Prion protein gene polymorphisms in classical scrapie-affected flocks of sheep in Central Macedonia

Για την έρευνα αυτή πραγματοποιήθηκε αιμοληψία σε 1.456 πρόβατα, ηλικίας 1-7 ετών, από 7 θετικές εκτροφές της Ημαθίας και της Θεσσαλονίκης, ως προς την κλασσική τρομώδη νόσο, στα πλαίσια της εφαρμογής του Εθνικού Προγράμματος Επιτήρησης, Ελέγχου και Εξάλειψης των Μεταδοτικών Σπογγωδών Εγκεφαλοπαθειών (ΜΣΕ) στα μικρά μηρυκαστικά του Υπουργείου Αγροτικής Ανάπτυξης & Τροφίμων, για τα έτη 2009-2013. Τα αιμοδείγματα συλλέχθηκαν από κρατικούς κτηνιάτρους των κτηνιατρικών υπηρεσιών της Ημαθίας και Θεσσαλονίκης και οι γονοτυπικές εξετάσεις πραγματοποιήθηκαν στο Εθνικό Εργαστήριο Αναφοράς για τις ΜΣΕ, Κτηνιατρικό Εργαστήριο Λάρισας με τη μέθοδο της αλυσιδωτής αντίδρασης της πολυμεράσης σε πραγματικό χρόνο (Real-time PCR). Από τα 1.456 πρόβατα που εξετάστηκαν, τα 340 ήταν φυλής Χίου, τα 633 ημίαιμα Χίου και τα 483 ημίαιμα. Τα εξεταζόμενα ζώα παρουσίασαν μεγάλη γενετική ποικιλομορφία, καθώς βρέθηκαν 7 διαφορετικά αλληλόμορφα και 23 διαφορετικοί γονότυποι. Ο κυρίαρχος γονότυπος σε όλες τις φυλές ήταν ο ARQ/ARQ,ακολουθούμενος από τον ARR/ARQ. Ο απλότυπος TRQ ήταν συχνός στα Χιώτικα και στα ημίαιμα Χίου πρόβατα, ενώ ο απλότυπος VRQ ήταν σπάνιος σε όλες τις φυλές. Είναι αξιοσημείωτο ότι 3 γονότυποι (ARH/TRQ, ARR/ARKκαι ARK/VRQ) ανιχνεύθηκαν για πρώτη φορά στην Ελλάδα. Επιπλέον, 2 από αυτούς τους γονότυπους (ARH/TRQ, και ARK/VRQ) δεν έχουν, καθ’ όσον γνωρίζουμε, αναφερθεί ποτέ μέχρι σήμερα. Επίσης, δίνεται έμφαση στο γεγονός ότι στη χώρα μας παρατηρούνται τα περισσότερα κρούσματα κλασσικής τρομώδους νόσου των προβάτων κάθε χρόνο σε όλη την Ευρώπη. Συνεπώς είναι επιτακτική ανάγκη να μειωθεί η επίπτωση της νόσου στην Ελλάδα, μέσω της εφαρμογής προγραμμάτων επιλογής/αναπαραγωγής ανθεκτικών προβάτων (selective breeding programs) ως προς την κλασσική τρομώδη νόσο. Σε αυτό συνηγορεί το γεγονός ότι ο επιπολασμός της νόσου στην Ελλάδα συνδέεται με τον γονότυπο ARQ/ARQ, ο οποίος είναι ο πιο συχνός γονότυπος. Άρα η δραστική μείωση του ARQ/ARQ και άλλων ευαίσθητων γονότυπων θα προκαλέσει δραματική μείωση της επίπτωσης της νόσου, προς όφελος της προβατοτροφίας και της εθνικής μας οικονομίας.The allele and genotype frequency distributions of the prion protein gene polymorphisms at codons 136, 154 and 171 were determined by real-time PCR for 1,456 sheep from 7 classical scrapie-affected flocks of Thessaloniki and Imathia, Central Macedonia, Greece. The blood samples were collected by official veterinarians and were examined by the National Reference Laboratory (NRL) for TSEs, Veterinary Laboratory of Larisa, Greece, in the framework of the National Program for Scrapie Surveillance and Control between 2009 and 2013. Among the 1,456 sheep, 340 were of Chios breed, 633 Chios crossbred and 483 crossbred. The examined sheep showed high genotype variability, as a total of 7 haplotypes and 23 different genotypes were found. The predominant allele and the predominant genotype were ARQ and ARQ/ARQ respectively, in all breeds studied, followed by the ARR allele and the ARR/ARQ genotype. The TRQ allele was frequent in Chios and Chios crossbred, while the VRQ allele was rare for all the breeds. Interestingly, 3 genotypes (ARH/TRQ, ARR/ARK and ARK/VRQ) were detected for the first time in Greece and two of them (ARH/TRQ and ARK/VRQ) have, to our knowledge, never been previously reported. Furthermore, it is emphasized that our country outnumbers all European countries in classical scrapie cases of sheep every year. Therefore, there is an urgent need to reduce the incidence of classical scrapie through the implementation of selective breeding programs. This is supported by the fact that the prevalence of classical scrapie in the Greek sheep population is highly associated with the predominant genotype ARQ/ARQ. Therefore, the elimination of the ARQ/ARQ and the other susceptible genotypes (belonging to Risk Groups 3 and 5, according to the National Scrapie Plan of Great Britain) would reduce dramatically the incidence of classical scrapie in Greece

