Potent Sensitisation of Cancer Cells to Anticancer Drugs by a Quadruple Mutant of the Human Deoxycytidine Kinase.

Identifying enzymes that, once introduced in cancer cells, lead to an increased efficiency of treatment constitutes an important goal for biomedical applications. Using an original procedure whereby mutant genes are generated based on the use of conditional lentivector genome mobilisation, we recently described, for the first time, the identification of a human deoxycytidine kinase (dCK) mutant (G12) that sensitises a panel of cancer cell lines to treatment with the dCK analogue gemcitabine. Here, starting from the G12 variant itself, we generated a new library and identified a mutant (M36) that triggers even greater sensitisation to gemcitabine than G12. With respect to G12, M36 presents an additional mutation located in the region that constitutes the interface of the dCK dimer. The simple presence of this mutation halves both the IC50 and the proportion of residual cells resistant to the treatment. Furthermore, the use of vectors with self-inactivating LTRs leads to an increased sensitivity to treatment, a result compatible with a relief of the transcriptional interference exerted by the U3 promoter on the internal promoter that drives the expression of M36. Importantly, a remarkable effect is also observed in treatments with the anticancer compound cytarabine (AraC), for which a 10,000 fold decrease in IC50 occurred. By triggering the sensitisation of various cancer cell types with poor prognosis to two commonly used anticancer compounds M36 is a promising candidate for suicide gene approaches

Experimental tests of Quantum Mechanics: from Pauli Exclusion Principle Violation to spontaneous collapse models

The Pauli exclusion principle (PEP) and, more generally, the spin-statistics connection, is at the very basis of our understanding of matter. The PEP spurs, presently, a lively debate on its possible limits, deeply rooted in the very foundations of Quantum Field Theory. Therefore, it is extremely important to test the limits of its validity. Quon theory provides a suitable mathematical framework of possible violation of PEP, where the q violation parameter translates into a probability of violating PEP. Experimentally, setting a bound on PEP violation means confining the q-parameter to a value very close to either 1 (for bosons) or -1 (for fermions). The VIP (Violation of the Pauli exclusion principle) experiment established a limit on the probability that PEP is violated by electrons, using the method of searching for PEP forbidden atomic transitions in copper. We describe the experimental method, the obtained results, both in terms of the q-parameter and as probability of PEP violation, we briefly discuss the results and present future plans to go beyond the actual limit by upgrading the experimental technique using vetoed new spectroscopic fast Silicon Drift Detectors. We mention as well the possibility of using a similar experimental technique to search for eventual X-rays generated as a signature of the spontaneous collapse of the wave function, predicted by continuous spontaneous localization type theories

Multidifferential study of identified charged hadron distributions in ZZ-tagged jets in proton-proton collisions at s=\sqrt{s}=13 TeV

Jet fragmentation functions are measured for the first time in proton-proton collisions for charged pions, kaons, and protons within jets recoiling against a $Z$ boson. The charged-hadron distributions are studied longitudinally and transversely to the jet direction for jets with transverse momentum 20 $< p_{\textrm{T}} < 100$ GeV and in the pseudorapidity range $2.5 < \eta < 4$. The data sample was collected with the LHCb experiment at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 1.64 fb$^{-1}$. Triple differential distributions as a function of the hadron longitudinal momentum fraction, hadron transverse momentum, and jet transverse momentum are also measured for the first time. This helps constrain transverse-momentum-dependent fragmentation functions. Differences in the shapes and magnitudes of the measured distributions for the different hadron species provide insights into the hadronization process for jets predominantly initiated by light quarks.Comment: All figures and tables, along with machine-readable versions and any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-013.html (LHCb public pages

Study of the B−→Λc+Λˉc−K−B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-} decay

