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Purpose Assessment of inflammatory bowel disease (IBD) currently relies on aspecific clinical signs of bowel inflammation. Specific imaging of the diseased bowel regions is still lacking. Here, we investigate mucosal addressin cell adhesion molecule 1 (MAdCAM-1) as a reliable and specific endothelial target for engineered nanoparticles delivering imaging agents to obtain an exact mapping of diseased bowel foci. Materials and Methods We generated a nanodevice composed of PLGA-PEG coupled with anti-MAdCAM-1 antibody half-chains and loaded with quantum dots (P@QD-MdC NPs). Bowel localization and systemic biodistribution of the nanoconjugate were analyzed upon injection in a murine model of chronic IBD obtained through repeated administration of dextran sulfate sodium salt. Specificity for diseased bowel regions was also assessed ex vivo in human specimens from patients with IBD. Potential for development as contrast agent in magnetic resonance imaging was assessed by preliminary study on animal model. Results Synthesized nanoparticles revealed good stability and monodispersity. Molecular targeting properties were analyzed in vitro in a cell culture model. Upon intravenous injection, P@QD-MdC NPs were localized in the bowel of colitic mice, with enhanced accumulation at 24 h post-injection compared to untargeted nanoparticles (p

One of the most important Staphylococcus aureus virulence factors is Panton–Valentine leukocidin (PVL). We describe an outbreak of recurrent cutaneous PVL infections in different members of three family clusters. Molecular investigations were performed to confirm the presence of the mecA and PVL genes and to assign the SCCmec type, sequence type (ST) and clonal relatedness. A strain of PVL-producing methicillin-resistant S. aureus (MRSA) was responsible for infection in two related families (A and B), and a third family (C) was infected with PVL-producing methicillin-sensitive S. aureus (MSSA). Molecular investigations revealed the same clone of community-acquired (CA)-MRSA, PVL positive ST8, and SCCmec IV in families A and B and CA-MSSA PVL positive ST15 in family C. S. aureus PVL may give rise to recurrent uncontrolled infections that are difficult to eradicate, and close family contacts are at high risk for transmission. Keywords: Panton–Valentine leukocidin, Community-acquired methicillin-resistant Staphylococcus aureus

Objective To study pathogenic features of the somatic testicular microenvironment associated with idiopathic germ cell aplasia. Design Cross-sectional study. Setting Tertiary referral center for reproductive medicine. Patient(s) Testicular specimens from men with idiopathic nonobstructive azoospermia (iNOA) prospectively submitted to microdissection testicular sperm extraction. Of 20 specimens used for histology, 10 were also available for proteomic analysis. Primary Sertoli cells with normal karyotype and phenotype were also used. Intervention(s) Patients with iNOA were dichotomized according to a positive versus negative sperm retrieval at microdissection testicular sperm extraction, and on the isolated extracellular matrix (ECM) the proteomic analysis was performed. Main Outcome Measure(s) Proteomic analysis of the ECM from testicular specimens with positive versus negative sperm retrieval. Gene ontology enrichment was used to identify upstream regulators based on the 11 deregulated ECM proteins, which were validated by immunohistochemistry and quantitative polymerase chain reaction. Continuous variables were expressed as medians and interquartile range. Result(s) Germ cell aplasia was characterized by an increased signaling of the retinoic acid in Sertoli cells and associated with decreased expression of the basal membrane markers nidogen-2 and heparan sulfate proteoglycan-2. Decreased levels of the interstitial matrisome-associated factor IX and its regulator VKORC1 were, instead, coupled with decreased signaling of vitamin K in Leydig cells. An altered expression of a further eight ECM proteins was also found, including laminin-4 and laminin-5. Peripheral levels of the two vitamins were within the reference range in the two cohorts of iNOA men. Conclusion(s) We identified the pathogenetic signature of the somatic human testicular microenvironment, providing two vitamin-related mechanistic insights related to the molecular determinants of the idiopathic germ cell aplasia.

Background Clinical experience highlights the wide heterogeneity of primary prostate cancer (PPCa), even when potentially related to the same grade and stage. Currently available prediction tools and biomarkers do not always allow for early recognition of PPCa aggressive phenotype, sometimes making it impossible to distinguish among men harbouring indolent tumours or life-threatening disease. Objective To establish a novel ex vivo/in vitro model suitable to estimate the invasive phenotype of PPCa cells (PPCaC). Design, setting, and participants The ability of PPCaC to infiltrate the prostate extracellular matrix (ECM) was used as an index of invasion. ECM was obtained by decellularising 24 NT-prostate specimens from radical prostatectomy. PPCaC were obtained from six tumours with different Gleason patterns and pathological stages. Invasion ability was estimated in direct-cocolture experiments. Outcome measurements and statistical analysis The extent of ECM invasion by PPCaC was quantified by counting the number of infiltrated cells. Mann–Whitney test was utilised for statistical comparisons. Results and limitations Samples of ECM resulted to be free of cells and DNA and with a preserved three-dimensional structure and stromal protein content. The system resulted to be reliable since well characterised normal-, benign-, and malignant-prostate cell lines either re-epitheliased or invaded the matrices, according to their specific nature. Similarly, PPCaC invaded the ECMs consistently with their stage and biochemical recurrence. Of notice, this model was able to identify a different invasive phenotype even among tumours with equal Gleason patterns and pathological stages. The small sample size represents a limitation. Conclusions We developed an ex vivo/in vitro model able to reproduce the original PPCa-microenvironment and suitable to recognise the inherent invasive behaviour of PPCaC. Patient summary We developed a novel ex vivo/in vitro system which enables us to uncover which prostate tumours host potentially aggressive cancer cells. The identification of cancer cells with different invasive abilities will likely lead to the identification of new biomarkers to safely predict disease progression.

