Laboratory-based surveillance of invasive listeriosis in Northern Italy over a fourteen-year period: epidemiological and clinical results.

INTRODUCTION: Invasive listeriosis is a rare foodborne disease with a significant impact on public health worldwide, because of the severity of its clinical manifestations and high fatality rate. In this study, we provide a snapshot of epidemiology of listeriosis in Lombardy Region, Northern Italy, reviewing enhanced surveillance data collected over fourteen years, after the implementation of a voluntary laboratory-based surveillance system for the referral of clinical isolates of Listeria monocytogenes to a regional reference laboratory, since 2005. METHODS: Invasive listeriosis cases data from 2005 to 2018 were extracted from the regional laboratory-based surveillance system database and compared with the regional mandatory notification disease system data. RESULTS: Over the fourteen-year period under study, 533 Listeria monocytogenes isolates were detected by the laboratory surveillance system, 55 of which from pregnancy-related cases. The median age of non-pregnancy-associated patients was 71 years, with 64.6% of cases observed in the elderly. Cases with underlying medical risk conditions accounted for 92.1%, and the fatality rate was 26.2%. By integrating data from the mandatory notification system and the laboratory-based surveillance system, a total of 935 cases were recorded. The collection of data through the laboratory surveillance system allowed to increase the surveillance sensitivity by 18%. CONCLUSIONS: Our results documented the growing epidemiological relevance of listeriosis through the analysis of two information sources, the regional mandatory notification system and the regional laboratory-based surveillance system. The data we obtained were consistent with the literature, except for pregnancy-related cases, which are often underdiagnosed. This study highlighted the importance of laboratory-based surveillance system, which led to a significant increase in the sensitivity of the mandatory notification system.

Genetic characterisation of Measles virus variants identified during a large epidemic in Milan, Italy, March-December 2017.

In 2017, Italy experienced a large measles epidemic with 5408 cases and four deaths. As Subnational Reference Laboratory of the Measles and Rubella surveillance NETwork (MoRoNET), the EpiSoMI (Epidemiology and Molecular Surveillance of Infections) Laboratory (University of Milan) set up rapid and active surveillance for the complete characterisation of the Measles virus (Mv) responsible for the large measles outbreak in Milan and surrounding areas (Lombardy, Northern Italy). The aims of this study were to describe the genetic profile of circulating viruses and to track the pathway of measles transmission. Molecular analysis was performed by sequencing the highly variable 450 nucleotides region of the N gene (N-450) of Mv genome. Two-hundred and ninety-nine strains of Mv were analysed. The phylogenetic analysis showed five different variants, two not previously described in the studied area, belonging to D8 and B3 genotypes. Three events of continuous transmission of autochthonous variants (D8-Osaka, D8-London and B3-Milan variants) and two events of continuous transmission of imported variants (B3-Dublin and D8-Hulu Langat) tracked five different transmission pathways. These pathways outlined two epidemic peaks: the first in April and the second in July 2017. The correlation between Mv variant and the epidemiological data may enable us to identify the sources of virus importation and recognise long-lasting virus transmission pathways.

Measles surveillance activities in the Metropolitan Area of Milan during 2017-2018.

INTRODUCTION: In Italy, the transmission of measles is still endemic, and 7,919 cases were reported to the National Surveillance System between January 2017 and December 2018. Aim of this study is to report the results of the measles surveillance activities in the Metropolitan City of Milan from March 2017 to December 2018, and to evaluate the surveillance performance WHO indicators. METHODS: The Local Health Units (LHUs) carried out case investigations and collected specimens to send to the EpiSoMI Lab (Subnational Reference Laboratory, SRL) of the University of Milan for cases/outbreaks confirmation and genotyping performed according to the WHO Guidelines. RESULTS: Overall, 610 suspected measles cases were reported by the surveillance system of the Metropolitan City of Milan. A total of 439 out of 540 cases with adequate specimens were laboratory-confirmed by molecular and/or serological assays. Two-hundred and thirty-six cases were notified as sporadic and 203 as related to 94 outbreaks. The most confirmed cases were aged 15-39 years, almost all not vaccinated. Overall, 282 cases were genotype D8 and 118 genotype B3.The evaluation of a set of indicators to monitor the quality of surveillance activities demonstrated the proficiency of the EpiSoMI Lab. CONCLUSIONS: A well-done investigation of cases and outbreaks by the surveillance local system, in a timely manner, in order to notify and investigate suspected cases and to laboratory confirm or discard cases is fundamental to reduce morbidity, to prevent further virus transmission and to achieve measles elimination.

