Inhibition of human glutathione transferases by pesticides: Development of a simple analytical assay for the quantification of pesticides in water

Glutathione transferases (GSTs; EC 2.5.1.18) form a group of multifunctional enzymes that are involved in phase II cellular detoxification mechanism. Here, screening of the inhibition potency of a wide range of pesticides toward selected human GST isoenzymes (hGSTA1-1, hGSTP1-1, hGSTT2-2 and hGSTO1-1) was carried out. hGSTA1-1 was found more susceptible to inhibition by pesticides than other isoenzymes. The insecticides dieldrin and spiromesifen were identified as potent reversible inhibitors toward hGSTA1- 1 with IC50 values equal to 17.9 ± 1.7 M and 12.1 ± 3.4 M, respectively. Based on in silico docking analysis and kinetic inhibition studies it was concluded that dieldrin and spiromesifen bind specifically to the enzyme presumably at a distinct position that partially overlaps with both the G- and H-site. The ability of dieldrin and spiromesifen to inhibit hGSTA1-1 activity was exploited for the development of analytical quantification assays for these two pesticides. Linear calibration curves were obtained for dieldrin and spiromesifen, with useful concentration in the range of 0–10 M. The reproducibility of the assay response, expressed by relative standard deviation, was in the order of 4.1% (N = 28). The method was successfully applied to the determination of these pesticides in real water samples without sample preparation steps

HLA-DPB1*03 as Risk Allele and HLA-DPB1*04 as Protective Allele for Both Early- and Adult-Onset Multiple Sclerosis in a Hellenic Cohort

Background: Human Leucocyte Antigens (HLA) represent the genetic loci most strongly linked to Multiple Sclerosis (MS). Apart from HLA-DR and HLA–DQ, HLA-DP alleles have been previously studied regarding their role in MS pathogenesis, but to a much lesser extent. Our objective was to investigate the risk/resistance influence of HLA-DPB1 alleles in Hellenic patients with early- and adult-onset MS (EOMS/AOMS), and possible associations with the HLA-DRB1*15:01 risk allele. Methods: One hundred MS-patients (28 EOMS, 72 AOMS) fulfilling the McDonald-2010 criteria were enrolled. HLA genotyping was performed with standard low-resolution Sequence-Specific Oligonucleotide techniques. Demographics, clinical and laboratory data were statistically processed using well-defined parametric and nonparametric methods and the SPSSv22.0 software. Results: No significant HLA-DPB1 differences were found between EOMS and AOMS patients for 23 distinct HLA-DPB1 and 12 HLA-DRB1 alleles. The HLA-DPB1*03 allele frequency was found to be significantly increased, and the HLA-DPB1*02 allele frequency significantly decreased, in AOMS patients compared to controls. The HLA-DPB1*04 allele was to be found significantly decreased in AOMS and EOMS patients compared to controls. Conclusions: Our study supports the previously reported risk susceptibility role of the HLA-DPB1*03 allele in AOMS among Caucasians. Additionally, we report for the first time a protective role of the HLA-DPB1*04 allele among Hellenic patients with both EOMS and AOMS

One-year infliximab administration for the treatment of chronic refractory pouchitis

Abstract Aim To determine the efficacy of infliximab in the treatment of chronic refractory pouchitis, following ileo-pouch anal anastomosis (IPAA) for ulcerative colitis (UC). Methods Seven patients (4 females, 3 males) with chronic refractory pouchitis were included in an open study. Pouchitis was diagnosed by clinical plus endoscopic and histological criteria. Three patients also had fistulae (pouch-bladder in 1 and perianal in 2). Extraintestinal manifestations were also present in 4 patients (erythema nodosum in 2, arthralgiae in 2). All patients were refractory to standard therapy. Crohn's disease was carefully excluded in all patients after re-evaluation of the history and examination of the small bowel with enteroclysis or small bowel capsule endoscopy. Patients received Infliximab 5 mg/kg at 0, 2 and 6 weeks and thereafter every 2 months for 1 year. Clinical response was classified as complete, partial, and no response. Fistulae closure was classified as complete, partial, and no closure. The pouchitis disease activity index (PDAI) was also used as an outcome measure. Results Clinically, all patients improved. After 1 year of follow-up, 5 of the 7 patients had a complete clinical response, and 2 of the 3 patients with a fistula had complete fistulae closure. At the end of the follow-up period the median PDAI dropped from 11 (baseline) (range, 10-14) to 5 (range, 3-8). Extraintestinal manifestations were in complete remission at the end of the followup period as well. Conclusions Our results indicate that infliximab may be recommended for the treatment of chronic refractory pouchitis complicated or not by fistulae following IPAA for UC

