La sorveglianza del morbillo per la Regione Liguria negli ultimi cinque anni.

Introduzione: Il virus del morbillo appartiene al genere Morbillivirus della famiglia dei Paramixoviridae. Il virione contiene un RNA non\u2013segmentato, a singolo strand, di senso negativo di circa 16 kb. Il genoma contiene 6 geni che codificano per 6 proteine: proteina del nucleocapside (N), fosfoproteina (P), proteina della matrice (M), proteina di fusione (F), emoagglutinina (H) e proteina grande (L). Il virus del morbillo \ue8 nato come una zoonosi da quello della peste bovina (bovini peste). Ad oggi l'uomo \ue8 l'unico serbatoio naturale del virus. Il contagio avviene per via aerea, con il virus trasportato dalle goccioline di Flugge ovvero le micro gocce di saliva che vengono emesse attraverso il respirare lo starnutire o il tossire, e per contatto diretto o indiretto con i fluidi corporei di una persona malata (saliva, sperma, muco). Il virus penetra attraverso le mucose respiratorie e la congiuntiva e raggiunge i linfonodi dove si moltiplica e si diffonde cos\uec per via sistemica. L'incubazione dura circa 10 giorni. La viremia ha un picco in corrispondenza del 12\ub0 giorno dopo il contagio. E' una patologia particolarmente contagiosa per cui rientra tra le patologie da notifica obbligatoria. E' disponibile un vaccino. Il PNPV 2017-2019 prevede il Vaccino anti Morbillo-Parotite-Rosolia. Il sistema di sorveglianza WHO si pone l\u2019obiettivo di eliminare la trasmissione endemica mantenendo alti i livelli delle vaccinazioni e implementando il sistema di sorveglianza in particolar modo nell\u2019ambito del WHO Global Measles and Rubella laboratory Network . A livello nazionale \ue8 stata istituita una rete di laboratori afferenti alla rete MoRoNet con obbligo di notifica all'Istituto Superiore di Sanit\ue0 dei casi confermati positivi. Oggetto di questa tesi \ue8 la descrizione dell'attivit\ue0 di sorveglianza per il morbillo della Regione Liguria negli ultimi cinque anni di lavoro. Materiali e metodi: Il laboratorio dell\u2019UO Igiene dell\u2019Ospedale Policlinico San Martino IRCCS, Universit\ue0 di Genova, raccoglie urine e tampone faringeo di pazienti con probabile o possibile morbillo, i casi da confermare e notificare su piattaforma MoRoNet quindi vengono analizzati come segue: estrazione degli acidi nucleici, amplificazione e sequenziamento del gene NP. Ottenuta la sequenza \ue8 possibile determinare il genotipo del virus circolante e confrontarlo con altre sequenze di riferimento disponibili o con altri casi di morbillo. Questa informazione si ottiene costruendo un albero filogenetico con programmi bioinformatici di allineamento di sequenze e analisi di omologia tra sequenze. Risultati: Nel laboratorio di riferimento della Regione Liguria nel periodo 2015-2019 abbiamo raccolto 114 casi di morbillo da confermare. Di questi 57 casi sono stati confermati positivi. La media dell'et\ue0 sul periodo \ue8 risultata essere 26,8 anni e la mediana 32 anni. In particolare abbiamo riscontrato la presenza dei focolai epidemici nella provincia di Imperia tra la fine 2017 e l' 2018. I risultati dei test di caratterizzazione molecolare hanno permesso di identificare nei campioni raccolti nel 2018 un\u2019unica variante di genotipo B3 circolante, mentre nel 2019 si \ue8 osservata la circolazione di diverse varianti appartenenti al genotipo D8 clade Manchester.UNK/3. Precedentemente le stagioni epidemiologiche sono state caratterizzate dalla circolazione della variante B3, quindi si conclude considerando attualmente una alternanza quasi annuale delle varianti circolanti di virus del morbillo. Conclusioni e discussione: Il morbillo continua a circolare in Italia e causare epidemie per le coperture vaccinali inadeguate nel corso degli anni, che hanno portato all\u2019accumulo di ampie quote di popolazione suscettibili all\u2019infezione. Le adesioni alla prima e alla seconda dose di vaccino MPR sono in aumento ma ancora inferiori al target del 95% e con una rilevante variabilit\ue0 tra regioni. Inoltre l'elevata et\ue0 mediana dei casi indica che esistono ampie sacche di giovani adulti suscettibili, mentre i casi tra gli operatori sanitari Imperiesi evidenziano il problema della bassa copertura vaccinale tra questi ultimi. E' infine evidente la necessit\ue0 di individuare nel Piano nazionale per l'eliminazione del morbillo e della rosolia congenita nuove azioni rispetto a quanto previsto nei precedenti Piani, per esempio rafforzare la copertura nella popolazione adulta. Si conclude ricordando che l'obiettivo generale da raggiungere entro il 2023 \ue8: incidenza <1 caso di morbillo / 1,000,000 popolazione, e tra gli