Sviluppo di una piattaforma per la diagnosi prenatale non invasiva di malattie genetiche in epoca gestazionale precoce

Prenatal diagnosis of aneuploidies and monogenic diseases is usually performed by amniocentesis or chorionic villous sampling. However, these procedures are associated with 0.5%-2% risk of miscarriage. The discovery of cell free fetal DNA (cffDNA) in maternal plasma in 1997 has provided a new source of fetal genetic material that can be safely obtained from maternal blood and successfully processed for non invasive genetic diagnosis (NIPD). In this study is described a new approach for non invasive prenatal diagnosis of β- thalassemia which is based on semiconductor sequencing (Ion Torrent PGM) and fetal haplotype inference. In particular, the approach is based on target sequencing of the mutation site, the β°39 non sense mutation of the HBB gene, and several informative SNPs spread in the β-globin gene cluster. The data analysis of each cffDNA sample and the inference of the most likely inherited haplotypes were determined by an automated pipeline which firstly constructs the parental haplotypes, using the sequencing data from the parental DNAs and an haplotype reference panel previously created. The pipeline, then, quantifies the allele counts observed in each site sequenced in the corresponding cffDNA sample and finally predicts the two haplotypes most likely inherited by the fetus using a hidden Markov Model (HMM). The results were finally compared with the sequencing data of the fetal DNA obtained by villocentesis. Using these approaches we have analyzed 30 out of 37 cffDNA samples; in seven samples, in fact, the pipeline could not proceed because of the lack of informative sites or of other parameters useful for downstream analysis. The fetal β°39 genotype was correctly predicted in 24/30 (80%) samples, while it was incorrectly defined in 6/30 (20%) cases. The incorrect results obtained in these last samples was due to the erroneus inference of the maternal haplotype. On the contrary, the paternal haplotype was correctly detected in all 30 samples processed. In the next future we are planning to improve the protocol by increasing the number of potentially informative SNPs and to process a higher number of Sardinian β°39 carriers in order to expand the haplotype reference panel. This haplotype-based approach has given encouraging results and we think that it could be a starting point for eventual future application of NIPD also to other monogenic disorders

Fatality rate and predictors of mortality in an Italian cohort of hospitalized COVID-19 patients

Clinical features and natural history of coronavirus disease 2019 (COVID-19) differ widely among different countries and during different phases of the pandemia. Here, we aimed to evaluate the case fatality rate (CFR) and to identify predictors of mortality in a cohort of COVID-19 patients admitted to three hospitals of Northern Italy between March 1 and April 28, 2020. All these patients had a confirmed diagnosis of SARS-CoV-2 infection by molecular methods. During the study period 504/1697 patients died; thus, overall CFR was 29.7%. We looked for predictors of mortality in a subgroup of 486 patients (239 males, 59%; median age 71 years) for whom sufficient clinical data were available at data cut-off. Among the demographic and clinical variables considered, age, a diagnosis of cancer, obesity and current smoking independently predicted mortality. When laboratory data were added to the model in a further subgroup of patients, age, the diagnosis of cancer, and the baseline PaO2/FiO2 ratio were identified as independent predictors of mortality. In conclusion, the CFR of hospitalized patients in Northern Italy during the ascending phase of the COVID-19 pandemic approached 30%. The identification of mortality predictors might contribute to better stratification of individual patient risk

Goodbye Hartmann trial: a prospective, international, multicenter, observational study on the current use of a surgical procedure developed a century ago

Background: Literature suggests colonic resection and primary anastomosis (RPA) instead of Hartmann's procedure (HP) for the treatment of left-sided colonic emergencies. We aim to evaluate the surgical options globally used to treat patients with acute left-sided colonic emergencies and the factors that leading to the choice of treatment, comparing HP and RPA. Methods: This is a prospective, international, multicenter, observational study registered on ClinicalTrials.gov. A total 1215 patients with left-sided colonic emergencies who required surgery were included from 204 centers during the period of March 1, 2020, to May 31, 2020. with a 1-year follow-up. Results: 564 patients (43.1%) were females. The mean age was 65.9 ± 15.6 years. HP was performed in 697 (57.3%) patients and RPA in 384 (31.6%) cases. Complicated acute diverticulitis was the most common cause of left-sided colonic emergencies (40.2%), followed by colorectal malignancy (36.6%). Severe complications (Clavien-Dindo ≥ 3b) were higher in the HP group (P < 0.001). 30-day mortality was higher in HP patients (13.7%), especially in case of bowel perforation and diffused peritonitis. 1-year follow-up showed no differences on ostomy reversal rate between HP and RPA. (P = 0.127). A backward likelihood logistic regression model showed that RPA was preferred in younger patients, having low ASA score (≤ 3), in case of large bowel obstruction, absence of colonic ischemia, longer time from admission to surgery, operating early at the day working hours, by a surgeon who performed more than 50 colorectal resections. Conclusions: After 100 years since the first Hartmann's procedure, HP remains the most common treatment for left-sided colorectal emergencies. Treatment's choice depends on patient characteristics, the time of surgery and the experience of the surgeon. RPA should be considered as the gold standard for surgery, with HP being an exception

