Engineered Sleeping Beauty Transposon as Efficient System to Optimize Chimp Adenoviral Production

Sleeping Beauty (SB) is the first DNA transposon employed for efficient transposition in vertebrate cells, opening new applications for genetic engineering and gene therapies. A transposon-based gene delivery system holds the favourable features of non-viral vectors and an attractive safety profile. Here, we employed SB to engineer HEK293 cells for optimizing the production of a chimpanzee Adenovector (chAd) belonging to the Human Mastadenovirus C species. To date, chAd vectors are employed in several clinical settings for infectious diseases, last but not least COVID-19. A robust, efficient and quick viral vector production could advance the clinical application of chAd vectors. To this aim, we firstly swapped the hAd5 E1 with chAd-C E1 gene by using the CRISPR/Cas9 system. We demonstrated that in the absence of human Ad5 E1, chimp Ad-C E1 gene did not support HEK293 survival. To improve chAd-C vector production, we engineered HEK293 cells to stably express the chAd-C precursor terminal protein (ch.pTP), which plays a crucial role in chimpanzee Adenoviral DNA replication. The results indicate that exogenous ch.pTP expression significantly ameliorate the packaging and amplification of recombinant chAd-C vectors thus, the engineered HEK293ch.pTP cells could represent a superior packaging cell line for the production of these vectors

Integrated Project 1: Gender and Population-level Variables in Language Learning and Use of Digital Tools

In the first integrated project, a versatile questionnaire was designed and successfully implemented to capture crucial data on children’s digital media exposure, household composition, language backgrounds, gender, and other population-level variables across different countries. With completed data collection in Germany and Latvia and ongoing efforts in Norway, Spain, and the United Kingdom, the project is contributing relevant insights into the diverse factors shaping language and cognitive development in children, exploring possible hidden correlations between the different collected measures. This information can then be effectively used to inform the development of policies and recommendations aimed at guiding children's digital media use, considering factors like consumption duration, age, and language habits

Detection of RET/PTC, TRK and BRAF mutations in preoperative diagnosis of thyroid nodules with indeterminate cytological findings.

Fine-needle aspiration biopsy (FNAB) is the primary means to distinguish benign from malignant nodules and select patients for surgery. However, adjunctive diagnostic tests are needed because in 20-40% of cases the FNAB result is uncertain. OBJECTIVE: We investigated whether a search for the oncogenes RET/PTC, TRK and BRAF(V600E) in thyroid aspirates could refine an uncertain diagnosis. PATIENTS AND METHODS: A total of 132 thyroid aspirates, including colloid nodules, inadequate samplings, indeterminate and suspicious for malignancy were analysed by reverse transcription polymerase chain reaction (RT-PCR) and mutant allele-specific amplification techniques for the presence of oncogenes. RESULTS: No oncogenes were detected in 48 colloid nodules, 46 inadequate and 19 indeterminate FNABs, then confirmed to be benign at histology. No oncogenes were detected in one follicular thyroid cancer (FTC) with indeterminate cytology. Five out of six papillary thyroid cancers (83%) with FNAB suspicious for malignancy were correctly diagnosed by the presence of oncogenes. Among these, four (67%) contained the BRAF mutation and one (17%) contained RET/PTC-3. On final analysis, no false-positive results were reported in 131 samples and five out of seven carcinomas (71%) were correctly diagnosed. The finding of oncogenes in FNAB specimens suspicious for malignancy guided the extent of surgical resection, changing the surgery from diagnostic to therapeutic in five cases. CONCLUSIONS: Detection of RET/PTC, TRK and BRAF(V600E) in FNAB specimens is proposed as a diagnostic adjunctive tool in the evaluation of thyroid nodules with suspicious cytological findings

Integrated Project 2: App Development and Education

In the second integrated project, a prototype of a tablet-based application was developed to promote early vocabulary learning among migrant preschool-aged children. Across Europe, half of the population is either multilingual or lives in a multicultural and multilingual environment. Meanwhile, a portion of children do not speak the language of school instruction at home. Insufficient language skills in the early years can potentially have a far-reaching and long-lasting impact on children’s educational trajectory. The goal of this interdisciplinary project was to address this issue by developing the prototype of a vocabulary learning app, following the latest literature regarding vocabulary learning and design principles to design apps for second language acquisition (L2)