presentation. Neurological manifestations of dengue encephalitis can range from fever and headache to altered levels of consciousness and seizures. Although the cerebrospinal fluid may be normal in up to a third of cases, it usually presents increased protein concentration and pleocytosis. Regarding neuroimaging methods, the findings are usually varied and nonspecific, and can even be normal in up to 40-50% of cases. We present three cases of dengue encephalitis diagnosed in a university hospital in Buenos Aires, Argentina, where the clinical presentation varied from temporal-spatial disorientation to refractory convulsive status with different presentations in the cerebrospinal fluid but all with positive PCR for dengue in it and with normal neuroimaging.

El virus dengue es un virus endémico en Argentina que, si bien inicialmente se consideró que no era neurotrópico, actualmente se reconoce que tiene neuroinvasión, condicionando así una prevalencia de manifestaciones neurológicas de hasta el 15% entre los enfermos. Aun siendo considerados síntomas de gravedad, existe subdiagnóstico de encefalitis por dengue debido a su variada forma de presentación clínica. Las manifestaciones neurológicas de la encefalitis por dengue pueden abarcar desde fiebre y cefalea hasta alteraciones del nivel de conciencia y convulsiones. Si bien el líquido cefalorraquídeo (LCR) puede hallarse normal en hasta un tercio de los casos, lo habitual es que presente aumento de concentración de proteínas y pleocitosis. En cuanto a los métodos de neuroimagen, los hallazgos suelen ser variados e inespecíficos, e incluso pueden ser normales hasta en 40-50% de los casos. Se presentan 3 casos de encefalitis por dengue diagnosticados en un hospital universitario de Buenos Aires, Argentina, en donde la presentación clínica varió desde desorientación témporo-espacial hasta estatus convulsivo refractario con diferentes presentaciones en el LCR pero todos con PCR positivo para dengue y con neuroimágenes sin alteraciones.

Effects of whole body vibration training in patients with severe chronic obstructive pulmonary disease.

BACKGROUND AND OBJECTIVE: To determine if whole body vibration training (WBVT) improves muscular force and modifies functional capacity parameters in patients with severe chronic obstructive pulmonary disease (COPD). METHODS: We performed a randomized controlled trial in the outpatient Physical Medicine and Rehabilitation Department of a general hospital. Sixty stable male patients with COPD and mean forced expiratory volume in 1 s (FEV1) 34.3% (predicted) were enrolled with 51 patients completing the study. Participants were randomized into two groups: Whole Body Vibration Training Group (WBVTG), (n = 26) undergoing three sessions per week for a total of 6 weeks and a Control Group (CG) (n = 25) without intervention. The main outcome measures were isokinetic knee flexor and extensor testing in a concentric-concentric regime and exercise capacity measured by the 6-min walking test (6MWT). Secondary outcomes were pulmonary muscular assessment with maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP). RESULTS: No significant differences were observed between groups in maximum moment of isokinetic knee flexor/extensor testing force in a concentric-concentric regime. In contrast, WBVTG patients showed a significant increase in the 6MWT (81.2 ± 9.2) meters; mean ± SD; P < 0.001). There was also a significant decrease in maximum oxygen desaturation in the 6MWT after 6 weeks of training (3.1 ± 1.1; P = 0.01). There were significant differences between groups in MIP and MEP at the end of the study in favour of the WBVTG. CONCLUSIONS: WBVT provided significant improvements in functional capacity in severe COPD patients without changes in muscular force.

Feasibility and impact of online HIV/STI screening addressed to men who have sex with men and transgender women users of pre-exposure prophylaxis (PrEP) in Spain (TESTATE PrEP): a study protocol for a non-blinded randomised controlled trial.

