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.

Hepatitis C virus outbreak in a haemodialysis unit: learning from failures.

The investigation of an outbreak of hepatitis C virus in an Italian haemodialysis (HD) centre showed that three patients acquired infection with the same strain, affecting a chronically hepatitis C virus (HCV)-infected patient receiving HD in the same room and during the same shifts. Through our observational analysis many possible modes of transmission were identified, but none could be definitively identified as the route of HCV spread in this small cluster. This outbreak confirms that repeated opportunities for nosocomial HCV transmission may occur among HD patients due to several breaches in the standard precautions for bloodborne infections by healthcare staff.

Tuberculosis-related stigma leading to an incomplete contact investigation in a low-incidence country.

A contact investigation following a case of infectious tuberculosis (TB) reported in a call centre in Milan (Italy) led to the identification of three additional cases that had occurred in employees of the same workplace during the previous 5 years, one of whom was the probable source case. Thirty-three latent infections were also identified. At the time of diagnosis, the source case, because of fear of stigma related to TB, claimed to be unemployed and a contact investigation was not performed in the workplace. Cases were linked through genotyping of Mycobacterium tuberculosis. TB stigma has been described frequently, mainly in high-incidence settings, and is known to influence health-seeking behaviours and treatment adherence. The findings in this report highlight that TB-associated stigma may also lead to incomplete contact investigations. Little is known about the causes and impact of TB-related stigma in low-incidence countries and this warrants further exploration. Research is also needed to evaluate the effectiveness of specific interviewing techniques and training interventions for staff in reducing feelings of stigma in TB patients. Finally, the outbreak emphasizes the importance of integrating routine contact investigations with genotyping.

Maternal mortality in Italy: a record-linkage study.

OBJECTIVE: To detect maternal deaths, analyse associated causes and compute absolute and specific maternal mortality ratio among five Italian regions in response to a recent ranking of Italy by the Lancet as having the lowest maternal mortality ratio among 181 countries. DESIGN: Record-linkage study. SETTING: Five Italian regions. POPULATION: All women aged 15-49 years resident in the participating regions, with one or more hospitalisations for pregnancy or any pregnancy outcome between 2000 and 2007. METHODS: Maternal deaths have been identified by record linkage between the Death Registry and the Hospital Discharge Database. Different time periods were analysed according to local data availability. Cases have been selected and causes of death have been classified according to the 10th International Classification of Diseases. MAIN OUTCOME MEASURE: Maternal mortality ratio. RESULTS: Underreporting of official figures based on death certification in the participating regions is 63%. A total of 118 maternal deaths have been identified resulting in a maternal mortality ratio of 11.8, compared with the official figure of 4.4, per 100,000 live births. Haemorrhage, thromboembolism, and hypertensive disorders of pregnancy are the leading causes of direct deaths. CONCLUSIONS: This study implies that only 37% of all maternal deaths are included in the official data. Our analysis shows a predominance of direct obstetric deaths, which implies that emphasis is needed on improvements of obstetric care.

[Colon recanalization after Hartmann's procedure. A challenge for the surgeon or a strategy to be changed?]. / La ricanalizzazione del colon dopo l'intervento di Hartmann. Un impegno per il chirurgo o una strategia da cambiare?

AIM: Restoration of colorectal continuity is the second step of the procedure originally described by H. Hartmann for the treatment of sigmoid colon cancers. It is a safe, but complex procedure, that needs a meticulous surgical technique and is associated with a significant morbidity in as many as 40% of cases. For this reason, many patients are not submitted to reversal of the colostomy. METHODS: The authors report their experience: 32 patients underwent colostomy closure after Hartmann's procedure during the last 8 years. RESULTS: The mortality rate was nil, but post-operative complications occurred in 12 patients (37.5%), namely: wound infection (11 patients) and anastomotic leak (2 patients). CONCLUSION: In light of this, it could be wiser to perform, in the patients affected with benign pathology, and in those affected with limited malignant pathology (T3), resection of the diseased segment of colon and primary anastomosis with temporary ileostomy, reserving Hartmann's procedure to patients unsuitable to one step treatment because of their poor general and local conditions.

Cloning and expression of a novel hepatitis B virus-binding protein from HepG2 cells.

A direct involvement of the hepatitis B virus (HBV) preS1-(21-47) sequence in virus attachment to cell membrane receptor(s) and the presence on the plasma membranes of HepG2 cells of protein(s) with receptor activity for HBV have been suggested by many previous experiments. In this study, by using a tetravalent derivative of the preS1-(21-47) sequence, we have isolated by affinity chromatography from detergent-solubilized HepG2 plasma membranes a 44-kDa protein (HBV-binding protein; HBV-BP), which was found to closely correspond to the human squamous cell carcinoma antigen 1 (SCCA1), a member of the ovalbumin family of serine protease inhibitors. Comparison of SCCA1 sequence with the sequence of the corresponding HBV-BP cDNA, cloned by polymerase chain reaction starting from RNA poly(A)(+) fractions extracted from HepG2 cells, indicated the presence of only four nucleotide substitutions in the coding region, leading to three amino acid changes. Intact recombinant HBV-BP lacked inhibitory activity for serine proteases such as alpha-chymotrypsin and trypsin but inhibited with high potency cysteine proteases such as papain and cathepsin L. Direct binding experiments confirmed the interaction of recombinant HBV-BP with the HBV preS1 domain. HepG2 cells overexpressing HBV-BP after transfection of corresponding cDNA showed a virus binding capacity increased by 2 orders of magnitude compared with untransfected cells, while Chinese hamster ovary cells, which normally do not bind to HBV, acquired susceptibility to HBV binding after transfection. Native HBV particle entry was enhanced in transfected cells. Both recombinant HBV-BP and antibodies to recombinant HBV-BP blocked virus binding and internalization in transfected cells as well as in primary human hepatocytes in a dose-dependent manner. Our findings suggest that this protein plays a major role in HBV infection.

