Identification of Ixodes ricinus blood meals using an automated protocol with high resolution melting analysis (HRMA) reveals the importance of domestic dogs as larval tick hosts in Italian alpine forests

Background In Europe, Ixodes ricinus L. is the main vector of a variety of zoonotic pathogens, acquired through blood meals taken once per stage from a vertebrate host. Defining the main tick hosts in a given area is important for planning public health interventions; however, until recently, no robust molecular methods existed for blood meal identification from questing ticks. Here we improved the time- and cost-effectiveness of an HRMA protocol for blood meal analysis and used it to identify blood meal sources of sheep tick larvae from Italian alpine forests. Methods Nine hundred questing nymphs were collected using blanket-dragging in 18 extensive forests and 12 forest patches close to rural villages in the Province of Trento. Total DNA was either extracted manually, with the QIAamp DNA Investigator kit, or automatically using the KingFisher™ Flex Magnetic Particle Processors (KingFisher Cell and Tissue DNA Kit). Host DNA was amplified with six independent host group real-time PCR reactions and identified by means of HRMA. Statistical analyses were performed in R to assess the variables important for achieving successful identification and to compare host use in the two types of forest. Results Automating DNA extraction improved time- and cost-effectiveness of the HRMA protocol, but identification success fell to 22.4% (KingFisher™) from 55.1% (QIAamp), with larval hosts identified in 215 of 848 questing nymphs; 23 mixed blood meals were noted. However, the list of hosts targeted by our primer sets was extended, improving the potential of the method. Host identification to species or genus level was possible for 137 and 102 blood meals, respectively. The most common hosts were Rodentia (28.9%) and, unexpectedly, Carnivora (28.4%), with domestic dogs accounting for 21.3% of all larval blood meals. Overall, Cetartiodactyla species fed 17.2% of larvae. Passeriformes (14.6%) fed a significantly higher proportion of larvae in forest patches (22.3%) than in extensive forest (9.6%), while Soricomorpha (10.9%) were more important hosts in extensive forest (15.2%) than in forest patches (4.3%). Conclusions The HRMA protocol for blood meal analysis is a valuable tool in the study of feeding ecology of sheep ticks, especially with the cost- and time- reductions introduced here. To our knowledge, we show for the first time that domestic dogs are important larval hosts in the Alps, which may have possible implications for tick-borne disease cycles in urbanized area

La pernice bianca alpina (Lagopus muta helvetica): una sottospecie endemica in declino. Filogeografia, variabilità genetica e aspetti biologici.

The alpine rock ptarmigan (Lagopus muta helvetica) is considered in the Alps as a relictual glacial species. It is nowadays threaten by global warming, that reduces suitable habitats, and by human disturbance, increasingly frequent at high altitude. On the Pyrenees limited genetic variability was already documented; on the Alps a number of populations display a decreasing census size. We analyzed sequence variation in the mitochondrial DNA D-loop for 148 individuals and ten microsatellite loci for 152 individuals in order to investigate evolutionary history, levels of genetic variability and structure of alpine rock ptarmigan. The samples came from different localities of the Alps

Teratoma with a malignant somatic component in pediatric patients: The Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) experience

Abstract Background. Teratoma with a malignant somatic component (TMSC) is rare but described in adults, whereas information on pediatric presentation is sparse Procedure. The Associazione Italiana Ematologia Oncologia Pediatrica identified 14 cases of TMSC Clinical files and pathology specimens were reviewed Results. The series (9 female, 5 male) showed the following disease testis (2), sacrococcygeal (3), ovary (3), retroperitoneum (3), mediastinum (2), and foot soft tissue (1) Distribution of the somatic component vas carcinoma (4), pancreatic neuroendocrine tumor (1), neuroblastoma (3), rhabdomyosarcoma (3), rhabdomyosarcoma plus liposarcoma, chondrosarcoma, neurogenic sarcoma (1), chondrosarcoma plus neuroectodermal sarcoma (1), malignant peripheral nerve sheath tumor(1) Three patients where in stage I, four in stage II, three in stage III, and four in stage IV All but one patient patient underwent surgery and only showed carcinoma components Nine patients relapsed or progressed and eight died Six patients are alive and disease-free Two patients underwent complete resection and four were treated based on transformed histologies Relapse-lice and overall survival rates were 35.7% and 42.8%, respectively (median follow-up, 3 1 months) Conclusions. Prognosis for germ cell tumors (GCTs) containing MSC is worse than that for GCTs The pediatric disease appears to be more heterogeneous in tumor site distribution and MSC histology than in adults Our series suggests no effects of age, histology, or gender on outcome Surgery has an essential role in localized disease, with complete resector highly desirable Chemotherapy optimized for histology should include reagents directed to the somatic malignancy, if chemosensitive Malignant GCT warrants GCT-directed chemotherapy Pediatr Blood Cancer 2010,54 532-537 (C) 2010 Wiley-Liss, In