INTRODUCTION: The objectives of the study are: to design and implement a pilot intervention to offer self-sampling kits to detect HIV, Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Treponema pallidum (TP) among gay, bisexual and other men who have sex with men and transgender women users of pre-exposure prophylaxis (PrEP) as part of PrEP follow-up. To evaluate if the pilot intervention causes a reduction of the retention to PrEP follow-up among the target population. To analyse the capacity of the intervention to reduce the healthcare burden on the PrEP service. To evaluate the acceptability of the intervention among PrEP users and PrEP service healthcare workers and; to validate dried blood samples for treponemal and non-treponemal antibody detection using the Dual Path Platform syphilis screening and confirmatory assay compared with blood drawn by venous puncture. METHODS AND ANALYSIS: We will perform a non-blinded randomised controlled non-inferiority trial among PrEP users on follow-up. Participants on the control arm will follow the usual follow-up protocol with quarterly face-to-face visits where they will be tested for HIV and sexually transmitted infections (STIs). Participants in the experimental arm will alternate face-to-face meetings with online screening of HIV and STIs. The website https://testate.org/ will include a module for online follow-up visits of participants. Participants of the experimental arm will order self-sampling kits for HIV, CT, NG and TP through the website, will send the samples to the laboratory and check their results online. We will compare the retention to follow up and the healthcare burden in both arms. The acceptability of the intervention among participants and healthcare workers will be assessed. ETHICS AND DISSEMINATION: The project has been approved by the CEIC-HUGTIP (Reference: PI-22-051). Subjects will be included after giving their informed consent. Final conclusions and recommendations will be shared with stakeholders. Two publications in peer-reviewed journals are expected. TRIAL REGISTRATION NUMBER: NCT05752643.

Trans-cinnamaldehyde-related overproduction of benzoic acid and oxidative stress on Arabidopsis thaliana.

Introduction: Trans-cinnamaldehyde is a specialised metabolite that naturally occurs in plants of the Lauraceae family. This study focused on the phytotoxic effects of this compound on the morphology and metabolism of Arabidopsis thaliana seedlings. Material and methods: To evaluate the phytotoxicity of trans-cinnamaldehyde, a dose-response curve was first performed for the root growth process in order to calculate the reference inhibitory concentrations IC50 and IC80 (trans-cinnamaldehyde concentrations inducing a 50% and 80% inhibition, respectively). Subsequently, the structure and ultrastructure of the roots treated with the compound were analysed by light and electron microscopy. Based on these results, the following assays were carried out to in depth study the possible mode of action of the compound: antiauxinic PCIB reversion bioassay, determination of mitochondrial membrane potential, ROS detection, lipid peroxidation content, hormone quantification, in silico studies and gene expression of ALDH enzymes. Results: Trans-cinnamaldehyde IC50 and IC80 values were as low as 46 and 87 µM, reducing the root growth and inducing the occurrence of adventitious roots. At the ultrastructural level, the compound caused alterations to the mitochondria, which were confirmed by detection of the mitochondrial membrane potential. The morphology observed after the treatment (i.e., appearance of adventitious roots) suggested a possible hormonal mismatch at the auxin level, which was confirmed after PCIB bioassay and hormone quantification by GC-MS. The addition of the compound caused an increase in benzoic, salicylic and indoleacetic acid content, which was related to the increased gene expression of the aldehyde dehydrogenase enzymes that can drive the conversion of trans-cinnamaldehyde to cinnamic acid. Also, an increase of ROS was also observed in treated roots. The enzyme-compound interaction was shown to be stable over time by docking and molecular dynamics assays. Discussion: The aldehyde dehydrogenases could drive the conversion of trans-cinnamaldehyde to cinnamic acid, increasing the levels of benzoic, salicylic and indoleacetic acids and causing the oxidative stress symptoms observed in the treated seedlings. This would result into growth and development inhibition of the trans-cinnamaldehyde-treated seedlings and ultimately in their programmed-cell-death.

Political Populism, Institutional Distrust and Vaccination Uptake: A Mediation Analysis.

Politics is ubiquitous in public health, but vaccines had never been weaponized to instill distrust to gain political advantage. In pandemic and post-pandemic scenarios, populist political parties could use vaccine-related issues to generate distrust in evidence-based knowledge. Therefore, some questions arise. What impact could populist political parties impinge on vaccination uptake rates through sowing political discontent? What could the medical institutions do to avoid the adverse effects that these political strategies could infringe? For answering these research questions, we first hypothesized that vaccine uptake was negatively associated with distrust in the institutions. Furthermore, we analyzed whether populism mediates this relationship. In doing so, we hypothesized a positive association between distrust and populism, because populists, mainly fueled by politically discontent citizens, offer hope of a better future, blaming their misfortune on the actions of the elite. Additionally, we hypothesized that those citizens with a higher level of political dissatisfaction, following the claims of the populist political parties, will have lower vaccine uptake results, because they will be discouraged from making the efforts to counter the pandemic. Based on a survey carried out by the European Commission that covered 27 E.U. + U.K. countries (totaling 27,524 respondents), this paper proves that an individual's political discontent fully mediates the relationship between distrust in institutions and vaccine uptake. Targeting the vaccine-hesitant population is quite convenient for populists because they only need to convince a minority of citizens not to be vaccinated to achieve their destabilizing goals. New outbreaks will appear if the minimum herd immunity coverage is not reached, reinforcing a vicious circle of distrust in elites, in consequence. For tackling this matter, recommendations are given to institutional managers from a social marketing standpoint.

