Search for heavy neutrinos in $D_s$ decays with the CMS detector at LHC

This thesis presents a search for a sterile right-handed neutrino $N$ produced in $D_s$ meson decays, using proton-proton collisions collected by the CMS experiment at the LHC. The data set used for the analysis, the B-Parking data set, corresponds to an integrated luminosity of $41.7\,\textrm{fb}^{-1}$ and was collected during the 2018 data-taking period. The analysis is targeting the $D_s^+\rightarrow N(\rightarrow\mu^{\pm}\pi^{\mp})\mu^{+}$ decays, where the final state muons can have the same electric charge allowing for a lepton flavor violating decay. To separate signal from background, a cut-based analysis is optimized using requirements on the sterile neutrino vertex displacement, muon and pion impact parameter, and impact parameter significance. The expected limit on the active-sterile neutrino mixing matrix parameter $|V_{\mu}|^2$ is extracted by performing a fit of the $\mu\pi$ invariant mass spectrum for two sterile neutrino mass hypotheses, 1.0 and 1.5 GeV. The analysis is currently blinded, following the internal CMS review process. The expected limit ranges between approximately $10^{-4}$ for a 1.0 GeV neutrino to $7\times10^{-5}$ for a 1.5 GeV neutrino. This is competitive with the best existing results from collider experiments over the same mass range.

Transcriptional functions of DNA Topoisomerases at a genome-wide scale and a single gene levels

The DNA topology is an important modifier of DNA functions. Torsional stress is generated when right handed DNA is either over- or underwound, producing structural deformations which drive or are driven by processes such as replication, transcription, recombination and repair. DNA topoisomerases are molecular machines that regulate the topological state of the DNA in the cell. These enzymes accomplish this task by either passing one strand of the DNA through a break in the opposing strand or by passing a region of the duplex from the same or a different molecule through a double-stranded cut generated in the DNA. Because of their ability to cut one or two strands of DNA they are also target for some of the most successful anticancer drugs used in standard combination therapies of human cancers. An effective anticancer drug is Camptothecin (CPT) that specifically targets DNA topoisomerase 1 (TOP 1). The research project of the present thesis has been focused on the role of human TOP 1 during transcription and on the transcriptional consequences associated with TOP 1 inhibition by CPT in human cell lines. Previous findings demonstrate that TOP 1 inhibition by CPT perturbs RNA polymerase (RNAP II) density at promoters and along transcribed genes suggesting an involvement of TOP 1 in RNAP II promoter proximal pausing site. Within the transcription cycle, promoter pausing is a fundamental step the importance of which has been well established as a means of coupling elongation to RNA maturation. By measuring nascent RNA transcripts bound to chromatin, we demonstrated that TOP 1 inhibition by CPT can enhance RNAP II escape from promoter proximal pausing site of the human Hypoxia Inducible Factor 1 (HIF-1) and c-MYC genes in a dose dependent manner. This effect is dependent from Cdk7/Cdk9 activities since it can be reversed by the kinases inhibitor DRB. Since CPT affects RNAP II by promoting the hyperphosphorylation of its Rpb1 subunit the findings suggest that TOP 1inhibition by CPT may increase the activity of Cdks which in turn phosphorylate the Rpb1 subunit of RNAP II enhancing its escape from pausing. Interestingly, the transcriptional consequences of CPT induced topological stress are wider than expected. CPT increased co-transcriptional splicing of exon1 and 2 and markedly affected alternative splicing at exon 11. Surprisingly despite its well-established transcription inhibitory activity, CPT can trigger the production of a novel long RNA (5’aHIF-1) antisense to the human HIF-1 mRNA and a known antisense RNA at the 3’ end of the gene, while decreasing mRNA levels. The effects require TOP 1 and are independent from CPT induced DNA damage. Thus, when the supercoiling imbalance promoted by CPT occurs at promoter, it may trigger deregulation of the RNAP II pausing, increased chromatin accessibility and activation/derepression of antisense transcripts in a Cdks dependent manner. A changed balance of antisense transcripts and mRNAs may regulate the activity of HIF-1 and contribute to the control of tumor progression After focusing our TOP 1 investigations at a single gene level, we have extended the study to the whole genome by developing the “Topo-Seq” approach which generates a map of genome-wide distribution of sites of TOP 1 activity sites in human cells. The preliminary data revealed that TOP 1 preferentially localizes at intragenic regions and in particular at 5’ and 3’ ends of genes. Surprisingly upon TOP 1 downregulation, which impairs protein expression by 80%, TOP 1 molecules are mostly localized around 3’ ends of genes, thus suggesting that its activity is essential at these regions and can be compensate at 5’ ends. The developed procedure is a pioneer tool for the detection of TOP 1 cleavage sites across the genome and can open the way to further investigations of the enzyme roles in different nuclear processes.

