A reverse transcription loop-mediated isothermal amplification (LAMP) assay for the detection of feline Coronavirus

Loop-mediated isothermal amplification (LAMP) is a molecular method that amplifies DNA underisothermal conditions. It relies on the use of 4 different primers recognizing 6 regions of the templatesequence and on the use of a DNA polymerase with strand displacement activity (Notomi et al., 2000).The addition of two loop primers allows the reaction time to be of one hour only (Nagamine et al., 2002).The aim of this study was to develop a reverse transcription LAMP assay for an easy and inexpensivedetection of feline Coronavirus (FCoV). Six primers binding the conserved 3’UTR region of the FCoVwere designed with the Primer Explorer software. Thirty-two samples of RNA (11 feces, 8 effusions, 9blood samples and 4 tissues) on which a reverse transcription polymerase chain reaction (RT-PCR) forthe 3’UTR region was performed were used. The reaction was carried out in 25μL reaction volume andthe mixture was incubated in a thermocycler at 63°C for 1 hour followed by 10 minutes at 80°C. LAMPproducts were visualized under UV after electrophoresis migration on a 1.5% agarose gel stained withethidium bromide, where they produce a ladder-like pattern if positive. Results where compared withthose obtained on standard PCR. Sensitivity and specificity were respectively 60% and 100% on feces,40% and 100% on effusions, 25% and 100% on blood, and 100% and 100% on tissues. The overall sensitivityand specificity of this method were of 57.1% and 100%, thus limiting a clinical application of this method,except for tissues

The gut microbiome and mucosal defenses in cats with coronaviruses: a pilot study

Feline Infectious Peritonitis (FIP) develops from a mutation of enteric feline coronaviruses (FCoVs) and an imbalance of the host immune response. The wide polymorphism of FCoVs is associated with the viral replication rate (Licitra et al. 2013).  Changes in the composition of the gut microbiota may induce quali-quantitative modifications in FCoVs and/or different immune profiles (Weese et al., 2015). Few information is available on feline gut microbiome and the association between microbiota and the predisposition to pathological conditions (Ramadan et al., 2014).The aim of this study is to provide preliminary data about the composition of gut microbiota in healthy cats compared with FCoV infected cats (with and without  FIP), in order to evaluate whether changes of gut microbiota may induce changes in FCoV, in its genetic polymorphism and in the mucosal immunity.Screening analyses have been performed on 22 cats:- Routine hematology and biochemistry on EDTA and serum (included electrophoresis and alpha-1-acid glycoprotein measurement for cats suspected with FIP)- Nested RT-PCR-3’UTR on frozen faeces- Effusion evaluation- FIV/FeLV serologyDue to strict inclusion criteria (cats younger than 2.5 years old, indoor and not assuming antibiotics in the previous two months) and based on the results obtained from the complete set of analysis, only 15 cats, specifically 5 cats for each of the following 3 groups: FIP- affected, healthy negative and positive for FCoV, have been recruited to perform the following analyses: - microbiota analysis through NGS of 16S rRNA gene (V4 region) amplicons followed by bioinformatic analysis -  evaluation of secretory IgA (ELISA kit)- phylogenetic analysis of FCoVs S gene sequencesInnovative results will be provided on the feline gut microbiota composition. These will be correlated with the presence and genetic polymorphisms of FCoV and mucosal defenses to establish significant correlations between the analysed factors

Activities Carried Out during the First COVID-19 Lockdown by Italian Citizens

: The SARS-CoV-2 pandemic has altered how citizens engage in activities. This study describes the new activities citizens engaged in during the first lockdown, factors that helped them cope with the confinement, the supports they used the most, and which supports they would have liked to receive. This cross-sectional study consists of an online survey made of 49 questions that was completed by the citizens of the province of Reggio Emilia (Italy) from 4 May until 15 June 2020. The outcomes of this study were explored by focusing on four of the survey questions. Of the 1826 citizens who responded, 84.2% had started new leisure activities. Males, participants who lived in the plain or foothills, and those who experienced nervousness engaged less in new activities, while those whose employment status changed, whose lifestyle worsened, or whose use of alcohol increased engaged in more activities. The support of family and friends, leisure activities, continuing to work, and an optimistic attitude were perceived to be of help. Grocery delivery and hotlines providing any type of information and mental health support were used frequently; a lack of health and social care services and of support in reconciling work with childcare was perceived. Findings may help institutions and policy makers to better support citizens in any future circumstances requiring prolonged confinement

Comparison between the diagnostic accuracy of clinico-pathological and molecular tests for feline infectious peritonitis (FIP)

