Lyme borreliosis in Europe.

Despite improvements in prevention, diagnosis and treatment, Lyme borreliosis (LB) is still the most common arthropod-borne disease in temperate regions of the northern hemisphere, with risk of infection associated with occupation (e.g. forestry work) and certain outdoor recreational activities (e.g. mushroom collecting). In Europe, LB is caused by infection with one or more pathogenic European genospecies of the spirochaete Borrelia burgdorferi sensu lato, mainly transmitted by the tick Ixodes ricinus. Recent surveys show that the overall prevalence of LB may be stabilising, but its geographical distribution is increasing. In addition, much remains to be discovered about the factors affecting genospecific prevalence, transmission and virulence, although avoidance of tick bite still appears to be the most efficient preventive measure. Uniform, European-wide surveillance programmes (particularly on a local scale) and standardisation of diagnostic tests and treatments are still urgently needed, especially in the light of climate change scenarios and land-use and socio-economic changes. Improved epidemiological knowledge will also aid development of more accurate risk prediction models for LB. Studies on the effects of biodiversity loss and ecosystem changes on LB emergence may identify new paradigms for the prevention and control of LB and other tick-borne diseases

Vectors as Epidemiological Sentinels: Patterns of Within-Tick Borrelia burgdorferi Diversity

Hosts including humans, other vertebrates, and arthropods, are frequently infected with heterogeneous populations of pathogens. Within-host pathogen diversity has major implications for human health, epidemiology, and pathogen evolution. However, pathogen diversity within-hosts is difficult to characterize and little is known about the levels and sources of within-host diversity maintained in natural populations of disease vectors. Here, we examine genomic variation of the Lyme disease bacteria, Borrelia burgdorferi (Bb), in 98 individual field-collected tick vectors as a model for study of within-host processes. Deep population sequencing reveals extensive and previously undocumented levels of Bb variation: the majority (~70%) of ticks harbor mixed strain infections, which we define as levels Bb diversity pre-existing in a diverse inoculum. Within-tick diversity is thus a sample of the variation present within vertebrate hosts. Within individual ticks, we detect signatures of positive selection. Genes most commonly under positive selection across ticks include those involved in dissemination in vertebrate hosts and evasion of the vertebrate immune complement. By focusing on tick-borne Bb, we show that vectors can serve as epidemiological and evolutionary sentinels: within-vector pathogen diversity can be a useful and unbiased way to survey circulating pathogen diversity and identify evolutionary processes occurring in natural transmission cycles

Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells.

One of the main goals of nanomedicine in cancer is the development of effective drug delivery systems, primarily nanoparticles. Survivin, an overexpressed anti-apoptotic protein in cancer, represents a pharmacological target for therapy and a Molecular Beacon (MB) specific for survivin mRNA is available. In this study, the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to promote survivin MB uptake in human A549 cells was investigated. Fluorescent and positively charged core PMMA-NPs of nearly 60nm, obtained through an emulsion co-polymerization reaction, and the MB alone were evaluated in solution, for their analytical characterization; then, the MB specificity and functionality were verified after adsorption onto the PMMA-NPs. The carrier ability of PMMA-NPs in A549 was examined by confocal microscopy. With the optimized protocol, a hardly detectable fluorescent signal was obtained after incubation of the cells with the MB alone (fluorescent spots per cell of 1.90±0.40 with a mean area of 1.04±0.20µm2), while bright fluorescent spots inside the cells were evident by using the MB loaded onto the PMMA-NPs. (27.50±2.30 fluorescent spots per cell with a mean area of 2.35±0.16µm2). These results demonstrate the ability of the PMMA-NPs to promote the survivin-MB internalization, suggesting that this complex might represent a promising strategy for intracellular sensing and for the reduction of cancer cell proliferation

Whole genome capture of vector-borne pathogens from mixed DNA samples: a case study of Borrelia burgdorferi

Background: Rapid and accurate retrieval of whole genome sequences of human pathogens from disease vectors or animal reservoirs will enable fine-resolution studies of pathogen epidemiological and evolutionary dynamics. However, next generation sequencing technologies have not yet been fully harnessed for the study of vector-borne and zoonotic pathogens, due to the difficulty of obtaining high-quality pathogen sequence data directly from field specimens with a high ratio of host to pathogen DNA. Results: We addressed this challenge by using custom probes for multiplexed hybrid capture to enrich for and sequence 30 Borrelia burgdorferi genomes from field samples of its arthropod vector. Hybrid capture enabled sequencing of nearly the complete genome (~99.5 %) of the Borrelia burgdorferi pathogen with 132-fold coverage, and identification of up to 12,291 single nucleotide polymorphisms per genome. Conclusions: The proprosed culture-independent method enables efficient whole genome capture and sequencing of pathogens directly from arthropod vectors, thus making population genomic study of vector-borne and zoonotic infectious diseases economically feasible and scalable. Furthermore, given the similarities of invertebrate field specimens to other mixed DNA templates characterized by a high ratio of host to pathogen DNA, we discuss the potential applicabilty of hybrid capture for genomic study across diverse study systems. Keywords: Hybrid capture Whole-genome sequencing SNPs Tick-borne pathogens Lyme diseas