Survivors of war in the Northern Kosovo (II): baseline clinical and functional assessment and lasting effects on the health of a vulnerable population

<p>Abstract</p> <p>Background</p> <p>This study documents torture and injury experience and investigates emotional well-being of victims of massive violence identified during a household survey in Mitrovicë district in Kosovo. Their physical health indicators such as body mass index (BMI), handgrip strength and standing balance were also measured. A further aim is to suggest approaches for developing and monitoring rehabilitation programmes.</p> <p>Methods</p> <p>A detailed assessment was carried out on 63 male and 62 female victims. Interviews and physical examination provided information about traumatic exposure, injuries, and intensity and frequency of pain. Emotional well-being was assessed using the "WHO-5 Well-Being" score. Height, weight, handgrip strength and standing balance performance were measured.</p> <p>Results</p> <p>Around 50% of victims had experienced at least two types of torture methods and reported at least two injury locations; 70% had moderate or severe pain and 92% reported constant or periodic pain within the previous two weeks. Only 10% of the victims were in paid employment. Nearly 90% of victims had experienced at least four types of emotional disturbances within the previous two weeks, and many had low scores for emotional well-being. This was found to be associated with severe pain, higher exposure to violence and human rights violations and with a low educational level, unemployment and the absence of political or social involvement.</p> <p>Over two thirds of victims were overweight or obese. They showed marked decline in handgrip strength and only 19 victims managed to maintain standing balance. Those who were employed or had a higher education level, who did not take anti-depressant or anxiety drugs and had better emotional well-being or no pain complaints showed better handgrip strength and standing balance.</p> <p>Conclusions</p> <p>The victims reported a high prevalence of severe pain and emotional disturbance. They showed high BMI and a reduced level of physical fitness. Education, employment, political and social participation were associated with emotional well-being. Interventions to promote physical activity and social participation are recommended. The results indicate that the rapid assessment procedure used here offers an adequate tool for collecting data for the monitoring of health interventions among the most vulnerable groups of a population exposed to violence.</p

Immobilized enzymes as on-line probes in biochemistry and new drug discovery : biosynthesis of catecholamines

The use of immobilized enzymes has steadily increased in recent years. Based upon the advantages that immobilized enzymes possess over soluble enzymes, numerous applications have emerged in medical and analytical fields. This work demonstrates the applicability of a liquid chromatographic system based upon coupled on-line immobilized enzyme reactors (IMERs) to organic synthesis, biochemistry and pharmacology. It is envisioned that the model system will grow into a modular process where synthetic chemists can add or subtract the enzymes necessary for their particular synthetic goal. The system allows for on-line chromatographic purification and structural identification of products and could greatly reduce time required to discover new synthetic pathways. In addition, the construction of a coupled enzyme system provides a number of approaches to basic research into synthetic and metabolic pathways as well as a rapid method for the discovery of new pharmaceutical substances.A coupled system using extremely different enzymes with incompatible cofactors and reaction conditions has been constructed. The significance of the proposed project not only lies in the development of the liquid chromatographic on-line enzyme cascade but also in the biosynthetic pathway chosen for this study. The biosynthetic pathway involving dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase encompass the synthesis of the key transmitters, norepinephrine and epinephrine. The results demonstrate for the first time the immobilization of dopamine beta-hydroxylase and phenylethanolamine N-methyltransferase. The IMERs are active and can be used in a liquid chromatographic format for qualitative and quantitative determinations. Studies with the IMER-HPLC systems have also shown that the activity of the immobilized enzymes reflects the non-immobilized enzymes. Thus, the IMER-HPLC system can be used to carry out standard Michaelis-Menten enzyme kinetic studies and to quantitatively determine enzyme kinetic constants, identify specific enzyme inhibitors, provide information regarding the mode of inhibition and the inhibitor constants (Ki). The immobilized enzyme reactors used independently or as a combination will provide a unique opportunity to explore the interrelationships between these enzymes, to investigate the source of catecholamine-related disorders and to design new drug entities for identified clinical syndromes