The aim of this study was to compare the diagnostic accuracy for feline infectious peritonitis (FIP) of conventional clinic-pathological tests with that of molecular tests such as routine PCR and PCR followed by the sequencing of the Spike (S) gene. Blood, effusion and tissues specimens were collected from 21 FIP suspected cats. In vivo examination consisted of CBC, serum protein electrophoresis, AGP measurement, cytological and biochemical examination and the evaluation of the ΔTNC on effusions, and of molecular tests such the screening PCR (target: 3’UTR region) and the PCR directed towards the S gene followed by the amplification products sequencing in order to detect the aminoacidic substitution recently considered diagnostic for FIP1. These molecular techniques were applied to tissues collected during necropsy, which also allowed forming an FIP group (13 cats) and a non-FIP group (5 cats) based on histology and immunohistochemistry. The best test on tissues was immunohistochemistry (sens: 92.3%; spec: 100%), while the screening PCR suffered of low specificity (spec: 33.3%) and the S gene sequencing showed low sensitivity (sens: 69.2%).On effusions, the best tests resulted screening PCR and cytology (sens and spec: 100%) in comparison with the ΔTNC measurement (sens: 85.7 %; spec: 100%) and the S gene sequencing (sens: 42.8%; spec: 100%).On blood, the best test resulted AGP measurement (sens: 81.8%; spec: 100%), while serum protein electrophoresis showed a surprisingly low sensitivity (sens: 41.7%). Screening PCR (sens: 55.6%; spec: 100%) and S gene sequencing (sens: 33.3%; spec: 100%) proved again low accuracy.

Factors influencing return to work of cancer survivors: a population-based study in Italy

Little is known about return to work (RTW) of cancer survivors (CSs) in Central and Southern Europe. This study investigates the RTW rate of Italian CSs, describes their sick leave (SL) pattern, and explores factors affecting their RTW process. Methods A population-based cross-sectional survey involving CSs registered at the Cancer Registry of Reggio Emilia Province (Italy) was launched in July 2016. Eligibility was restricted to individuals with first diagnosis of cancer in 2012 (stages I–III), aged 20–59, and employed at the time of diagnosis. Results Of the 266 individuals interviewed, 140 (52.6%) were reintegrated without difficulty, 113 (42.5%) returned to work with some difficulty, and 13 did not RTW (4.9%). The majority of CSs (56%) took SL for some periods during treatment. Age > 50 years and higher income seemed to facilitate RTW (RR = 0.65, 95% CI 0.49–0.88 and RR = 0.72, 95% CI 0.54–0.97, respectively), while being divorced acted as a barrier compared to being married (RR = 1.45, 95% CI 1.04–2.01). Individuals uncertain about the type of company they were working for reported greater difficulty in RTW (RR = 1.68, 95% CI 1.03–2.72). Individuals who had undergone chemotherapy and those perceiving physical limitations had a higher risk of difficulty in the RTW process (RR = 1.79, 95% CI 1.42–2.24 and RR = 1.59, 95% CI 1.25–2.02, respectively). Conclusions Most CSs did RTW, with 2/3 combining work and treatment. However, almost half reported difficulty in RTW process. Factors affecting this process should be addressed throughout context-specific social and healthcare pathways aimed at preventing difficulties and potential job loss in this population

Patient-reported outcome measure to implement routine assessment of cancer survivors' unmet needs: An overview of reviews and COSMIN analysis

: As the number of cancer survivors (CSs) is increasing worldwide, providing services relevant to the specific, unmet needs of these individuals is crucial. There are currently various patient-reported outcome measures (PROMs) whose aim is to identify the unmet needs of CSs. Still, limited guidance supports healthcare providers in choosing the most valid and reliable PROMs for this purpose. We conducted this overview of systematic reviews (SRs) on the psychometric properties of PROMs addressing the unmet needs of adult CSs suffering from non-cutaneous cancers. We searched databases for SRs published between 2012 and January 2023. Two SRs were included, covering 14 PROMs tested on 19,151 CSs. These were assessed according to the COSMIN methodology for SRs of PROMs for the quality of their measurement properties and risk of bias, thus providing guidance in selecting PROMs that appropriately reflect the unmet needs of CSs

Comment on "How green is blue hydrogen?"

This paper is written in response to the paper “How green is blue hydrogen?” by R. W. Howarth and M. Z. Jacobson. It aims at highlighting and discussing the method and assumptions of that paper, and thereby providing a more balanced perspective on blue hydrogen, which is in line with current best available practices and future plant specifications aiming at low CO2 emissions. More specifically, in this paper, we show that: (i) the simplified method that Howarth and Jacobson used to compute the energy balance of blue hydrogen plants leads to significant overestimation of CO2 emissions and natural gas (NG) consumption and (ii) the assumed methane leakage rate is at the high end of the estimated emissions from current NG production in the United States and cannot be considered representative of all-NG and blue hydrogen value chains globally. By starting from the detailed and rigorously calculated mass and energy balances of two blue hydrogen plants in the literature, we show the impact that methane leakage rate has on the equivalent CO2 emissions of blue hydrogen. On the basis of our analysis, we show that it is possible for blue hydrogen to have significantly lower equivalent CO2 emissions than the direct use of NG, provided that hydrogen production processes and CO2 capture technologies are implemented that ensure a high CO2 capture rate, preferably above 90%, and a low-emission NG supply chain