Report on the Verification of the Performance of 1507, 59122, MON 810 and NK603 Event-specific PCR-based Methods applied to DNA extracted from Stack Maize 1507 x 59122 x MON 810 x NK603

An application was submitted by Pioneer Overseas Corporation to request the authorization of the genetically modified maize stack 1507 x 59122 x MON 810 x NK603, resistant against certain lepidopteran pests, protected against corn rootworm larvae, and glufosinate-ammonium and glyphosate tolerant, and all sub-combinations of the individual events as present in the segregating progeny, for food and feed uses, and import and processing, in accordance with articles 5 and 17 of Regulation (EC) N° 1829/2003 GM Food and GM Feed. The unique identifier assigned to 1507 x 59122 x MON 810 x NK603 maize is DAS-Ø15Ø7-1xDAS-59122-7xMON-ØØ81Ø-6xMON-ØØ6Ø3-6. The genetically modified maize line 1507 x 59122 x MON 810 x NK603 has been obtained by conventional crossing of four genetically modified single maize events: 1507, 59122, MON 810 and NK603 without any new genetic modification. The EU-RL GMFF has previously validated, and declared fit for purpose, the detection methods for the single events 1507, 59122, MON 810 and NK603 (see: http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm). In line with the approach defined by the ENGL (http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf) the EU-RL GMFF therefore has carried out only an in-house verification of the performance of each validated method when applied to DNA extracted from 1507 x 59122 x MON 810 x NK603. The herewith reported in-house verification study lead to the conclusion that the individual methods meet the ENGL performance criteria also when applied to DNA extracted from the GM maize stack 1507 x 59122 x MON 810 x NK603.JRC.I.3-Molecular Biology and Genomic

Event-specific Method for the Quantification of Soybean DAS-68416-4 Using Real-time PCR: Validation report

In line with its mandate the European Union Reference Laboratory for GM Food and Feed (EU RL GMFF), in collaboration with the European Network of GMO Laboratories (ENGL), has validated an event-specific polymerase chain reaction (PCR) method for detecting and quantifying soybean event DAS-68416-4 (unique identifier DAS-68416-4). The validation study was conducted according to the EU-RL GMFF validation procedure (http://gmo-crl.jrc.ec.europa.eu/guidancedocs.htm) and internationally accepted guidelines. In accordance with current EU legislation , Dow AgroSciences LLC provided the detection method and the positive and negative control samples (genomic DNA extracted from soybean kernels harbouring the DAS-68416-4 event as positive control DNA, genomic DNA extracted from conventional soybean kernels as negative control DNA). The EU-RL GMFF prepared the validation samples (calibration samples and blind samples at different GM percentage [DNA/DNA]), organised an international collaborative study and analysed the results. The study confirms that the method meets the method performance requirements as established by the EU-RL GMFF and the ENGL and according to Annex I-2.C.2 to Regulation (EC) No 641/2004 and it fulfils the analytical requirements of Regulation (EU) No 619/2011JRC.I.3-Molecular Biology and Genomic

Changing trend of caries from 1989 to 2004 among 12-year old Sardinian children

Background. During the past decades, the prevalence of caries disease in the population of Western industrialized countries has decreased markedly. In children also, a reduction of dental caries experience has been reported by many authors. The aim of this paper was to evaluate the trend of dental caries prevalence in 12-year-old children living in the city of Sassari, (Italy), by five cross-sectional studies conducted in 1989, 1992, 1995, 1998 and 2004. Methods. In all cohorts, dental caries (DMFT and SiC Index according to WHO indications), was measured. For each variable measured (DMFT and sub-indices, SiC Index), differences in proportions among the five cohorts during the fifteen years were tested using χ2-square test. Results. The mean DMFT index decreased from 4.3 ± 3.1 in 1989 to 0.8 ± 1.5 in 2004. The prevalence of untreated caries (DT) had a notable decrease between 1992 and 1995, increased slightly between 1995 and 1998 and had the greatest decrease in 2004. The number of filled teeth remains low. The percentage of caries-free children increased from 10% to 64%, whereas the percentage of untreated caries changed from 44% in 1989 to 62% in 2004. SiC Index decreased from 7.8 in 1989 to 3.9 in 2004. Conclusion. On the basis of the results of DMFT and SiC Index, caries experience has been reduced. The vigilance and the promotion of a higher standard of personal oral hygiene and dental check-ups are necessary to obtain an improvement of oral status in the future adult population and to reach the new WHO global goals

Report on the Verification of the Performance of MON 87705 and MON 89788 Event-specific PCR-based Methods applied to DNA Extracted from GM Stack Soybean MON 87705 x MON 89788