obiettivi specifici rimane fondamentale raggiungere e mantenere una copertura vaccinale maggiore o uguale al 95% per la prima dose di morbillo.Introduction: Measles virus belongs to the genus Morbillivirus of the Paramixoviridae family. The virion contains a non-segmented RNA, a single strand, of a negative sense of approximately 16 kb. The genome contains 6 genes that code for 6 proteins: nucleocapsid protein (N), phosphoprotein (P), matrix protein (M), fusion protein (F), hemagglutinin (H) and large protein (L). Measles virus originated as a zoonosis from that of the bovine plague ( Rinderpest) . To date, man is the natural reservoir user of the virus. Contagion occurs by air, with the virus carried by the droplets of Flugge or the micro drops of saliva that are emitted through breathing, sneezing or coughing, and by direct or indirect contact with the body fluids of a sick person (saliva, sperm, mucus). The virus penetrates through the respiratory mucous membranes and the conjunctiva and observes the lymph nodes where it multiplies and spreads systemically. Incubation lasts approximately 10 days. The viraemia peaks in the correspondence on the 12th day after the infection. It is a particularly contagious pathology for which it falls under the pathologies of mandatory notification. A vaccine is available. The PNPV 2017-2019 provides for the measles-mumps-rubella vaccine.nThe WHO surveillance system aims to eliminate endemic transmission by keeping vaccination levels high and by implementing the surveillance system especially within the WHO Global Measles and Rubella laboratory Network. At national level, a network of laboratories has been set up belonging to the MoRoNet network with the obligation to notify the Istituto Superiore di Sanit\ue0 of confirmed positive cases. The subject of this thesis is the description of the surveillance activity for measles of the Liguria Region in the last five years of work. Materials and methods: The laboratory of the Hygiene Unit of the Policlinico San Martino IRCCS Hospital, University of Genoa, collects urine and pharyngeal swab of patients with probable or possible measles, the cases to be confirmed and notified on the MoRoNet platform are therefore analyzed as follows: extraction of nucleic acids , amplification and sequencing of the NP gene. Once the sequence is obtained, it is possible to determine the genotype of the circulating virus and compare it with other available reference sequences or with other cases of measles. This information is obtained by building a phylogenetic tree with bioinformatic programs of sequence alignment and homology analysis between sequences. Results: In the reference laboratory of the Liguria Region in the period 2015-2019 we collected 114 cases of measles to be confirmed. Of these 57 cases were confirmed positive. The average age over the period was 26.8 years and the median 32 years. In particular, we found the presence of epidemic outbreaks in the province of Imperia between the end of 2017 and 2018. The results of the molecular characterization tests made it possible to identify a single circulating variant of genotype B3 in the samples collected in 2018, while in 2019 the circulation of several variants belonging to the D8 clade Manchester genotype was observed.UNK / 3. Previously the epidemiological seasons have been characterized by the circulation of the B3 variant, therefore it concludes considering at present an almost annual alternation of the circulating variants of measles virus. Conclusions and discussion: Measles continues to circulate in Italy and cause epidemics for inadequate vaccination coverage over the years, which have led to the accumulation of large portions of the population susceptible to infection. The adhesions to the first and second dose of MPR vaccine are increasing but still lower than the target of 95% and with a significant variability between regions. Furthermore, the high median age of the cases indicates that there are large pockets of susceptible young adults, while the cases among Imperia health workers highlight the problem of low vaccination coverage among the latter. Lastly, there is a clear need to identify new actions in the National Plan for the elimination of measles and congenital rubella compared to the previous Plans, for example to strengthen coverage in the adult population. It concludes by recalling that the general objective to be achieved by 2023 is: incidence <1 case of measles / 1,000,000 population, and among the specific objectives it remains essential to achieve and maintain a vaccination coverage greater than or equal to 95% for the first dose of measles