The decay $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ is studied in proton-proton collisions at a center-of-mass energy of $\sqrt{s}=13$ TeV using data corresponding to an integrated luminosity of 5 $\mathrm{fb}^{-1}$ collected by the LHCb experiment. In the $\Lambda_{c}^+ K^{-}$ system, the $\Xi_{c}(2930)^{0}$ state observed at the BaBar and Belle experiments is resolved into two narrower states, $\Xi_{c}(2923)^{0}$ and $\Xi_{c}(2939)^{0}$, whose masses and widths are measured to be $$m(\Xi_{c}(2923)^{0}) = 2924.5 \pm 0.4 \pm 1.1 \,\mathrm{MeV}, \\ m(\Xi_{c}(2939)^{0}) = 2938.5 \pm 0.9 \pm 2.3 \,\mathrm{MeV}, \\ \Gamma(\Xi_{c}(2923)^{0}) = \phantom{000}4.8 \pm 0.9 \pm 1.5 \,\mathrm{MeV},\\ \Gamma(\Xi_{c}(2939)^{0}) = \phantom{00}11.0 \pm 1.9 \pm 7.5 \,\mathrm{MeV},$$ where the first uncertainties are statistical and the second systematic. The results are consistent with a previous LHCb measurement using a prompt $\Lambda_{c}^{+} K^{-}$ sample. Evidence of a new $\Xi_{c}(2880)^{0}$ state is found with a local significance of $3.8\,\sigma$, whose mass and width are measured to be $2881.8 \pm 3.1 \pm 8.5\,\mathrm{MeV}$ and $12.4 \pm 5.3 \pm 5.8 \,\mathrm{MeV}$, respectively. In addition, evidence of a new decay mode $\Xi_{c}(2790)^{0} \to \Lambda_{c}^{+} K^{-}$ is found with a significance of $3.7\,\sigma$. The relative branching fraction of $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ with respect to the $B^{-} \to D^{+} D^{-} K^{-}$ decay is measured to be $2.36 \pm 0.11 \pm 0.22 \pm 0.25$, where the first uncertainty is statistical, the second systematic and the third originates from the branching fractions of charm hadron decays.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-028.html (LHCb public pages

Measurement of the ratios of branching fractions R(D∗)\mathcal{R}(D^{*}) and R(D0)\mathcal{R}(D^{0})

The ratios of branching fractions $\mathcal{R}(D^{*})\equiv\mathcal{B}(\bar{B}\to D^{*}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(\bar{B}\to D^{*}\mu^{-}\bar{\nu}_{\mu})$ and $\mathcal{R}(D^{0})\equiv\mathcal{B}(B^{-}\to D^{0}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(B^{-}\to D^{0}\mu^{-}\bar{\nu}_{\mu})$ are measured, assuming isospin symmetry, using a sample of proton-proton collision data corresponding to 3.0 fb${ }^{-1}$ of integrated luminosity recorded by the LHCb experiment during 2011 and 2012. The tau lepton is identified in the decay mode $\tau^{-}\to\mu^{-}\nu_{\tau}\bar{\nu}_{\mu}$. The measured values are $\mathcal{R}(D^{*})=0.281\pm0.018\pm0.024$ and $\mathcal{R}(D^{0})=0.441\pm0.060\pm0.066$, where the first uncertainty is statistical and the second is systematic. The correlation between these measurements is $\rho=-0.43$. Results are consistent with the current average of these quantities and are at a combined 1.9 standard deviations from the predictions based on lepton flavor universality in the Standard Model.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-039.html (LHCb public pages

Measurement of the B0s→μ+μ− Branching Fraction and Effective Lifetime and Search for B0→μ+μ− Decays