The COVID-19 Sentinel Schools Network of Catalonia (CSSNC) project: Associated factors to prevalence and incidence of SARS-CoV-2 infection in educational settings during the 2020-2021 academic year.

The Sentinel Schools project was designed to monitor and evaluate the epidemiology of COVID-19 in Catalonia, gathering evidence for health and education policies to inform the development of health protocols and public health interventions to control of SARS-CoV-2 infection in schools. The aim of this study was to estimate the prevalence and incidence of SARS-CoV-2 infections and to identify their determinants among students and staff during February to June in the academic year 2020-2021. We performed two complementary studies, a cross-sectional and a longitudinal component, using a questionnaire to collect nominal data and testing for SARS-CoV-2 detection. We describe the results and perform a univariate and multivariate analysis. The initial crude seroprevalence was 14.8% (95% CI: 13.1-16.5) and 22% (95% CI: 18.3-25.8) for students and staff respectively, and the active infection prevalence was 0.7% (95% CI: 0.3-1) and 1.1% (95% CI: 0.1-2). The overall incidence for persons at risk was 2.73 per 100 person-month and 2.89 and 2.34 per 100 person-month for students and staff, respectively. Socioeconomic, self-reported knowledge, risk perceptions and contact pattern variables were positively associated with the outcome while sanitary measure compliance was negatively associated, the same significance trend was observed in multivariate analysis. In the longitudinal component, epidemiological close contact with SARS-CoV-2 infection was a risk factor for SARS-CoV-2 infection while the highest socioeconomic status level was protective as was compliance with sanitary measures. The small number of active cases detected in these schools suggests a low transmission among children in school and the efficacy of public health measures implemented, at least in the epidemiological scenario of the study period. The major contribution of this study was to provide results and evidence that help analyze the transmission dynamic of SARS-CoV-2 and evaluate the associations between sanitary protocols implemented, and measures to avoid SARS-CoV-2 spread in schools.

Monitoring of SARS-CoV-2 seroprevalence among primary healthcare patients in the Barcelona Metropolitan Area: the SeroCAP sentinel network protocol.

INTRODUCTION: SARS-CoV-2 seroprevalence studies are currently being recommended and implemented in many countries. Forming part of the COVID-19 monitoring and evaluation plan of the Catalan Government Health Department, our network aims to initiate a primary healthcare sentinel monitoring system as a surrogate of SARS-CoV-2 exposure in the Barcelona Metropolitan Area. METHODS AND ANALYSIS: The seroCAP is a serial cross-sectional study, which will be performed in the Barcelona Metropolitan Area to estimate antibodies against SARS-CoV-2. From February 2021 to March 2022, the detection of serum IgG antibodies against SARS-CoV-2 trimeric spike protein will be performed on a monthly basis in blood samples collected for diverse clinical purposes in three reference hospitals from the three Barcelona healthcare areas (BCN areas). The samples (n=2588/month) will be from patients attended by 30 primary healthcare teams at 30 basic healthcare areas (BHA). A lab software algorithm will systematically select the samples by age and sex. Seroprevalence will be estimated and monitored by age, sex, BCN area and BHA. Descriptive and cluster analysis of the characteristics and distribution of SARS-CoV-2 infections will be performed. Sociodemographic, socioeconomic and morbidity-associated factors will be determined using logistic regression. We will explore the association between seroprevalence, SARS-CoV-2 confirmed cases and the implemented measures using interrupted time series analysis. ETHICS AND DISSEMINATION: Ethical approval was obtained from the University Institute Foundation for Primary Health Care Research Jordi Gol i Gurina ethics committee. An informed consent is not required regarding the approval of the secondary use of biological samples within the framework of the COVID-19 pandemic. A report will be generated quarterly. The final analysis, conclusions and recommendations will be shared with the stakeholders and communicated to the general public. Manuscripts resulting from the network will be submitted for publication in peer-reviewed journals.

Histone Deacetylase Inhibitors Increase the Embryogenic Potential and Alter the Expression of Embryogenesis-Related and HDAC-Encoding Genes in Grapevine (Vitis vinifera L., cv. Mencía).