Measuring Incompatible Observables by Exploiting Sequential Weak Values.

One of the most intriguing aspects of quantum mechanics is the impossibility of measuring at the same time observables corresponding to noncommuting operators, because of quantum uncertainty. This impossibility can be partially relaxed when considering joint or sequential weak value evaluation. Indeed, weak value measurements have been a real breakthrough in the quantum measurement framework that is of the utmost interest from both a fundamental and an applicative point of view. In this Letter, we show how we realized for the first time a sequential weak value evaluation of two incompatible observables using a genuine single-photon experiment. These (sometimes anomalous) sequential weak values revealed the single-operator weak values, as well as the local correlation between them.

Experiment Investigating the Connection between Weak Values and Contextuality.

Weak value measurements have recently given rise to a great amount of interest in both the possibility of measurement amplification and the chance for further quantum mechanics foundations investigation. In particular, a question emerged about weak values being proof of the incompatibility between quantum mechanics and noncontextual hidden variables theories (NCHVTs). A test to provide a conclusive answer to this question was given by Pusey [Phys. Rev. Lett. 113, 200401 (2014)], where a theorem was derived showing the NCHVT incompatibility with the observation of anomalous weak values under specific conditions. In this Letter we realize this proposal, clearly pointing out the connection between weak values and the contextual nature of quantum mechanics.

[The effective Communication in Vaccination Program as a Patient Safety tool: the Lombardy region experience]. / La comunicazione in campo vaccinale quale strumentodi implementazione della Patient safety e della partecipazione responsabile del Cittadino: l'esperienza della Direzione Generale Sanità della Regione Lombardia.

The communication is strategic in Public Health because it is at the heart of who we are as human beings. It is our way of exchanging information and it also ensures the effectiveness of empowerment strategies. In this semantic context, the effective communication/education/information can be a Patient Safety tool because can reduce the adverse events by failure of information or education. In the multicultural collectivity, the risk of miscommunication and unsafe care is very high. This paper describes the design of a multilingual information/education tools to reduce the information failure in vaccination and the related adverse events by information failure or deficiency of adherence in the paediatric vaccination.

Epidemiological surveillance of West Nile neuroinvasive diseases in Italy, 2008 to 2011.

We describe the geographical and temporal distribution of West Nile neuroinvasive diseases (WNND) cases in Italy from 2008 to 2011. The increasing number of confirmed human cases from eight in 2008 to 18 in 2009 and the occurrence of the virus in a larger geographical area in 2009 (moving from east to west) prompted the Ministry of Health to publish, in spring 2010, a national programme for WNND human surveillance, comprising veterinary and vector surveillance. Subsequently, in 2011, a new national plan on integrated human surveillance of imported and autochthonous vector-borne diseases (chikungunya, dengue and West Nile disease) was issued. Between 2008 and 2011, 43 cases of WNND were reported from five regions in Italy with a case fatality rate of 16%. The incidence of WNND during the entire study period was 0.55 per 100,000 population (range: 0.06­0.23 per 100,000). During 2011, two new regions (Friuli-Venezia Giulia and Sardinia) reported confirmed cases in humans. Integrated human, entomological and animal surveillance for West Nile virus is a public health priority in Italy and will be maintained during 2012.

Self consistent, absolute calibration technique for photon number resolving detectors.

Well characterized photon number resolving detectors are a requirement for many applications ranging from quantum information and quantum metrology to the foundations of quantum mechanics. This prompts the necessity for reliable calibration techniques at the single photon level. In this paper we propose an innovative absolute calibration technique for photon number resolving detectors, using a pulsed heralded photon source based on parametric down conversion. The technique, being absolute, does not require reference standards and is independent upon the performances of the heralding detector. The method provides the results of quantum efficiency for the heralded detector as a function of detected photon numbers. Furthermore, we prove its validity by performing the calibration of a Transition Edge Sensor based detector, a real photon number resolving detector that has recently demonstrated its effectiveness in various quantum information protocols.

Testing ultrafast two-photon spectral amplitudes via optical fibres.

We test two-dimensional TPSA of biphoton light emitted via ultrafast spontaneous parametric down-conversion (SPDC) using the effect of group-velocity dispersion in optical fibres. Further, we apply this technique to demonstrate the engineering of biphoton spectral properties by acting on the pump pulse shape.