A search for the rare decays Bs0→μ+μ- and B0→μ+μ- is performed at the LHCb experiment using data collected in pp collisions corresponding to a total integrated luminosity of 4.4  fb-1. An excess of Bs0→μ+μ- decays is observed with a significance of 7.8 standard deviations, representing the first observation of this decay in a single experiment. The branching fraction is measured to be B(Bs0→μ+μ-)=(3.0±0.6-0.2+0.3)×10-9, where the first uncertainty is statistical and the second systematic. The first measurement of the Bs0→μ+μ- effective lifetime, τ(Bs0→μ+μ-)=2.04±0.44±0.05  ps, is reported. No significant excess of B0→μ+μ- decays is found, and a 95% confidence level upper limit, B(B0→μ+μ-)<3.4×10-10, is determined. All results are in agreement with the standard model expectations.A search for the rare decays $B^0_s\to\mu^+\mu^-$ and $B^0\to\mu^+\mu^-$ is performed at the LHCb experiment using data collected in $pp$ collisions corresponding to a total integrated luminosity of 4.4 fb$^{-1}$. An excess of $B^0_s\to\mu^+\mu^-$ decays is observed with a significance of 7.8 standard deviations, representing the first observation of this decay in a single experiment. The branching fraction is measured to be ${\cal B}(B^0_s\to\mu^+\mu^-)=\left(3.0\pm 0.6^{+0.3}_{-0.2}\right)\times 10^{-9}$, where the first uncertainty is statistical and the second systematic. The first measurement of the $B^0_s\to\mu^+\mu^-$ effective lifetime, $\tau(B^0_s\to\mu^+\mu^-)=2.04\pm 0.44\pm 0.05$ ps, is reported. No significant excess of $B^0\to\mu^+\mu^-$ decays is found and a 95 % confidence level upper limit, ${\cal B}(B^0\to\mu^+\mu^-)<3.4\times 10^{-10}$, is determined. All results are in agreement with the Standard Model expectations

A snapshot on HIV-1 evolution through the identification of phylogenetic-specific properties of HIV-1 integrases M/O.

Transmissions of simian viruses to humans has originated the different groups of HIV-1. We recently identified a functional motif (CLA), in the C-terminal domain of the integrase, essential for integration in HIV-1 group M. Here, we found that the motif is instead dispensable in group O isolates, because of the presence, in the N-terminal domain of HIV-1 O of a specific sequence, Q7G27P41H44, that we define as the NOG motif. Alterations of reverse transcription and of 3' processing observed by mutating the CLA motif of IN M are fully rescued to wt levels by inserting the sequence of the NOG motif in the N-ter of the protein. These results indicate that the two motifs (CLA and NOG) functionally complement each other and a working model accounting for these observations is proposed. The establishment of these two alternative motifs seems to be due to the different phylogenetic origin and history of these two groups. Indeed, the NOG motif is already present in the ancestor of group O (SIVgor) while it is absent from SIVcpzPtt, the ancestor of group M. The CLA motif, instead, seems to have emerged after SIVcpzPtt has been transferred to humans, since no conservation is found at the same positions in these simian viruses. These results show the existence of two-group specific motifs in HIV-1 M and O integrases. In each group, only one of the motifs is functional, potentially leading the other motif to diverge from its original function and, in an evolutionary perspective, assist other functions of the protein, further increasing HIV genetic diversity

Potent sensitisation of cancer cells to anticancer drugs by a quadruple mutant of the human deoxycytidine kinase

Identifying enzymes that, once introduced in cancer cells, lead to an increased efficiency of treatment constitutes an important goal for biomedical applications. Using an original procedure whereby mutant genes are generated based on the use of conditional lentivector genome mobilisation, we recently described, for the first time, the identification of a human deoxycytidine kinase (dCK) mutant (G12) that sensitises a panel of cancer cell lines to treatment with the dCK analogue gemcitabine. Here, starting from the G12 variant itself, we generated a new library and identified a mutant (M36) that triggers even greater sensitisation to gemcitabine than G12. With respect to G12, M36 presents an additional mutation located in the region that constitutes the interface of the dCK dimer. The simple presence of this mutation halves both the IC50 and the proportion of residual cells resistant to the treatment. Furthermore, the use of vectors with self-inactivating LTRs leads to an increased sensitivity to treatment, a result compatible with a relief of the transcriptional interference exerted by the U3 promoter on the internal promoter that drives the expression of M36. Importantly, a remarkable effect is also observed in treatments with the anticancer compound cytarabine (AraC), for which a 10,000 fold decrease in IC50 occurred. By triggering the sensitisation of various cancer cell types with poor prognosis to two commonly used anticancer compounds M36 is a promising candidate for suicide gene approaches

UN MODELLO OPERATIVO PER LA VALUTAZIONE DEL RISCHIO AMBIENTALE APPLICATA AI SITI ITALIANI DI IMPORTANZA COMUNITARIA: IDENTIFICAZIONE DEI POTENZIALI EFFETTI SUL SUOLO

