4 Years of EU Cookie Law: Results and Lessons Learned

Personalized advertisement has changed the web. It lets websites monetize the content they offer. The downside is the continuous collection of personal information with significant threats to personal privacy. In 2002, the European Union (EU) introduced a first set of regulations on the use of online tracking technologies. It aimed, among other things, to make online tracking mechanisms explicit to increase privacy aware- ness among users. Amended in 2009, the EU Directive mandates websites to ask for informed consent before using any kind of profiling technology, e.g., cookies. Since 2013, the ePrivacy Directive became mandatory, and each EU Member State transposed it in national legislation. Since then, most of European websites embed a “Cookie Bar”, the most visible effect of the regulation. In this paper, we run a large-scale measurement campaign to check the current implementation status of the EU cookie directive. For this, we use CookieCheck, a simple tool to automatically verify legislation violations. Results depict a shady picture: 49 % of websites do not respect the Directive and install profiling cookies before any user’s consent is given. Beside presenting a detailed picture, this paper casts lights on the difficulty of legislator attempts to regulate the troubled marriage between ad-supported web services and their users. In this picture, online privacy seems to be continuously at stake, and it is hard to reach transparency

INTRACELLULAR DNA DELIVERY USING POLYMERIC NANOPARTICLES FOR LYSOSOMAL STORAGE DISORDERS GENE THERAPY

INTRODUCTION: Gene therapy can be currently performed by viral or non-viral vectors. Although viral ones have already come to a clinical translation, concerns remain about both safety and the industrial applicability of this system. We therefore started the study of a transposon-based gene transfer system. We used nanoparticles (NPs) as vector to carry the transposon within the blood stream, then in cells. These had to be able to hold large plasmids, such as transposon. The NPs must then be able to reach the tissue most affected by the genetic disorder. Coating the NPs with specific peptides, they acquire the ability to accumulate in the target tissue, in our case the Central Nervous System (CNS) penetrating the blood-brain barrier (BBB). We started to develop a non-viral gene therapy approach in order to treat a neurological disease, globoid cell leukodystrophy in (GLD), an inherited, rapidly fatal disorder affecting myelin. METHODS: We tested the ability of a transposon-based system called Sleeping Beauty (SB), to convey the luciferase marker gene in the control cells. We then verified the ability of NPS of chitosan to contain large plasmids. We then verified the quality (size, shape) of the NPS obtained and their ability to integrate the gene of interest (GALC) within the cellular genome. Lastly, we verified the ability of a heptameric synthetic glycopeptide, of opioid origin, g7, to convey inside the CNS NPs of Poly Lactic-Co-Glycolic Acid (PLGA). We injected PLGA NPs coated with g7 in tail vein\u2019s Balb/c mice and followed through fluorescence spectroscopy their distribution. RESULTS: We verified the high SB transduction efficiency on control cells by their expression of luciferase marker gene. We then assessed the copies number of the integrated gene. We demonstrated the high transduction efficiency of the transgene within the genome of fibroblasts grown from biopsies of patients with GLD. We assessed the quality and features of chitosan-SB NPs. We could observe that g7 coated PLGA NPs accumulate in the CNS 10 minutes after infusion, with accumulation even and up to 2 hours. DISCUSSION: SB demonstrated high efficiency in stable transduction of control cells. So we thought to encapsulate SB in chitosan NPs. These have proven to be highly efficient in stably transducing deficient fibroblasts. We can confirm that g7 is today the best way to address the NPs to the CNS. CONCLUSIONS: Future studies will test the ability of g7 to coat the NPs of chitosan, verify its efficiency in conveying the chitosan NPs to the CNS as well as they did for PLGA NPs. Finally, by cloning into SB the gene whose mutation causes GLD, can be assessed the therapeutic efficacy of the system in the mouse model of the disease

The decline in muscle strength and muscle quality in relation to metabolic derangements in adult women with obesity

