Generation of iPSC lines derived from skin fibroblasts of two healthy controls using non-transmissible form of Sendai Virus.

We established two iPSC lines starting from skin fibroblasts of two healthy individuals using Sendai-virus-based technique. The obtained iPSCs were characterized showing same STR profile as starting fibroblasts, normal karyotype, loss of stemness vectors, expression of stemness markers, both through real-time PCR and immunofluorescence, (OCT4, SOX2, TRA-1-60, NANOG and SSEA4) and in vitro differentiation into three germ layers.

Generation of an iPSC line from skin fibroblasts of a patient with Joubert syndrome carrying the homozygous loss of function variant c.787dupC in the AHI1 gene.

We produced an iPSC line from a patient with Joubert syndrome carrying the homozygous c.787dupC variant in the AHI1 gene. The iPSC line was obtained by reprogramming skin fibroblasts, mycoplasma-free, using Sendai-virus-based technique. Characterization of iPSCs showed the same Short Tandem Repeats profile than fibroblasts, normal karyotype, expression of staminal markers (OCT4, SOX2, SSEA4 and NANOG) and ability to differentiate into three germ layers in vitro.

Efeito da suplementação de suínos em terminação com diferentes associações entre minerais sobre o desempenho, as características de carcaça e a viabilidade econômica / Effect of supplementation of finishing swines with different associations between minerals on performance, carcass characteristics and economic viability

Objetivou-se, com este estudo, verificar a influência da suplementação com diferentes associações entre minerais orgânicos sobre o desempenho e as características de carcaça, bem como avaliar sua viabilidade econômica, para suínos em terminação. Foram utilizados 88 machos castrados, divididos entre quatro tratamentos: controle com dieta basal e suplementação com CrFe, MgSe e CrFeMgSe, durante os 28 dias que antecederam o abate. O delineamento experimental foi em blocos ao acaso, com parcela experimental representada por dois animais. Os animais suplementados com CrFeMgSe apresentaram piores índices de peso final, ganho de peso diário e consumo durante todo o período, com consequente redução no peso de carcaça, menor profundidade de lombo, área de olho-de-lombo e menor quantidade de carne magra na carcaça. Houve um aumento no custo da dieta para os grupos CrFe, MgSe e CrFeMgSe, respectivamente. O uso de cromo mais ferro e magnésio mais selênio, associados dois a dois, não altera os parâmetros de desempenho e as características de carcaça. A associação dos quatro minerais, no entanto, promove piora nessas características e aumenta o custo da dieta.(AU)

This study aimed to verify the influence of supplementation with different associations between organic minerals on performance, carcass characteristics and economic viability for finishing swine. 88 castrated males were divided in four treatments: control with basal diet and supplementation with CrFe, MgSe and CrFeMgSe during the 28 days prior to slaughter. The experimental design was randomized blocks with an experimental plot represented by two animals. The animals supplemented with CrFeMgSe had worse final weight, daily weight gain and consumption throughout the period, with consequent reduction in carcass weight, lower loin depth, loin eye area and amount of lean meat. There was an increase in the effective operational cost of diet with CrFe, MgSe and CrFeMgSe groups, respectively. The use of chromium plus iron and magnesium plus selenium, associated two by two, did not alter the performance parameters and carcass. However, the four minerals association promote a worsening in these characteristics and increase the cost of production.(AU)

Detection of influenza A(H1N1)pdm09 virus in a patient travelling from Shanghai to Italy in July 2018: an uncommon clinical presentation in a non-seasonal period.

Influenza is one of the most common infectious diseases in travellers, especially in those returning from subtropical and tropical regions. In late June 2018 an influenza A(H1N1)pdm09 virus infection was diagnosed in a 36-years-old man, returned from a travel in Shanghai and hospitalized at the Ospedale Policlinico San Martino, Genoa, Italy, with a diagnosis of fever and an uncommon clinical presentation characterised by a persistent leukopenia. Phylogenetic analysis revealed a closeness with influenza A(H1N1)pdm09 strains circulating in the US in May-June 2018. Prompt recognition of influenza infection led to a proper case management, demonstrating the crucial role of the continuous influenza surveillance programme.

