Comparative Testing Report on the Detection and Quantification of Maize Events GA21, TC1507 and MIR604

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This preliminary report describes the outcome of the fourth comparative testing round ILC-EURL-GMFF-CT-02/11. Participants were required to screen two test items denoted maize powder levels 1 and 2, for the presence of maize events 3272, Bt11, Bt176, 59122, GA21, MIR604, MON 810, MON 863, NK603 and TC1507. Any events detected were then to be quantified. This comparative testing round was organised in collaboration with the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize test items were produced in-house. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 159 laboratories were invited to participate in ILC-EURL-GMFF-CT-02/11. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. In this fourth comparative testing round greater than 86 % of participants gained a satisfactory z-score in the range of -2 to +2 for the results expressed in mass/mass % for both maize powder levels 1 and 2 regardless of the GM event. However, a lower percentage (43–86 %) of z-scores within the working range of -2 to +2 was calculated for those participants that expressed the results in copy/copy %.JRC.I.3-Molecular Biology and Genomic

Comparative Testing Report on the Detection and Quantification of Maize Event MON 810 - Comparative testing round: ILC-CRL-GMFF-CT-02/10

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the second comparative testing round ILC-CRL-GMFF-CT-02/10. Participants had to determine the GM content in two test items denoted maize powder levels 1 and 2, containing different GM percentages of maize event MON 810. This comparative testing round was organised in collaboration with the Reference Materials Unit and the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize event MON 810 test items were produced by the Reference Materials Unit. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 136 laboratories were invited to participate in ILC-CRL-GMFF-CT-02/10. Six National Reference Laboratories declined participation, of which two were no longer a National Reference Laboratory. Ninety laboratories from 41 countries returned results, of which 65 were National Reference Laboratories, six were members of the European Network of GMO Laboratories only and 19 were laboratories from third countries. Two National Reference Laboratories, two Official control laboratories and nine laboratories from a third country did not submit any results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. The outcome of this second comparative testing round was in general positive, with 82-100 % of participants gaining a z-score in the range of -2 to +2 for both maize powder levels 1 and 2 regardless of the calibration method, the measurement unit and the approach used for calculating the z-score.JRC.I.3-Molecular Biology and Genomic

THE EXPRESSION PROFILES OF SELECTED GENES IN DIFFERENT BEAN SPECIES (PHASEOLUS SPP.) AS RESPONSE TO WATER DEFICIT

The aim of this study was to compare expression profiles of a number of transcripts from leaves of different Phaseolus species under drought stress, in order to ascertain whether changes in their expression in Phaseolus spp. are part of a general or a species specific response to drought. Relative gene expression analysis using quantitative PCR were carried out in P. coccineus, P. lunatus and P. acutifolius for 13 transcripts previously identified as up- or down-regulated in leaves of P. vulgaris. The mode of expression was found consistent within Phaseolus spp., despite the fact that the four species differ in their responses to drought at the physiological and morphological levels. The present results suggest that this is a common feature of the response of Phaseolus spp. The majority of the genes shown here to be influenced by water deficit in beans have been reported in other plant species under similar conditions, suggesting that they play a role in the general response to drought stress

Comparative Testing Report on the Detection and Quantification of Maize Event NK603 - Comparative testing round: ILC-CRL-GMFF-CT-01/10

In the frame of Regulation EC (No) 882/2004, the European Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the first comparative testing round ILC-CRL-GMFF-CT-01/10. Participants had to determine the GM content in two test items denoted maize powder level 1 and level 2, containing different GM percentages of maize event NK603. This comparative testing round was organised in collaboration with the Reference Materials Unit and the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize event NK603 test items were produced by the Reference Materials Unit. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 110 laboratories were invited to participate in ILC-CRL-GMFF-CT-01/10. Five National Reference Laboratories declined participation. Ninety-three results were returned from 84 laboratories from 36 countries, of which 66 were National Reference Laboratories, seven were members of the European Network of GMO Laboratories only and 11 were laboratories from third countries. Two National Reference Laboratories, two European Network of GMO Laboratories only members and one laboratory from a third country did not submit any results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. The outcome of this first comparative testing round was in general positive with a share of 86-96 % and 94-100 % of participants exhibiting a z-score in the range of -2 to +2 for maize powder level 1 and level 2, respectively.JRC.DG.I.4-Molecular biology and genomic

