Key Role of Sequencing to Trace Hepatitis A Viruses Circulating in Italy During a Large Multi-Country European Foodborne Outbreak in 2013

Background

Foodborne Hepatitis A Virus (HAV) outbreaks are being recognized as an emerging public health problem in industrialized countries. In 2013 three foodborne HAV outbreaks occurred in Europe and one in USA. During the largest of the three European outbreaks, most cases occurred in Italy (>1,200 cases as of March 31, 2014). A national Task Force was established at the beginning of the outbreak by the Ministry of Health. Mixed frozen berries were early demonstrated to be the source of infection by the identity of viral sequences in patients and in food. In the present study the molecular characterization of HAV isolates from 355 Italian cases is reported.

Methods

Molecular characterization was carried out by PCR/sequencing (VP1/2A region), comparison with reference strains and phylogenetic analysis.

Results

A unique strain was responsible for most characterized cases (235/355, 66.1%). Molecular data had a key role in tracing this outbreak, allowing 110 out of the 235 outbreak cases (46.8%) to be recognized in absence of any other link. The data also showed background circulation of further unrelated strains, both autochthonous and travel related, whose sequence comparison highlighted minor outbreaks and small clusters, most of them unrecognized on the basis of epidemiological data. Phylogenetic analysis showed most isolates from travel related cases clustering with reference strains originating from the same geographical area of travel.

Conclusions

In conclusion, the study documents, in a real outbreak context, the crucial role of molecular analysis in investigating an old but re-emerging pathogen. Improving the molecular knowledge of HAV strains, both autochthonous and circulating in countries from which potentially contaminated foods are imported, will become increasingly important to control outbreaks by supporting trace back activities, aiming to identify the geographical source(s) of contaminated food, as well as public health interventions.     

Biophysical and antisense properties of oligodeoxynucleotides containing 7-propynyl-, 7-iodo- and 7-cyano-7-deaza-2-amino-2'-deoxyadenosines.

The synthesis of 7-propynyl-, 7-iodo- and 7-cyano-7-deaza-2-amino-2'-deoxyadenosines is described. The nucleosides were synthesized, functionalized into the phosphoramidites and incorporated into oligodeoxynucleotides. Spectroscopic melting experiments against complementary RNA showed increases of 3-4 degreesC per modification for single substitutions and smaller increases per incorporation for multiple substitutions relative to unmodified control sequences. The 7-propyne and 7-iodo nucleosides were incorporated into antisense sequences targeting the 3'-UTR of murine C- raf mRNA. Both nucleosides demonstrated substitution-dependent potency. The sequences with three and four substitutions of the 7-propyne-7-deaza-2-amino-2'-deoxyadenosine exhibited a 2-3-fold increase in potency over unmodifed controls.     

Phaeodactylum tricornutum: An established model species for diatom molecular research and an emerging chassis for algal synthetic biology

Diatoms are prominent and highly diverse microalgae in aquatic environments. Compared with other diatom species, Phaeodactylum tricornutum is an “atypical diatom” displaying three different morphotypes and lacking the usual silica shell. Despite being of limited ecological relevance, its ease of growth in the laboratory and well-known physiology, alongside the steady increase in genome-enabled information coupled with effective tools for manipulating gene expression, have meant it has gained increased recognition as a powerful experimental model for molecular research on diatoms. We here present a brief overview of how over the last 25 years P. tricornutum has contributed to the unveiling of fundamental aspects of diatom biology, while also emerging as a new tool for algal process engineering and synthetic biology.     

Renewable Energy and Carbon Management in the Cradle‐to‐Cradle Certification: Limitations and Opportunities

As part of the Cradle to Cradle^® (C2C) certification program, the C2C certification criterion, Renewable Energy and Carbon Management (RE&CM), focuses on use of electricity from renewable energy (RE) and direct greenhouse gas offsets in the manufacturing stage and, to a limited extent, on the cradle to gate only at the highest level of certification. The aim of this study is to provide decision makers with a quantified overview of possible limitations of that C2C certification requirement and potential gains by introducing a full life cycle assessment (LCA) perspective to the scheme. Scenario analysis was used to perform an LCA of an aluminum can system representing different levels of the C2C certification criterion, RE&CM, considering different strategies to achieve 100% RE in the manufacturing stage. The adoption of a broader life cycle RE perspective was considered through the implementation of electricity from renewable sources from cradle to grave. Our results show that compliance with the current RE&CM certification framework offers limited benefits, that is, significant reduction for climate change, but negligible reductions for other environmental impacts (e.g., particulate matter and acidification). However, increasing the share of RE in the primary aluminum production from a full life cycle perspective can greatly increase the environmental benefits brought up by the C2C certification not only for climate change, but also for the broader range of impact categories. In our striving toward environmental sustainability, which often cannot be approximated by climate‐change impacts alone, we therefore recommend decision makers in industries to combine the C2C certification with LCA when they define strategies for the selection of RE and raw materials suppliers.     