Additional file 2: Table S2. of Identification of Ixodes ricinus blood meals using an automated protocol with high resolution melting analysis (HRMA) reveals the importance of domestic dogs as larval tick hosts in Italian alpine forests

BLASTn search results and alignment for Ovis spp. amplicons obtained with Cervidae primer set. (PDF 323 kb

Additional file 1: Table S1. of Identification of Ixodes ricinus blood meals using an automated protocol with high resolution melting analysis (HRMA) reveals the importance of domestic dogs as larval tick hosts in Italian alpine forests

Melting temperatures for amplicons obtained from questing tick extracted using the KingFisher Cell and Tissue kit. (PDF 415 kb

Identification of an Actionable Mutation of KIT in a Case of Extraskeletal Myxoid Chondrosarcoma

Extraskeletal myxoid chondrosarcoma (EMC) is an extremely rare soft tissue sarcoma, marked by a translocation involving the NR4A3 gene. EMC is usually indolent and moderately sensitive to anthracycline-based chemotherapy. Recently, we reported on the therapeutic activity of sunitinib in a series of EMC cases, however the molecular target of sunitinib in EMC is unknown. Moreover, there is still the need to identify alternative therapeutic strategies. To better characterize this disease, we performed whole transcriptome sequencing in five EMC cases. Peculiarly, in one sample, an in-frame deletion (c.1735_1737delGAT p.D579del) was identified in exon 11 of KIT. The deletion was somatic and heterozygous and was validated both at DNA and mRNA level. This sample showed a marked high expression of KIT at the mRNA level and a mild phosphorylation of the receptor. Sanger sequencing of KIT in additional 15 Formalin Fixed Paraffin Embedded (FFPE) EMC did not show any other mutated cases. In conclusion, exon 11 KIT mutation was detected only in one out of 20 EMC cases analyzed, indicating that KIT alteration is not a recurrent event in these tumors and cannot explain the EMC sensitivity to sunitinib, although it is an actionable mutation in the individual case in which it has been identified

Sexual assault and abuse committed against family members: An analysis of 1342 legal outcomes and their motivations

Background Over the past years medical centres specifically addressed in gender-based violence have developed protocols for the collections of evidence useful in the courtroom, including accurate documentation of physical and psychological states of the victim and collection of samples. Previous studies showed an association between documented physical trauma and conviction but unfortunately, few studies in the recent literature analysed the factors that influence the legal outcome and final judgement. The present study focused on the elements that appeared of significance in the legal outcome, including medico-legal evaluation, source of the crime report and circumstance of the assault. Methods It was conducted a retrospective analysis of all the judgments issued by the Public Prosecutor's Office at a Court of a Metropolitan Italian city regarding sexual and domestic violence, from January 1st 2011 to 31st December 31st 2015. Examination regarded the demographic information of the victim and of the defendant, information on the crime, the circumstances of the aggression and medical information retrieved. Sentences were subsequently divided into two categories based on the legal outcome (conviction vs acquittal) and the different characteristics of the two sub-populations were compared to verify if there were variables significantly associated to the judge's final judgment. Results Over the 5 years taken into consideration, there have been 1342 verdicts regarding crimes of sexual violence (374 cases) and regarding abuses against family members or cohabitants (875): Other 93 cases regarded both sexual violence and abuse. 66.3% ended in conviction of the offender and 33.7% in acquittal of the accused. Cases of conviction were more frequent when they involved: Use of a weapon by the assailant, as well as if the assailant had a criminal record and had a history of drug abuse or other addictions; duration of proceeding less 22 months and a civil party involved; presence of clinical documentation together with other deposition in addition to victim's deposition; also frequent episodes of violence and application of precautionary measures were associated to conviction. Conclusions Many factors seem able to influence the judge's judgment, although clearly each case must be singularly evaluated. The mere presence of medical documentation, without the support of other sources of evidence, such as the victim's statement or further declarations, however, is almost always not definitive for the verdict. Despite so, in cases where there are multiple sources of evidence, clinical documentation can provide useful elements and can give clues on the consistency between the history told and injuries observed

Coherent wavepackets in the Fenna-Matthews-Olson complex are robust to excitonic-structure perturbations caused by mutagenesis

The photosynthetic FMO complex is probed by femtosecond pump-probe spectroscopy and compared with a series of genetically modified mutants with distinct excitonic interactions. These experiments allow us to identify vibrational contributions to the coherence