Vaccination coverage in healthcare workers: a multicenter cross-sectional study in Italy.

INTRODUCTION: In recent years, a phenomenon known as "vaccine hesitancy" has spread throughout the world, even among health workers, determining a reduction in vaccination coverage (VC).A study aimed at evaluating VC among healthcare workers (HCWs) in 10 Italian cities (L'Aquila, Genoa, Milan, Palermo, Sassari, Catanzaro, Ferrara, Catania, Naples, Messina) was performed. MATERIALS AND METHODS: Annex 3 of the Presidential Decree n. 445 of 28 December 2000 was used to collect information on the vaccination status of HCWs. The mean and standard deviation (SD) were calculated with regard to the quantitative variable (age), while absolute and relative frequencies were obtained for categorical data (sex, professional profile, working sector, vaccination status). The connection between VC and the categorical variables was evaluated by chi-square method (statistical significance at p < 0.05). The statistical analyses were performed by SPSS and Stata software. RESULTS: A total of 3,454 HCWs participated in the project: 1,236 males and 2,218 females.The sample comprised: physicians (26.9%), trainee physicians (16.1%), nurses (17.2%) and other professional categories (9.8%). Low VC was generally recorded. Higher VC was found with regard to polio, hepatitis B, tetanus and diphtheria, while coverage was very low for measles, mumps, rubella, pertussis, chickenpox and influenza (20-30%). CONCLUSIONS: This study revealed low VC rates among HCWs for all the vaccinations. Measures to increase VC are therefore necessary in order to prevent HCWs from becoming a source of transmission of infections with high morbidity and/or mortality both within hospitals and outside.

How should vaccination services be planned, organized, and managed? Results from a survey on the Italian vaccination services.

BACKGROUND: Quality improvement is an increasingly recognized approach to maximize service effectiveness and minimize costs in public health. However, the Italian law never provided for the institutional accreditation of vaccination services. Furthermore, a recently approved law added six more compulsory vaccinations to the original four, which has led to a considerable increase in vaccination efforts, without any previous resources evaluation. The aim of the study was to investigate structural, organizational and managerial characteristics of the Italian vaccination services, in order to suggest the adoption of adequate quality standards. STUDY DESIGN: A survey involving the representatives of the Italian Regions and Autonomous Provinces was performed between September 2017 and September 2018. METHODS: An online questionnaire, including 26 items, designed to evaluate the structural, organizational and managerial characteristics of vaccination services was administered. The correlation between the number of vaccination centres and the coverage for each region was used to evaluate the performance of the vaccination services. RESULTS: Respondents from seven Regions, totaling >15,000,000 inhabitants, answered the questionnaire. Overall, each vaccination service was potentially accessed by an average of 519 children aged zero to 24 months, with a ß-coefficient of -0.87 (p = 0.01) for infant vaccination coverage in 2016. Eighty-five percent of vaccination services were provided with architectural features to accommodate the disabled but only 49% provided reserved parking lots. An average of 0.4 physicians and 0.6 other healthcare workers per 10,000 inhabitants were employed in vaccination services, with complete computerization in 74% of them. CONCLUSION: The inverse relation between vaccination services' spatial accessibility and vaccination coverage suggests that distance and accessibility of vaccination services should be considered in planning. This survey constitutes a baseline data for Italian vaccination services that could be useful for decision makers in establishing minimum requirements to provide high-quality preventive healthcare service.

Hexavalent vaccines: characteristics of available products and practical considerations from a panel of Italian experts.

Combination vaccines represent a valuable technological innovation in the field of infectious disease prevention and public health, because of their great health and economic value from the individual, societal, and healthcare system perspectives. In order to increase parents' and healthcare professionals' confidence in the vaccination programs and maintain their benefits to society, more information about the benefits of innovative vaccination tools such as combination vaccines is needed. Purpose of this work is an examination of available hexavalent vaccines, that protect against Diphtheria, Tetanus, Pertussis, Poliomyelitis, Hepatitis B and Haemophilus influenzae type b infections. From the epidemiological updates of vaccine preventable diseases to the vaccine development cycle, from the immunogenicity of antigenic components to the safety and co-administration with other vaccines, several aspects of available hexavalent vaccines are discussed and deepened. Also a number of practical considerations on schedules, age of employment, strategies for vaccination recovery, vaccination in at-risk births are issued, based on the recommendations of Italian Ministry of Health, Italian Society of Pharmacology (SIF), Italian Society for Pediatrics (SIP), Italian Federation of Family Paediatricians (FIMP) and Italian Society of Hygiene, Preventive Medicine and Public Health (SItI).

