Targeted mutagenesis using CRISPR-Cas9 in the chelicerate herbivore Tetranychus urticae

The use of CRISPR-Cas9 has revolutionized functional genetic work in many organisms, including more and more insect species. However, successful gene editing or genetic transformation has not yet been reported for chelicerates, the second largest group of terrestrial animals. Within this group, some mite and tick species are economically very important for agriculture and human health, and the availability of a gene-editing tool would be a significant advancement for the field. Here, we report on the use of CRISPR-Cas9 in the spider mite Tetranychus urticae. The ovary of virgin adult females was injected with a mix of Cas9 and sgRNAs targeting the phytoene desaturase gene. Natural mutants of this laterally transferred gene have previously shown an easy-to-score albino phenotype. Albino sons of injected virgin females were mated with wild-type females, and two independent transformed lines where created and further characterized. Albinism inherited as a recessive monogenic trait. Sequencing of the complete target-gene of both lines revealed two different lesions at expected locations near the PAM site in the target-gene. Both lines did not genetically complement each other in dedicated crosses, nor when crossed to a reference albino strain with a known genetic defect in the same gene. In conclusion, two independent mutagenesis events were induced in the spider mite T. urticae using CRISPR-Cas9, hereby providing proof-of-concept that CRISPR-Cas9 can be used to create gene knockouts in mites

INVESTIGATING STUDENTS' - FUTURE TEACHERS' VIEWS ON THEIR INTERNSHIP: BENEFITS AND CHALLENGES

In several countries worldwide, the internship is part of teacher education programs and aims at the professional and personal development of teachers. In Greece, the Pedagogical Departments of Primary Education (PPT) include in their curriculum the students’ practical education which is an internship supervised and supported by teachers - mentors. Through this process, students gain the opportunity to acquire more knowledge of teaching and improve the needed skills, attitudes and values associated with the efficient performance of their job. This case study seeks to unearth the benefits of the internship program and also intends to explore the potential obstacles encountered by students - future teachers during their internship. The qualitative method of research using the semi-directed interview tool was adopted to study the opinion of students - future teachers about the internship program. The survey involved eleven students - future teachers of the Department of Education and Social Work of the University of Patras, who participated in internships in schools in the region of Patras. The results showed that students - future teachers benefited from the development of integrated teaching skills with the proper guidance and support of mentors, yet they simultaneously indicated some potential inhibitory aspects and challenges which arise through their relationship with mentors.  Article visualizations

Η χρήση της δεξμεδετομιδίνης στους περιφερικούς νευρικούς αποκλεισμούς

Η χρήση των περιφερικών νευρικών αποκλεισμών διευρύνεται χάρη στα πλεονεκτήματα που προσφέρουν ως προς την μετεγχειρητική αναλγησία συγκριτικά με τις συμβατικές τεχνικές, όπως η ταχύτερη ανάνηψη και η μείωση των αναγκών σε οπιοειδή αναλγητικά. Ωστόσο η διάρκεια της αναλγησίας μετά από έναν περιφερικό νευρικό αποκλεισμό που χορηγείται μόνο τοπικό αναισθητικό είναι περιορισμένη χρονικά. Προκειμένου να αντιμετωπιστεί ο περιορισμός αυτός έχουν χρησιμοποιηθεί και δοκιμαστεί αρκετοί παράγοντες ως επικουρικά φάρμακα που θα μπορούσαν να παρατείνουν την μετεγχειρητική αναλγησία μετά από έναν περιφερικό νευρικό αποκλεισμό. Σκοπός της συγκεκριμένης μελέτης είναι μετά από ανασκόπηση της βιβλιογραφίας να παρουσιάσει τα πιο πρόσφατα δεδομένα σχετικά με τη χρήση της δεξμεδετομιδίνης, ενός εκλεκτικού α2 αγωνιστή, ως επικουρικό φάρμακο στα χορηγούμενα τοπικα αναισθητικά σε διαφορετικούς τύπους περιφερικών νευρικών αποκλεισμών. Τα ιδιαίτερα φαρμακολογικά χαρακτηριστικά της, ο μηχανισμός δράσης της, η αποτελεσματικότητα αλλά και η ασφάλεια της περινευρικής χορήγησής της, περιγράφονται με βάση την υπάρχουσα βιβλιογραφία (τυχαιοποιημένες μελέτες, ανασκοπήσεις και μετα-αναλύσεις) επιβεβαιώνοντας ως συμπέρασμα το χαρακτηρισμό της ως ένα από τα πλέον υποσχόμενα επικουρικά φάρμακα στην περιοχική αναισθησία.Peripheral nerve blocks (PNBs) are widely used for postoperative pain management offering several advantages over conventional analgesics such as quicker recovery after surgery and sparing the use of medications like opiates. However the duration of analgesia provided by nerve blocks is limited if local anaesthetics are administered alone. Therefore several additives have been investigated in order to prolong postoperative analgesia following single shot peripheral nerve blocks. The aim of this study is to review and present the current knowledge related to the use of dexmedetomidine, an a2 adrenergic agonist, as an additive drug to local aneasthetics in various peripheral nerve blocks. The pharmacological profile, mechanism of action, efficacy and safety of dexmedetomidine are described based on existing literature (RCTs, reviews and meta-analyses) justifying in conclusion its title as one of the most promising adjuvants in regional anaesthesia

