Diagnosing congenital Cytomegalovirus infection: Don't get rid of dried blood spots

Background: Congenital Cytomegalovirus (cCMV) is a serious global public health issue that can cause irreversible fetal and neonatal congenital defects in symptomatic or asymptomatic newborns at birth. In absence of universal cCMV screening, the retrospective diagnosis of cCMV infection in children is only possible by examining Dried Blood Spot (DBS) samples routinely collected at birth and stored for different time spans depending on the newborn screening regulations in force in different countries. In this article, we summarize the arguments in favor of long-term DBS sample storage for detecting cCMV infection. Main text: CMV infection is the most common cause of congenital infection resulting in severe defects and anomalies that can be apparent at birth or develop in early childhood. Sensorineural hearing loss is the most frequent consequence of cCMV infection and may have a late onset and progress in the first years of life. The virological diagnosis of cCMV is essential for clinical research and public health practices. In fact, in order to assess the natural history of CMV infection and distinguish between congenital or acquired infection, children should be diagnosed early by analyzing biological samples collected in the first weeks of life (3 weeks by using viral culture and 2 weeks by molecular assays), which, unfortunately, are not always available for asymptomatic or mildly symptomatic children. It now seems possible to overcome this problem since the CMV-DNA present in the blood of congenitally infected newborns can be easily retrieved from the DBS samples on the Guthrie cards routinely collected and stored within 3 days from birth in the neonatal screening program for genetic and congenital diseases. Early collection and long-term storage are inexpensive methods for long-term bio-banking and are the key points of DBS testing for the detection of cCMV. Conclusion: DBS sampling is a reliable and inexpensive method for long-term bio-banking, which enables to diagnose known infectious diseases - including cCMV - as well as diseases not jet recognized, therefore their storage sites and long-term storage conditions and durations should be the subject of political decision-making

Measurement of surface velocity in open channels using a lightweight remotely piloted aircraft system

ABSTRACTIn this paper, a low-cost remotely piloted aircraft system (RPAS) technique is proposed for measurement of the surface velocity in rivers or channels with low surface velocity and small discharge. To verify the reliability of the results obtained with the RPAS, we simultaneously measured the surface velocity with other methods based on total stations and close range photogrammetry. The RPAS was used both with ground control points (GCPs) for orientation of the photographic images and without GCPs. The data analysis showed that the RPAS provides valid results even without GCPs. Use of a RPAS without GCPs, relying solely on flight altitude to determine the water velocity, opens the way for its utilization in emergency conditions when it is impossible to access the river banks for the realization and survey of GCPs

Contribution of respiratory syncytial virus (RSV) among patients <15 years hospitalized with severe acute respiratory infection (SARI) in Milan, 2014-2017

Aim: Respiratory syncytial virus (RSV) infections may range from cold to severe acute respiratory infection (SARI) and are responsible for substantial pediatric morbidity. We describe the results of RSV molecular detection in respiratory samples collected from children <15 years hospitalized with SARI in Milan (Italy) during four consecutive years. Method: From January 1st, 2014, to December 31st, 2017, 3013 respiratory samples (2826 upper-respiratory-tract [UTR] and 187 lower-respiratory-tract [LTR] specimens) collected from as many children <15 years hospitalized with SARI at an University hospital in Milan were analysed. After DNA/RNA extraction, samples were tested by a multiplex real-time PCR to detect RSV and other respiratory viruses. Results: 571 (19%) respiratory samples tested RSV-positive. RSV-positivity rate by sample type was similar (URT vs LRT: 19.2% vs 14.4%; p=0.09). The median age of RSV-positive cases was 6.6 months (inter-quartile range: 17.2 months); 52.2% were males. 62.2% (355/571) of RSV-positive samples were identified in children <1 year and 12.4% (71/571) in those <1 month. RSV was detected throughout the study period; 59.2% (338/571) cases were identified during seasonal peaks (December-February). In 49.9% (285/571) of RSV-positive samples at least another virus (mainly Rhinovirus: 45.9%) was detected, particularly (60%; 171/285) in samples collected from children >1 year. Conclusions: Accordingly to other studies, RSV was detected in 19% of hospitalized-SARI cases <15 years, mainly in children <1 year and in December-February. Sampling of upper or lower airways resulted in similar RSV-positivity rate. Routine molecular testing to detect RSV is warranted to improve clinical management of pediatric patients

