Full genome sequence of a novel circo-like virus detected in an adult European eel Anguilla anguilla showing signs of cauliflower disease.

An adult European eel Anguilla anguilla, showing typical signs of the so-called cauliflower disease, was subjected to pathological and molecular virological examinations. Samples taken from internal organs and the polypoid proliferative tissue from the mouth were examined by PCR for the detection of several viruses. Positive results were obtained with a nested PCR targeting the rep gene of circoviruses. Analysis of the partial rep sequence indicated the presence of a putative novel circovirus, but attempts to isolate it remained unsuccessful. The missing part of the genome was acquired by an inverse nested PCR with 2 specific primer pairs, designed from the newly determined rep sequence, followed by genome walking. The circular full genome was found to consist of 1378 nt (GenBank accession no. KC469701). Two oppositely oriented open reading frames (ORFs) were present, of which one was unambiguously identified as a circoviral rep gene. However, the predicted product of the other ORF, though it is a clear positional counterpart of the cap genes, showed no obvious homology to any known circoviral capsid proteins. A stem-loop-like element in the intergenic region between the 5' ends of the ORFs was also found. Phylogenetic calculations indicated that the novel virus belongs to the genus Circovirus in the family Circoviridae. The relative amount of the viral DNA in the organ samples was estimated by quantitative real-time PCR. The results suggested that the examined fish was caught in an active viremic state, although the role of this circovirus in the etiology of the cauliflower diseases could not be ascertained

A high abdominal aortic calcification score by dual X-ray absorptiometry is associated with cardiovascular events after kidney transplantation

Background: Aortic calcification is associated with an increased risk for cardiovascular events in renal transplant recipients. This study focused on the association of abdominal aortic calcification (AAC) and cardiovascular events assessed using a dual-energy X-ray absorptiometry (DXA) scoring methodology for AAC. Methods: From 2008 to 2014, renal transplant recipients referred for a DXA procedure within 6 months after transplantation were included in a retrospective, single-centre study. The primary endpoint was the occurrence of cardiovascular events, defined as myocardial infarction, cerebrovascular accident or transient ischaemic attack, after transplantation. AAC was quantified using an 8-point scoring system and patients were divided into three groups; a control group (AAC = 0), a low AAC group (AAC = 1-3) and a high AAC group (AAC = 4-8). Results: We evaluated 701 patients, 267 (38.1%) had detectable calcifications (low AAC 190 patients, high AAC 77 patients) and 434 (61.9%) had no calcifications. Cardiovascular events were seen in 37 (8.5%) patients in the control group, in 18 (9.5%) in the low AAC group and in 20 (26.0%) in the high AAC group. Univariate Cox proportional hazards analysis of the high AAC score showed a hazard ratio (HR) of 4.23 [95% confidence interval (CI) 2.44-7.33; P < 0.01] for cardiovascular events, while results were not significant for the low AAC score. Multivariate analysis showed an independent significant association between a high AAC score and cardiovascular events [HR 2.78 (95% CI 1.05-7.64); P = 0.04]. Assessment of the continuous net reclassification index (NRI), comparing the combined clinical variables with a model of both AAC scoring and clinical variables, showed an NRI of 0.76 (95% CI 0.65-0.86; P < 0.01). Conclusions: An independent association between a high AAC score, assessed by DXA, and cardiovascular events was identified and provides an opportunity for early cardiovascular risk stratification in renal transplant recipients

Identification of Disease-Associated Cryptococcal Proteins Reactive With Serum IgG From Cryptococcal Meningitis Patients

Cryptococcus neoformans, an opportunistic fungal pathogen ubiquitously present in the environment, causes cryptococcal meningitis (CM) mainly in immunocompromised patients, such as AIDS patients. We aimed to identify disease-associated cryptococcal protein antigens targeted by the human humoral immune response. Therefore, we used sera from Colombian CM patients, with or without HIV infection, and from healthy individuals living in the same region. Serological analysis revealed increased titers of anti-cryptococcal IgG in HIV-negative CM patients, but not HIV-positive CM patients, compared to healthy controls. In contrast, titers of anti-cryptococcal IgM were not affected by CM. Furthermore, we detected pre-existing IgG and IgM antibodies even in sera from healthy individuals. The observed induction of anti-cryptococcal IgG but not IgM during CM was supported by analysis of sera from C. neoformans-infected mice. Stronger increase in IgG was found in wild type mice with high lung fungal burden compared to IL-4Ra-deficient mice showing low lung fungal burden. To identify the proteins targeted by human anti-cryptococcal IgG antibodies, we applied a quantitative 2D immunoproteome approach identifying cryptococcal protein spots preferentially recognized by sera from CM patients or healthy individuals followed by mass spectrometry analysis. Twenty-three cryptococcal proteins were recombinantly expressed and confirmed to be immunoreactive with human sera. Fourteen of them were newly described as immunoreactive proteins. Twelve proteins were classified as disease-associated antigens, based on significantly stronger immunoreactivity with sera from CM patients compared to healthy individuals. The proteins identified in our screen significantly expand the pool of cryptococcal proteins with potential for (i) development of novel anticryptococcal agents based on implications in cryptococcal virulence or survival, or (ii) development of an anti-cryptococcal vaccine, as several candidates lack homology to human proteins and are localized extracellularly. Furthermore, this study defines preexisting anti-cryptococcal immunoreactivity in healthy individuals at a molecular level, identifying target antigens recognized by sera from healthy control persons