[Investigation of potential risk factors for Salmonella enterica subsp enterica serotype Napoli: a nested case-control study in Lombardia region]. / Identificazione di potenziali fattori di rischio per i casi di infezione da Salmonella enterica subsp enterica sierotipo Napoli attraverso uno studio caso-controllo nidificato con- dotto in Regione Lombardia.

Enter-net surveillance system has detected the re-emergence of Salmonella enterica subsp enterica serotype Napoli (S. Napoli) since 2002. Preliminary data show that food vehicle could not be the only one implicated in the transmission of this serotype. A nested case-control study has been conducted using data form a prospective cohort: for each salmonellosis notification in Lombardia from May 2004 to December 2005 an ad hoc epidemiological investigation has been performed. Cases have been defined as subjects affected by salmonellosis with a positive faecal sample for Salmonella serotype Napoli and controls as subjects affected by salmonellosis with a positive faecal sample for Salmonella serotype not Napoli. Among 2185 salmonellosis notifications collected by 12 of 15 Local Health Authorities, we identified 67 cases and 867 controls. Infection due to S. Napoli were significantly more common among subjects with exposure to surface water (adjusted Odds Ratio (OR) = 3,82; 95% Confidence Interval (C.I.)=1,03-14,19), to domestic animals (adjusted OR = 3,22; 95% C.I. = 1,28-8,10) and among person who practice activities connected to surface water (adjusted OR 3,88; 95% C.I.=1,15-13,05) compared to subjects not exposed to these factors. Hyperendemia of S. Napoli could be attributable to an animal reservoir not yet identified: surface water contamination may be a direct (waterborne infection) or indirect (foodborne infection) vehicle for transmission of S. Napoli. behaviour. Regarding the relations of the neuroendocrine parameters and driving behaviour, positive correlations were observed between dopamine levels and frequency of driving violations while a negative relationship was found between adrenaline levels and frequency of driving errors. In conclusion the identification of psycho-physiological variables related to driving risky behaviour might be a useful instrument to design traffic safety programs tailored to high risk subjects.

A comparison of three diagnostic techniques for malaria: a rapid diagnostic test (NOW Malaria), PCR and microscopy.

Malaria is a common, life-threatening infection in endemic tropical areas and one that presents a diagnostic challenge to laboratories in most non-endemic countries. A rapid and accurate diagnosis is a prerequisite for effective treatment, especially for the potentially fatal cases of Plasmodium falciparum infection. In the present, multi-centre study, the performances of a rapid diagnostic test (NOW) Malaria) and several, commercial, PCR-based assays (AMS61, AMS42, AMS43, AMS4 and AMS45) were compared against the results of microscopical examination of bloodsmears (the current 'gold standard'). The subjects were either non-European immigrants (N=135) or international travellers (N=171). There was good concordance between the results of all the detection methods, with kappa values of >0.8. Although the NOW Malaria rapid test was both sensitive (100%) and specific (100%) in detecting P. falciparum infections, it was less specific (93.1%) and sensitive (90.7%) in identifying the other Plasmodium species. The results from the AMS61 assay, designed to detect any malarial infection, generally parallelled those of the microscopy (kappa = 0.89), giving a specificity of 98.2% and a sensitivity of 91.0%. Although the use of species-specific molecular primers to identify pure infections with P. falciparum and P. vivax gave results that were in good agreement with those of the microscopy, the subjects who had apparently pure infections with P. ovale or P. malariae were always found PCR-negative. Compared with the standard microscopy, both the NOW Malaria test and the PCR-based assays were therefore poor at identifying mixed infections. The NOW Malaria test and the PCR-based assays clearly need to be improved, particularly for the correct identification of infections with Plasmodium spp. other than P. falciparum, including mixed infections. For now, expert microscopy must remain the mainstay of the laboratory diagnosis of malaria.

Dispersion spreading of biphotons in optical fibers and two-photon interference.

We present the first observation of two-photon polarization interference structure in the second-order Glauber correlation function of two-photon light generated via type-II spontaneous parametric down-conversion. In order to obtain this result, two-photon light is transmitted through an optical fiber and the coincidence distribution is analyzed by means of the start-stop method. Beyond the experimental demonstration of an interesting effect in quantum optics, these results also have considerable relevance for quantum communications.

Diagnosis of visceral leishmaniasis: the sensitivities and specificities of traditional methods and a nested PCR assay.