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

Potential Onco-Suppressive Role of miR122 and miR144 in Uveal Melanoma through ADAM10 and C-Met Inhibition

Uveal melanoma (UM) is a rare tumor of the eye that leads to deadly metastases in about half of the patients. ADAM10 correlates with c-Met expression in UM and high levels of both molecules are related to the development of metastases. MiR122 and miR144 modulate ADAM10 and c-Met expression in different settings. We hypothesized a potential onco-suppressive role for miR122 and miR144 through modulation of ADAM10 and c-Met in UM. We analyzed the UM Cancer Genome Atlas data portal (TCGA) dataset, two other cohorts of primary tumors and five human UM cell lines for miR122 and miR144 expression by miR microarray, RT-qPCR, Western blotting, miR transfection and luciferase reporter assay. Our results indicate that miR122 and miR144 are expressed at low levels in the UM cell lines and in the TCGA UM dataset and were down-modulated in a cohort of seven UM samples, compared to normal choroid. Both miR122 and miR144 directly targeted ADAM10 and c-Met. Overexpression of miR122 and miR144 led to reduced expression of ADAM10 and c-Met in the UM cell lines and impaired cell proliferation, migration, cell cycle and shedding of c-Met ecto-domain. Our results show that miR122 and miR144 display an onco-suppressive role in UM through ADAM10 and c-Met modulation. View Full-Tex

Mda-9/Syntenin Is Expressed in Uveal Melanoma and Correlates with Metastatic Progression

Uveal melanoma is an aggressive cancer that metastasizes to the liver in about half of the patients, with a high lethality rate. Identification of patients at high risk of metastases may provide indication for a frequent follow-up for early detection of metastases and treatment. The analysis of the gene expression profiles of primary human uveal melanomas showed high expression of SDCBP gene (encoding for syndecan-binding protein-1 or mda-9/syntenin), which appeared higher in patients with recurrence, whereas expression of syndecans was lower and unrelated to progression. Moreover, we found that high expression of SDCBP gene was related to metastatic progression in two additional independent datasets of uveal melanoma patients. More importantly, immunohistochemistry showed that high expression of mda-9/syntenin protein in primary tumors was significantly related to metastatic recurrence in our cohort of patients. Mda-9/syntenin expression was confirmed by RT-PCR, immunofluorescence and immunohistochemistry in cultured uveal melanoma cells or primary tumors. Interestingly, mda-9/syntenin showed both cytoplasmic and nuclear localization in cell lines and in a fraction of patients, suggesting its possible involvement in nuclear functions. A pseudo-metastatic model of uveal melanoma to the liver was developed in NOD/SCID/IL2Rγ null mice and the study of mda-9/syntenin expression in primary and metastatic lesions revealed higher mda-9/syntenin in metastases. The inhibition of SDCBP expression by siRNA impaired the ability of uveal melanoma cells to migrate in a wound–healing assay. Moreover, silencing of SDCBP in mda-9/syntenin-high uveal melanoma cells inhibited the hepatocyte growth factor (HGF)-triggered invasion of matrigel membranes and inhibited the activation of FAK, AKT and Src. Conversely syntenin overexpression in mda-9/syntenin-low uveal melanoma cells mediated opposite effects. These results suggest that mda-9/syntenin is involved in uveal melanoma progression and that it warrants further investigation as a candidate molecular marker of metastases and a potential therapeutic target

