Evaluation of the Possible Transmission of BSE and Scrapie to Gilthead Sea Bream (Sparus aurata)

In transmissible spongiform encephalopathies (TSEs), a group of fatal neurodegenerative disorders affecting many species, the key event in disease pathogenesis is the accumulation of an abnormal conformational isoform (PrPSc) of the host-encoded cellular prion protein (PrPC). While the precise mechanism of the PrPC to PrPSc conversion is not understood, it is clear that host PrPC expression is a prerequisite for effective infectious prion propagation. Although there have been many studies on TSEs in mammalian species, little is known about TSE pathogenesis in fish. Here we show that while gilthead sea bream (Sparus aurata) orally challenged with brain homogenates prepared either from a BSE infected cow or from scrapie infected sheep developed no clinical prion disease, the brains of TSE-fed fish sampled two years after challenge did show signs of neurodegeneration and accumulation of deposits that reacted positively with antibodies raised against sea bream PrP. The control groups, fed with brains from uninfected animals, showed no such signs. Remarkably, the deposits developed much more rapidly and extensively in fish inoculated with BSE-infected material than in the ones challenged with the scrapie-infected brain homogenate, with numerous deposits being proteinase K-resistant. These plaque-like aggregates exhibited congophilia and birefringence in polarized light, consistent with an amyloid-like component. The neurodegeneration and abnormal deposition in the brains of fish challenged with prion, especially BSE, raises concerns about the potential risk to public health. As fish aquaculture is an economically important industry providing high protein nutrition for humans and other mammalian species, the prospect of farmed fish being contaminated with infectious mammalian PrPSc, or of a prion disease developing in farmed fish is alarming and requires further evaluation

STREAMLINE HUB: a high capacity hub for research of neurodevelopmental disorders in the Western Balkan region

Neurodevelopmental disorders (NDDs) are caused by alterations in early brain development. They are a group of geographically dispersed, complex and heterogeneous disorders that give rise to the psychiatric conditions such as autism spectrum disorders, intellectual disability, schizophrenia and bipolar disorder. In order to build global research activity for study of NDDs, the main goals of the Twinning project STREAMLINE are to enhanced strategic networking and reinforce research and innovation potential of the Institute of Molecular Genetics and Genetic Engineering, University of Belgrade (IMGGE) in order to develop IMGGE as a high capacity hub for research of NDDs in the Western Balkans. This will be achieved by twinning IMGGE with three top-class research institutions in Europe (Cardiff University, University of Maastricht and Centre for Research and Technology Hellas) with an exceptional expertise in the stem cells based research of NDDs, -OMICS technologies, bioinformatics data analysis and drug testing and through staff exchanges, training, and organization of summer schools, Industry Open Days, symposia and workshops

22q11.2 Deletion syndrome as a tool for modelling and research of neurodevelopmental disorders

Introduction: Neurodevelopmental disorders (NDDs) are a group of complex and heterogeneous disorders that include autism spectrum disorders, intellectual disability, schizophrenia and bipolar disorder. However, underlying pathophysiological mechanisms are mostly unknown. In order to get better understanding of the underlying mechanisms and to discover potential therapeutics we have focused our research on 22q11.2 Deletion Syndrome (22q11.2DS), caused by microdeletion of the region q11.2 of chromosome 22 and associated with a high risk for NDDs. Methods: To study molecular mechanisms underlying intrafamilial phenotypic variability, we have identified families with the inherited form of 22q11.2DS with the aim of conducting the following analyses: whole genome sequencing in order to detect additional genetic variation(s) present in the affected child; generation of induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells; analysis of the effects of 22q11.2 microdeletion on neural differentiation including organoids as 3D model system; transcriptome analysis of iPSC-derived neurons and astrocytesto determine differentially expressed gene sets and dysregulated pathways; and testing the metabolic changes and drug responsiveness of neurons and astrocytes by high-throughput cell-based assays. Results: Peripheral blood mononuclear cells of the families with inherited form of 22q11.2DS were reprogrammed and established iPSCs were characterized. Generated iPSCs will be subjected to the further analyses. Conclusion: Currently, most of the treatments of NDDs are symptom-based due to limited understanding of underlying pathophysiological mechanisms. It is expected that patient-derived iPSCs will enable a deeper understanding of unique disease mechanisms and may also provide a significant contribution in preclinical drug development