Background & aims: The metabolic and functional characteristics related to sarcopenic obesity have not been thoroughly explored in the earlier stages of the aging process. The aim of the present study was to examine the phenotype of sarcopenic obesity, in terms of lean body mass, muscle strength and quality, in adult women with and without the Metabolic Syndrome (MetS), and its relationship with the features of myosteatosis. Methods: Study participants were enrolled at the Sapienza University, Rome, Italy. Body composition was assessed by DXA. The Handgrip strength test (HGST) was performed. HGST was normalized to arm lean mass to indicate muscle quality; intermuscular adipose tissue (IMAT) and intramyocellular lipid content (IMCL) were measured by magnetic resonance imaging and spectroscopy, as indicators of myosteatosis. Different indices of sarcopenia were calculated, based on appendicular lean mass (ALM, kg) divided by height squared, or weight. The NCEP-ATPIII criteria were used to diagnose the MetS. HOMA-IR was calculated. The physical activity level (PAL) was assessed through the IPAQ questionnaire. Results: 54 women (age: 48 ± 14 years, BMI: 37.9 ± 5.4 kg/m 2 ) were included. 54% had the MetS (metabolically unhealthy, MUO). HGST/arm lean mass was lower in MUO women than women without the MetS (6.3 ± 1.8 vs. 7.8 ± 1.6, p = 0.03). No differences emerged in terms of absolute ALM (kg) or other indices of sarcopenia (ALM/h 2 or ALM/weight) between metabolically healthy (MHO) vs. MUO women (p > 0.05). Muscle quality was negatively associated with HOMA-IR (p = 0.02), after adjustment for age, body fat, hs-CRP levels, and PAL. IMAT, but not IMCL, was significantly higher in obese women with the MetS compared to women without the MetS (p > 0.05). No association emerged between HGST/arm lean mass and IMAT or IMCL when HOMA-IR was included in the models. Conclusion: Insulin resistance, and not sarcopenia or myosteatosis per se, was associated with muscle weakness, resulting in the phenotype of “dynapenic obesity” in middle-aged women with the metabolic syndrome

Impact of standardization in tissue processing: the performance of different fixatives

Most tissues in clinical practice are formalin-fixed and paraffin-embedded for histological as well as molecular analyses. The reproducibility and uniformity of molecular analyses is strictly dependent on the quality of the biomolecules, which is highly influenced by pre-analytical processes. In this study, the effect of different fixatives was compared, including formalin, Bouin's solution, RCL2® and TAG-1™ fixatives, by stringent application of ISO standards in mouse liver tissue processing, including formalin-free transport of tissues and tissue grossing in a refrigerated environment. The effect of fixatives was studied in terms of nucleic acid quality at the time of tissue processing and after one year of tissue storage at room temperature in the dark. Furthermore, a microcomputed tomography (CT) scan analysis was applied to investigate the paraffin embedding. The results show that the application of ISO standards in tissue processing allows analysis of 400 bases amplicons from RNA and 1000 bases from DNA, even in extracts from formalin-fixed and paraffin-embedded tissues. However, after one year storage at room temperature in the dark, a degradation of the nucleic acids was observed. Nevertheless, extracts can still be analyzed, but for metachronous tests it is highly recommended to repeat the quantitation of housekeeping genes in order to standardize the extent of nucleic acid degradation

Self-efficacy for coping. Utility of the Cancer behavior inventory (Italian) for use in palliative care