Η παρουσίαση έργων προστατευόμενων από τις διατάξεις πνευματικής ιδιοκτησίας στον ψηφιακό χώρο, ιδίως μέσω υπερσυνδέσμων

Καθώς μεταβαίνουμε από τον αναλογικό στον ψηφιακό χώρο, οι αρχές που ίσχυαν για το υποκείμενο και αντικείμενο προστασίας της πνευματικής ιδιοκτησίας εφαρμόζονται ακόμα. Μεταξύ των δικαιωμάτων πνευματικής ιδιοκτησίας και των συγγενικών βρίσκεται το δικαίωμα παρουσίασης του έργου στο κοινό. Η νομολογία (ITV Broadcasting, Svensson, BestWater, GS Media, Stichting Brein I, Stichting Brein II κ.ά.) διασαφηνίζει το περιεχόμενο αυτού του δικαιώματος, ιδίως με χρήση υπερσυνδέσμων. Τα κριτήρια που αναδείχθηκαν, για το αν μια πράξη συνιστά παρουσίαση στο κοινό προστατευόμενου περιεχομένου μέσω συνδέσμων, είναι ο χρησιμοποιούμενος τεχνικός τρόπος παρουσίασης, ύπαρξη νέου κοινού, γνώση του παροχέα συνδέσμων για το παράνομο της αρχικής παρουσίασης, κερδοσκοπικός σκοπός και η απαραίτητη παρέμβαση του παροχέα για την πρόσβαση σε προστατευόμενο περιεχόμενο. Οι σύνδεσμοι διακρίνονται σε τέσσερις κατηγορίες και έκαστη έχει διαφορετική νομική αξιολόγηση. Επίσης, διαφορετικά ισχύουν, όταν ένα έργο είναι ήδη αναρτημένο με ή χωρίς άδεια του δικαιούχου, για την ευθύνη του παρόχου μεμονωμένων συνδέσμων, των μηχανών αναζήτησης και των ISPs. Τέλος, τα μέσα προστασίας των δικαιούχων απέναντι στους παραβάτες παρόχους συνδέσμων προς τα έργα τους είναι οι εκ του νόμου αστικές κυρώσεις (κυρίως αποζημίωση), τα τεχνολογικά μέτρα, η αποκάλυψη των επί μέρους προσβολέων - χρηστών του Διαδικτύου ή διακοπή παροχής υπηρεσιών προς τους προσβολείς.In the context of transition from the analogue to the digital era, the principles of intellectual property concerning the protected subject and object still apply. Among the IP and related rights is the right of communication to the public. The case law (ITV Broadcasting, Svensson, BestWater, GS Media, Stichting Brein I, Stichting Brein II, etc.) clarifies the scope of same, especially by use of hyperlinks. Criteria that have emerged as to whether an act is communication to the public via links are the technical means of transmission of works, the new public, the knowledge of the link provider about the illegal initial transmission, a profit-making nature and the essential intervention of the link provider. The links are categorized into four types, each undergoing different legal assessment. Additionally, it is different to link to works already available online with or without permission, attribute liability to providers of individual links, search engines and ISPs. Finally, the protection of copyright holders against offenders includes penalties by civil law (mainly compensation), technological measures, disclosure of individual offenders or disruption in Internet services

Evaluation of tolerance/resistance to phosphine of stored product beetle populations from Europe, by using different diagnostic methods: Presentation