The low induction rates of somatic embryogenesis are one of the main limitations in its routine application in the grapevine (Vitis vinifera L.). The use of an induction medium containing histone deacetylase inhibitors (trichostatin A and, mainly, sodium butyrate) resulted in an improvement of the embryogenic responses in grapevine (cv. Mencía) cotyledonary and recently germinated somatic embryos. The relative expression of several grapevine genes related to embryogenic competence or encoding histone deacetylase enzymes was studied in cotyledonary somatic embryos that were cultured in the presence of 0.5 mM sodium butyrate. The results showed a significant overexpression of the BBM and VvSERK2 genes after 24 h of culture, whereas the VvWOX2 gene was underexpressed less in treated versus untreated explants. The results suggest that the inhibitor may trigger a molecular response related to an increase in embryogenic competence and changes in the expression of associated genes. The treatment with sodium butyrate also produced significant variations in the expression of several histone deacetylase enzyme-encoding genes. These results may enhance the possibility of obtaining somatic embryos, reducing the seasonal constraints associated with the use of floral explants in grapevines.

siRNA Therapeutics in Ocular Diseases.

The field of RNAi therapeutics has quickly adapted to the treatment of ocular diseases. Although the eye provides a unique system for the delivery of siRNAs, its complex structure and composition fostered the development of novel strategies for efficient gene silencing in the target compartment. Moreover, anterior and posterior segments differ in their multiple drug barriers and clearance mechanisms. This chapter summarizes the recent achievements in terms of routes of administration, chemical modifications, and delivery systems for siRNAs that specifically apply to eye disorders. Methods employed for siRNA detection/quantitation in ocular tissues are also described, together with safety concerns that need to be addressed to fulfill regulatory requirements of new drug approval. Even though RNAi therapies for ocular diseases have not yet translated into patient care, we document herein the rising number of candidate drugs currently under preclinical or clinical development.

Global Vaccine Hesitancy Segmentation: A Cross-European Approach.

Vaccine-preventable diseases are global mainly in a globalized world that is characterized by a continuous movement of people and goods across countries. Vaccine hesitancy, the reluctance or refusal to vaccinate despite the availability of vaccines, is rising worldwide. What if the problem of vaccine hesitancy could be most effectively managed when treated globally rather than on a national or regional basis? What if a global vaccine-hesitant segment exists and the differences among countries are not so significant? Based on the Global Marketing Strategy paradigm, this paper shows that seven different cross-European segments exist based on the beliefs, attitudes, and behaviors collected in 28 European countries. These pan-European segments are differentiable (people in those segments have similar characteristics that are visibly dissimilar from the ones in other segments) and actionable (organizations would be able to propose interventions to the hesitant segments based on their profiles). With segmentation being the starting point of many public health intervention strategies for avoiding vaccine-hesitancy, the results recommend moderating the full-adaptation strategy that follows the "context matters" principle suggested by several political and public health international organizations. Embracing a more standardized strategy will allow the development of better services and strategies that support and enable desirable vaccination behaviors.

Vaccine Hesitancy and Political Populism. An Invariant Cross-European Perspective.

Vaccine-hesitancy and political populism are positively associated across Europe: those countries in which their citizens present higher populist attitudes are those that also have higher vaccine-hesitancy rates. The same key driver fuels them: distrust in institutions, elites, and experts. The reluctance of citizens to be vaccinated fits perfectly in populist political agendas because is a source of instability that has a distinctive characteristic known as the "small pockets" issue. It means that the level at which immunization coverage needs to be maintained to be effective is so high that a small number of vaccine-hesitants have enormous adverse effects on herd immunity and epidemic spread. In pandemic and post-pandemic scenarios, vaccine-hesitancy could be used by populists as one of the most effective tools for generating distrust. This research presents an invariant measurement model applied to 27 EU + UK countries (27,524 participants) that segments the different behaviours found, and gives social-marketing recommendations for coping with the vaccine-hesitancy problem when used for generating distrust.

Prognostic significance of lymphangiogenesis in pharyngolaryngeal carcinoma patients.