Classroom climate and academic achievement with and without using comics in the curriculum subjects of Ancient and Modern Greek Language in Model Experimental Gymnasia of Greece: comparative study

Classroom climate has been identified as a key factor determining school effectiveness. Positive classroom climate is about nurturing an atmosphere that promotes learning, improves the relationships of pupils with their teachers and classmates, and cultivates attachment to curriculum subjects. The scope of this doctoral project is to examine the impact of comics and teaching with the help of comics on classroom climate, as well as on the cognitive understanding and academic achievements of pupils. Teaching interventions implemented in this study were based on Vygotsky’s principles of constructivism, the discovery learning model of Bruner and Gardner’s theory of multiple intelligences. The sample of research consisted of 651 pupils and 13 Greek teachers (nine women and four men) from 12 Model Experimental Schools. The data of research were collected in the academic year 2013-14 during the teaching of Ancient Greeks in the 2nd year of the Gymnasium. In particular, 150 lessons were observed (75 without using comics and 75 with the help of comics) aiming at giving answers to the project’s research questions. To this end, a variety of quantitative and qualitative approaches were selected and combined, i.e. teaching observation protocols, tests and questionnaires for pupils and interviews for teachers, in order to ensure the validity and reliability of research and to reach safe and valid conclusions about the impact of comics on learning, and more generally, classroom climate. The analysis of the data points to a significant difference in classroom climate and the cognitive understanding and academic achievements of pupils. Specifically, comics improved the educational atmosphere of classrooms, making lessons more interesting, lively and interactive. Teaching with comics also created positive attitudes and feelings in classrooms, affecting positively the mood of both pupils and teachers, as well as their academic performance. At the same time, teaching with the help of comics motivated pupils to take an active part in lessons, who ceased to be passive receivers of knowledge. Apart from cultivating pupils’ eagerness to participate in lessons, the contribution of comics was also crucial in increasing their levels of concentration throughout the lesson, thus promoting a better and deeper understanding of the new content that was being taught. Furthermore, pupils’ academic achievement in lessons where comics were used increased significantly (17,83) compared to teaching without comics (14,47). Moreover, by increasing classroom workload, teaching with comics provided not only more opportunities to pupils to collaborate with classmates. It also gave more opportunities to teachers to observe and evaluate pupils in carrying out various tasks and activities related to comics, and also, to give them more feedback about the progress of their work, thus cultivating amongst pupils a strong sense of satisfaction and acceptance. To wrap things up, the diversification of teaching methodology with the utilisation of ICT in general and comics in particular affected positively pupils and facilitated teachers in carrying out their teaching task, highlighting that comics can be an effective supplementary learning tool. For these reasons, all stakeholders of education (Ministry of Education, Institute of Education Policy, teachers, etc.) ought to act in the direction of incorporating technology in teaching and to further enhance pupils’ learning experience.