We evaluated the susceptibility to phosphine in different populations originated from 14 European countries, by following different diagnostic protocols. In total, more than 200 populations were screened during these tests, classified to 9 beetle species: Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), Sitophilus oryzae (L.) (Coleoptera: Curculionidae), Sitophilus granarius (L.) (Coleoptera: Curculionidae), Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) and Lasioderma serricorne (F.) (Coleoprtera: Anobiidae). The different bioassay-related diagnostic protocols that were followed were based on different exposure intervals and phosphine concentrations, ranging between 90 min and 4 d, and between 30 and 3000 ppm, respectively. Our results indicated that one of the populations that had been sampled from Europe was strongly resistant to phosphine. Moreover, the different protocols provide comparable results, which means that a standardized diagnostic can be further designed and adopted. Moreover, molecular assays indicated that the mutations P49S in R. dominica and P45S in T. castaneum are common among different populations, regardless of the degree of resistance to phosphine. Our results suggest that there are reliable quick tools for the evaluation of resistance to phosphine and that insect sampling in target areas should be conducted on a regular basis.We evaluated the susceptibility to phosphine in different populations originated from 14 European countries, by following different diagnostic protocols. In total, more than 200 populations were screened during these tests, classified to 9 beetle species: Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), Sitophilus oryzae (L.) (Coleoptera: Curculionidae), Sitophilus granarius (L.) (Coleoptera: Curculionidae), Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae), Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae) and Lasioderma serricorne (F.) (Coleoprtera: Anobiidae). The different bioassay-related diagnostic protocols that were followed were based on different exposure intervals and phosphine concentrations, ranging between 90 min and 4 d, and between 30 and 3000 ppm, respectively. Our results indicated that one of the populations that had been sampled from Europe was strongly resistant to phosphine. Moreover, the different protocols provide comparable results, which means that a standardized diagnostic can be further designed and adopted. Moreover, molecular assays indicated that the mutations P49S in R. dominica and P45S in T. castaneum are common among different populations, regardless of the degree of resistance to phosphine. Our results suggest that there are reliable quick tools for the evaluation of resistance to phosphine and that insect sampling in target areas should be conducted on a regular basis

The relative contribution of target-site mutations in complex acaricide resistant phenotypes as assessed by marker assisted backcrossing in Tetranychus urticae

The mechanisms underlying insecticide and acaricide resistance in insects and mites are often complex, including additive effects of target-site insensitivity, increased metabolism and transport. The extent to which target-site resistance mutations contribute to the resistance phenotype is, however, not well studied. Here, we used marker-assisted backcrossing to create 30 congenic lines carrying nine mutations (alone, or in combination in a few cases) associated with resistance to avermectins, pyrethroids, mite growth inhibitors and mitochondrial complex III inhibitors (QoI) in a polyphagous arthropod pest, the spider mite Tetranychus urticae. Toxicity tests revealed that mutations in the voltage-gated sodium channel, chitin synthase 1 and cytochrome b confer high levels of resistance and, when fixed in a population, these mutations alone can result in field failure of acaricide treatment. In contrast, although we confirmed the implication of mutations in glutamate-gated chloride channels in abamectin and milbemectin insensitivity, these mutations do not lead to the high resistance levels that are often reported in abamectin resistant strains of T. urticae. Overall, this study functionally validates reported target-site resistance mutations in T. urticae, by uncoupling them from additional mechanisms, allowing to finally investigate the strength of the conferred phenotype in vivo

THE COMPLEXITY OF THE CONCEPT OF FRAGILITY IN MEDICAL PRACTICE. CLINICAL IMPLICATIONS OF FRAGILITY SYNDROME IN ELDERLY PATIENTS ASSOCIATING CARDIOVASCULAR DISEASES

Frailty, in association with aging and chronic disorders, is a key predictive factor of the quality of life and accelerating aging, which contributes to disability and, therefore, has important social-economic implications. The frailty nosological specific features outline to elderly people must be emphasized with its correlative specificities. Without considering a gold standard, we consider that it is useful to make a few clear delimitations and correlations of various frailty concepts and of assessment methods and biomarkers that help easy identify fragile patients. Frailty is a result of the decreasing quality of life in the elderly, that could be prevented, postponed or even improved by well-defined multi-factorial interventions

Abdominal shotgun trauma: A case report

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

The relationship between quality of life and functionality in patients with schizophrenia – A preliminary report

This research aimed to investigate the relationship between quality of life and level of functioning in a group of Romanian patients with schizophrenia. A cross-sectional, observational, prospective study on 47 patients with schizophrenia was conducted. Socio-demographic and clinical data were documented and WHOQOL-BREF and Life Skills Profile-16 instruments were further administered. To examine the correlation between variables, Person correlation test was employed. The mean age of the sample was 38.32±12.32 years and 66% of the patients were males. Significant correlations were found between all aspects of the quality of life (physical health, psychological, social relationships and environmental health) and total score of LSP-16 (r=-0.426,