Combined Inhibition of Hedgehog and HDAC6: In Vitro and In Vivo Studies Reveal a New Role for Lysosomal Stress in Reducing Glioblastoma Cell Viability.

Glioblastoma multiforme (GBM) is the most common and malignant brain tumor in adults. The invasiveness and the rapid progression that characterize GBM negatively impact patients' survival. Temozolomide (TMZ) is currently considered the first-choice chemotherapeutic agent. Unfortunately, over 50% of patients with GBM do not respond to TMZ treatment, and the mutation-prone nature of GBM enables the development of resistance mechanisms. Therefore, efforts have been devoted to the dissection of aberrant pathways involved in GBM insurgence and resistance in order to identify new therapeutic targets. Among them, sphingolipid signaling, Hedgehog (Hh) pathway, and the histone deacetylase 6 (HDAC6) activity are frequently dysregulated and may represent key targets to counteract GBM progression. Given the positive correlation between Hh/HDAC6/sphingolipid metabolism in GBM, we decided to perform a dual pharmacological inhibition of Hh and HDAC6 through cyclopamine and tubastatin A, respectively, in a human GMB cell line and zebrafish embryos. The combined administration of these compounds elicited a more significant reduction of GMB cell viability than did single treatments in vitro and in cells orthotopically transplanted in the zebrafish hindbrain ventricle. We demonstrated, for the first time, that the inhibition of these pathways induces lysosomal stress which results in an impaired fusion of lysosomes with autophagosomes and a block of sphingolipid degradation in GBM cell lines. This condition, which we also recapitulated in zebrafish embryos, suggests an impairment of lysosome-dependent processes involving autophagy and sphingolipid homeostasis and might be instrumental in the reduction of GBM progression.

CT colonography: patient tolerance of laxative free fecal tagging regimen versus traditional cathartic cleansing.

PURPOSE: The aim of our prospective study was to compare patient tolerance of laxative free fecal tagging regimen (LFT) versus traditional cathartic cleansing (TC). MATERIALS AND METHODS: 264 patients, at average risk for development of colorectal cancer (105 men and 159 women; mean age 62 years ± 5 SD), underwent 32 rows CT colonography. Patients were alternatively placed into 2 study groups: Group 1 (n = 132) followed TC and Group 2 (n = 132) LFT. TC protocol consisted of no fiber diet and Phospho-lax(®) 80 mL in 2 L of water the day before imaging. LFT protocol consisted of no fiber diet and ingestion with meals of 30 mL of water-soluble iodinated contrast agent (Gastrografin(®)) for 2 days before imaging. No frank laxative drugs were administered. All studies were reviewed in a combined fashion, primary 2D followed by 3D endoluminal and dissected views. After the examination all patients were asked to provide a feedback about tolerance to the each bowel preparation. The first 30 patients of each group were also investigated with optical colonoscopy (OC) used as gold standard to confirm our diagnosis (Group 1* and Group 2*). CONCLUSIONS: LFT reduces discomfort and seems to improve diagnostic accuracy of CTC.

A survey of fleas on dogs in southern Italy.

A survey aimed at studying the presence and distribution of fleas on dogs was conducted in an area of southern Italy. Between February 2005 and 2006, dogs were examined for fleas at four private veterinary clinics, with a twice-weekly frequency. Fleas were detected on 246 (17.9%) out of the 1376 tested dogs. A total of 960 fleas were sampled and two species were identified, namely Ctenocephalides felis felis (16.3% of the tested dogs) and Ctenocephalides canis (1.5% of the tested dogs). The results of the logistic regression model showed a significant association between the flea positivity and the following independent variables: housed with other dogs or cats and utilization, i.e. increasing prevalence from pets to guard, hunting, and stray dogs. Clinical symptoms (pruritus, alopecia, and flea allergic dermatitis) were also observed in some of the flea positive dogs. Flea infestation was detected throughout the year, although the prevalence was higher during the period between June and October.

Mapping the main Leishmania phlebotomine vector in the endemic focus of the Mt. Vesuvius in southern Italy.

Geographical information systems and remote sensing were used to analyze the distribution of the Leishmania infantum-Phlebotomus perniciosus parasite-vector system in relation to environmental features of two opposite sides (coastal and Apennine) of Mt. Vesuvius, an area of intense transmission of human and canine leishmaniasis in southern Italy. Weekly phlebotomine collections were carried out during two consecutive warm seasons (2004- 2005) in 24 and 25 sites of the coastal and Apennine sides, respectively. Sandflies were caught using over one-thousand and seven hundred 20 x 20 cm-sticky traps placed in different environments. A total of 873 sandflies were collected, of which 284 (32.5%) were identified as P. perniciosus. The cumulative density (number of specimens/m2 of sticky trap/two nights) of this vector species was 3.9. P. perniciosus was significantly more abundant in the coastal side (5.8) as compared to the Apennine side (1.4). The main environmental differences between the two sides were the aspect (south-west for the coastal and north-east for the Apennine side) and land use. The predominance of green vegetated environments (forest, semi-natural and agricultural areas) in the coastal side, in contrast with the predominance of artificial surfaces (namely urban environment) in the Apennine side, could be responsible for the different P. perniciosus densities between the two surveyed areas.

Canine faecal contamination and parasitic risk in the city of Naples (southern Italy).

BACKGROUND: Dogs are associated with more than 60 zoonotic diseases among which, parasitosis and, in particular, helminthosis, can pose serious public-health concerns worldwide. Many canine gastrointestinal parasites eliminate their dispersion elements (eggs, larvae, oocysts) by the faecal route. The quantity of canine faeces deposited on public and private property in cities worldwide is both a perennial nuisance and an important health issue. Public sites such as playgrounds, parks, gardens, public squares and sandpits may be an important source of human infection. The aim of this study was to evaluate the extent of both canine faecal contamination in the city of Naples (southern Italy), and presence of canine parasitic elements, with particular regard to those which are potential agents of zoonosis. A regular grid of sub-areas (1 km x 700 m) was overlaid on the city map using a Geographical Information System (GIS). In each sub-area the straightest 1 km transect was drawn and digitalized on-screen in the GIS. Between February and May 2005 canine faeces were counted along the 1 km transects in 143 sub-areas, and 415 canine faecal samples were collected and submitted to coprological examinations. Negative binomial regression models and Gaussian random effects models were used to analyze the association between faeces count and human population density taking into account for extraPoisson variability. Logistic regression model was used to evaluate the association between positivity to parasitic elements and number of canine faeces. RESULTS: Out of the 143 studied sub-areas, 141 (98.6%) contained canine faeces. There was a strong spatial gradient with 48% of the total variability accounted by between neighbourhood variability; a positive association between the number of faeces and the human population density was found. Seventy (over 415, 16.9%) canine faecal samples were positive for parasitic elements. There was no association between positivity to parasitic elements and the number of canine faeces. Eggs of Toxocara canis, Toxascaris leonina, Ancylostoma caninum and Trichuris vulpis were found, as well as oocysts of Isospora canis. CONCLUSION: In conclusion, the results of the present study, conducted using GIS both for planning and sampling and for evaluation and presentation of findings, showed the presence of canine faecal contamination in the city of Naples, and the presence of canine parasitic elements, some of which are potential agents of zoonosis.