In the present study, 67 patients suspected to be cases of visceral leishmaniasis (VL) were each checked for leishmanial infection by the microscopical evaluation of various biological specimens, in-vitro culture, serology and an assay based on nested PCR. Most (35) of the subjects were immunocompetent (IC) but 32 were immunodeficient (ID) as the result of HIV infection (18 cases), treatment to prevent transplanted organs being rejected (six) or haematological malignancies (eight). Forty-one (61.2%) of the subjects (19 IC subjects, 12 HIV-positive patients, four transplant patients and six patients with malignancies) were considered true cases of VL. For the IC subjects, only the production and microscopical examination of leucocytoconcentrates and cultures of Buffy coats gave sensitivities of <80%, the results of the other methods showing higher sensitivities and almost perfect agreement with the 'gold-standard' diagnoses. For the ID subjects, however, only the serological tests and the PCR gave reasonable sensitivities (of >80%). For the initial diagnosis of leishmaniasis in ID patients, IFAT and western blots may be useful, as, among the present ID patients, they gave sensitivities (of 80.9% and 88.2%, respectively) that were almost as high as that for the PCR, and specificities of 100%. In the diagnosis of VL in either IC or ID patients, the assay based on a nested PCR appeared to be particularly reliable, with sensitivities of 88.9% and 95.2%, respectively, and a specificity of 100% in both groups of patients. The testing of bone-marrow aspirates by PCR revealed very few VL cases who were not found positive when samples of their peripheral blood were checked in the same assay. For both IC and ID subjects therefore, the use of the PCR-based method to test samples of peripheral blood (which can be collected much more easily than bone-marrow aspirates and with much less pain for the subject) is recommended.

Evidence of Epstein-Barr virus infection in ulcerative colitis.

BACKGROUND AND AIM: The aetiology of ulcerative colitis is still controversial, however, recent studies have emphasised the possible role of infectious agents or ingested substances and their breakdown products, which might activate immune-mediated mechanisms eventually leading to tissue damage. Aim of this investigation was to ascertain the occurrence and the potential role of Epstein-Barr virus infection in large bowel mucosa of ulcerative colitis patients. PATIENTS AND METHODS: Twenty-three biopsies and six total colectomies from 17 patients were analysed for the expression of Epstein-Barr virus proteins and RNAs. Polymerase chain reaction experiments were also carried out to detect Epstein-Barr virus DNA. For comparison, ten biopsies from patients with Crohn's disease, ten biopsies from patients with different types of colitis, seven biopsies and five surgical margins of normal colonic mucosa from the small and large bowels were studied (controls). RESULTS: Six biopsies and four colectomies from seven ulcerative colitis patients showed scattered lymphocytes expressing nuclear EBER 1-2 and harbouring polymerase chain reaction-amplifiable Epstein-Barr virus-DNA. In some cases, linear viral DNA (typical of lytic Epstein-Barr virus infection) was also found. Epithelial cells were invariably negative in all cases. All control tissues from non-ulcerative colitis patients were also invariably non-reactive. CONCLUSION: Evidence of Epstein-Barr virus infection in the mucosal inflammatory cells of ulcerative colitis patients suggests a possible role of this virus in the chronicity of ulcerative colitis.

Immunoreactivity for latent membrane protein 1 of Epstein-Barr virus in nevi and melanomas is not related to the viral infection.

Epstein-Barr virus (EBV) is a human herpes virus with oncogenic potential, associated with several malignancies. The EBV-encoded latent membrane protein 1 (LMP1) is one of nine proteins regularly expressed in virally infected and immortalised B lymphocytes. We now document the consistent immunoreactivity for LMP1 in 90% of 65 nevi and melanomas, using the monoclonal antibody cocktail CS1-4. The immunocytochemical findings, however, were not confirmed using reverse-transcription polymerase chain reaction (RT-PCR) experiments, which failed to demonstrate any actual expression of LMP1 mRNA. In situ hybridisation for EBV-encoded RNAs (EBERs 1 and 2) and PCR amplification of EBV genomic sequences also failed to document any viral infection. Several normal and neoplastic human tissues have also been immunostained for LMP1, without any positive staining, with the exception of a minor percentage of skin melanocytes and of normal blasts of the myeloid and erythroid lineages. We conclude that the vast majority of nevi and melanomas express a still uncharacterised molecule, cross-reacting with anti-LMP1 (CS1-4) antibodies, which may be considered a consistent marker of melanocytic proliferations. The immunoreactivity of normal and neoplastic human tissues for the anti-LMP1 reagent should not be taken as evidence of EBV infection.

The tumor-suppressor gene FHIT is involved in the regulation of apoptosis and in cell cycle control.