Regional point prevalence study of healthcare-associated infections and antimicrobial use in acute care hospitals in Liguria, Italy.

BACKGROUND: Given the importance of monitoring healthcare-associated infections (HCAIs) and the consumption of antibiotics, a regional point prevalence survey was conducted in Liguria between March and April 2016. AIM: To measure the overall prevalence of HCAI and describe the use of antibiotics in all public hospitals. METHODS: Data on risk factors and use of antibiotics were collected for each hospitalized patient. To define the variables significantly associated with HCAI, univariate and multivariate analyses were conducted. Standardized infection ratio and standardized antimicrobial use ratio were measured for each participating hospital. FINDINGS: A total of 3647 patients were enrolled. In all, 429 HCAIs were diagnosed in 376 patients, giving a prevalence of HCAI of 10.3%. Respiratory tract (21.7%) and urinary tract (20%) were the most frequent sites of infection. High rates of meticillin-resistant Staphylococcus aureus (47.4%) and Enterobacteriaceae resistant to carbapenems (26.3%) were isolated. Forty-six percent of patients received at least one antibiotic. Combinations of penicillins including ß-lactamase inhibitors (24.1%) were the most widely used; the main indication (46.7%) was the treatment of a community-acquired infection. CONCLUSION: There was an increase in HCAI prevalence compared to a similar survey conducted in 2007; however, the performance of overlapping investigations will enable more reliable considerations. Nevertheless, data on antimicrobial resistance and use of antibiotics are consistent with the national trend. Despite methodological limitations, prevalence studies are useful to monitor HCAI over time and encourage greater awareness of the problem by all stakeholders.

Available influenza vaccines: immunization strategies, history and new tools for fighting the disease.

The main public health strategy for containing influenza-related disease is annual vaccination, which is recommended for the elderly and others belonging to risk-factor categories, who present the highest morbidity and mortality, as reported by the World Health Organization (WHO) Recommendations. The availability of different influenza vaccine formulations makes the choice of the best immunization strategy a challenge for stakeholders and public health experts. Heterogeneity in at-risk categories included in national influenza vaccine recommendations still exists, in particular among European countries. Broader consensus is expected, which should positively impact on influenza vaccination coverage. The availability of quadrivalent vaccines, containing both influenza B lineages, offers the potential to improve protection by overcoming the drawbacks of wrongly predicting which B lineage will predominate in a given year.

Increasing incidence of Clostridium difficile infections: results from a 5-year retrospective study in a large teaching hospital in the Italian region with the oldest population.

Limited information is available on the incidence of Clostridium difficile infections (CDIs) in Italian hospitals. In this study, we assessed the changes in the incidence of CDI over a 5-year period in a teaching hospital in Liguria, the Italian region with the oldest population. Secondary endpoints were the development of severe CDI and 30-day mortality. The annual incidence of CDI/10000 patient-days significantly increased from 0·54 in 2010 to 3·04 in 2014 (χ 2 for trend, P < 0·001). The median age of patients with CDI was 81 years. As many as 81% and 89% of these patients had comorbid conditions and previous exposure to antibiotics, respectively. In the multivariate analysis of risk factors for severe CDI, previous therapy with histamine 2 blockers and low serum albumin were associated with severe CDI, while diabetes appeared to be protective. In the multivariate model of risk factors for 30-day mortality, high leukocyte count, low serum albumin, and increased serum creatinine were unfavourably associated with outcome. Strict adherence to infection control measures was of utmost importance to counteract the increasing incidence of CDI in our hospital, particularly because of the advanced age of the patients and their very high frequency of chronic conditions and use of antibiotics, which readily predispose them to the development of CDI.

Determinants of adherence to seasonal influenza vaccination among healthcare workers from an Italian region: results from a cross-sectional study.

