Protection of Animals during Transport: Analysis of the Infringements Reported from 2009 to 2013 during On-Road Inspections in Italy

Council Regulation (EC) No 1/2005 requires that vehicles that are transporting animals be subjected to checks conducted by competent authorities. Yearly, each member state sends a report to the European government on the infringements that have been discovered during on-road inspections. The reports that were published by the Italian Ministry of Public Health from 2009 to 2013 were analyzed. Possible associations between the type of infringement (related to animal welfare (AW), vehicle (V) and accompanying documents (D)), year, season, transported species, place of inspection, and competent authorities were identified. A total of 985 infringements were analyzed, with some vehicles receiving more than one (mean: 1.58; max: 9). A score (from 1 to 3) that was related to the severity of the infringements was created. In 2009 and 2010, there was a 50% higher probability of encountering penalties of a lower severity (D or V) than in 2011 (p < 0.0001). Vehicles that were transporting pigs showed the highest probability of committing animal welfare-related infringements (odds ratio (OR) = 3.85, 95% confidence interval (95%CI) = 1.82-8.76, p < 0.0001). Vehicles were four times more likely to suffer animal welfare-related penalties when traffic police worked in synergy with veterinary services (OR = 4.12, 95%CI = 1.70-11.13, p = 0.0005). Vehicles that were transporting Equidae and "other species," including pets, for commercial purposes were more likely to be fined for a lack or incompleteness of the veterinary documents than those transporting cattle (p = 0.002 and p = 0.004, respectively). This study gives statistical evidence of the implementation of EC 1/2005. The training of transporters and drivers on how to manage transport in an animal welfare-friendly manner and a standardized method on how to conduct road inspections among competent authorities are recommended

Sorbitol dehydrogenase overexpression and other aspects of dysregulated protein expression in human precancerous colorectal neoplasms: a quantitative proteomics study

Colorectal adenomas are cancer precursor lesions of the large bowel. A multitude of genomic and epigenomic changes have been documented in these preinvasive lesions, but their impact on the protein effectors of biological function has not been comprehensively explored. Using shotgun quantitative MS, we exhaustively investigated the proteome of 30 colorectal adenomas and paired samples of normal mucosa. Total protein extracts were prepared from these tissues (prospectively collected during colonoscopy) and from normal (HCEC) and cancerous (SW480, SW620, Caco2, HT29, CX1) colon epithelial cell lines. Peptides were labeled with isobaric tags (iTRAQ 8-plex), separated via OFFGEL electrophoresis, and analyzed by means of LC-MS/MS. Nonredundant protein families (4325 in tissues, 2017 in cell lines) were identified and quantified. Principal component analysis of the results clearly distinguished adenomas from normal mucosal samples and cancer cell lines from HCEC cells. Two hundred and twelve proteins displayed significant adenoma-related expression changes (q-value < 0.02, mean fold change versus normal mucosa ±1.4), which correlated (r = 0.74) with similar changes previously identified by our group at the transcriptome level. Fifty-one (∼25%) proteins displayed directionally similar expression changes in colorectal cancer cells (versus HCEC cells) and were therefore attributed to the epithelial component of adenomas. Although benign, adenomas already exhibited cancer-associated proteomic changes: 69 (91%) of the 76 protein up-regulations identified in these lesions have already been reported in cancers. One of the most striking changes involved sorbitol dehydrogenase, a key enzyme in the polyol pathway. Validation studies revealed dramatically increased sorbitol dehydrogenase concentrations and activity in adenomas and cancer cell lines, along with important changes in the expression of other enzymes in the same (AKR1B1) and related (KHK) pathways. Dysregulated polyol metabolism might represent a novel facet of metabolome remodeling associated with tumorigenesis

West Nile virus transmission. results from the integrated surveillance system in Italy, 2008 to 2015

IIn Italy a national Plan for the surveillance of imported and autochthonous human vector-borne diseases (chikungunya, dengue, Zika virus disease and West Nile virus (WNV) disease) that integrates human and veterinary (animals and vectors) surveillance, is issued and revised annually according with the observed epidemiological changes. Here we describe results of the WNV integrated veterinary and human surveillance systems in Italy from 2008 to 2015. A real time data exchange protocol is in place between the surveillance systems to rapidly identify occurrence of human and animal cases and to define and update the map of affected areas i.e. provinces during the vector activity period from June to October. WNV continues to cause severe illnesses in Italy during every transmission season, albeit cases are sporadic and the epidemiology varies by virus lineage and geographic area. The integration of surveillance activities and a multidisciplinary approach made it possible and have been fundamental in supporting implementation of and/or strengthening preventive measures aimed at reducing the risk of transmission of WNV trough blood, tissues and organ donation and to implementing further measures for vector control

PIK3CA Mutation in the ShortHER Randomized Adjuvant Trial for Patients with Early HER2\ufe Breast Cancer: Association with Prognosis and Integration with PAM50 Subtype.