IL-27 induces the expression of IDO and PD-L1 in human cancer cells

none9IL-27 is a member of the IL-12 family that is produced by macrophages and dendritic cells. IL-27 inhibits the growth and invasiveness of different cancers and therefore represents a potential anti-tumor agent. By contrast, it may exert immune-regulatory properties in different biological systems. We reported that IL-27 induces the expression of the IL-18 inhibitor IL-18BP, in human Epithelial Ovarian Cancer (EOC) cells, thus potentially limiting the immune response. Here, we tested whether IL-27 may modulate other immune-regulatory molecules involved in EOC progression, including Indoleamine 2,3-dioxygenase (IDO) and Programmed Death-Ligand (PD-L)1. IDO and PD-L1 were not constitutively expressed by EOC cells in vitro, but IL-27 increased their expression through STAT1 and STAT3 tyrosine phosphorylation. Differently, cells isolated from EOC ascites showed constitutive activation of STAT1 and STAT3 and IDO expression. These findings, together with the expression of IL-27 in scattered leukocytes in EOC ascites and tissues, suggest a potential role of IL-27 in immune-regulatory networks of EOC. In addition, IL-27 induced IDO or PD-L1 expression in monocytes and in human PC3 prostate and A549 lung cancer cells. A current paradigm in tumor immunology is that tumor cells may escape from immune control due to "adaptive resistance" mediated by T cell-secreted IFN-γ, which induces PD-L1 and IDO expression in tumor cells. Our present data indicate that also IL-27 has similar activities and suggest that the therapeutic use of IL-27 as anti-cancer agent may have dual effects, in some tumors.Carbotti, Grazia; Barisione, Gaia; Airoldi, Irma; Mezzanzanica, Delia; Bagnoli, Marina; Ferrero, Simone; Petretto, Andrea; Fabbi, Marina; Ferrini, SilvanoCarbotti, Grazia; Barisione, Gaia; Airoldi, Irma; Mezzanzanica, Delia; Bagnoli, Marina; Ferrero, Simone; Petretto, Andrea; Fabbi, Marina; Ferrini, Silvan

Different subcellular localization of ALCAM molecules in neuroblastoma: Association with relapse

Background: The Activated Leukocyte Cell Adhesion Mol. (ALCAM/CD166), involved in nervous system development, has been linked to tumor progression and metastasis in several tumors. No information is available on ALCAM expression in neuroblastoma, a childhood neoplasia originating from the sympathetic nervous system. Methods: ALCAM expression was analyzed by immunofluorescence and immunohistochem. on differentiated neuroblastoma cell lines and on archival specimens of stroma-poor, not MYCN amplified, resectable neuroblastoma tumors, resp. Results: ALCAM is variously expressed in neuroblastoma cell lines, is shed by metalloproteases and is cleaved by ADAM17/TACE in vitro. ALCAM is expressed in neuroblastoma primary tumors with diverse patterns of subcellular localization and is highly expressed in the neuropil area in a subgroup of cases. Tumor specimens showing high expression of ALCAM at the membrane of the neuroblast body or low levels in the neuropil area are assocd. with relapse (P=0.044 and P<0.0001, resp.). In vitro differentiated neuroblastoma cells show strong ALCAM expression on neurites, suggesting that ALCAM expression in the neuropil is related to a differentiated phenotype. Conclusion: Assessment of ALCAM localization by immunohistochem. may help to identify patients who, in the absence of neg. prognostic factors, are at risk of relapse and require a more careful follow-up

Different subcellular localization of ALCAM molecules in neuroblastoma: Association with relapse

Background: The Activated Leukocyte Cell Adhesion Mol. (ALCAM/CD166), involved in nervous system development, has been linked to tumor progression and metastasis in several tumors. No information is available on ALCAM expression in neuroblastoma, a childhood neoplasia originating from the sympathetic nervous system. Methods: ALCAM expression was analyzed by immunofluorescence and immunohistochem. on differentiated neuroblastoma cell lines and on archival specimens of stroma-poor, not MYCN amplified, resectable neuroblastoma tumors, resp. Results: ALCAM is variously expressed in neuroblastoma cell lines, is shed by metalloproteases and is cleaved by ADAM17/TACE in vitro. ALCAM is expressed in neuroblastoma primary tumors with diverse patterns of subcellular localization and is highly expressed in the neuropil area in a subgroup of cases. Tumor specimens showing high expression of ALCAM at the membrane of the neuroblast body or low levels in the neuropil area are assocd. with relapse (P=0.044 and P<0.0001, resp.). In vitro differentiated neuroblastoma cells show strong ALCAM expression on neurites, suggesting that ALCAM expression in the neuropil is related to a differentiated phenotype. Conclusion: Assessment of ALCAM localization by immunohistochem. may help to identify patients who, in the absence of neg. prognostic factors, are at risk of relapse and require a more careful follow-up

Doxorubicin impairs the insulin-like growth factor-1 system and causes insulin-like growth factor-1 resistance in cardiomyocytes.

