Prevalence and Genetic Diversity of Human Enteroviruses in the Context of Poliovirus Eradication

Viruses belonging to the family Picornaviridae are small, non-enveloped viruses with a single-stranded, positive-sense RNA genome. Up to present, thirteen genera within this large family have been designated by the International Committee on Taxonomy of Viruses: Aphtovirus, Erbovirus, Teschovirus, Sapelovirus, Senecavirus, Tremovirus, Avihepatovirus, Cardiovirus, Hepatovirus, Cosavirus, Parechovirus, Kobuvirus and Enterovirus (Fig. 1). Members within the latter six genera have been reported to cause human disease. Enteroviruses are, next to viruses of the Herpesviridae family, the major viral cause of neurologic disease with a known etiology in humans, including meningitis, encephalitis and acute flaccid paralysis (24, 38, 47). By that they form a serious threat for human health. A well known representative of these is poliovirus (species Human enterovirus C), which has inextricably been associated with large outbreaks of neurologic disease in children. This thesis will focus on prevalence and geneti

Genetic engineering of vaccine manufacturing cell lines enhances poliovirus and enterovirus 71 production

Vaccine manufacturing costs and production limitations represent two fundamental challenges facing researchers, public health officials and vaccine manufacturers committed to global health solutions. To address these issues, we have investigated whether the cell lines employed by vaccine manufacturers can be engineered to enhance vaccine virus production. As a first step in a proof-of-principle study, a genome-wide RNA Interference (RNAi) screen was conducted to identify host gene modulation events that increased Sabin 2 poliovirus (PV) replication. Primary screen hits were validated in a Vero vaccine manufacturing cell line using both attenuated and wild type poliovirus strains. This approach identified multiple single and dual gene knockdown events that increased PV titers \u3e20-fold and \u3e50-fold, respectively. Top candidate genes did not affect virus antigenicity, cell viability, or cell doubling times. Moreover, CRISPR/Cas9-mediated knockout (KO) of the top three targets created stable cell substrates with improved viral vaccine strain production. Please click Additional Files below to see the full abstract

High frequency of Polio-like Enterovirus C strains with differential clustering of CVA-13 and EV-C99 subgenotypes in a cohort of Malawian children

Enteroviruses (EVs) are among the most commonly detected viruses infecting humans worldwide. Although the prevalence of EVs is widely studied, the status of EV prevalence in sub-Saharan Africa remains largely unknown. The objective of our present study was therefore to increase our knowledge on EV circulation in sub-Saharan Africa. We obtained 749 fecal samples from a cross-sectional study conducted on Malawian children aged 6 to 60 months. We tested the samples for the presence of EVs using real time PCR, and typed the positive samples based on partial viral protein 1 (VP1) sequences. A large proportion of th

A prediction model for underestimation of invasive breast cancer after a biopsy diagnosis of ductal carcinoma in situ: based on 2892 biopsies and 589 invasive cancers

Background: Patients with a biopsy diagnosis of ductal carcinoma in situ (DCIS) might be diagnosed with invasive breast cancer at excision, a phenomenon known as underestimation. Patients with DCIS are treated based on the risk of underestimation or progression to invasive cancer. The aim of our study was to expand the knowledge on underestimation and to develop a prediction model. Methods: Population-based data were retrieved from the Dutch Pathology Registry and the Netherlands Cancer Registry for DCIS between January 2011 and June 2012. Results: Of 2892 DCIS biopsies, 21% were underestimated invasive breast cancers. In multivariable analysis, risk factors were high-grade DCIS (odds ratio (OR) 1.43, 95% confidence interval (CI): 1.05–1.95), a palpable tumour (OR 2.22, 95% CI: 1.76–2.81), a BI-RADS (Breast Imaging Reporting and Data System) score 5 (OR 2.36, 95% CI: 1.80–3.09) and a suspected invasive component at biopsy (OR 3.84, 95% CI: 2.69–5.46). The predicted risk for underestimation ranged from 9.5 to 80.2%, with a median of 14.7%. Of the 596 invasive cancers, 39% had unfavourable features. Conclusions: The risk for an underestimated diagnosis of invasive breast cancer after a biopsy diagnosis of DCIS is considerable. With our prediction model, the individual risk of underestimation can be calculated based on routinely available preoperatively known risk factors (https://www.evidencio.com/models/show/1074)

Specific In Vivo Staining of Astrocytes in the Whole Brain after Intravenous Injection of Sulforhodamine Dyes