Metagenomic Profile of the Bacterial Communities Associated with Ixodes ricinus Ticks

Assessment of the microbial diversity residing in arthropod vectors of medical importance is crucial for monitoring endemic infections, for surveillance of newly emerging zoonotic pathogens, and for unraveling the associated bacteria within its host. The tick Ixodes ricinus is recognized as the primary European vector of disease-causing bacteria in humans. Despite I. ricinus being of great public health relevance, its microbial communities remain largely unexplored to date. Here we evaluate the pathogen-load and the microbiome in single adult I. ricinus by using 454- and Illumina-based metagenomic approaches. Genomic DNA-derived sequences were taxonomically profiled using a computational approach based on the BWA algorithm, allowing for the identification of known tick-borne pathogens at the strain level and the putative tick core microbiome. Additionally, we assessed and compared the bacterial taxonomic profile in nymphal and adult I. ricinus pools collected from two distinct geographic regions in Northern Italy by means of V6-16S rRNA amplicon pyrosequencing and community based ecological analysis. A total of 108 genera belonging to representatives of all bacterial phyla were detected and a rapid qualitative assessment for pathogenic bacteria, such as Borrelia, Rickettsia and Candidatus Neoehrlichia, and for other bacteria with mutualistic relationship or undetermined function, such as Wolbachia and Rickettsiella, was possible. Interestingly, the ecological analysis revealed that the bacterial community structure differed between the examined geographic regions and tick life stages. This finding suggests that the environmental context (abiotic and biotic factors) and host-selection behaviors affect their microbiome

Diffusion-Weighted MR imaging: Clinical applications of kurtosis analysis to prostate cancer

Magnetic resonance imaging technique known as DWI (diffusion-weighted imaging) allows measurement of water diffusivity on a pixel basis for evaluating pathology throughout the body and is now routinely incorporated into many body MRI protocols, mainly in oncology. Indeed water molecules motion reflects the interactions with other molecules, membranes, cells, and in general the interactions with the environment. Microstructural changes as e.g. cellular organization and/or integrity then affect the motion of water molecules, and consequently alter the water diffusion properties measured by DWI. Then DWI technique can be used to extract information about tissue organization at the cellular level indirectly from water motion. In general the signal intensity in DWI can be quantified by using a parameter known as ADC (Apparent Diffusion Coefficient) emphasizing that it is not the real diffusion coefficient, which is a measure of the average water molecular motion. In the simplest models, the distribu- tion of a water molecule diffusing in a certain period of time is considered to have a Gaussian form with its width proportional to the ADC. However, water in biological structures often displays non-Gaussian diffusion behavior, consequently the DWI signal shows a more complex behavior that need to be modeled following different approaches. In this work we explore the possibility to quantify the degree to which water diffusion in biologic tissues is non-Gaussian introducing the AKC parameter (Apparent Kurtosis Coefficient). In this work we have realized DWI non-Gaussian diffusion maps to be used in the clinical routine along with standard ADC maps, giving to the radiologist another tool to explore how much structure inside a voxel is organized. In particular in this work some prostate DWI examples have been analyzed and will be shown

Within-tick <i>Bb</i> diversity in single infected ticks.

<p>(a) The minor allele frequency (MAF) spectrum of a representative “singly” infected tick sample. The <i>Bb</i> population infecting Bbcap22 (600 X coverage) harbors few variants with MAF < 5%, likely accrued via <i>in situ</i> evolution. Histogram bars represent the count of genomic sites with a minor allele frequency that falls within the MAF frequency bin on the x-axis. Note the differing y-axes for (a) and (c, d, and e). (b) MAF spectrum of a “multiply” infected tick sample, Bbcap5 (109 X coverage). The <i>Bb</i> population infecting Bbcap5 harbors a high number of intermediate frequency minor alleles, a level of diversity likely preexisting within a diverse inoculum. (c) MAF spectrum of a “multiply” infected tick sample, Bbcap17 (221 X coverage). (d) MAF spectrum of a “multiply” infected tick sample, Bbcap31 (309 X coverage).</p