Virological surveillance of SARS-CoV-2 in an Italian northern area: comparison of Real Time RT PCR cycle threshold (Ct) values in three epidemic periods.

Aim of the study was to investigate the differences in Ct values in nasopharingeal swabs collected in three SARS-CoV-2 epidemic periods: first one from February 23 to March 25 (14 days from lockdown started on March 11); the second one from  March  26 to May 18 (14 days  from the end of strict lockdown on May 4) and the third one from May 19 until June 15. Viral RNA was detected in nasopharyngeal swabs obtained both from inpatients and outpatients. COVID-19 infection was confirmed according to the Ct values for N1 and N2 genes ascertained by Real-Time RT-PCR assay as described by the CDC. We calculated the prevalence of nasopharyngeal swabs tested positive for SARS-CoV-2, the mean and median of the Cts and the percentage of samples equal or below the Ct value of 25 in the 3 periods considered. The average value of Ct increased, going from 24.80 in the first epidemic period to 26.64 in the second period to 28.50 in the third period (p <0.001). The percentage of samples with Ct lower than or equal to 25 also decreased sharply from 54.7% to 20.0%. These findings need to be integrated with epidemiological and clinical data.

Influenza vaccine effectiveness in children: a retrospective study on eight post-pandemic seasons with trivalent inactivated vaccine.

BACKGROUND AND AIM OF THE WORK: The global burden of disease attributable to seasonal influenza virus in children is difficult to quantify. Children with chronic medical conditions and healthy children may experience severe or fatal complications. Aim of the study was to estimate the influenza vaccine effectiveness (VE) in a cohort of outpatient children. METHODS: From 2010 to 2018, a Pediatrician of Parma from the InfluNet network of Emilia-Romagna Region, performed nasal/throat swabs on every child with Influenza-like illness at least 14 days from the vaccination with trivalent vaccine. VE estimates against influenza season, virus type and subtype and age group were evaluated using a test-negative design. RESULTS: 2,480 swabs were performed. The 57.6% of the analyzed swabs were positive for influenza viruses. Type A (57%) and type B viruses (43%) co-circulated. The 37.1% of type A viruses belonged to subtype A(H3N2), 19.4% to subtype A(H1N1)pdm09. The subtype A(H3N2) was prevalent among children up to 23 months (42.4%) while the type B in the 2-4 (40.7%) and 5-16 year old age groups (49.4%). Overall, 19.9% of the children were vaccinated. The highest prevalence of vaccinated subjects was found in children aged 5-16 (30.5%). The VE against subtype A(H1N1)pdm09 was 63% (95%CI 42.6-76.0), against type B 27.5% (95%CI 7.9-42.9) and against subtype A(H3N2) -14.3%  (95%CI - 46.0-10.7). CONCLUSIONS: Our findings represent a useful contribution to the ongoing debate about the appropriateness of including influenza vaccination for healthy children, 6 months and older, in the updating National Vaccine Prevention Plan (PNPV).

Immunity status against tetanus in young migrants: a seroprevalence study.

BACKGROUND AND AIM OF THE WORK: Thanks to the highly effective vaccine, tetanus became sporadic in high-income countries with well-established primary childhood immunization programs, but it is common in low-income countries. The migrants, leaving countries with poor immunization programs or where vaccinations have been interrupted, may represent a new risk group for tetanus in host countries. A seroprevalence study was conducted to estimate the immunological status against tetanus in young migrants without vaccination documentation. METHODS: After a careful assessment by vaccination services of the Local Health Authority, all migrants recently arrived in Italy were included in the serosurvey. Titers of anti-tetanus toxoid were measured using a commercial ELISA kit. Subjects were stratified by age and by WHO region. Antibody titers <0.10 IU/ml were considered to be seronegative, between 0.10 and 1.00 IU/ml as intermediate protection, and >1.00 IU/ml high protection. RESULTS: From January 2004 to December 2019, 2,326 blood samples were collected. Mean age was 13.9 years with no differences between WHO regions. The percentage of the subjects without protective antibodies was 22.3%, with an intermediate level was 45.2%, with high titer was 32.5%. Among migrant coming from African and Eastern Mediterranean WHO regions, the highest percentages of seronegative titers and, at the same time, the low percentages of high protective levels were found. Titers decreased with age. CONCLUSIONS: The significant proportion of seronegative migrants and the decrease of protective titers increasing age, confirm the importance of the evaluation of the immunological status to employ the appropriate vaccination strategy.