Background: Newer models of palliative and supportive cancer care view the person as an active agent in managing physical and psychosocial challenges. Therefore, personal efficacy is an integral part of this model. Due to the lack of instruments in Italian to assess coping self-efficacy, the present study included the translation and validation of the Italian version of the Cancer Behavior Inventory-Brief (CBI-B/I) and an initial analysis of the utility of self-efficacy for coping in an Italian sample of palliative care patients. Methods: 216 advanced cancer patients who attended palliative care clinics were enrolled. The CBI-B/I was administered along with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 30 (EORTC QLQ-C30), the Mini Mental Adjustment to Cancer Scale (Mini-MAC), the Cancer Concerns Checklist (CCL), and the Hospital Anxiety and Depression Scale (HADS). The Eastern Cooperative Oncology Group Performance Status (ECOG-PS) ratings of functional capacity were completed by physicians. Results: Factor analysis confirmed that the structure of the CBI-B/I was consistent with the English version. Internal consistency reliability and significant correlations with the EORTC QLQ-C30, Mini-MAC, and HADS supported the concurrent validity of the CBI-B/I. Differences in CBI-B/I scores for high versus low levels of the CCL and ECOG-PS supported the clinical utility of the CBI-B/I. Conclusions: The CBI-B/I has strong psychometric properties and represents an important addition to newer model of palliative and supportive care. In order to improve clinical practice, the CBI-B/I could be useful in identifying specific self-efficacy goals for coping in structured psychosocial intervention

Vitamin B6 rescues insulin resistance and glucose-induced DNA damage caused by reduced activity of Drosophila PI3K

: The insulin signaling pathway controls cell growth and metabolism, thus its deregulation is associated with both cancer and diabetes. Phosphatidylinositol 3-kinase (PI3K) contributes to the cascade of phosphorylation events occurring in the insulin pathway by activating the protein kinase B (PKB/AKT), which phosphorylates several substrates, including those involved in glucose uptake and storage. PI3K inactivating mutations are associated with insulin resistance while activating mutations are identified in human cancers. Here we show that RNAi-induced depletion of the Drosophila PI3K catalytic subunit (Dp110) results in diabetic phenotypes such as hyperglycemia, body size reduction, and decreased glycogen content. Interestingly, we found that hyperglycemia produces chromosome aberrations (CABs) triggered by the accumulation of advanced glycation end-products and reactive oxygen species. Rearing PI3KRNAi flies in a medium supplemented with pyridoxal 5'-phosphate (PLP; the catalytically active form of vitamin B6) rescues DNA damage while, in contrast, treating PI3KRNAi larvae with the PLP inhibitor 4-deoxypyridoxine strongly enhances CAB frequency. Interestingly, PLP supplementation rescues also diabetic phenotypes. Taken together, our results provide a strong link between impaired PI3K activity and genomic instability, a crucial relationship that needs to be monitored not only in diabetes due to impaired insulin signaling but also in cancer therapies based on PI3K inhibitors. In addition, our findings confirm the notion that vitamin B6 is a good natural remedy to counteract insulin resistance and its complications

Top-down structures of mafic enclaves within the Valle Fértil magmatic complex (Early Ordovician, San Juan, Argentina)

Magmatic structures related to the mechanical interaction between mafic magmas and granitoids have been studied in the Valle Fértil calc-alkaline igneous complex, Argentina. Excepcional outcrops with vertical walls of more than 300 m high allow us the study of three-dimensional geometries of individual blobs of mafic magma as well as the geometry of pipe-like structures in which mafic microgranular enclaves are concentrated in more than 50 times the normal abundance in the granodiorite mass. The shape of enclaves and pipe-like structures are interpreted as the ressult of top-to-down intrusions of a mafic magma into a granodiorite-tonalite mass. These sinking structures are the result of a reverselly stratified magma chamber with gabbros and diorites at the top and granodiorite-tonalite at the bottom. They may account for most of the structures found in microgranular enclaves and magma mingling zones that characterize calc-alkaline batholiths. Synplutonic intrusions from the top is the only plausible mechanism to account for the observed structures. The model may be of general application to calc-alkaline batholiths characterized by the presence of mafic microgranular enclaves. An implication of these reverselly stratified magma chambers is the presence of a petrological inversion which may be the consequence of cold diapirs emplaced below the mantle wedge in a suprasubduction setting

An update on microbiological causes of canine otitis externa in Campania Region – Italy.