BACKGROUND: Lymphatic vessel spread is considered a major route for head and neck squamous cell carcinoma metastasis. Formation of new lymphatic vessels could facilitate the process, raising the malignant potential of these tumours. Recent identification of lymphatic markers allows the study of the lymphangiogenesis phenomenon. We searched for molecular events involved in the lymphangiogenic process that could have prognostic value in laryngeal/pharyngeal carcinoma patients. METHODS: 104 paraffin-embedded pharyngeal/laryngeal tumour samples were studied. Immunohistochemical analysis of podoplanin and double immunofluorescence analysis of Ki-67 and D2-40 were performed. Lymph vessel density (inside the tumour mass, at its periphery or considered as a whole) and the presence of tumour emboli inside lymphatics were recorded. The proliferative state of endothelial lymphatic cells was evaluated. RESULTS: Lymphatic vessels were detected inside the tumour mass (75%) and in the surrounding tissue (80%); some of them in a proliferative state. Tumour emboli were detected in a high proportion of the cases (45%). Lymphatic vessel density was higher in the pharyngeal cases (p = 0.0029), in greater size (p = 0.039), more advanced stage primary tumours (p = 0.006) and in carcinomas of patients with affected nodes (p = 0.019). The presence of tumour emboli and a high global vessel density were indicators of poor prognosis (recorded as death from tumour) in the laryngeal group (p = 0.015 and p = 0.027, respectively), but notably not in the pharyngeal one. Interestingly, high global vessel density showed a negative prognostic value among pathologically staged N0 laryngeal carcinomas (p = 0.03). CONCLUSIONS: The lymphangiogenic process correlated with aggressive tumour features (pN category, tumour size, tumour stage), but might play different roles in tumours arising from different anatomic sites. Our results suggest that detection of tumour emboli and assessment of global vessel density using the D2-40 antibody, may be useful in the clinical practice, as predictors of reduced survival among pN0 laryngeal carcinoma patients.

Food Reward and Food Choice. An Inquiry Through The Liking and Wanting Model.

What if consumers are getting obese because eating less calories is more difficult for persons that have a higher pleasure and desire towards food (Ikeda et al., 2005) and food companies do not help given only a two extreme option choice to satisfy their needs (i.e., low calories vs. high calories or healthy vs. unhealthy)? Reward systems are being described with a new conceptual approach where liking-the pleasure derived from eating a given food-and wanting-motivational value, desire, or craving-can be seen as the significant forces guiding eating behavior. Our work shows that pleasure (liking), desire (wanting), and the interaction between them influence and are good predictors of food choice and food intake. Reward responses to food are closely linked to food choice, inducing to caloric overconsumption. Based on the responses given to a self-administered questionnaire measuring liking and wanting attitudes, we found three different segments named 'Reward lovers,' 'Half epicurious,' and 'Non indulgents'. Their behavior when choosing food is quite different. Results show differential effects on caloric consumption depending on segments. The introduction of more food choices that try to balance their content is a win-win strategy for consumers, companies, and society.

Encefalitis por dengue en Argentina / Dengue encephalitis in Argentina

Resumen El virus dengue es un virus endémico en Argentina que, si bien inicialmente se consideró que no era neu rotrópico, actualmente se reconoce que tiene neuroinva sión, condicionando así una prevalencia de manifesta ciones neurológicas de hasta el 15% entre los enfermos. Aun siendo considerados síntomas de gravedad, existe subdiagnóstico de encefalitis por dengue debido a su variada forma de presentación clínica. Las manifesta ciones neurológicas de la encefalitis por dengue pueden abarcar desde fiebre y cefalea hasta alteraciones del nivel de conciencia y convulsiones. Si bien el líquido cefalorraquídeo (LCR) puede hallarse normal en hasta un tercio de los casos, lo habitual es que presente aumento de concentración de proteínas y pleocitosis. En cuanto a los métodos de neuroimagen, los hallazgos suelen ser variados e inespecíficos, e incluso pueden ser normales hasta en 40-50% de los casos. Se presentan 3 casos de encefalitis por dengue diag nosticados en un hospital universitario de Buenos Ai res, Argentina, en donde la presentación clínica varió desde desorientación témporo-espacial hasta estatus convulsivo refractario con diferentes presentaciones en el LCR pero todos con PCR positivo para dengue y con neuroimágenes sin alteraciones.

Abstract Dengue virus is an endemic virus in Argentina that, although it was initially considered to be non-neuro tropic, it is currently recognized to be neuroinvasive; thus conditioning a prevalence of neurological mani festations of up to 15% among patients. Even being considered severe symptoms, there is underdiagnoses of dengue encephalitis due to its varied clinical presenta tion. Neurological manifestations of dengue encephalitis can range from fever and headache to altered levels of consciousness and seizures. Although the cerebrospinal fluid may be normal in up to a third of cases, it usually presents increased protein concentration and pleocyto sis. Regarding neuroimaging methods, the findings are usually varied and nonspecific, and can even be normal in up to 40-50% of cases. We present three cases of dengue encephalitis di agnosed in a university hospital in Buenos Aires, Ar gentina, where the clinical presentation varied from temporal-spatial disorientation to refractory convulsive status with different presentations in the cerebrospinal fluid but all with positive PCR for dengue in it and with normal neuroimaging.