OBJECTIVES: Notwithstanding decades of efforts to increase the uptake of seasonal influenza (flu) vaccination among European healthcare workers (HCWs), the immunisation rates are still unsatisfactory. In order to understand the reasons for the low adherence to flu vaccination, a study was carried out among HCWs of two healthcare organisations in Liguria, a region in northwest Italy. METHODS: A cross-sectional study based on anonymous self-administered web questionnaires was carried out between October 2013 and February 2014. Through univariate and multivariate regression analysis, the study investigated the association between demographic and professional characteristics, knowledge, beliefs and attitudes of the study participants and (i) the seasonal flu vaccination uptake in the 2013/2014 season and (ii) the self-reported number of flu vaccination uptakes in the six consecutive seasons from 2008/2009 to 2013/2014. RESULTS: A total of 830 HCWs completed the survey. Factors statistically associated with flu vaccination uptake in the 2013/2014 season were: being a medical doctor and agreeing with the statements 'flu vaccine is safe', 'HCWs have a higher risk of getting flu' and 'HCWs should receive flu vaccination every year'. A barrier to vaccination was the belief that pharmaceutical companies influence decisions about vaccination strategies. DISCUSSION: All the above-mentioned factors, except the last one, were (significantly) associated with the number of flu vaccination uptakes self-reported by the respondents between season 2008/2009 and season 2013/2014. Other significantly associated factors appeared to be level of education, being affected by at least one chronic disease, and agreeing with mandatory flu vaccination in healthcare settings. CONCLUSIONS: This survey allows us to better understand the determinants of adherence to vaccination as a fundamental preventive strategy against flu among Italian HCWs. These findings should be used to improve and customise any future promotion campaigns to overcome identified barriers to immunisation.

Influenza vaccination in the elderly: why are the overall benefits still hotly debated?

The exact magnitude of the benefit of influenza vaccine among elderly individuals is subject of considerable debated. Existing vaccine effectiveness estimates come mostly from observational studies, which may be biased because of difficulties in identifying and adjusting for confounders. In this paper, we examine the potential sources of bias in observational studies of influenza vaccine effectiveness in the elderly and we discuss available evidence regarding the efficacy and effectiveness of licensed influenza vaccines. Although several methodological criticisms among the available analyses on seasonal vaccines for elderly were identified, overall seasonal influenza vaccines showed relevant efficacy/effectiveness in reducing the risk of influenza and its complications in the elderly, considering different measure of outcome.

Pneumococcal vaccination strategies in adult population: perspectives with the pneumococcal 13 - valent polysaccharide conjugate vaccine.

Streptococcus pneumoniae (Sp) is a leading cause of infection in people of all ages worldwide, determining a significant impact because of its relatively high incidence rate, the associated economic costs, and the high case-fatality rates. More attention has to be paid for elderly and people with one or more risk factors, in order to reduce health costs and pneumococcal hospital admissions. Moreover, the increasing incidence of antibiotic-resistant Sp strains is a source of concern for its relevant clinical importance in health-care settings. At present, the 23-valent polysaccharide (PPV23) vaccine has shown some limits in terms of protection in the elderly population and against invasive diseases, among adults affected with chronic diseases, non-bacteriemic pneumonias and with immune suppression, in particular in adults with HIV. In December 2011, FDA licensed the 13-valent pneumococcal conjugate vaccine (PCV13) for prevention of pneumonia and invasive diseases (IPDs) in adults aged ≥50 years. The same decision was concomitantly assumed in Europe by EMA. PCV13 has shown superior results both in terms of immunogenicity and of adequate stimulation of a stable and long-lasting immunological memory. European recommendations for PCV13 vaccination in adults are still heterogeneous between Countries. The availability of PCV13 for adults offer a new and promising tool against Sp IPDs and non-IPDs, especially in elderly and at risk populations.

Polychlorinated dibenzodioxins, dibenzofurans, and biphenyls in fresh water fish from Campania Region, southern Italy.