Examination of Genetic Diversity of Common Bean from the Western Balkans

In this study, genetic diversity of 119 accessions of common bean (Phaseolus vulgaris) from five former Yugoslav republics constituting the western Balkans was assessed by 13 microsatellite markers. This set of markers has proven before to efficiently distinguish between bean genotypes and assign them to either the Andean or the Mesoamerican gene pool of origin. In this study, 118 alleles were detected or 9.1 per locus on average. Four groups (i.e., Slovene, Croatian, Bosnian, and Serbian) showed similarly high levels of genetic diversity as estimated by the number of different alleles, number of effective alleles, Shannon's information index, and expected heterozygosity. Mildly narrower genetic diversity was identified within a group of Macedonian accessions; however, this germplasm yielded the highest number of private alleles. All five germplasms share a great portion of genetic diversity as indicated by the analysis of molecular variance (AMOVA). On the basis of the scored number of migrants, we concluded that the most intensive gene flow in the region exists in Bosnia and Herzegovina. Cluster analysis based on collected molecular data classified the accessions into two large clusters that corresponded to two gene pools of origin (i.e., Andean and Mesoamerican). We found that Andean genotypes are more prevalent than Mesoamerican in all studied countries, except Macedonia, where the two gene pools are represented evenly. This could indicate that common bean was introduced into the western Balkans mainly from the Mediterranean Basin. Bayesian cluster analysis revealed that in the area studied an additional variation exists which is related to the Andean gene pool. Different scenarios of the origin of this variation are discussed in the article

Genetic diversity of the autochtonous Phaseolus bean germplasm originating from five South east European countries

Genetic differentiation of 167 Phaseolus vulgaris L. accessions originating from five South east European countries was performed using 13 SSR markers. Analysis included 21 accessions from Bosnia and Herzegovina, 15 accessions from Croatia, 73 accessions from Macedonia, 38 accessions from Serbia and 20 accessions from Slovenia. Calculated mean number of alleles per locus was 9.8. The average polymorphic information content over all loci reached value of 0.710 where the most informative locus was GATS91 (0.926). These results showed that selected set of SSR markers are highly informative and applicable for studies of genetic diversity within germplasm collected in the five South east European countries, which was also confirmed with the calculated value of probability of identity (1.831x10). High mean value of expected heterozygosity (0.731) and Shannon’s information index (1.486) reflects high genetic diversity within accessions from five countries, where accessions from Macedonia are genetically the most uniform, due to 17completely identical accessions included

The Immune Response to Herpes Simplex Virus Encephalitis in Mice Is Modulated by Dietary Vitamin E

Herpes simplex virus encephalitis (HSE) is the most common fatal sporadic encephalitis in humans. HSE is primarily caused by herpes simplex virus (HSV)-1 infection of the brain. HSE results in increased levels of oxidative stress, including the production of reactive oxygen species, free radicals, and neuroinflammation. The most biologically active form of vitamin E (VE) is α-tocopherol (α-TOC). In cellular membranes, α-TOC prevents lipid peroxidation by scavenging free radicals and functioning as an antioxidant. Supplementation with VE has been shown to decrease immunosenescence, improve immune function, and may be neuroprotective. To determine how VE deficiency and VE supplementation would alter the pathogenesis of HSE, we placed weanling male BALB/cByJ mice on VE-deficient (VE-D), VE-adequate (VE-A), or 10× VE-supplemented diets for 4 wk, and then infected the mice intranasally with HSV-1. VE-D mice had more severe symptoms of encephalitis than VE-A mice, including weight loss, keratitis, hunched posture, and morbidity. VE-D mice had increased cytokine and chemokine expression in the brain and increased viral titers. In contrast, VE supplementation failed to decrease cytokine production and had no effect on viral titer. We demonstrated that adequate levels of VE are important in limiting HSE pathology and that 10× supplementation does not enhance protection

