Birefringent Fourier filtering for single molecule Coordinate and Height super-resolution Imaging with Dithering and Orientation

Super-resolution imaging based on single molecule localization allows accessing nanometric-scale information in biological samples with high precision. However, complete measurements including molecule orientation are still challenging. Orientation is intrinsically coupled to position in microscopy imaging, and molecular wobbling during the image integration time can bias orientation measurements. Providing 3D molecular orientation and orientational fluctuations can offer new ways to assess the degree of alignment of protein structures, which cannot be monitored by pure localization. Here we demonstrate that by adding polarization control to phase control in the Fourier plane of the imaging path, all parameters can be determined unambiguously from single molecules: 3D spatial position, 3D orientation and wobbling or dithering angle. The method, applied to fluorescent labels attached to single actin filaments, provides precisions within tens of nanometers in position and few degrees in direction

Can Stimulus Valence Modulate Task-Switching Ability? A Pilot Study on Primary School Children

Executive functions and emotional processes follow a time-dependent development that reflects the brain’s anatomo-functional maturation. Though the assessment of these cognitive functions is largely examined, in children the role of emotions in the mental set-shifting is still rarely investigated. The aim of this study was to assess how attention shifting can be modulated by the valence of emotional stimuli. To this end, sixty-two primary school children were tested with a new emotional task-switching paradigm obtained by manipulating the emotional valence and physical features of the stimulus pool. Thus, two tasks were alternatively presented: the Valence task and the Color task. Based on executive performance results, we found a lengthening of response times and a lower accuracy in the emotionally connoted task (Valence task), compared to the neutral task (Color task). The data demonstrate that the processing of emotional stimuli modulates the task-switching performance during development. These findings could help in the implementation of teaching strategies that can promote the development of executive functions and, therefore, functionally improve the overall academic performance of children. Finally, a better understanding of the developmental trajectories of executive functions can help neuropsychologists both in the early diagnosis and treatment of potential executive alterations

Rotavirus Quantification in Children with Acute Gastroenteritis

Introduction: Most intractable diarrheas are treated with antibiotics, irrespective of the causative agent. This study aimed to quantify rotavirus (RV) with TaqMan real-time PCR in fecal samples of children with acute gastroenteritis (AGE) in accordance with the program of reduction of drug resistance and use of antibiotics. Methods: A total of 190 fecal specimens were collected from under-5-year-old children with AGE in pediatric Hospital Regina Margherita of Turin in Italy in 2017. A total of 38 out of 67 (56.7%) episodes of AGE were associated with RV genomic detection with a multiplex commercial kit. Results: All fecal specimens were tested for the presence of RV and other GE viruses. The most commonly detected virus was norovirus (41%), astrovirus (15.8%), human bocavirus (8.9%), sapovirus (7.9%), human parechovirus (5.8%), rhinovirus (4.2%), and adenovirus (1%). In 66 out of 190 (34.7%), RV was detectable with the median viral load 7.2E + 11 ± 60E + 11 genomes/mg fecal specimens. Discussion/Conclusions: Our results showed that RV was present in around 34.7% of children hospitalized for AGE, a rate similar to those reported in previous studies conducted elsewhere which were in the range of 33–75%. Our protocols are able to quantify the absolute number of viral particle/mg of feces. The clinical utility of quantitative molecular assays, such as RV viral load, will be markedly improved

Evaluation of qualitative and semi-quantitative cut offs for rapid diagnostic lateral flow test in relation to serology for the detection of SARS-CoV-2 antibodies: findings of a prospective study

There is limited information to compare the qualitative and semi-quantitative performance of rapid diagnostic tests (RDT) and serology for the assessment of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Therefore, the objective of the study was (a) to compare the efficacy of SARS-CoV-2 antibody detection between RDT and laboratory serology, trying to identify appropriate semi-quantitative cut-offs for RDT in relation with quantitative serology values and to (b) evaluate diagnostic accuracy of RDT compared to the NAAT gold standard in an unselected adult population

Il CNR e i risultati della ricerca scientifica_ Le nuove procedure di controllo interno Attività di AUDIT Progetti PRIN e FIRB

Nel presente volume sono evidenziate le procedure relative alla nuova disciplina dei controlli interni a valere sulle spese di progetto. Tale disciplina, dettata dal MIUR, ha fornito lo spunto per l’implementazione e la sperimentazione di un coordinamento interno CNR volto alla certificazione delle spese effettuate dalla Rete Scientifica

Convenzione CNR-MiSE fondo per la crescita sostenibile

Nel presente documento vengono illustrate in dettaglio le finalità del FCS e le attività rea- lizzate dal RTI nell’ambito del servizio realizzato per il primo intervento che il MiSE ha bandito sul Fondo con Decreto del 20 giugno 2013 e attuato con procedura valutativa “a sportello”, definita nel Decreto del 25 luglio 2014. In particolare, oltre alle caratteristiche principali dei progetti di ricerca che il MiSE intende finanziare attraverso tale intervento, nel presente documento viene altresì descritta l’attività che l’Ufficio Supporto alla Programmazione Operativa della Direzione Centrale alla Rete Scientifica e Infrastrutture del CNR ha condotto per coordinare e gestire il processo di valutazione delle proposte progettuali. Viene infatti descritta la compagine di valutazione del CNR, che è stata attivata grazie alla partecipazione dei ricercatori e tecnologi della rete scientifica dell’Ente, e le attività che questi, in veste di esperti valutatori, hanno condotto per determinare la qualità tecnico-scientifica delle proposte progettuali presentate