Twenty-eight fish muscle specimens from the main water bodies of the Campania Region were analyzed in our laboratory. On average, results showed a low contamination by PCDDs+PCDFs and a relatively more important presence of DL-PCBs. All specimens were compliant with EU regulatory maximum levels. Cumulative PCDD+PCDF+DL-PCB concentrations (TEQ(TOT)) were comprised in the range 0.223-11.4 pgWHO(97)-TEQ g(-1) fresh weight (fw). DL-PCB contribution to TEQ(TOT) was on average greater than 86% (range, 50.2-97.1%). The cumulative concentrations of 30 non-dioxin-like PCB congeners (Σ(30)(NDL-PCBs)) and of the six indicators (Σ(6)(NDL-PCBs)) were respectively in the ranges 3.30-515 and 1.30-195 ng g(-1) fw. The hybrid clustering approach adopted to analyze the sample-specific congener profiles indentified the main analytical patterns present in the database and, in particular, two main diverse exposure macro-areas that seem to exist north and south of the city of Naples. The distribution of PCDD and PCDF congeners among different species showed significant variations from chub (Leuciscus cephalus), characterized by a higher proportion of low-chlorinated congeners (e.g. 2,3,7,8-T(4)CDD), to eel (Anguilla anguilla), whose contamination consisted mainly of highly chlorinated congeners (e.g. O(8)CDD). To have a more complete perspective in relation to the contaminants present in the environment, the study suggestion is to use benthic as well as pelagic species to obtain an integrated characterization of fish tissue contamination.

HPV related diseases in males: a heavy vaccine-preventable burden.

Human Papillomavirus (HPV) has a significant impact in male's health, as cause of clinical manifestations ranging from genital warts to several cancers of the anogenital and aero-digestive tract. HPV types which most frequently affect men are 6, 11, 16 and 18, included in the HPV quadrivalent vaccine, recently approved for use in males by Food and Drug Administration (FDA) and European Medicines Agency (EMA). Although several data about the safety and efficacy of quadrivalent vaccine are available, the implementation of proper immunization plans dedicate to male's population cannot ignore the knowledge of the characteristics of the disease in men, which in some aspects should be clarify, in particular clearance of type-specific HPV infections and transmission dynamics. Purpose of this review is to summarise the main information about the burden and the natural history of the HPV related disease in males.

Influenza vaccination in HIV-positive subjects: latest evidence and future perspective.

Annual influenza vaccination is recommended for persons with human immunodeficiency virus (HIV) infection. Recent reports indicate that immunizations may increase IIeplication in infected individuals. Generally, vaccination against influenza is well tolerated in both children and adult individuals with HNIVand does not induce significant changes in viral load and CD4+ cell counts. The observed increase in viral replication is usually transient and a clear, measurable progression of the underlying HIV disease is hard to be determined. Several studies reported immunogenicity data in HIV+ population, by comparing diferfent influenza vaccines, adjuvanted or not, and different administration routes. Data are encouraging because an adequate immune response is shown, although split/subunit vaccines do not elicite an efficient immune response in these subjects. New strategies have been evaluated to increase the immune response in immunocompromised patients.The aim of this review is to evaluate tolerability, safety, immunogenicity and efficacy of vaccines actually approved for human use and to consider latest evidence and future perspective in HIV positive subjects.

Dynamic and transient interactions of Atg9 with autophagosomes, but not membrane integration, are required for autophagy.

Autophagy is a catabolic process essential for cell homeostasis, at the core of which is the formation of double-membrane organelles called autophagosomes. Atg9 is the only known transmembrane protein required for autophagy and is proposed to deliver membrane to the preautophagosome structures and autophagosomes. We show here that mammalian Atg9 (mAtg9) is required for the formation of DFCP1-positive autophagosome precursors called phagophores. mAtg9 is recruited to phagophores independent of early autophagy proteins, such as ULK1 and WIPI2, but does not become a stable component of the autophagosome membrane. In fact, mAtg9-positive structures interact dynamically with phagophores and autophagosomes without being incorporated into them. The membrane compartment enriched in mAtg9 displays a unique sedimentation profile, which is unaltered upon starvation-induced autophagy. Correlative light electron microscopy reveals that mAtg9 is present on tubular-vesicular membranes emanating from vacuolar structures. We show that mAtg9 resides in a unique endosomal-like compartment and on endosomes, including recycling endosomes, where it interacts with the transferrin receptor. We propose that mAtg9 trafficking through multiple organelles, including recycling endosomes, is essential for the initiation and progression of autophagy; however, rather than acting as a structural component of the autophagosome, it is required for the expansion of the autophagosome precursor.