Demagnetisation procedures

U ovom radu je obrađena problematika demagnetizacije uzoraka nakon provedenih nerazornih ispitivanja magnetskom metodom. U radu je objašnjeno kako nastaje magnetizam, te kako različiti materijali reagiraju na vanjsko magnetsko polje. Detaljnije su obrađeni feromagnetski materijali koji su u ovom slučaju najzanimljiviji. Pojašnjeni su relevantni parametri potrebni za opisivanje magnetskog polja kako u okolini tako i u samom uzorku. Ukratko su obrađene vrste električne struje koje se koriste prilikom magnetizacije uzoraka pri nerazornim ispitivanjima magnetskom metodom. \Nakon kratkog uvoda u osnove magnetizma i načina na koji se materijali ponašaju prilikom izlaganja magnetskom polju, obrađena je problematika demagnetizacije uzoraka. Prikazano je u kojim slučajevima je postupak demagnetizacije obavezan odnosno u kojim slučajevima nije obavezan. Objašnjeni su postupci demagnetizacije, oprema kojom je moguće mjeriti jakost zaostalog magnetskog polja i oprema kojom je moguće provesti postupak demagnetizacije. U zadnjem poglavlju prikazana su provedena mjerenja jakosti zaostalog magnetizma na dva uzorka prije i nakon postupka demagnetizacije, te rezultati provedenih mjerenja. Na kraju ovog rada dan je zaključak

Demagnetisation procedures

U ovom radu je obrađena problematika demagnetizacije uzoraka nakon provedenih nerazornih ispitivanja magnetskom metodom. U radu je objašnjeno kako nastaje magnetizam, te kako različiti materijali reagiraju na vanjsko magnetsko polje. Detaljnije su obrađeni feromagnetski materijali koji su u ovom slučaju najzanimljiviji. Pojašnjeni su relevantni parametri potrebni za opisivanje magnetskog polja kako u okolini tako i u samom uzorku. Ukratko su obrađene vrste električne struje koje se koriste prilikom magnetizacije uzoraka pri nerazornim ispitivanjima magnetskom metodom. \Nakon kratkog uvoda u osnove magnetizma i načina na koji se materijali ponašaju prilikom izlaganja magnetskom polju, obrađena je problematika demagnetizacije uzoraka. Prikazano je u kojim slučajevima je postupak demagnetizacije obavezan odnosno u kojim slučajevima nije obavezan. Objašnjeni su postupci demagnetizacije, oprema kojom je moguće mjeriti jakost zaostalog magnetskog polja i oprema kojom je moguće provesti postupak demagnetizacije. U zadnjem poglavlju prikazana su provedena mjerenja jakosti zaostalog magnetizma na dva uzorka prije i nakon postupka demagnetizacije, te rezultati provedenih mjerenja. Na kraju ovog rada dan je zaključak

Demagnetisation procedures

U ovom radu je obrađena problematika demagnetizacije uzoraka nakon provedenih nerazornih ispitivanja magnetskom metodom. U radu je objašnjeno kako nastaje magnetizam, te kako različiti materijali reagiraju na vanjsko magnetsko polje. Detaljnije su obrađeni feromagnetski materijali koji su u ovom slučaju najzanimljiviji. Pojašnjeni su relevantni parametri potrebni za opisivanje magnetskog polja kako u okolini tako i u samom uzorku. Ukratko su obrađene vrste električne struje koje se koriste prilikom magnetizacije uzoraka pri nerazornim ispitivanjima magnetskom metodom. \Nakon kratkog uvoda u osnove magnetizma i načina na koji se materijali ponašaju prilikom izlaganja magnetskom polju, obrađena je problematika demagnetizacije uzoraka. Prikazano je u kojim slučajevima je postupak demagnetizacije obavezan odnosno u kojim slučajevima nije obavezan. Objašnjeni su postupci demagnetizacije, oprema kojom je moguće mjeriti jakost zaostalog magnetskog polja i oprema kojom je moguće provesti postupak demagnetizacije. U zadnjem poglavlju prikazana su provedena mjerenja jakosti zaostalog magnetizma na dva uzorka prije i nakon postupka demagnetizacije, te rezultati provedenih mjerenja. Na kraju ovog rada dan je zaključak