Epidemiological and molecular characteristics of HPEV infection in children<6 months hospitalized with symptoms of sepsie-like illness, northen Italy, 2015-2018

BACKGROUND. Human parechoviruses (HPeVs) are widespread pathogens belonging to the Picornaviridae family and currently divided into 19 genotypes. HPeV infections can be associated with severe clinical manifestations, such as sepsis-like illness, particularly in youngest children. The epidemiological and molecular characteristics of HPeV infections observed in children <6 months hospitalized with symptoms of sepsis-like illness were investigated. METHODS. From January 1st, 2015, to December 31st, 2018, clinical samples (cerebrospinal fluid samples and/or blood samples) were collected for diagnosis of HPeV infection from 193 patients (median age: 21 days, range: 1 day - 6 months) hospitalized with symptoms of sepsis-like illness, in two hospitals of Northern Italy. HPeV-RNA was detected by real-time RT-PCR (target 5\u2019UTR) and a portion of HPeV VP3/VP1 junction (nt. 2159\u20132458) was sequenced for typing and molecular characterization. RESULTS. 14% (27/193) of patients with symptoms of sepsis-like illness tested HPeV-positive. 26/27 (96.3%) HPeV-cases were <3 months and 20/27 (74.1%) <1 month. HPeV-positive cases were detected throughout the study period, mainly (12/27; 44.4%) during the summertime (June-August). 17/27 (63%) HPeV-positive samples were molecularly characterized: 16 resulted HPeV-3 and 1 HPeV-5. CONCLUSIONS. HPeV infection was identified in 14% of children <6 months with symptoms of sepsis-like illness. Almost all HPeV infections were detected in children <3 months and mainly during the summertime; almost all molecularly characterized HPeV belonged to type 3. Including HPeV molecular detection in routine diagnostic tests would allow estimating the burden of HPeV infection and improving clinical management of pediatric patients

Hospital discharges-based search of acute flaccid paralysis cases 2007-2016 in Italy and comparison with the National Surveillance System for monitoring the risk of polio reintroduction

Background: Acute flaccid paralysis (AFP) surveillance has been adopted globally as a key strategy for monitoring the progress of the polio eradication initiative. Hereby, to evaluate the completeness of the ascertainment of AFP cases in Italy, a hospital-discharges based search was carried out. Methods: AFP cases occurring between 2007 and 2016 among children under 15 years of age were searched in the Italian Hospital Discharge Records (HDR) database using specific ICD-9-CM diagnostic codes. AFP cases identified between 2015 and 2016 were then compared with those notified to the National Surveillance System (NSS). Results: Over a 10-year period, 4163 hospital discharges with diagnosis of AFP were reported in Italy. Among these, 956 (23.0%) were acute infective polyneuritis, 1803 (43.3%) myopathy, and 1408 (33.8%) encephalitis, myelitis and encephalomyelitis. During the study period, a decreasing trend was observed for all diagnoses and overall the annual incidence rate (IR) declined from 5.5 to 4.5 per 100,000 children. Comparing NSS with HDR data in 2015-2016, we found a remarkable underreporting, being AFP cases from NSS only 14% of those recorded in HDR. In particular, the acute infective polyneuritis cases reported to NSS accounted for 42.6% of those detected in HDR, while only 0.9% of myopathy cases and 13.1% of encephalitis/myelitis/encephalomyelitis cases have been notified to NSS. The highest AFP IRs per 100,000 children calculated on HDR data were identified in Liguria (17.4), Sicily (5.7), and Veneto (5.1) Regions; regarding the AFP notified to the NSS, 11 out of 21 Regions failed to reach the number of expected cases (based on 1/100,000 rate), and the highest discrepancies were observed in the Northern Regions. Overall, the national AFP rate was equal to 0.6, therefore did not reach the target value. Conclusions: AFP surveillance data are the final measure of a country's progress towards polio eradication. The historical data obtained by the HDR have been useful to assess the completeness of the notification data and to identify the Regions with a low AFP ascertainment rate in order to improve the national surveillance system