Copy number variants (CNVs): a powerful tool for iPSC-based modelling of ASD

Patients diagnosed with chromosome microdeletions or duplications, known as copy number variants (CNVs), present a unique opportunity to investigate the relationship between patient genotype and cell phenotype. CNVs have high genetic penetrance and give a good correlation between gene locus and patient clinical phenotype. This is especially effective for the study of patients with neurodevelopmental disorders (NDD), including those falling within the autism spectrum disorders (ASD). A key question is whether this correlation between genetics and clinical presentation at the level of the patient can be translated to the cell phenotypes arising from the neurodevelopment of patient induced pluripotent stem cells (iPSCs). Here, we examine how iPSCs derived from ASD patients with an associated CNV inform our understanding of the genetic and biological mechanisms underlying the aetiology of ASD. We consider selection of genetically characterised patient iPSCs; use of appropriate control lines; aspects of human neurocellular biology that can capture in vitro the patient clinical phenotype; and current limitations of patient iPSC-based studies. Finally, we consider how future research may be enhanced to maximise the utility of CNV patients for research of pathological mechanisms or therapeutic targets

Pengaruh proporsi tepung labu kuning dan tepung terigu terhadap sifat fisikokimia dan organoleptik cake kukus

Labu kuning kaya akan β-karoten, gula, dan serat. Upaya diversifikasi pengolahan labu kuning adalah tepung labu kuning. Tepung labu kuning dapat dimanfaatkan dalam pembuatan cake kukus sebagai bahan pensubstitusi tepung terigu. Penggunaan tepung labu kuning diharapkan dapat meningkatkan kadar β-karoten dalam cake kukus. Penelitian ini bertujuan untuk menentukan proporsi tepung labu kuning dan tepung terigu yang tepat untuk menghasilkan cake kukus dengan sifat fisikokimia dan organoleptik yang dapat diterima oleh konsumen. Rancangan penelitian yang digunakan adalah Rancangan Acak Kelompok dengan satu faktor, yaitu faktor proporsi tepung labu kuning dan tepung terigu dengan lima level, yaitu: 0%:100%; 10%:90%; 20%:80%; 30%:70%; 40%:60%, yang diulang sebanyak lima kali. Parameter pengujian meliputi volume spesifik, firmness, dan organoleptik kesukaan warna, kemudahan digigit, moistness, dan rasa. Data dianalisis dengan ANOVA (Analysis of Variance) pada α = 5% untuk mengetahui ada tidaknya pengaruh proporsi tepung labu kuning dan tepung terigu terhadap sifat fisikokimia dan organoleptik cake kukus. Apabila terdapat pengaruh nyata maka dilanjutkan dengan uji DMRT (Duncan's Multiple Range Test) pada α = 5%. Penentuan perlakuan terbaik dilakukan dengan uji pembobotan. Cake kukus dengan perlakuan terbaik diuji kadar β-karoten. Hasil penelitian menunjukkan bahwa perlakuan proporsi tepung labu kuning : tepung terigu memberikan pengaruh nyata (α = 5%) terhadap volume spesifik, firmness, dan tingkat kesukaan terhadap warna, kemudahan digigit, moistness, dan rasa. Semakin besar proporsi tepung labu kuning, semakin kecil nilai volume spesifik, firmness, dan kesukaan warna cake, namun semakin besar nilai kesukaan kemudahan digigit, moistness, dan rasa cake. Proporsi tepung labu kuning : tepung terigu 20%:80% merupakan perlakuan terbaik dengan kadar β-karoten sebesar 8,569 µg β- karoten/g cake kukus

Membranous nephropathy and lupus-like syndrome after hematopoietic cell transplantation: a case report

<p>Abstract</p> <p>Introduction</p> <p>The kidney is increasingly recognised as a target organ of chronic graft-versus-host disease after hematopoietic cell transplantation in the context of the development of the nephrotic syndrome. Chronic graft-versus-host disease is associated with autoimmune phenomena similar, but not identical, to those observed in various rheumatologic disorders, implicating autoimmunity as an important component of the disease.</p> <p>Case presentation</p> <p>We report the case of a 57-year-old Caucasian man who developed the nephrotic syndrome due to membranous nephropathy in association with recurrent chronic graft-versus-host disease, along with a lupus-like syndrome manifested with pancytopenia, hair loss, positive anti-DNA antibodies and sub-epithelial and mesangial immune deposits. To the best of our knowledge, this is the first case reported in the literature. The nephrotic syndrome subsided soon after he was treated with a short course of cyclosporin with steroids. Unfortunately he died seven months later due to a relapse of leukemia.</p> <p>Conclusions</p> <p>Our case report confirms the notion that chronic graft-versus-host disease is characterized by the appearance of autoimmune phenomena similar, but not identical, to those seen in autoimmune diseases. The decision for more immunosuppression has to be weighed against the need for preservation of the graft versus leukemia phenomenon.</p

Study on the molecular structure of the infections agent that causes spongiform encephalopathies in humans and animals