Effects of Anacetrapib in Patients with Atherosclerotic Vascular Disease

BACKGROUND: Patients with atherosclerotic vascular disease remain at high risk for cardiovascular events despite effective statin-based treatment of low-density lipoprotein (LDL) cholesterol levels. The inhibition of cholesteryl ester transfer protein (CETP) by anacetrapib reduces LDL cholesterol levels and increases high-density lipoprotein (HDL) cholesterol levels. However, trials of other CETP inhibitors have shown neutral or adverse effects on cardiovascular outcomes. METHODS: We conducted a randomized, double-blind, placebo-controlled trial involving 30,449 adults with atherosclerotic vascular disease who were receiving intensive atorvastatin therapy and who had a mean LDL cholesterol level of 61 mg per deciliter (1.58 mmol per liter), a mean non-HDL cholesterol level of 92 mg per deciliter (2.38 mmol per liter), and a mean HDL cholesterol level of 40 mg per deciliter (1.03 mmol per liter). The patients were assigned to receive either 100 mg of anacetrapib once daily (15,225 patients) or matching placebo (15,224 patients). The primary outcome was the first major coronary event, a composite of coronary death, myocardial infarction, or coronary revascularization. RESULTS: During the median follow-up period of 4.1 years, the primary outcome occurred in significantly fewer patients in the anacetrapib group than in the placebo group (1640 of 15,225 patients [10.8%] vs. 1803 of 15,224 patients [11.8%]; rate ratio, 0.91; 95% confidence interval, 0.85 to 0.97; P=0.004). The relative difference in risk was similar across multiple prespecified subgroups. At the trial midpoint, the mean level of HDL cholesterol was higher by 43 mg per deciliter (1.12 mmol per liter) in the anacetrapib group than in the placebo group (a relative difference of 104%), and the mean level of non-HDL cholesterol was lower by 17 mg per deciliter (0.44 mmol per liter), a relative difference of -18%. There were no significant between-group differences in the risk of death, cancer, or other serious adverse events. CONCLUSIONS: Among patients with atherosclerotic vascular disease who were receiving intensive statin therapy, the use of anacetrapib resulted in a lower incidence of major coronary events than the use of placebo. (Funded by Merck and others; Current Controlled Trials number, ISRCTN48678192 ; ClinicalTrials.gov number, NCT01252953 ; and EudraCT number, 2010-023467-18 .)

Laboratory Simulation of Transients in Looped Water Distribution Network

This paper analyses some tests carried out in a looped water distribution network (WDN) at the Water Engineering Laboratory of the University of Perugia. Specifically, the aim of such tests is to evaluate the effect of transients generated by a water consumption change, i. e., a complete and fast closure of a valve simulating an end-user maneuver. This end-user was located at the downstream end section of a service line placed in three different sections of the WDN. The tests allow examining the effect of the network topology and the location of the transient generation point

A Quick Survey of the Most Vulnerable Areas of a Water Distribution Network Due to Transients Generated in a Service Line: A Lagrangian Model Based on Laboratory Tests

This paper analyses the propagation and mechanisms of interaction of a pressure wave in a looped water distribution network by means of laboratory and numerical tests. Transients are generated by the complete and fast closure of a valve, simulating an end-user maneuver, located at the downstream end section of a service line. The adequate length of the service line allows capturing each single pressure wave inserted into the network. The executed tests and successive analysis by means of a Lagrangian model (LM) highlight the effect of the network topology and the location of the transient generation point but in a more expeditious way with respect to the use of a complete transient model. The most excited part of the system is the one in close proximity of the end-user and then the corresponding service line. Within the network, pressure waves accumulate in the areas with the smallest diameter pipes. By means of the refined LM—which is able to capture the pressure extreme values occurring in the first phases of the transient—the vulnerability maps of the network are provided. Such maps identify the nodes subjected to the most severe pressure waves in terms of both frequency and amplitude. The exposure level to transients of each node is synthesized by the value of the vulnerability index proposed in this paper

Monitoring a real service line under user activity

The present study is aimed at characterizing a real service line subjected to users’ activity. The minor system considered serves a holiday home in a seaside resort on the Adriatic coast, in northern Italy. The household service line was monitored by acquiring pressure sampled at a frequency of 500 Hz. Two experimental campaigns were executed: in a winter period when no people were living in the holiday home nor in nearby houses, and in a summer period during which both the selected holiday home and nearby houses were inhabited. In the first campaign, transients were generated by manoeuvring single devices within the plumbing system of the house (mixer taps, knob taps and toilet flushes), and pressure signals were acquired just during these manoeuvers. The aim of these tests was to characterize the effects of each single device manoeuver, in pressure terms. In the second campaign, pressure signals were acquired in continuous for more than one week. The analyses of the collected data highlighted that the service line is subjected to pressure variations which can be very significant and depend on the type and position of the operated devices within the plumbing system of the house, as observed in the first campaign, and also of the nearby users, as observed in the second campaign