The fast development of agro-biotechnologies asks for a harmonized approach in risk analysis of GMOs releases. An Italian experts group has elaborated an operating model for the environmental risk assessment (OMERA) based on the assumption that the occurring of a risk is related to the presence of four components: source, diffusion factors, dispersal routes, receptors. This model has been further developed to become a Decision Supporting System based on Fuzzy logic (FDSS) to assessors and notifiers. It is a web based Questionnaire that conducts the user through a decision tree from the source to the receptors and leads to the identification and assessment of the risks. The FDSS has been tested on case studies, simulating, as source, herbicide tolerant oilseed rape and insect resistant maize. The resulting identified potential effects on soil are changes to structure and microbial diversity.Le développement rapide des agro-biotechnologies demande une approche harmonisée en matière d'analyse des risques des GMO. Un groupe d'experts italien a élaboré un modèle opérationnel pour l'évaluation des risques pour l'environnement (OMERA), basé sur l'hypothèse que le survenant d'un risque est lié à la présence de quatre éléments: la source, les facteurs de diffusion, les voies de dispersion, les récepteurs. Ce modèle a été développé pour devenir une véritable système d'information décisionnel sur la base de la logique fuzzy (SIDF). Un questionnaire Web mène l'utilisateur à travers un arbre de décision à partir de la source aux récepteurs et conduit à l'identification et l'évaluation des risques. Le SID a été testé en simulant, en tant que source, colza tolérant à un herbicide et maïs résistant aux insectes. Pour le sol, les effets potentiels identifiés sont les changements à la biodiversité et aux pratiques agricoles.Il rapido sviluppo delle agrobiotecnologie richiede un approccio armonizzato nell’analisi del rischio dei rilasci di OGM. Un gruppo di esperti italiani ha elaborato un modello operativo per la valutazione dei rischi ambientali (OMERA), basato sul presupposto che l’insorgere di un rischio è legato alla presenza di quattro componenti: fonte, fattori di diffusione, vie di migrazione, recettori.. Questo modello è stato ulteriormente sviluppato fino a diventare Sistema di Supporto alle Decisioni basato sulla logica fuzzy (FDSS) per valutatori del rischio e notificanti. Un questionario web-based conduce l'utente attraverso un albero decisionale dalla sorgente ai ricettori e ha porta alla individuazione e valutazione dei rischi. . Il DSS è stato testato su casi-studio usando come fonti colza tollerante agli erbicidi e mais resistente agli insetti. I potenziali effetti identificati per il suolo sono i cambiamenti nella struttura e nella diversità della comunità microbica

Diversity of coccinellidae in ecological compensation areas of Italy and overlap with maize pollen shedding period

Ladybird beetles (Coleoptera Coccinellidae) are important predators of aphids and other crop pests, and there is great interest in their conservation in agroecosystems. Bt-maize, genetically engineered to express insecticidal Cry proteins, is regarded as a taxon-specific way of controlling pests, but some concerns have been raised about the possibility of unintended negative effects on non-target organisms, including coccinellids. One of the possible routes of exposure of ladybird beetles to Cry toxins is by feeding on maize pollen, as for many species pollen is an important integrative food source. In this study, coccinellid adults were sampled by sweep netting in Ecological Compensation Areas (ECAs) in three sites of Northern and Central Italy, where conventional maize cultivars are grown (Bt-maize is currently forbidden for commercial cultivation in Italy). The coccinellid communities were sampled during and around the typical flowering periods of maize in order to check their diversity and their overlap with pollen shedding. A total of 11 species were recorded. Hippodamia variegata (Goeze), Coccinella septempunctata L. and the exotic Harmonia axyridis (Pallas) were the most abundant species in Northern Italy, whereas Tytthaspis sedecimpunctata (L.) and Coccinula quatuordecimpustulata (L.) were dominant in Central Italy. The potential exposure to maize pollen was different in the two areas, since in Northern Italy the maize flowering coincided with a period of high coccinellid activity in the field, whereas in Central Italy the ladybird population peaks occurred roughly one month earlier than the anthesis. The collected data might be useful for exposure characterization of ladybird beetles in a possible future scenario of Bt-maize cropping in the studied areas