Insulin-like growth factor-1 (IGF-1) promotes the survival of cardiomyocytes by activating type 1 IGF receptor (IGF-1R). Within the myocardium, IGF-1 action is modulated by IGF binding protein-3 (IGFBP-3), which sequesters IGF-1 away from IGF-1R. Since cardiomyocyte apoptosis is implicated in anthracycline cardiotoxicity, we investigated the effects of the anthracycline, doxorubicin, on the IGF-1 system in H9c2 cardiomyocytes.Besides inducing apoptosis, concentrations of doxorubicin comparable to those observed in patients after bolus infusion (0.1-1 µM) caused a progressive decrease in IGF-1R and increase in IGFBP-3 expression. Exogenous IGF-1 was capable to rescue cardiomyocytes from apoptosis triggered by 0.1 and 0.5 µM, but not 1 µM doxorubicin. The loss of response to IGF-1 was paralleled by a significant reduction in IGF-1 availability and signaling, as assessed by free hormone levels in conditioned media and Akt phosphorylation in cell lysates, respectively. Doxorubicin also dose-dependently induced p53, which is known to repress the transcription of IGF1R and induce that of IGFBP3. Pre-treatment with the p53 inhibitor, pifithrin-α, prevented apoptosis and the changes in IGF-1R and IGFBP-3 elicited by doxorubicin. The decrease in IGF-1R and increase in IGFBP-3, as well as apoptosis, were also antagonized by pre-treatment with the antioxidant agents, N-acetylcysteine, dexrazoxane, and carvedilol.Doxorubicin down-regulates IGF-1R and up-regulates IGFBP-3 via p53 and oxidative stress in H9c2 cells. This leads to resistance to IGF-1 that may contribute to doxorubicin-initiated apoptosis. Further studies are needed to confirm these findings in human cardiomyocytes and explore the possibility of manipulating the IGF-1 axis to protect against anthracycline cardiotoxicity

Effect of exogenous IGF-1 on doxorubicin-induced apoptosis of H9c2 cells: TUNEL and caspase 3/7 activity.

<p>Frequency of apoptotic cells, as assessed by TUNEL (A; representative microphotographs are shown in B) and fluorescence (AUF) produced by the cleavage of a substrate of activated caspase 3/7 (C), 24 hours after no treatment (Ctr) or incubation of H9c2 cardiomyocytes with doxorubicin (Dox) ± IGF-1 at the indicated concentrations. ***, P <0.001 vs. Ctr. c, P <0.001 vs. Dox 0.1; d, P <0.001 vs. Dox 0.5; e, P <0.01 vs. Dox 0.1; f, P <0.05 vs. Dox 0.5. ¥, P <0.001 vs. Dox 0.1 + IGF-1 100; ¢, P <0.001 vs. Dox 0.1 + IGF-1 100 and Dox 0.5 + IGF-1 100.</p

Antioxidants reverse doxorubicin-initiated apoptosis and IGF-1R /IGFBP-3 perturbation.

<p>Frequency of apoptotic cells (A) and IGF-1R and IGFBP-3 expression (representative western blot in B and densitometry of western blot bands in C) 24 hours after no treatment (Ctr) or incubation of H9c2 cardiomyocytes with 1 μM doxorubicin (Dox) with or without pre-treatment with N-acetylcysteine (NAC), dexrazoxane (DEX), or carvedilol (CARV). *, P <0.05 vs. Ctr; **, P <0.01 vs. Ctr; ***, P <0.001 vs. Ctr; ****, P <0.0001 vs. Ctr. #, P <0.05 vs. Dox 1; ^, P <0.01 vs. Dox 1.</p