Reduction of low molecular weight protein-tyrosine phosphatase expression improves hyperglycemia and insulin sensitivity in obese mice

To investigate the role of low molecular weight protein-tyrosine phosphatase (LMW-PTP) in glucose metabolism and insulin action, a specific antisense oligonucleotide (ASO) was used to reduce its expression both in vitro and in vivo. Reduction of LMW-PTP expression with the ASO in cultured mouse hepatocytes and in liver and fat tissues of diet-induced obese (DIO) mice and ob/ob mice led to increased phosphorylation and activity of key insulin signaling intermediates, including insulin receptor-beta subunit, phosphatidylinositol 3-kinase, and Akt in response to insulin stimulation. The ASO-treated DIO and ob/ob animals showed improved insulin sensitivity, which was reflected by a lowering of both plasma insulin and glucose levels and improved glucose and insulin tolerance in DIO mice. The treatment did not decrease body weight or increase metabolic rate. These data demonstrate that LMW-PTP is a key negative regulator of insulin action and a potential novel target for the treatment of insulin resistance and type 2 diabetes.     

Some properties of lysozyme--lithium perfluorononanoate complexes

Mixtures containing lysozyme, LYSO, and a fully fluorinated surfactant, lithium perfluorononanoate, LiPFN, were investigated in a wide range of concentrations and mole ratios. To ensure consistency to the data, a comparison was made, when possible, with the more conventional SDS as surfactant. Molecular solutions, precipitates, and micellar phases have been observed. The region of existence for each phase depends on the LiPFN/LYSO mole ratios, r, and was determined by different experimental methods. Optical absorbance, CD, 19F NMR, viscosity, electrical conductivity, and dielectric relaxation methods were used. Some methods give information on the protein conformation, others on the state of the surfactant or on the collective system properties, respectively. Addition of LiPFN gives rise to a solution, a poly phase dispersion (at low surfactant to protein ratios) and to a micelle-mediated redissolution of the precipitates. Concomitant to the above macroscopic properties, peculiar effects in the state of LYSO are observed. Low amounts of surfactant reduce significantly the amount of alpha-helix in favor of the beta-sheet conformation of the protein. The former is almost completely regained once micelle-assisted redissolution of the complex occurs. The tertiary structure of the protein, conversely, is lost at low surfactant content and never recovered. Such evidence suggests the occurrence of a molten globule conformation for LYSO in micellar media.     

Characterization of anti-coagulant properties of prenylated coumarin ferulenol

We investigated the mechanisms underlying severe bleeding occurring upon consumption of Ferula communis. The prenylated coumarin ferulenol extracted from this plant did not directly affect blood coagulation but showed hepatocyte cytotoxicity and, at non-cytotoxic concentrations (<100 nM), impaired factor X biosynthesis (40% reduction). Studies with ferulenol derivatives indicated the prenyl residue as major determinant of ferulenol activity.     

Abundance and seasonal activity of questing Ixodes ricinus ticks in their natural habitats in southern Germany in 2011

Questing ticks were sampled monthly over a period of 11 months from February, 2011 to December, 2011 at 13 sites in southern Germany using the flagging method. The ticks were identified to species, gender, and stadium. Although both I. ricinus and D. reticulatus were sampled, this study concentrated on I. ricinus, since it was the most abundant tick to be found. Additional weather data (air and soil temperature, relative air humidity, precipitation, sunshine duration) were recorded on each sampling site and the local vegetation described. A total of 14, 394 ticks was collected (7,862 larvae, 5,568 nymphs, 964 adults) and their activity was recorded in order to determine the seasonal activity pattern over different periods of the year. In contrast to the widely accepted pattern of a bimodal seasonal activity in moderate areas with a dominant peak in spring and a minor peak in autumn, a unimodal activity pattern was found for all development stages on six of the 12 sampling sites. Tick abundance was compared to weather variables. Tick host‐seeking activity was found to be significantly dependent on the temperature at ground level, precipitation, and sunshine duration as well as relative air humidity. Adult ticks showed a positive correlation with the duration of sunshine, whereas nymphs were mostly unaffected by this phenomenon.     

The effects of disodium pamidronate on human polymorphonuclear leukocytes and platelets: An <Emphasis Type="Italic">in vitro</Emphasis> study

Recent reports have indicated that, as well as having antiresorptive effects, bisphosphonates could have an application as anti-inflammatory drugs. Our aim was to investigate whether this anti-inflammatory action could be mediated by the nitric oxide (NO) released by the leukocytes migrating to the site of inflammation. In particular, we investigated in vitro the intracellular calcium concentration ([Ca²⁺]_i), the level of NO released by PMN and platelets, and the PMN myeloperoxidase activity after incubation with disodium pamidronate, since there was a postulated modulatory effect of this aminosubstituted bisphosphonate on leukocytes both in vitro and in vivo. Our data shows that the pamidronate treatment provoked a significant increase in the [Ca²⁺]_i parallel to the enhancement in NO release, suggesting a possible activation of constitutive nitric oxide synthase, while the myeloperoxidase activity was significantly reduced. In conclusion, we hypothesized that treatment with pamidronate could stimulate NO-production by cells present near the bone compartment, thus constituting a protective mechanism against bone resorption occurring during inflammation. In addition, PMN- and platelet-derived NO could act as a negative feed-back signal to restrict the inflammatory processes.