Antimicrobial activity of apple cider vinegar against Escherichia coli, Staphylococcus aureus and Candida albicans; downregulating cytokine and microbial protein expression

The global escalation in antibiotic resistance cases means alternative antimicrobials are essential. The aim of this study was to investigate the antimicrobial capacity of apple cider vinegar (ACV) against E. coli, S. aureus and C. albicans. The minimum dilution of ACV required for growth inhibition varied for each microbial species. For C. albicans, a 1/2 ACV had the strongest effect, S. aureus, a 1/25 dilution ACV was required, whereas for E-coli cultures, a 1/50 ACV dilution was required (p < 0.05). Monocyte co-culture with microbes alongside ACV resulted in dose dependent downregulation of inflammatory cytokines (TNFα, IL-6). Results are expressed as percentage decreases in cytokine secretion comparing ACV treated with non-ACV treated monocytes cultured with E-coli (TNFα, 99.2%; IL-6, 98%), S. aureus (TNFα, 90%; IL-6, 83%) and C. albicans (TNFα, 83.3%; IL-6, 90.1%) respectively. Proteomic analyses of microbes demonstrated that ACV impaired cell integrity, organelles and protein expression. ACV treatment resulted in an absence in expression of DNA starvation protein, citrate synthase, isocitrate and malate dehydrogenases in E-coli; chaperone protein DNak and ftsz in S. aureus and pyruvate kinase, 6-phosphogluconate dehydrogenase, fructose bisphosphate were among the enzymes absent in C.albican cultures. The results demonstrate ACV has multiple antimicrobial potential with clinical therapeutic implications

Recommendations from the European Working Group for Value Assessment and Funding Processes in Rare Diseases (ORPH-VAL)