Η σχολική αποτελεσματικότητα συνδέεται άμεσα με το κλίμα που επικρατεί στη σχολική τάξη. Το θετικό και ευνοϊκό κλίμα της τάξης σχετίζεται με τη δημιουργία ενός περιβάλλοντος που προωθεί τη μάθηση, ενισχύοντας τις σχέσεις του μαθητή με τον εκπαιδευτικό, τους συμμαθητές του και το διδακτικό αντικείμενο. Σκοπός της παρούσας διδακτορικής διατριβής ήταν η διεξαγωγή μιας συγκριτικής μελέτης, που θα εξετάζει την πιθανή διαφοροποίηση του εκπαιδευτικού κλίματος της τάξης, της γνωστικής πρόσληψης και της σχολικής επίδοσης των μαθητών στις «παραδοσιακές» διδασκαλίες και τις διδασκαλίες με τη χρήση κινουμένων σχεδίων. Οι διδακτικές παρεμβάσεις της έρευνας βασίστηκαν στις αρχές του κονστρουκτιβισμού (Vygotsky), το διερευνητικό μοντέλο (Bruner) και τη θεωρία των πολλαπλών τύπων νοημοσύνης (Gardner). Το ερευνητικό δείγμα αποτέλεσαν 651 μαθητές και 13 εκπαιδευτικοί (εννέα γυναίκες και τέσσερις άνδρες) από 12 Πρότυπα Πειραματικά Γυμνάσια της Ελλάδας κατά τη διάρκεια του σχολικού έτους 2013-2014 στο πλαίσιο του μαθήματος της Αρχαίας Ελληνικής Γλώσσας από το πρωτότυπο στη Β’ Γυμνασίου τάξη στα Πειραματικά Γυμνάσια της χώρας. Αναλυτικότερα διεξήχθησαν από τους εκπαιδευτικούς 150 διδασκαλίες (75 χωρίς τη χρήση κινουμένων σχεδίων και 75 με τη χρήση κινουμένων σχεδίων) με σκοπό να δοθούν απαντήσεις στα ερευνητικά ερωτήματα. Για το λόγο αυτό επιλέχθηκε ένας συνδυασμός ερευνητικών μεθόδων (πολυμεθοδική προσέγγιση), αξιοποιώντας τα έντυπα παρατήρησης των διδασκαλιών, τα διαγνωστικά κριτήρια αξιολόγησης και τα ερωτηματολόγια των μαθητών, και την τεχνική των συνεντεύξεων των εκπαιδευτικών, ώστε να είναι όσο το δυνατόν πιο αξιόπιστες οι ερμηνείες για το υπό μελέτη αντικείμενο. Κατά τη συγκέντρωση και ανάλυση των δεδομένων έγινε τριγωνοποίηση με σκοπό την εξασφάλιση ασφαλών συμπερασμάτων για τη μαθησιακή διαδικασία με και χωρίς τη χρήση κινουμένων σχεδίων. Η ανάλυση των δεδομένων ανέδειξε ότι υπήρξε διαφοροποίηση στις διδασκαλίες χωρίς και με τη χρήση κινουμένων σχεδίων αναφορικά με το κλίμα της τάξης, την κατανόηση της νέας γνώσης από τους μαθητές και τη σχολική τους επίδοση. Συγκεκριμένα, φάνηκε ότι τα κινούμενα σχέδια βελτίωσαν το εκπαιδευτικό κλίμα, κάνοντας το μάθημα πιο ζωντανό, πιο δραστήριο και πιο διαδραστικό. Το θετικό εκπαιδευτικό κλίμα δημιούργησε στη σχολική αίθουσα ευχάριστα συναισθήματα, τα οποία επηρέαζαν θετικά την ψυχική διάθεση των μαθητών και των εκπαιδευτικών αναφορικά με την εκτέλεση των καθηκόντων τους, τη γενικότερη επίδοση των μαθητών και αυτό είχε ως αποτέλεσμα τη βελτίωση της αποτελεσματικότητας τους. Ταυτόχρονα ενεργοποιήθηκαν και διαμορφώθηκαν εσωτερικά κίνητρα στους μαθητές, οι οποίοι συμμετείχαν ενεργά στην εκπαίδευσή τους και δεν αποτελούσαν παθητικούς δέκτες πληροφοριών και γνώσης. Η συμβολή των κινουμένων σχεδίων υπήρξε καθοριστική και στην κατανόηση της νέας γνώσης των μαθητών, οι οποίοι με τη συμμετοχή τους, την επιθυμία τους για ενεργό ρόλο και την προσήλωση που επέδειξαν σε όλη τη διδακτική διαδικασία φάνηκαν να κατανοούν καλύτερα τη νέα πληροφορία. Επιπλέον, στις διδακτικές παρεμβάσεις με κινούμενα σχέδια οι μαθητές παρουσίασαν εμφανή βελτίωση της σχολικής τους επίδοσης, με το μέσο όρο των βαθμολογιών τους στις διδασκαλίες με κινούμενα σχέδια να έχει αυξηθεί σημαντικά (17,83) συγκριτικά με τις βαθμολογίες των ίδιων μαθητών στις διδασκαλίες χωρίς κινούμενα σχέδια (14,47). Παράλληλα, η αύξηση της σχολικής εργασίας έδωσε τη δυνατότητα στον εκπαιδευτικό της τάξης για συστηματικότερη παρακολούθηση και αξιολόγηση των μαθητών σε θέματα προόδου και επομένως περισσότερες ευκαιρίες ανατροφοδότησης, καλλιεργώντας στους μαθητές ένα αίσθημα ικανοποίησης και αποδοχής. Συνοψίζοντας, διαφαίνεται ότι η διαφοροποίηση του διδακτικού σχεδιασμού του μαθήματος και η ένταξη σ’ αυτό Ψηφιακών Τεχνολογιών και κινουμένων σχεδίων επέδρασε θετικά στους μαθητές και διευκόλυνε ουσιαστικά τους εκπαιδευτικούς, καταδεικνύοντας τα ως ένα αποτελεσματικό συμπληρωματικό εκπαιδευτικό εργαλείο στο πλαίσιο του μαθήματος. Για τους λόγους αυτούς καθίσταται αναγκαία η δράση όλων των εμπλεκόμενων φορέων (Υπουργείο Παιδείας, Ινστιτούτο Εκπαιδευτικής Πολιτικής, εκπαιδευτικοί κ.ά.) με σκοπό την ενσωμάτωση των ψηφιακών τεχνολογιών, παρέχοντας περισσότερες δυνατότητες ανάπτυξης και βελτίωσης στους μαθητές