The EU-RL GMFF has previously validated individually, and declared fit for purpose, the detection methods for the single line soybean events MON 87705 and MON 89788 and has published the corresponding reports (see http://gmo-crl.jrc.ec.europa.eu/StatusOfDossiers.aspx). In line with the approach defined by the ENGL (http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf) the EU-RL GMFF therefore has carried out only an in-house verification of the performance of each validated method when applied to DNA extracted from the GM stack MON 87705 x MON 89788 soybean. The hereby reported in-house verification study led to the conclusion that the individual methods meet the ENGL requirements also when applied to DNA extracted from the GM stack MON 87705 x MON 89788 soybean.JRC.I.3-Molecular Biology and Genomic

The impact of lockdown on sleep patterns of children and adolescents with ADHD

STUDY OBJECTIVES: The current study examined the impact of home confinement (lockdown) due to the COVID-19 pandemic on sleep patterns of children and adolescents with ADHD.METHODS: Nine hundred ninety-two parents of children and adolescents with ADHD filled out an anonymous online survey through the ADHD family association website. The survey investigated the sleep patterns and disturbances (using a modified version of the Sleep Disturbance Scale for Children) and screen exposure time before and during the lockdown.RESULTS: During the lockdown, 59.3% of children and 69.4% of adolescents with ADHD reported a change of bedtime with significant increase of ADHD patients that went to sleep at 11pm or later. Sleep duration, in contrast, resulted in two opposing processes with more children and adolescent sleeping either less than 6 hours/night or 10-11 hours/night. Among children and adolescents, respectively, 19.9% and 22% slept less than they did before lockdown, while 21.4% and 27.4% slept more hours. Bedtime delay and decreased sleep duration were associated with an increase in the screen time exposure. Moreover, ADHD patients reported an increase in sleep disturbances when compared to previous condition, including mainly difficulties falling asleep, anxiety at bedtime, night awakenings, nightmares and daytime sleepiness.CONCLUSIONS: The lockdown impacted on sleep-wake rhythms by strengthening the maladaptive sleep patterns reported in usual life conditions in ADHD children

Development and validation of the ID-EC - The ITALIAN version of the identify chronic migraine

Background: Case-finding tools, such as the Identify Chronic Migraine (ID-CM) questionnaire, can improve detection of CM and alleviate its significant societal burden. We aimed to develop and validate the Italian version of the ID-CM (ID-EC) in paper and as a smart app version in a headache clinic-based setting. Methods: The study investigators translated and adapted to the Italian language the original ID-CM questionnaire (ID-EC) and further implemented it as a smart app. The ID-EC was tested in its paper and electronic version in consecutive patients referring to 9 Italian tertiary headache centers for their first in-person visit. The scoring algorithm of the ID-EC paper version was applied by the study investigators (case-finding) and by patients (self-diagnosis), while the smart app provided to patients automatically the diagnosis. Diagnostic accuracy of the ID-EC was assessed by matching the questionnaire results with the interview-based diagnoses performed by the headache specialists during the visit according to the criteria of International Classification of Headache Disorders, III edition, beta version. Results: We enrolled 531 patients in the test of the paper version of ID-EC and 427 in the validation study of the smart app. According to the clinical diagnosis 209 patients had CM in the paper version study and 202 had CM in the smart app study. 79.5% of patients returned valid paper questionnaires, while 100% of patients returned valid and complete smart app questionnaires. The paper questionnaire had a 81.5% sensitivity and a 81.1% specificity for case-finding and a 30.7% sensitivity and 90.7% specificity for self-diagnosis, while the smart app had a 64.9% sensitivity and 90.2% specificity. Conclusions: Our data suggest that the ID-EC, developed and validated in tertiary headache centers, is a valid case-finding tool for CM, with sensitivity and specificity values above 80% in paper form, while the ID-EC smart app is more useful to exclude CM diagnosis in case of a negative result. Further studies are warranted to assess the diagnostic accuracy of the ID-EC in general practice and population-based settings

Extraction of DNA from Choline Chloride Feed Additive (CC) and from derived Pre-Mixes (PMCC) and Screening of CC and PMCC for (a) presence of rice and (b) presence of Bt63