Alteration of the FHIT (fragile histidine triad) gene occurs as an early and frequent event in lung carcinogenesis. FHIT gene transfer into lung cancer cell line H460 lacking Fhit protein expression resulted in reversion of tumorigenicity. To gain insight into the biological function of FHIT, we compared the H460 cell line with its Fhit transfectants (H460/FHIT). A significant inhibition of cell growth was observed in H460/FHIT cells. The analysis of apoptosis by in situ terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling revealed a high rate of apoptosis-induced DNA strand breaks in stable clones. In situ results were confirmed by FACScan analysis that showed an apoptotic rate of 44-47% compared with a 15% level in the control H460 cells. Analysis of cell cycle-phase distribution indicated a significant G(0)/G(1) arrest and the presence of a sub-G(1) peak in the stable clones. No significant changes in Bcl2, BclX, and Bax protein expression level were observed in the transfected clones as compared with the control H460 cells whereas a 2-fold increase in Bak protein levels was noticed. An increased level of p21(waf) protein paralleled by an up-regulation of p21(waf) transcripts also was found in Fhit-expressing clones compared with the H460 cell line. No differences in p53 levels were observed in the same cells, suggesting a p53-independent effect. These data suggest that the observed growth-inhibitory effect in FHIT-reexpressing cells could be related to apoptosis and cell cycle arrest and link the tumor-suppressor activity of FHIT to its proapoptotic function.

Development of a nested PCR assay for detection of feline infectious peritonitis virus in clinical specimens.

A diagnostic test for feline infectious peritonitis virus (FIPV) infection based on a nested PCR (nPCR) assay was developed and tested with FIPV, feline enteric coronavirus (FECV), canine coronavirus (CCV), and transmissible gastroenteritis virus (TGEV) and clinical fluid samples from cats with effusive feline infectious peritonitis (FIP). The target sequence for the assay is in the S1 region of the peplomer protein E2 gene. A vaccine strain of FIPV and two wild-type FIPV strains tested positive, but FECV, TGEV, and CCV tested negative. Preliminary tests with 12 cats with clinical evidence of effusive FIP and 11 cats with an illness associated with effusions, but attributed to other causes, were performed. Eleven of the 12 cats with effusive FIP tested positive, while 1 was negative. Ten of the 11 cats ill from other causes tested negative, while 1 was positive. On the basis of clinical laboratory and histopathologic criteria, the preliminary sensitivity and specificity of the assay were 91.6 and 94%, respectively.

Characterization of four microsatellites in an Italian population and their application to paternity testing.

Microsatellites have recently been used for linkage analysis of genetic diseases and for DNA fingerprinting in forensic medicine. In the present study the heterozygosity, PIC values and allele distributions of four microsatellites, D8S85, D8S88, D5S346 and D7S460, in an Italian population have been investigated. After amplification with primers specific for each locus, alleles were separated and detected by denaturing gel electrophoresis and ethidium bromide staining. High heterozygosity and PIC values were observed for all microsatellites in accordance with data in other Caucasian populations. However, different allele distributions for D8S85, D8S88 and D5S346, due to the presence of additional bands or to different frequencies, were found. D7S460, which has never been fully characterized before, appeared to have five alleles in the range 172 to 188 bp. When used for paternity testing, all microsatellites gave results which were consistent with those obtained with established markers, including apo B 3'HVR, D1S80 and COL2A1. This indicates that D8S85, D8S88, D5S346 and D7S460 may be useful as additional informative markers or for solving discrepancies in selected cases.

[Monitoring and prevention of hospital infections in neonatal intensive care]. / Monitoraggio e profilassi delle infezioni ospedaliere nella terapia intensiva neonatale.

The authors monitored newborns in the Intensive Care Unit and the equipment with culture exams for a period of 6 months in order to evaluate the incidence of microbic infections and eventual clinical manifestations in relation to risk factors and to test the efficacy of the prophylactic and therapeutic measurements adopted. The results evidenced a positivity in the culture exams of 24.4% at the 1st control, 14.3% at the 2nd and no positive results at the 3rd. Only two newborns presented sepsis that was resolved without any consequences. Aimed antimicrobic therapy was begun at the 1st clinical and/or haematochemical sign that could suspect the onset of infection. A careful control of the microbic flora, the use of aimed antibiotic therapy and respect of the hygienic conditions permitted us to obtain an excellent control of the infections the morbidity, mortality, the days of hospitalization and the onset of bacterial resistance.