SBI-fungicides : fungicidal effectiveness and resistance in «Botrytis cinerea»

In vitro fungitoxicity tests with 14 sterol biosynthesis inhibitors (SBIs) showed that the SBIs pyrifenox, flusilazol, propiconazole, triflumizole and fenpropimorph applied at the low concentration of 1 µg ml-1 inhibited the mycelial growth of wild-type and mutant strains of Botrytis cinerea that were resistant to the benzimidazoles, to the dicarboximides and to a mixture of benzimidazole+phenylcarbamate (carbendazim+diethofencarb). The SBIs tested exhibited higher effectiveness against the wild-type and mutant strains of B. cinerea compared with some widely used botryticides, such as the dicarboximides iprodione, procymidone and chlozolinate, the aromatic hydrocarbons quintozene, chloroneb and tolclofos-methyl, and the relatively new fungicides cyprodinil and fenhexamid. Only benomyl and fludioxonil presented higher effectiveness than the SBIs. In planta pot experiments with preventive applications of the commercial products Dorado 20 EC (pyrifenox), Punch 40 EC (flusilazol), Tilt 25 EC (propiconazole), Corbel 75 EC (fenpropimorph) and Trifmine 30 EC (triflumizole) showed that lesions of cucumber seedlings by all the abovementioned strains of B. cinerea were completely inhibited at low SBI concentrations of 0.05–0.1 g a.i. l-1. After chemical mutagenesis with N-methyl-N-nitro-N-nitrosoguanidine (MNNG), only strains with low resistance (Rf 6–9, based on MIC values) to triadimefon were isolated at a mutation frequency of 3.9 10-5. Cross-resistance studies with other SBIs showed that these triadimefon-resistant strains exhibited positive cross-resistance (Rf 2–10) to the other C-14 demethylase inhibitors (DMIs), but not to the morpholine fungicides fenpropimorph or tridemorph. Study of fitness of DMI-resistant strains showed that these mutation(s) were pleiotropic, with significant adverse effects on characteristics determining phytopathogenic fitness such as rate of mycelial growth, sporulation, conidial germination and pathogenicity on cucumber seedlings. The results indicate that some SBI-fungicides are suitable for use in resistance management programmes against grey mould

Psycho-neuro-endocrino-immunologic issues in multiple sclerosis: a critical review of clinical and therapeutic implications

Multiple sclerosis (MS) is a multifactorial, chronic, immune-mediated, and neurodegenerative disease, having a well-known hypothalamic-pituitary-adrenal (HPA) axis dysfunction. Several hormones have a great impact in the immune dysregulation, psychology, and cognitive status of patients with MS, as also in the fertility and response to treatment. In this comprehensive review, as an introduction, we mention basic data concerning MS: epidemiology, genetics, immunogenetics, epigenetics, pathophysiology, and neuroimmunology. Hormonal components of the disease cascade, mainly glucocorticoids (stress-related hormone), estrogens, prolactin and dehydroepiandrosterone (sex-related hormones), melatonin, and vitamin D, are discussed, aiming at focusing on core data regarding the impact of these hormones in MS pathophysiology, severity of the disease, correlation with comorbid mental disorders, and fertility. A great focus is given in the pre- and post-pregnancy period of MS patients, in the context of the disease-modifying treatments (DMTs) and HPA status, having in mind that there are only very limited knowledge and few papers on this specific life period of these women, having MS. All this data are presented in the main text and also in the workable tables, for the first time, suggesting targeted topics that need to be addressed in the near future. © 2020, Hellenic Endocrine Society