Non invasive assessment of right ventricle in patients with operated tetralogy of Fallot

The objective of this study was to evaluate right ventricular function in patients with right ventricular volume overload in patients with (tetralogy of Fallot, and pulmonary atresia + VSD ) underwent corrective surgery; with echocardiography measure that can be easily applied; and to study the relationship between ProBNP and the contractile function of the right ventricle, dilated right atrium, and the consequences of pulmonary insufficiency . Methods: The study included 50 patients (50% males, mean age 30.64 ± 13.30 years) with prior cardiac surgical intervention of TDF (90%) or pulmonary atresia + VSD (10%). (49 pz) have performed a cardiac MRI and clinical evaluation, (47 pz) echocardiogram, (48 pz) ECG, (34 pz) a cardiopulmonary exercise testing, (29 pz) a dosage of ProBNP. Results: The S-wave velocity (p <0.0001), the TAPSE (p <0.0001) correlated significantly with RVEF estimated by cardiac MRI. The VO2 max was 27.93 ± 12.91 ml / kg / min, 15% of patients had VE/VCO2 The peak> 35. ProBNP correlated positively and significantly with the area of the right atrium (p = 0.0001), and negative and significant with VO2 max (p = 0.04). Those who have increased pulmonary insufficiency (PVR fraction> 30%) have a significantly increased RVED volume (p = 0.01), reduced VO2 max (p = 0.04), and lower ejection fraction of LV (p = 0.02) than the group of patients with PVR ≤ 30. Conclusion: The TAPSE and S-wave velocity are fundamental and may become the technique of choice for routine assessment of RV systolic function in adult patients with TOF. The monitoring of the Pro BNP is probably a choice, given the simplicity and their information that correlate with the test cardiopulmonary. In view of the ventricular-ventricular interaction, so measures to maintain or restore the functioning of the pulmonary valve could preserve biventricular function.

The Right to Voice: Its Realization by Estonian and American Parents in Public Schools

Investigating parents’ formal engagement opportunities in public schools serves well to characterize the relationship between states and societies. While the relationship between parental involvement and students’ academic success has been thoroughly investigated, rarely has it been seen to indicate countries’ governing regimes. The researcher was curious to see whether and how does parents’ voice differ in different democracies. The hypothesis was that in mature regimes, institutional opportunities for formal parental engagement are plenty and parents are actively involved; while in young democracies there are less opportunities and the engagement is lower. The assumption was also that parental deliberation in expressing their dissatisfaction with schools differs across democracies: where it is more intense, there it translates to higher engagement. Parents’ informedness on relevant regulations and agendas was assumed to be equally average, and their demographic background to have similar effects on engagement. The comparative, most different systems design was employed where public middle schools last graders’ parents in Tartu, Estonia and in Huntsville, Alabama the United States served as a sample. The multidimensional study includes the theoretical review, country and community analyses, institutional analysis in terms of formal parental involvement, and parents’ survey. The findings revealed sizeable differences between parents’ engagement levels in Huntsville and Tartu. The results indicate passivity in both communities, while in Tartu the engagement seems to be alarmingly low. Furthermore, Tartu parents have much less institutional opportunities to engage. In the United States, multilevel efforts to engage parents are visible from local to federal level, in Estonia similar intentions seem to be missing and meaningful parental organizations do not exist. In terms of civic education there is much room for development in both countries. The road will be longer for a young democracy Estonia in transforming its institutional systems from formally democratic to inherently inclusive.