International audienceAbstractRare diseases are an important public health issue with high unmet need. The introduction of the EU Regulation on orphan medicinal products (OMP) has been successful in stimulating investment in the research and development of OMPs. Despite this advancement, patients do not have universal access to these new medicines. There are many factors that affect OMP uptake, but one of the most important is the difficulty of making pricing and reimbursement (P&R) decisions in rare diseases. Until now, there has been little consensus on the most appropriate assessment criteria, perspective or appraisal process. This paper proposes nine principles to help improve the consistency of OMP P&R assessment in Europe and ensure that value assessment, pricing and funding processes reflect the specificities of rare diseases and contribute to both the sustainability of healthcare systems and the sustainability of innovation in this field. These recommendations are the output of the European Working Group for Value Assessment and Funding Processes in Rare Diseases (ORPH-VAL), a collaboration between rare disease experts, patient representatives, academics, health technology assessment (HTA) practitioners, politicians and industry representatives. ORPH-VAL reached its recommendations through careful consideration of existing OMP P&R literature and through a wide consultation with expert stakeholders, including payers, regulators and patients. The principles cover four areas: OMP decision criteria, OMP decision process, OMP sustainable funding systems and European co-ordination. This paper also presents a guide to the core elements of value relevant to OMPs that should be consistently considered in all OMP appraisals. The principles outlined in this paper may be helpful in drawing together an emerging consensus on this topic and identifying areas where consistency in payer approach could be achievable and beneficial. All stakeholders have an obligation to work together to ensure that the promise of OMP’s is realised

Ispitivanje antidepresivnog, sedativnog i analgetskog djelovanja novih fuzioniranih derivata tiofena

This study was aimed at the synthesis of fused benzothiophene derivatives containing heterocyclic moiety. The reaction of the tetrahydrobenzo[b]thiophene derivatives 1a,b with ethoxycarbonylisothiocyanate afforded the thiourea derivatives 2a,b. Cyclization of the latter products gave the annulated benzo[b]thienopyrimidine derivatives 3a,b. Compounds 2a,b and 3a underwent a series of heterocyclization reactions through the reaction with some chemical reagents to give the new benzo[b]thienopyrimidine derivatives 5a,b to 8a-c. Also, this work was extended to study the potential role of the novel synthesized thiourea derivative 2a and benzo[b]thienopyrimidine derivatives 3a, 5b, 6a and 8b as antidepressant, sedative or analgesic agents at two doses (15 or 30 mg kg1 body mass). Some compounds (2a, 3a and 5b) showed mild antidepressant activity in the forced-swimming test. No compound showed sedative effect. Visceral pain evoked by i.p. injection of acetic acid in mice was significantly inhibited by all compounds at a high doses.U radu je opisana sinteza fuzioniranih derivata benzotiofena koji sadrže heterociklički ostatak bitan za farmakološko djelovanje. Tiourea derivati 2a,b dobiveni su reakcijom derivata tetrahidrobenzo[b]tiofena 1a,b s etoksikarbonilizotiocijanatom. Iz njih su dalje priređeni anulirani derivati benzo[b]tienopirimidina 3a,b. Spojevi 2a,b i 3a prevedeni su reakcijama heterociklizacije u benzo[b]tienopirimidine 5a,b-8a-c. Ispitivano je antidepresivno, sedativno i analgetsko djelovanje novosintetiziranih derivata tiouree 2a i benzo[b]tienopirimidina 3a, 5b, 6a i 8b u dvije doze (15 ili 30 mg kg1 tjelesne mase). Spojevi 2a, 3a i 5b pokazali su blago antidepresivno djelovanje u testu forsiranog plivanja, dok sedativni učinak nije pokazao niti jedan ispitivani spoj. Visceralna bol inducirana i.p. injekcijom octene kiseline u miševa značajno je inhibirana sa svim spojevima, ali u visokim dozama

Systemic immunity shapes the oral microbiome and susceptibility to bisphosphonate-associated osteonecrosis of the jaw