["Activate your Wait" project : pilot study carried out at a thermal centre]

Physical activity is recognised as a major health determinant, with positive effects on health, environmental sustainability and economy. National surveillance data show that one out of three Italians - adult and elderly alike - declares to be sedentary, with a progressively increasing trend. From the urgent need to implement strategies to promote physical activity the "Activate your Wait" ("Attiva l'Attesa") project was born, aimed at transforming the waiting pauses during day to day life into opportunities to perform simple stretching and active mobilization exercises. The pilot study was carried out at the Terme S. Egidio, Suio Terme Castelforte (Latina). The results of the questionnaire distributed in the pre-intervention phase, aimed at assessing the users' interest in the project and physical activity in general, are reported. The questionnaire was administrated in the waiting rooms in September 2018 and September 2019. A total of 129 subjects responded to the questionnaire: 43% declared themselves sedentary, 73% reported willingness to perform simple physical exercises while waiting, and 76% believed that physical activity during waiting moments could have a positive impact on health. The project's subsequent goal is to identify suitable exercises to be proposed during the intervention period, which can be easily reproduced independently by users in their everyday life.

Local Excision of a Mucinous Adenocarcinoma of the Anal Margin (Extramammary Paget's Disease) and Reconstruction with a Bilateral V-Y Flap.

The case of a 75-year-old female with invasive extramammary Paget's disease of the anal margin, without involvement of the anal canal, is reported. The patient underwent wide local excision of the lesion with reconstruction with a double V-Y flap, a biopsy of the inguinal sentinel node, and a laparoscopic temporary colostomy. No guidelines exist on the treatment of this rare disease, and both wide local excision and abdominoperineal resection have been proposed. In the present case, the absence of invasion of the anal canal, also confirmed by intraoperative biopsies on the resection margins, and of local lymph node metastasis, as confirmed by the sentinel lymph node biopsy, allowed a sphincter-sparing operation with good functional and oncological results.

On field vaccine effectiveness in three periods of 2018/2019 influenza season in Emilia-Romagna Region.

BACKGROUND AND AIM OF THE WORK: Epidemic influenza is associated with significant morbidity and mortality, particularly in people at risk. The vaccine reduces complications, hospitalization and mortality excess, as well as health care and social costs. Aim of the study was to estimate the influenza vaccine effectiveness (VE) in Emilia-Romagna Region during the 2018/2019 season. METHODS: Within the context of virological surveillance conducted at the Regional Reference Laboratory of Parma, nasal/throat swabs were performed by sentinel practitioners and clinicians, on patients with ILI (Influenza-like illness). VE estimates, overall and against subtype A(H1N1)pdm09 and A(H3N2), were evaluated in three periods of the season, using a test-negative case-control design. RESULTS: From November 2018 to April 2019, 2,230 specimens were analyzed: 1,674 (75.1%) performed by clinicians and 556 (24.9%) by sentinel practitioners of the regional network. The season was characterized by the predominant circulation of influenza type A viruses: 57.4% belonged to subtype A(H3N2), 41.2% to subtype A(H1N1)pdm09. 23.5% of patients was vaccinated against influenza with quadrivalent or adjuvate vaccine. The overall VE was -5% (95% CI -33% - 18%) with a decreasing trend during the season. The overall VE against subtype A(H1N1)pdm09 was 39% (95% CI 11% - 58%) and remained stable during the season. The overall VE against subtype A(H3N2) was -43% (95% CI -89% - -9%), and showed an important decreasing trend. CONCLUSIONS: The possibility to make accurate and continuous VE estimates during the season will help to better define the composition of the vaccine for the following season.

Virological Surveillance of Influenza in the eight epidemic seasons after the 2009 pandemic in Emilia-Romagna (Northern Italy).

BACKGROUND AND AIM OF THE WORK: Influenza virological surveillance is essential for monitoring the evolution of influenza viruses (IVs) as well as for annual updating of the vaccine composition. The aim of this study is to analyse IVs circulation in Emilia-Romagna during the eight epidemic seasons after the 2009 pandemic and to evaluate their match with seasonal vaccine strains. METHODS: A total of 7882 respiratory specimens from patients with influenza-like illness (ILI), were collected by regional sentinel practitioners and hospital physicians. Viral investigations were conducted by rRT-PCR assay. Genetic characterization was performed for a spatial-temporal representative number of influenza laboratory-confirmed specimens. RESULTS: Influenza-positive samples per season ranged between 28.9% (2013-2014) and 66.8% (2012-2013). Co-circulation of IVs type A and type B was observed in all seasons, although with a different intensity. In all seasons, the highest number of positive samples was recorded in younger patients aged 5-14 years with relative frequencies ranging from 40% in the 2013-2014 season and 78% in the 2012-2013 season. Since the 2009 pandemic, A/H1N1pdm09 IVs circulating were closely related to the vaccine strain A/California/7/2009. Antigenic mismatch between vaccine strain and A/H3N2 IVs was observed in the 2011-2012 and 2014-2015 seasons. During 2015-2016, 2016-2017 and 2017-2018 seasons a complete or nearly complete mismatch between the predominant influenza B lineage of IVs type B circulating and vaccine B lineage occurred. CONCLUSIONS: This analysis confirms the importance of the virological surveillance and highlights the need of a continuous monitoring of IVs circulation, to improve the most appropriate vaccination strategies. (www.actabiomedica.it).

Scarless outpatient ablation of pilonidal sinus: a pilot study of a new minimally invasive treatment.