The Fall in Antibody Response to SARS-CoV-2: a Longitudinal Study of Asymptomatic to Critically Ill Patients Up to 10 Months after Recovery

The aim of this study was to assess the long-term dynamics and factors associated with the serological response against the severe acute respiratory syndrome coronavirus 2 after primary infection. A prospective longitudinal study was conducted with monthly serological follow-up during the first 4 months, and then at 6, 8, and 10 months after the disease onset of all recovered adult in- and outpatients with coronavirus disease 2019 (COVID-19) attending Udine Hospital (Italy) during the first wave (from March to May 2020). A total of 546 individuals were included (289 female, mean age 53.1 years), mostly with mild COVID-19 (370, 68.3%). Patients were followed for a median of 302 days (interquartile range, 186 to 311). The overall seroconversion rate within 2 months was 32% for IgM and 90% for IgG. Seroreversion was observed in 90% of patients for IgM at 4 months and in 47% for IgG at 10 months. Older age, number of symptoms at acute onset, and severity of acute COVID-19 were all independent predictors of long-term immunity both for IgM (beta, linear regression coefficient, 1.10, P = 0.001; beta 5.15 P = 0.014; beta 43.84 P = 0.021, respectively) and for IgG (beta 1.43 P < 0.001; beta 10.46 P < 0.001; beta 46.79 P, 0.001, respectively), whereas the initial IgG peak was associated only with IgG duration (beta 1.12, P < 0.001). IgM antibodies disappeared at 4 months, and IgG antibodies declined in about half of patients 10 months after acute COVID-19. These effects varied depending on the intensity of the initial antibody response, age, and burden of acute COVID-19

MR-proADM as prognostic factor of outcome in COVID-19 patients

17siMid Regional pro-ADM (MR-proADM) is a promising novel biomarker in the evaluation of deteriorating patients and an emergent prognosis factor in patients with sepsis, septic shock and organ failure. It can be induced by bacteria, fungi or viruses. We hypothesized that the assessment of MR-proADM, with or without other inflammatory cytokines, as part of a clinical assessment of COVID-19 patients at hospital admission, may assist in identifying those likely to develop severe disease. A pragmatic retrospective analysis was performed on a complete data set from 111 patients admitted to Udine University Hospital, in northern Italy, from 25th March to 15th May 2020, affected by SARS-CoV-2 pneumonia. Clinical scoring systems (SOFA score, WHO disease severity class, SIMEU clinical phenotype), cytokines (IL-6, IL-1b, IL-8, TNF-α), and MR-proADM were measured. Demographic, clinical and outcome data were collected for analysis. At multivariate analysis, high MR-proADM levels were significantly associated with negative outcome (death or orotracheal intubation, IOT), with an odds ratio of 4.284 [1.893–11.413], together with increased neutrophil count (OR = 1.029 [1.011–1.049]) and WHO disease severity class (OR = 7.632 [5.871–19.496]). AUROC analysis showed a good discriminative performance of MR-proADM (AUROC: 0.849 [95% Cl 0.771–0.730]; p < 0.0001). The optimal value of MR-proADM to discriminate combined event of death or IOT is 0.895 nmol/l, with a sensitivity of 0.857 [95% Cl 0.728–0.987] and a specificity of 0.687 [95% Cl 0.587–0.787]. This study shows an association between MR-proADM levels and the severity of COVID-19. The assessment of MR-proADM combined with clinical scoring systems could be of great value in triaging, evaluating possible escalation of therapies, and admission avoidance or inclusion into trials. Larger prospective and controlled studies are needed to confirm these findings.openopenSozio E.; Tascini C.; Fabris M.; D'Aurizio F.; De Carlo C.; Graziano E.; Bassi F.; Sbrana F.; Ripoli A.; Pagotto A.; Giacinta A.; Gerussi V.; Visentini D.; De Stefanis P.; Merelli M.; Saeed K.; Curcio F.Sozio, E.; Tascini, C.; Fabris, M.; D'Aurizio, F.; De Carlo, C.; Graziano, E.; Bassi, F.; Sbrana, F.; Ripoli, A.; Pagotto, A.; Giacinta, A.; Gerussi, V.; Visentini, D.; De Stefanis, P.; Merelli, M.; Saeed, K.; Curcio, F

Biologically driven cut-off definition of lymphocyte ratios in metastatic breast cancer and association with exosomal subpopulations and prognosis

High neutrophil to lymphocyte ratio (NLR) and monocyte to lymphocyte ratio (MLR) are respectively associated with systemic inflammation and immune suppression and have been associated with a poor outcome. Plasmatic exosomes are extracellular vesicles involved in the intercellular communication system that can exert an immunosuppressive function. Aim of this study was to investigate the interplay between the immune system and circulating exosomes in metastatic breast cancer (MBC). A threshold capable to classify patients according to MLR, NLR and PLR, was computed through a receiving operator curve analysis after propensity score matching with a series of female blood donors. Exosomes were isolated from plasma by ExoQuick solution and characterized by flow-cytometry. NLR, MLR, PLR and exosomal subpopulations potentially involved in the pre-metastatic niche were significantly different in MBC patients with respect to controls. MLR was significantly associated with number of sites at the onset of metastatic disease, while high levels of MLR and NLR were found to be associated with poor prognosis. Furthermore, exosomal subpopulations varied according to NLR, MLR, PLR and both were associated with different breast cancer subtypes and sites of distant involvement. This study highlights the nuanced role of immunity in MBC spread, progression and outcome. Moreover, they suggest potential interaction mechanisms between immunity, MBC and the metastatic niche