CtBP/BARS induces fission of Golgi membranes by acylating lysophosphatidic acid.

Membrane fission is essential in intracellular transport. Acyl-coenzyme As (acyl-CoAs) are important in lipid remodelling and are required for fission of COPI-coated vesicles. Here we show that CtBP/BARS, a protein that functions in the dynamics of Golgi tubules, is an essential component of the fission machinery operating at Golgi tubular networks, including Golgi compartments involved in protein transport and sorting. CtBP/BARS-induced fission was preceded by the formation of constricted sites in Golgi tubules, whose extreme curvature is likely to involve local changes in the membrane lipid composition. We find that CtBP/BARS uses acyl-CoA to selectively catalyse the acylation of lysophosphatidic acid to phosphatidic acid both in pure lipidic systems and in Golgi membranes, and that this reaction is essential for fission. Our results indicate a key role for lipid metabolic pathways in membrane fission.

Tissue polypeptide antigen (TPA) in urinary bladder cancer cytology: a follow-up study.

Tissue polypeptide antigen (TPA) has been detected by immunocytochemical assay on urine cytological samples from 28 asymptomatic patients, who previously had a histological diagnosis of papillary transitional cell carcinoma (PTCT) of the urinary bladder (UB), in order to evaluate its role in follow-up controls. TPA staining intensity (SI) in urothelial cells was evaluated to improve the diagnostic accuracy of cytology. Differentiated tumor cells were strongly stained for TPA, heavier than normal urothelial cells. Undifferentiated neoplastic cells were less stained for TPA with a wide range of SI. TPA detection revealed positive cytology in 21 (75%) of the considered cases. The accuracy of our cytological findings compared with both routine examinations and subsequent histopathological diagnosis was of 95.2%. Follow-up urinary cytology limits could be reduced by TPA searching in differentiated tumor cells, deriving from low-grade neoplasms.

Neuroendocrine carcinoma of the skin (Merkel cell carcinoma): immunocytochemical study of a case.

The immunocytochemical phenotype was evaluated in a case of Merkel cell carcinoma of the skin. Intermediate filaments, i.e. neurofilament, glial fibrillary acid protein, cytokeratins, keratin and panfilament as well as S-100 protein, calcitonin and epithelial membrane antigen were detected by immunoperoxidase methods. Nodular positivity for neurofilament was observed. The remaining intermediate filaments and other markers were negative. Thus the origin of Merkel cell carcinoma appears uncertain and this tumor probably has neuroendocrine activity.

Increased serum calcitonin, reduced serum 1,25-(OH)2-vitamin D and normal parathormone concentrations in idiopathic hypercalciuria.

Twelve children (5 males and 7 females, mean age 8.5 years) with idiopathic hypercalciuria (IH), 9 with absorptive IH (1 type I and 8 type II) and 3 with renal IH, were followed in our outpatient hospital from September 1981 to March 1987. Mean (+/- 1 SD) free diet calciuria was 5.9 +/- 1.66 mg/kg/day. Diagnosis was made measuring urinary Ca/Cr ratio after controlled diet and calcium loading test. Serum parathormone (PTH), 1,25-(OH)2-D3 and calcitonin (TCT) values and renal threshold phosphate concentration (TmPO4/GFR) were also studied. Serum PTH was normal in all children. Serum TCT levels were significantly increased in the patients with IH compared with controls (p less than 0.001) while serum 1,25-(OH)2-D3 levels were significantly reduced compared with controls (p less than 0.001).

New phenotypic aspects in a family with Lynch syndrome II.

Increasing attention has been given to hereditary nonpolyposis colorectal cancer (HNPCC). This report provides medical genetic/pathologic findings on an HNPCC kindred from southern Italy that shows criteria consistent with Lynch syndrome II. An international collaborative effort led to extension of this kindred with disclosure of a potentially new spectrum of phenotypic findings: an excess of gastric carcinoma; complete intestinal metaplasia and chronic atrophic gastritis restricted to the antrum; an apparent excess of colonic mucosal macrophagia, which by special stain appeared to be positive for mucin, with a constant content of both sialo and sulfomucin, a lack of iron, and an inconstant positivity for lysozyme obtained by immunoperoxidase technique; and findings of crypt atrophy of the colonic mucosa. During the relatively short period of investigation of this family, an intensive educational and surveillance program has been mounted in the interest of improving cancer control through direct application of knowledge of natural history and the risk factor evidence through pedigree assessment.

Tissue polypeptide antigen in normal and neoplastic urinary bladder. Preliminary reports.

Tissue polypeptide antigen (TPA) both in normal and neoplastic urinary bladders has been studied by immunocytochemistry. A comparison of TPA with epithelial membrane antigen (EMA), keratins and carcinoembryonic antigen (CEA) in various tumor grades and stages has been performed better to define TPA role in bladder carcinomas. Well-differentiated tumors were strongly stained for TPA with a uniform staining intensity. Undifferentiated tumors were weakly stained for TPA with an uneven staining intensity. There was no relation between TPA findings and stages of invasion. However, TPA seems to be a very helpful diagnostic tool for tumor grading and staging.

Lymphocyte subpopulations in Lynch syndrome II: an immunocytochemical study on gastrointestinal biopsies. Preliminary report.

Gastrointestinal biopsies from 18 members of a family with Lynch Syndrome II were evaluated and immunocytochemical studies were made to characterize the phenotypic expression of the tissue's immune populations. The intestinal findings suggest polyclonal B-cell activation related to the T-helper distribution. Our evaluation provides no specific information so far on the management of patients with Lynch Syndrome II.