Influenza Vaccination in Italian Healthcare Workers (2018-2019 Season): Strengths and Weaknesses. Results of a Cohort Study in Two Large Italian Hospitals

Background: Annual vaccination is the most effective way to combat influenza. As influenza viruses evolve, seasonal vaccines are updated annually. Within the European project Development of Robust and Innovative Vaccine Effectiveness (DRIVE), a cohort study involving Italian healthcare workers (HCWs) was carried out during the 2018-2019 season. Two aims were defined: to measure influenza vaccine effectiveness (IVE) against laboratory-confirmed influenza cases and to conduct an awareness-raising campaign to increase vaccination coverage. Methods: Each subject enrolled was followed up from enrollment to the end of the study. Each HCW who developed ILI was swabbed for laboratory confirmation of influenza. Influenza viruses were identified by molecular assays. A Cox regression analysis, crude and adjusted for confounding variables, was performed to estimate the IVE. Results: Among the 4483 HCWs enrolled, vaccination coverage was 32.5%, and 308 ILI cases were collected: 23.4% were positive for influenza (54.2% A(H1N1) pdm09; 45.8% A(H3N2)). No influenza B viruses were detected. No overall IVE was observed. Analyzing the subtypes of influenza A viruses, the IVE was estimated as 45% (95% CI: -59 to 81) for A(H1N1) pdm09. Conclusions: Vaccination coverage among HCWs increased. Study difficulties and the circulation of drifted variants of A(H3N2) could partly explain the observed IVE

Sphingolipids and glycerophospholipids - The "ying and yang" of lipotoxicity in metabolic diseases

Sphingolipids in general and ceramides in particular, contribute to pathophysiological mechanisms by modifying signalling and metabolic pathways. Here, we present the available evidence for a bidirectional homeostatic crosstalk between sphingolipids and glycerophospholipids, whose dysregulation contributes to lipotoxicity induced metabolic stress. The initial evidence for this crosstalk originates from simulated models designed to investigate the biophysical properties of sphingolipids in plasma membrane representations. In this review, we reinterpret some of the original findings and conceptualise them as a sort of "$\textit{ying/yang}$" interaction model of opposed/complementary forces, which is consistent with the current knowledge of lipid homeostasis and pathophysiology. We also propose that the dysregulation of the balance between sphingolipids and glycerophospholipids results in a lipotoxic insult relevant in the pathophysiology of common metabolic diseases, typically characterised by their increased ceramide/sphingosine pools.This work was funded by the Medical Research Council Programme grants (MC_UU_12012/2) “Lipotoxicity and obesity-associated metabolic complications”. FP7-ETHERPATHS (KBBE-2007-2A-222639-2), FP7-MITIN (223450), BBSRC (BB/H002731/1)

Defective extracellular matrix remodeling in brown adipose tissue is associated with fibro-inflammation and reduced diet-induced thermogenesis.

The relevance of extracellular matrix (ECM) remodeling is reported in white adipose tissue (AT) and obesity-related dysfunctions, but little is known about the importance of ECM remodeling in brown AT (BAT) function. Here, we show that a time course of high-fat diet (HFD) feeding progressively impairs diet-induced thermogenesis concomitantly with the development of fibro-inflammation in BAT. Higher markers of fibro-inflammation are associated with lower cold-induced BAT activity in humans. Similarly, when mice are housed at thermoneutrality, inactivated BAT features fibro-inflammation. We validate the pathophysiological relevance of BAT ECM remodeling in response to temperature challenges and HFD using a model of a primary defect in the collagen turnover mediated by partial ablation of the Pepd prolidase. Pepd-heterozygous mice display exacerbated dysfunction and BAT fibro-inflammation at thermoneutrality and in HFD. Our findings show the relevance of ECM remodeling in BAT activation and provide a mechanism for BAT dysfunction in obesity