Pain and secretion of purulent materials are symptoms that are often associated with the pilonidal sinus. Generally, these symptoms are neglected by patients for a long time. Patients seek medical attention too late, fearing a prolonged period of pain and inability after surgery. The optimal therapy for pilonidal sinus should have the following characteristics: high healing rate, low recurrence rate, minimal postoperative pain and low cost. The aim of this study is to explore the effectiveness and safety of a new minimally invasive treatment: the scarless outpatient ablation of pilonidal sinus (SOAP). A total of 31 consecutive patients were enrolled; 27 of 31 patients were declared healed after the surgery (87·09%). After 1 year, only 4 of 27 patients (14·81%) reported symptoms related to recurrence of the disease. The average duration of the operation was 7·47 minutes. Patients reported the disappearance of painful symptoms after approximately 2·62 days and had been away from work for 0·53 days. No complications were recorded during the period of study. The advantages of the procedure examined in this study are its simplicity and rapidity of execution, its outpatient setting, its low cost and lack of complications. In our opinion, a randomised controlled trial should be conducted to validate the results related to this technique.

MicroRNA profiling of hepatocarcinogenesis identifies C19MC cluster as a novel prognostic biomarker in hepatocellular carcinoma.

BACKGROUND AND AIMS: Progressive hepatocarcinogenesis is a stepwise process that drives liver transformation. However, the molecular mechanisms of early liver transformation are far from clear. A role for microRNAs (miRNA) as diagnostic and prognostic factors in human tumours, including hepatocellular carcinoma (HCC), is promising. We aimed to identify novel miRNA as biomarkers for differential diagnosis and predictors of disease progression. METHODS: We used a low-density array platform to profile the expression of 664 mature miRNA in a cohort of 60 hepatitis C virus-positive liver lesions representative of all stages of progressive hepatocarcinogenesis. We validated selected miRNA in two independent patient series by qPCR and we characterized the genomic status of the miRNA cluster C19MC by fluorescent in situ hybridization and copy-number variation analyses. RESULTS: A 18-miRNA signature distinguished cirrhosis, dysplastic nodules and HCC lesions. Four miRNAs overexpressed in HCCs belonged to chromosome 19 miRNA cluster (C19MC). Significant overexpression of C19MC in early HCC compared to dysplastic nodules could be confirmed in a second series of hepatitis B virus-related liver lesions (n = 30). In a third series of 61 HCCs, C19MC cluster was overexpressed in HCCs compared to matched cirrhotic parenchyma and regardless of the type of viral infection. High C19MC miRNA levels were correlated with poor clinico-pathological features, increased risk of tumour recurrence and shorter overall survival time. HCCs overexpressing the C19MC cluster showed genetic amplification of the corresponding locus. CONCLUSIONS: C19MC cluster is a novel molecular alteration characteristic of liver cancer and predictor of poor prognosis. C19MC is an attractive candidate for novel HCC therapies.

Inhibitors of apoptosis proteins (IAPs) expression and their prognostic significance in hepatocellular carcinoma.

BACKGROUND: Similarly to other tumor types, an imbalance between unrestrained cell proliferation and impaired apoptosis appears to be a major unfavorable feature of hepatocellular carcinoma (HCC). The members of IAP family are key regulators of apoptosis, cytokinesis and signal transduction. IAP survival action is antagonized by specific binding of Smac/DIABLO and XAF1. This study aimed to investigate the gene and protein expression pattern of IAP family members and their antagonists in a series of human HCCs and to assess their clinical significance. METHODS: Relative quantification of IAPs and their antagonist genes was assessed by quantitative Real Time RT-PCR (qPCR) in 80 patients who underwent surgical resection for HCC. The expression ratios of XIAP/XAF1 and of XIAP/Smac were also evaluated. Survivin, XIAP and XAF1 protein expression were investigated by immunohistochemistry. Correlations between mRNA levels, protein expression and clinicopathological features were assessed. Follow-up data were available for 69 HCC patients. The overall survival analysis was estimated according to the Kaplan-Meier method. RESULTS: Survivin and Livin/ML-IAP mRNAs were significantly over-expressed in cancer tissues compared to non-neoplastic counterparts. Although Survivin immunoreactivity did not correlate with qPCR data, a significant relation was found between higher Survivin mRNA level and tumor stage, tumor grade and vascular invasion.The mRNA ratio XIAP/XAF1 was significantly higher in HCCs than in cirrhotic tissues. Moreover, high XIAP/XAF1 ratio was an indicator of poor prognosis when overall survival was estimated and elevated XIAP immunoreactivity was significantly associated with shorter survival. CONCLUSION: Our study demonstrates that alterations in the expression of IAP family members, including Survivin and Livin/ML-IAP, are frequent in HCCs. Of interest, we could determine that an imbalance in XIAP/XAF1 mRNA expression levels correlated to overall patient survival, and that high XIAP immunoreactivity was a poor prognostic factor.