Fluorescent staining of astrocytes without damaging or interfering with normal brain functions is essential for intravital microscopy studies. Current methods involved either transgenic mice or local intracerebral injection of sulforhodamine 101. Transgenic rat models rarely exist, and in mice, a backcross with GFAP transgenic mice may be difficult. Local injections of fluorescent dyes are invasive. Here, we propose a non-invasive, specific and ubiquitous method to stain astrocytes in vivo. This method is based on iv injection of sulforhodamine dyes and is applicable on rats and mice from postnatal age to adulthood. The astrocytes staining obtained after iv injection was maintained for nearly half a day and showed no adverse reaction on astrocytic calcium signals or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows to quantify 3D morphological parameters of the astrocytes and to characterize their network. Our method may become a reference for in vivo staining of the whole astrocytes population in animal models of neurological disorders

Un instrument pour inventorier les aspects culturels contenus dans des manuels d'enseignement de langues étrangères

What about culture in foreign language textbooks? Since the end of the sixties, it is usually asserted that culture should be part of the foreign language curriculum. If we want to communicate effectively with the members of other communities, we have to understand their language and their culture taken in a broad sense. Every year many language textbooks are published and used by various learners and teachers in different countries. Given his role in the classroom, it seems particularly important to have a good instrument to describe the cultural content of a textbook. Although culture is considered as a necessary part of the teaching of a foreign language, we still don't know how exactly (in which way) cultural information should be integrated into textbooks. The aim of our study was to provide foreign language teachers and authors of foreign language textbooks with an instrument for describing the cultural content of textbooks. In this study we have developed an instrument that is based on two models which have been used to describe the cultural content in recent editions of textbooks for French and Dutch as foreign languages. First, we generated a list of 14 cultural themes and 99 topics, which is based on three different sources. Secondly, we developed a model that analyses how the vocabulary is used in teaching culture in textbooks. The results with the first model show very little differences between the cultural content in French and Dutch textbooks. With the second model, the results show significant differences between the cultural content in the eight selected textbooks. Therefore it seems appropriate to argue that the second model is a better instrument to describe the cultural content in foreign language textbook

Epidemiology of Enterovirus 71 in The Netherlands, 1963 to 2008▿

The incidence of enterovirus 71 (EV71) infection has greatly increased in the Asian Pacific region since 1997. Several large outbreaks, caused by different subgenogroups of EV71, occurred with high rates of morbidity and a substantial number of deaths. In 2007, 58 cases of EV71 infection requiring hospitalization were reported in The Netherlands after a period of low endemicity of 21 years. These events triggered a study on the epidemiology of EV71 in The Netherlands. Genetic analysis of the VP1 capsid region of 199 EV71 isolates collected from 1963 to 2008 as part of enterovirus surveillance activities revealed a change in the prevailing subgenogroups over time. From 1963 to 1986 infections were caused by three different and successive lineages belonging to subgenogroup B (the novel lineage designated B0, as well as B1 and B2). In 1987, following a major epidemic the previous year, the B genogroup was replaced by genogroup C strains of lineages C1 and, later, C2. Analyses of the clinical data suggested that there were differences between infection with genogroup B and with genogroup C strains in terms of the age groups affected and the severity of illness. From comparative analysis with genomic data available in the public domain, we concluded that EV71 strain evolution shows a global pattern, which leads to the question of whether the recently emerged C4 lineage strains will also spread outside of Asia

Evolutionary trajectory of the VP1 gene of human enterovirus 71 genogroup B and C viruses

From 1963 to 1986, human enterovirus 71 (HEV71) infections in the Netherlands were successively caused by viruses of subgenogroups B0, B1 and B2. A genogroup shift occurred in 1987, after which viruses of subgenogroups C1 and C2 were detected exclusively. This is in line with HEV71 typing in Australia, Europe and the USA, but is distinct from that in the Asian Pacific region, where HEV71 subgenogroups B3-B5 and C4-C5 have caused large outbreaks since 1997. To understand these observations in HEV71 epidemiology, the VP1-encoding regions of 199 HEV71 strains isolated in the Netherlands between 1963 and 2008 were used to study the detailed evolutionary trajectory and population dynamics of HEV71. Genogroup B viruses showed an epochal evolution, whereas genogroup C viruses evolved independently, which is in line with the co-circulation of C1 and C2 viruses in the Netherlands since 1997. Considering that strains from the Netherlands are interspersed phylogenetically with Gen Bank reference strains, the evolution of B1-B2, C1-C2 viruses has a global nature. Phylodynamic analysis confirmed that increased reporting of HEV71 infections in 1986 and 2007 reflected true epidemics of B2 and C2 viruses, respectively. Sequence analysis of the complete capsid region of a subset of isolates revealed several (sub)genogroup-specific residues. Subgenogroup B2-specific rabbit antiserum showed cross-neutralization of B0, B1 and B2 viruses, but not of subgenogroup C1 or C2 viruses, probably explaining the global shift to genogroup C in 1987 following a B2 epidemic. Anti-C1 rabbit serum neutralized both genogroup B and C viruses. Global herd immunity against C1 and C2 viruses possibly explains why epidemics with subgenogroups B4 and 04 are restricted to the Asian Pacific region