Objective: To update the recent knowledge of the microbiological causes of canine otitis externa in Campania Region (Italy) and the antibiotic susceptibility patterns of the isolated strains. Methods: A total of 122 dogs were examined by otoscopy, and auricular swab samples were collected from both ears in 74 dogs presenting clinical bilateral otitis and from single ears in 48 dogs displaying clinical unilateral otitis. Cytological examination, bacteriological analysis and antimicrobial susceptibility tests were performed. Results: Thirty-one out of 122 dogs were positive for yeast species (25.4%, 95% confidence interval (CI): 18.2%–34.2%) with a higher prevalence of Malassezia pachydermatis (21/31 isolates, 67.7%, CI: 48.5%–82.7%), and a total of 91 out of 122 dogs were positive for bacterial species (74.6%; CI: 65.8%–81.8%) with a higher prevalence of Staphylococcus pseudintermedius (45/143 isolates, 31.5%, CI: 24.1%–39.8%). These results are the first description of Streptococcus agalactiae-associated otitis. The yeasts isolated showed high levels of susceptibility to all antifungal agents tested; on the contrary all the isolated bacterial strains were highly resistant to at least four out of ten antimicrobial classes. Both Gram-positive and Gram-negative bacteria showed high resistance to amoxicillin/clavulanate and kanamycin hence they are not recommended as initial empirical therapy for the otitis treatment. Conclusions: This update illustrates an increase in antibiotic resistances providing an insight into the current knowledge of the therapeutic procedures followed on canine otitis externa in Italy. It also emphasizes the importance of considering the results of the microbiological and sensitivity tests to decide on an appropriate antibiotic therapy

Single step multiple genotyping by MALDI-TOF mass spectrometry, for evaluation of minor histocompatibility antigens in patients submitted to allogeneic stem cell transplantation from HLA-matched related and unrelated donor

The outcome of patients underwent to allogeneic stem cell transplantation (allo- SCT) is closely related to graft versus host disease (GvHD) and graft versus leukemia (GvL) effects which can be mediated by mHAgs. 23 mHAgs have been identified and reported to be differently correlated with GVHD or GVL and the aim of this work was develop a method to genotype the mHAgs described so far. For this study we used MALDI-TOF iPLEX Gold Mass Array technology. We tested 46 donor/recipient matched pairs that underwent allo-SCT because of Philadelphia positive (Ph+) chronic myeloid leukemia (n=29) or Ph+ acute lymphoblastic leukemia (n=17). Our data show that sibling pairs had a lesser number of mHAgs mismatches compared to MUD pairs. Notably, donor/recipient genomic mismatch on DPH1 was correlated with an increased risk of acute GvHD and LB-ADIR-1R mismatch on graft versus host direction was correlated with a better RFS with no increase of GvHD risk. Our work provides a simple, accurate and highly automatable method for mHAgs genotyping and suggest the role of mHAgs in addressing the immune reaction between donor and host

Health conditions of inmates in Italy

Abstract Background Several studies have shown that prison is characterized by a higher prevalence of chronic diseases than unconfined settings. The aim of this study was to describe the characteristics and health of inmates, focusing on internal diseases. Methods We designed a specific clinical record using the Python programming language. We considered all of the diagnoses according to the ICD-9-CM. Results Of a total of 17,086 inmates, 15,751 were enrolled in our study (M = 14,835; F = 869), corresponding to 92.2% of the entire inmate population (mean age of 39.6 years). The project involved a total of 57 detention facilities in six Italian regions (for a total of 28% of all detainees in Italy), as counted in a census taken on February 3, 2014. From the entire study sample, 32.5% of prisoners did not present any disorders, while 67.5% suffered from at least one disease. The most frequent pathologies were psychiatric (41.3%), digestive (14.5%), infectious (11.5%), cardiovascular (11.4%), endocrine, metabolic, and immune (8.6%), and respiratory (5.4%). Conclusion The findings showed that a large number of detainees were affected by several chronic conditions such as hypertension, dyslipidemia and type 2 diabetes mellitus, with an unusually high prevalence for such a young population. Therefore, a series of preventive measures is recommended to strengthen the entire care process and improve the health and living conditions of prisoners