Purpose: We explored the prognostic effect of PIK3CA mutation in HER2\ufe patients enrolled in the ShortHER trial. Patients and Methods: The ShortHER trial randomized 1,253 patients with HER2\ufe breast cancer to 9 weeks or 1 year of adjuvant trastuzumab combined with chemotherapy. PIK3CA hotspot mutations in exon 9 and 20 were analyzed by pyrosequencing. Expression of 60 genes, including PAM50 genes was measured using the nCounter platform. Results: A mutation of the PIK3CA gene was detected in 21.7% of the 803 genotyped tumors. At a median follow-up of 7.7 years, 5-year disease-free survival (DFS) rates were 90.6% for PIK3CA mutated and 86.2% for PIK3CA wild-type tumors [HR, 0.84; 95% confidence interval (CI), 0.56\u20131.27; P \ubc 0.417]. PIK3CA mutation showed a favorable prognostic impact in the PAM50 HER2-enriched subtype (n \ubc 232): 5-year DFS 91.8% versus 76.1% (log-rank P \ubc 0.049; HR, 0.46; 95% CI, 0.21\u20131.02). HER2-enriched/PIK3CA mutated versus wild-type tumors showed numerically higher tumor-infiltrating lymphocytes (TIL) and significant upregulation of immune-related genes (including CD8A, CD274, PDCD1, and MYBL2, a proliferation gene involved in immune processes). High TILs as well as the upregulation of PDCD1 and MYBL2 were associated with a significant DFS improvement within the HER2-enriched subtype (HR, 0.82; 95% CI, 0.68\u20130.99; P \ubc 0.039 for 10% TILs increment; HR, 0.81; 95% CI, 0.65\u20130.99; P \ubc 0.049 for PDCD1 expression; HR, 0.72; 95% CI, 0.53\u20130.99; P \ubc 0.042 for MYBL2 expression). Conclusions: PIK3CA mutation showed no prognostic impact in the ShortHER trial. Within the HER2-enriched molecular subtype, patients with PIK3CA mutated tumors showed better DFS versus PIK3CA wild-type, which may be partly explained by upregulation of immune-related genes

Journeys, Journey Conditions, and Welfare Assessment of Broken (Handled) Horses on Arrival at Italian Slaughterhouses

During horse transportation, the journey conditions are considered a welfare risk. This study aimed to document journeys, journey conditions, and welfare status of handled horses on arrival at two different slaughterhouses in Northern and Southern Italy, to find possible associations between journey conditions and welfare problems. The welfare status of 613 draft-breed and light-breed horses from 32 different journeys was evaluated on arrival at the slaughterhouses with a standardized protocol, using animal-based (ABMs) and environmental-based (EBMs) measures. The drivers&rsquo; skills and vehicle characteristics were found to be mostly compliant with EC 1/2005. The horses traveled in single bays, 90&deg; to the direction of travel for an average journey duration of 26.5 &plusmn; 14 h. On arrival at the slaughterhouses, the horses were unloaded by handlers, via halter and rope. The prevalence of reluctance to unload, injuries, nasal, and lacrimal discharge was 22.2%, 24.6%, 11.6%, and 10%, respectively. Journey duration, unloading duration, vehicle changes, long stops, handlers/drivers&rsquo; skills, temperature, season, and horse individual characteristics were associated with horses&rsquo; welfare and health status (all p &lt; 0.05). Our study confirms the hypothesis that appropriate journey conditions are of crucial importance to safeguard the welfare of broken/handled horses transported over long distances for slaughter

Targeted proteomics for multiplexed verification of markers of colorectal tumorigenesis

Targeted proteomic methods can accelerate the verification of multiple tumor marker candidates in large series of patient samples. We utilized the targeted approach known as selected / multiple reaction monitoring (SRM/MRM) to verify potential protein markers of colorectal adenoma identified by our group in previous transcriptomic and quantitative shotgun proteomic studies of a large cohort of precancerous colorectal lesions. We developed SRM assays to reproducibly detect and quantify 25 (62.5%) of the 40 selected proteins in an independent series of precancerous and cancerous tissue samples (19 adenoma/normal mucosa pairs; 17 adenocarcinoma/normal mucosa pairs). Twenty-three proteins were significantly upregulated (n=17) or downregulated (n=6) in adenomas and/or adenocarcinomas, as compared with normal mucosa (linear fold changes ≥ ±1.3, adjusted P value <0.05). Most changes were observed in both tumor types (upregulation of ANP32A, ANXA3, SORD, LDHA, LCN2, NCL, S100A11, SERPINB5, CDV3, OLFM4, and REG4; downregulation of ARF6 and PGM5), and a five-protein biomarker signature distinguished neoplastic tissue from normal mucosa with a maximum area under the receiver operating curve greater than 0.83. Other changes were specific for adenomas (PPA1 and PPA2 upregulation; KCTD12 downregulation) or adenocarcinoma (ANP32B, G6PD, RCN1, and SET upregulation; downregulated AKR1B1, APEX1, and PPA1). Some changes significantly correlated with a few patient- or tumor-related phenotypes. Twenty-two (96%) of the 23 proteins have a potential to be released from the tumors into the bloodstream, and their detectability in plasma has been previously reported. The proteins identified in this study expand the pool of investment-worthy biomarker candidates that can be used to develop a standardized pre-colonoscopy blood test for the early detection of colorectal tumors