This thesis consists of two main parts. The main goal of this research project was the isolation and identification of brain proteins interacting with either cellular (PrPC) or infectious (PrPSc) prion protein. To achieve this goal we performed co-immunoprecipitation assays, in the presence or absence of chemical cross-linkers and PrP/affinity chromatography experiments. The results showed that PrPC is related to β4 subunit of nicotinic acetylcholine receptor (nAChR) and other proteins of the membrane and cytoskeleton. The possible participation of β4 subunit nAChR in the pathogenesis of prion diseases was further studied in bioassays, using β4 knockout mice. In parallel, we characterized proteins co-purifying with the infectious prion protein in PrPSc enriched brain preparations. At the second part, we studied the proteolytic activity of the tissue plasminogen activator (tPA)/plasminogen system in normal and pathological brain tissue. The experimental procedures showed that tPA/plasminogen digests cellular PrPC but not the infectious PrPSc. These data, along with bioassays in tPA and plasminogen knockout animals, suggest that these molecules play a neuroprotective role in the propagation of prion diseases.Η παρούσα διατριβή έχει δύο σκέλη. Το κύριο σκέλος υπήρξε η απομόνωση και ταυτοποίηση πρωτεϊνών που αλληλεπιδρούν με τη φυσιολογική ή τη παθολογική PrP. Γι’ αυτό το λόγο, διερευνήθηκε η αλληλεπίδραση της PrPC με πρωτεΐνες του εγκεφαλικού ιστού. Πιο συγκεκριμένα, εκτελέστηκαν πειράματα διπλής ανοσοκατακρήμνισης, παρουσία ή απουσία χημικών συνδετών, καθώς και χρωματογραφίας αγχιστείας ομογενοποιημάτων εγκεφάλου σε στήλες ακινητοποιημένης και αναδιπλωμένης PrP. Τα αποτελέσματα έδειξαν ότι η φυσιολογική prion πρωτεΐνη αλληλεπιδρά με τη β4 υπομονάδα του νικοτινικού υποδοχέα της ακετυλοχολίνης (nAChR) καθώς και με πρωτεΐνες της πλασματικής μεμβράνης και του κυτταροσκελετού. H συμμετοχή της β4 υπομονάδας nAChR στην παθογένεση των ασθενειών prion μελετήθηκε με πειράματα βιοδοκιμασιών σε διαγονιδιακά ποντίκια. Ταυτόχρονα, μελετήθηκε η πρωτεϊνική σύσταση παθολογικών παρασκευασμάτων, εμπλουτισμένα στην PrPSc ισομορφή, προκειμένου να ταυτοποιηθούν πρωτεΐνες που συγκαθαρίζουν με τη μολυσματική PrP. Στο δεύτερο σκέλος μελετήθηκε η πρωτεολυτική δράση του ιστοειδικού ενεργοποιητή του πλασμινογόνου (tPA) σε φυσιολογικούς και παθολογικούς ιστούς. Οι πειραματικές δοκιμασίες έδειξαν ότι το σύμπλοκο tPA/πλασμινογόνου μπορεί να πέψει τη φυσιολογική PrPC αλλά όχι την παθολογική PrPSc. Αυτά τα δεδομένα, σε συνδυασμό με πειράματα μόλυνσης σε μοντέλα διαγονιδιακών πειραματόζωων, δείχνουν ότι το σύμπλοκο tPA/πλασμινογόνου πιθανόν να έχει νευροπροστατευτικό ρόλο στην εξέλιξη των ασθενειών prion

Protein Interaction Networks in Health and Disease

The identification and mapping of protein-protein interactions (PPIs) is a major goal in systems biology. Experimental data are currently produced in large scale using a variety of high-throughput assays in yeast or mammalian systems. Analysis of these data using computational tools leads to the construction of large protein interaction networks, which help researchers identify novel protein functions. However, our current view of protein interaction networks is still limited and there is an active field of research trying to further develop this concept to include important processes: the topology of interactions and their changes in real time, the effects of competition for binding to the same protein region, PPI variation due to alternative splicing or post-translational modifications, etc. In particular, a clinically relevant topic for development of the concept of protein interactions networks is the consideration of mutant isoforms, which may be responsible for a pathological condition. Mutations in proteins may result in loss of normal interactions and appearance of novel abnormal interactions that may affect a protein’s function and biological cycle. This Research Topic presents novel findings and recent achievements in the field of protein interaction networks with a focus on disease. Authors describe methods for the identification and quantification of PPIs, the annotation and analysis of networks, considering PPIs and protein complexes formed by mutant proteins associated with pathological conditions or genetic diseases