Background Osteonecrosis of the jaw (ONJ) is a rare but serious adverse drug effect linked to long-term and/or high-dose exposure to nitrogen-bisphosphonates (N-BP), the standard of care for the treatment of bone fragility disorders. The mechanism leading to bisphosphonate-associated ONJ (BAONJ) is unclear and optimal treatment strategies are lacking. Recent evidence suggests that BAONJ may be linked to drug-induced immune dysfunction, possibly associated with increased susceptibility to infections in the oral cavity. The objective of this investigation was to comprehensively assess the relationship linking immune function, N-BP exposure, the oral microbiome and ONJ susceptibility. Methods Leukocyte gene expression of factors important for immunity, wound healing and barrier function were assessed by real-time quantitative PCR and the oral microbiome was characterized by 454 pyrosequencing of the 16S rRNA gene in 93 subjects stratified by N-BP exposure and a history of ONJ. Results There were marked differences in the systemic expression of genes regulating immune and barrier functions including RANK (p = 0.007), aryl hydrocarbon receptor (AHR, p < 0.001), and FGF9 (p < 0.001), which were collectively up-regulated in individuals exposed to N-BP without ONJ relative to treatment controls. In contrast, the expression levels of these same genes were significantly down-regulated in those who had experienced BAONJ. Surprisingly, the oral microbiome composition was not directly linked to either BAONJ or N-BP exposure, rather the systemic leukocyte expression levels of RANK, TNFA and AHR each explained 9% (p = 0.04), 12% (p = 0.01), and 7% (p = 0.03) of the oral bacterial beta diversity. Conclusions The oral microbiome is unlikely causative of ONJ, rather individuals with BAONJ lacked immune resiliency which impaired their capacity to respond adequately to the immunological stress of N-BP treatment. This may be the common factor linking N-BP and anti-RANK agents to ONJ in at-risk individuals. Preventive and/or therapeutic strategies should target the wound healing deficits present in those with ONJ

Two N-substituted 3,5-diphenyl-2-pyrazoline-1-thiocarboxamides

The structures of N-ethyl-3-(4-fluorophenyl)-5-(4-methoxyphenyl)-2-pyrazoline-1-thiocarboxamide, C19H20FN3OS, (I), and 3-(4-fluorophenyl)-N-methyl-5-(4-methylphenyl)-2-pyrazoline-1-thiocarboxamide, C18H18FN3S, (II), have similar geometric parameters. The methoxy/methyl-substituted phenyl groups are almost perpendicular to the pyrazoline (pyraz) ring [interplanar angles of 89.29 (8) and 80.39 (10)degrees for (I) and (II), respectively], which is coplanar with the fluorophenyl ring [ interplanar angles of 5.72 ( 9) and 10.48 (10)degrees]. The pyrazoline ring approximates an envelope conformation in both structures, with the two-coordinate N atom involved in an intramolecular N-H center dot center dot center dot N-pyraz interaction. In (I), N-H center dot center dot center dot O and C-H center dot center dot center dot S intermolecular hydrogen bonds are the primary interactions, whereas in (II), there are no intermolecular hydrogen bonds

Synthesis and antimicrobial activity of some 1,3,4-oxadiazole derivatives

Six new 5-(1-/2-naphthyloxymethyl)-1,3,4-oxadiazole-2(3H)-thione, 2-amino-5-(1-/2-naphthyloxymethyl)-1,3,4-oxadiazole, 5-(1-/ 2-naphthyloxymethyl)-1,3,4-oxadiazole-2(3H)-one derivatives have been synthesized from 1-and/or 2-naphthol. The structures,of the compounds were confirmed by IR and H-1 NMR spectral data and microanalysis. The antimicrobial properties of the compounds were investigated against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa, Candida albicans, C. krusei and C. parapsilosis using microbroth dilution method. 2-Amino-5-(2-naphthytoxymethyl)-1,3,4-oxadiazole and 5-(2-naphthyloxymethyl)-1,3,4-oxadiazole-2(3H)-one show significantly (32 mug/ml), compounds 5-(1-/2-naphthyloxymethyl)-1,3,4-oxadiazole-2(3H)-thione, 2-amino-5-(1-naphthyloxymethyl)-1,3,4-oxadiazole and 5-(1-naphthyloxymethyl)-1,3,4-oxadiazole-2(3H)-one moderately (64 mug/ml) active against C. krusei. All the compounds were active against S. a ureus,. coli, P. aeruginosa, C. albicans, and C. parapsilosis at 64-256 mug/ml concentration. (C) 2002 Editions scientifiques et medicales Elsevier SAS. All rights reserved