Ultrastructure of the spermatozoa of the domestic duck (Anas platyrhynchos sp.).

The ultrastructure of the spermatozoa of the domestic duck (Anas platyrhynchos sp.) was analysed by transmission and scanning electron microscopies and compared with the results obtained in preliminary studies involving other non-passerine birds. The spermatozoa were characterised by the presence of a short head, short midpiece and long principal piece. The head consisted of a reduced acrosome that contained moderately electron-dense homogenous material. The implantation fossa was observed between the base of the nucleus and the proximal centriole. The midpiece contained electron-dense material associated with the proximal centriole and nuclear membrane, and a long distal centriole surrounded throughout its length by 11-12 elliptical mitochondria. A dense annulus separating the midpiece from the principal piece was visible. Posterior to the annulus, the axoneme was formed surrounded by a dense fibrous sheath, representing the principal piece or flagellum, which was a long segment with a smooth surface and a smaller diameter than the midpiece. The spermatozoon of the domestic duck resembles that of other non-passerine birds, corresponding to a basic type of spermatozoon similar to that of reptiles, called sauropsid type.

Genotoxicity testing of a Hoodia gordonii extract.

Hoodia gordonii extract consists of a mixture of steroid glycosides, fatty acids, plant sterols and alcohols. As part of the overall safety assessment H. gordonii extract was assessed for genotoxicity in two assays in vitro: a bacterial mutation assay; and a gene mutation assay using mouse lymphoma cells. H. gordonii extract showed no evidence of genotoxic activity in either of these assays. In addition, H. gordonii extract was assessed for mutagenic activity in a bone marrow micronucleus (MN) assay in the mouse, with 400mg/kg selected as the high-dose group, based on observations in a dose-range-finding study. The group mean frequencies of micronucleated polychromatic erythrocytes of treated animals were similar to those of the vehicle control group, indicating H. gordonii extract to be non-genotoxic under the conditions of this test. All assays were performed in compliance with the Good Laboratory Practice Regulations and in accordance with standard guidelines for genotoxicity tests. H. gordonii extract was shown to be non-genotoxic in 3 independent assays (a bacterial mutation test, a gene mutation assay using mouse lymphoma cells and a bone marrow micronucleus assay in the mouse).

Impact of influenza during the post-pandemic season: epidemiological picture from syndromic and virological surveillance.

INTRODUCTION: Following the observation that 1 or 2 pandemic peak due to the circulation ofAHINlv had occurred in most countries and in most World Health Organization (WHO) Regions, WHO declared on August 10"h, 2010 that the world was moving into the post-pandemic period, whose surveillance presents considerable interest both from epidemiological and clinical point of view. We described the epidemiological picture emerged from syndromic and virological surveillance during the post-pandemic season in Liguria, Italy. MATERIALS AND METHODS: An Emergency Department Syndrome surveillance system, based on data collected at "San Martino" and IRCCS "G. Gaslini" Liguria Regional Reference University Hospitals for adults and children is active since July 2007. Monitored syndromes include "Influenza-Like Illness" (ILl) and "Low Respiratory Tract Infections" (LRTI). The Ligurian Regional Reference laboratory for Influenza virological surveillance and diagnosis offers rapid detection of influenza viruses by real-time and block RT-PCR, viral culture and genetic characterization by entire sequence analysis of haemagglutinin- and neuraminidase-coding regions in accordance with the international standards established by the global laboratory network. RESULTS AND DISCUSSION: The integration of syndromic surveillance system and laboratory surveillance for rapid detection and characterization of the disease responsible agent represented a specific and sensitive tool for influenza surveillance. The post-pandemic season was characterized by early onset and by the heaviest impacts for ILI and LRTI among the recent epidemic seasons. In contrast to the picture observed during the pandemic season, the 2010/11 winter was characterized by the intensive circulation of pandemic AH1N1v coupled with sustained activity due to influenza B and Respiratory Syncytial Virus (RSV). Antigenic and molecular characterization of influenza strains confirmed the good matching between circulating and 2010/11 vaccine viruses.