Choline Chloride 60% (CC) is a feed additive that is imported in significant quantities from China. In a number of cases GM rice, harbouring the event Bt63, has been found in imported CC batches but not in derived pre-mixes. The Member States were asked to provide positive CC samples and derived pre-mixes to the EU-RL GMFF in order to allow the EU-RL GMFF establishing practical approaches to DNA extraction and the subsequent testing for presence of (a) rice and (b) the Bt63 rice event. This technical note has been derived based on data provided from National Control Laboratories and experience made by the EU-RL GMFF when re-testing nine feed additive samples and ten pre-mixes samples. The EU-RL GMFF found that the extraction of DNA from the feed additive Choline Chloride (CC) does not normally pose specific problems. It can be carried out following the available protocols or using available standard DNA extraction kits. The extraction of DNA from derived pre-mixes (PMCC) is, however, posing problems because of observed strong inhibition and the presence of DNA from additional sources than those present in the CC. The EU-RL GMFF has tried a number of protocols and extraction kits on PMCC samples. On this basis it is concluded that it is advisable to carefully purify the DNA, to verify the possible presence of inhibition effects and eventually to try to reduce any inhibition observed. Concerning testing the extracted DNA for rice and Bt63 rice event presence, this does not pose a problem in case of the CC while for PMCC rice was not anymore detectable or was detected in trace amounts. In any case testing for Bt63 presence was impossible. A possible explanation could be that much of the extracted DNA is from other plants than rice and hence the concentration of rice DNA is below the LOD of the rice-taxon specific method. In order to verify this, the EU-RL GMFF has analysed the available rice (taxon-) specific methods by means of bio-informatics and (partly) in the laboratory and concludes that they all should be suitable for specifically and reliably detecting rice from the Oryza sativa species. The re-testing by the EU-RL GMFF of CC and PMCC samples has confirmed the results of the initial tests.JRC.I.3-Molecular Biology and Genomic

Report on the Verification of the Performance of Bt11, DAS-59122-7, MIR604, TC 1507 and GA21 Event-specific PCR-based Methods Applied to DNA Extracted from GM Stack Bt11 x DAS-59122-7 x MIR604 x TC 1507 x GA21 Maize

An application was submitted by Syngenta Crop Protection AG to request the authorisation of genetically modified stack (GM stack) Bt11 x 59122 x MIR604 x TC1507 x GA21 maize (tolerant to herbicide products containing glufosinate ammonium/glyphosate and resistant to certain lepidopteran/coleopteran pests) and all sub-combinations of the individual events as present in the segregating progeny (except for 1507 x 59122), for food and feed uses, and import and processing, in accordance with articles 5 and 17 of Regulation (EC) No 1829/2003 on GM Food and Feed. The unique identifier assigned to GM stack Bt11 x 59122 x MIR604 x TC1507 x GA21 maize is SYN-BTØ11-1 × DAS-59122-7 × SYN-IR6Ø4-5 × DAS-Ø15Ø7-1 × MON-ØØØ21-9. The GM stack Bt11 x 59122 x MIR604 x TC1507 x GA21 maize has been obtained by conventional crossing of genetically modified maize events: Bt11, 59122, MIR604, TC1507 and GA21, without any new genetic modification. The European Union Reference Laboratory for GM Food and Feed (EU-RL GMFF) has previously validated individually, and declared fit for purpose, the detection methods for the single events Bt11, 59122, MIR604, TC1507 and GA21 (see http://gmo-crl.jrc.ec.europa.eu/StatusOfDossiers.aspx). In line with the approach defined by the European Network of GMO Laboratories (ENGL) (http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf) the EU-RL GMFF has carried out only an in-house verification of the performance of each validated method when applied to genomic DNA extracted from GM stack Bt11 x 59122 x MIR604 x TC1507 x GA21 maize. The results of the in-house verification led to the conclusion that the individual methods meet the ENGL performance criteria also when applied to genomic DNA extracted from the GM stack Bt11 x 59122 x MIR604 x TC1507 x GA21 maize.JRC.I.3-Molecular Biology and Genomic

SARS-CoV-2 Gamma and Delta Variants of Concern Might Undermine Neutralizing Activity Generated in Response to BNT162b2 mRNA Vaccination

The Delta variant raised concern regarding its ability to evade SARS-CoV-2 vaccines. We evaluated a serum neutralizing response of 172 Italian healthcare workers, three months after complete Comirnaty (BNT162b2 mRNA, BioNTech-Pfizer) vaccination, testing their sera against viral isolates of Alpha, Gamma and Delta variants, including 36 subjects with a previous SARS-CoV-2 infection. We assessed whether IgG anti-spike TRIM levels and serum neutralizing activity by seroneutralization assay were associated. Concerning Gamma variant, a two-fold reduction in neutralizing titres compared to the Alpha variant was observed, while a four-fold reduction of Delta virus compared to Alpha was found. A gender difference was observed in neutralizing titres only for the Gamma variant. The serum samples of 36 previously infected SARS-CoV-2 individuals neutralized Alpha, Gamma and Delta variants, demonstrating respectively a nearly three-fold and a five-fold reduction in neutralizing titres compared to Alpha variant. IgG anti-spike TRIM levels were positively correlated with serum neutralizing titres against the three variants. The Comirnaty vaccine provides sustained neutralizing antibody activity towards the Alpha variant, but it is less effective against Gamma and even less against Delta variants