Evaluation of acute colitis relies on invasive endoscopic examination to assess the presence and the severity of the disease, and on aspecific radiological signs of bowel inflammation, such as contrast enhancement of bowel wall thickening. Consequently, the current best clinical management only offers an approximation of the real clinical scenario, and has a negative impact on the quality of life of patients. An accurate stadiation of inflammatory bowel diseases for assessment of response to therapies or indication for surgery remains challenging. Here, mucosal addressin cell-adhesion molecule-1 (MAdCAM-1) was investigated as a reliable and specific target of active bowel inflammation, to be exploited for site-specific nanotheranostics in a preclinical murine model of colitis. MAdCAM-1 is a cell adhesion molecule, which is upregulated on gut endothelium in case of inflammation at very early stages of the disease, and is responsible for T cells recruitment in involved bowel tracts. Moreover, it is finely related to activity of inflammatory bowel diseases and correlates with response to therapy. Anti-MAdCAM-1 antibodies were conjugated to the surface of smart and safe delivery nanoparticles, consisting of a manganese oxide core endowed with paramagnetic properties. The aim of the study was to assess the in vivo targeting efficacy and the accuracy of these innovative low-toxic MAdCAM-1-targeted nanoparticles, specifically designed to detect active bowel inflammation foci at early stages of the disease. Manganese oxide nanoparticles revealed good stability and polydispersity. Their negligible citotoxicity was demonstrated on endothelial cell cultures by analysis of viability and apoptosis, and on blood samples by hemolysis assay. Colitis was induced in mice by oral treatment with dextran sulfate sodium, and MAdCAM-1 overexpression was assessed on the intestinal vascular endothelium by immunohistochemistry and western blotting. Fluorescent anti-MAdCAM-1 nanoparticles were intravenously injected in mice at the time of early acute phase of the disease, and their biodistribution was compared to that of untargeted nanoparticles. Anti-MAdCAM-1 nanoparticles localized in the inflamed bowel of colitic mice, with preferential accumulation in the proximal colon at 24 hours post-injection. Active targeting of the endothelium within the inflamed bowel was demonstrated by transmission electron microscopy, and allowed to identify the sites of bowel inflammation. By contrast, nanoparticles coupled with a control immunoglobulin were only transiently observed in the bowel, and underwent a faster bowel wash-out. Histopathological lesions were not observed in mice organs upon nanoparticles treatment. Our results indicated that MAdCAM-1-targeted nanoparticles uncovered active bowel inflammation foci, accurately following the expression profile of MAdCAM-1 in inflamed mucosal tissue. Biomedical application of this targeted strategy could provide a noninvasive and specific system to improve clinical care and management of inflammatory bowel diseases.

Aim: We investigate MAdCAM-1 as a reliable target to detect active bowel inflammation for selective noninvasive nanodiagnostics. Materials & methods: We coupled anti-MAdCAM-1 antibodies to manganese oxide nanoparticles, and analyzed nanoconjugate biodistribution and safety in murine model of inflammatory bowel disease by imaging and histology. Results: Nanoparticles were stable and nontoxic. Upon administration in colitic mice, anti-MAdCAM-1 functionalized nanoparticles preferentially localized in the inflamed bowel, whereas untargeted nanoparticles were more rapidly washed out. Nanoparticles did not induce lesions in nontarget organs. Conclusion: Anti-MAdCAM-1 functionalized nanoparticles detected active bowel inflammation foci, accurately following MAdCAM-1 expression pattern. These nanoconjugates could be a promising noninvasive imaging system for an early and accurate follow-up in patients affected by acute colitis.

Merkel cell carcinoma (MCC) is a skin cancer that can also rarely arise in extracutaneous sites including mucosal surfaces. About 80% of MCCs harbor the Merkel cell polyomavirus (MCPyV). All cases of gastric MCCs so far reported were metastases from cutaneous sources. In the present article, we describe for the first time a primary gastric MCC harboring MCPyV. A 72-year-old man presented to clinical observation due to epigastric pain. Upper endoscopy revealed an ulcerated gastric tumor. The patient underwent total gastrectomy. The tumor was composed of mitotically active monomorphic small cells showing round nuclei with finely dispersed chromatin arranged in sheets and nests with large areas of necrosis. Tumor cells were positive for neuroendocrine markers and showed paranuclear dot immunoreactivity for cytokeratin 20. MCPyV was demonstrated with immunohistochemistry and electron microscopy, which showed intranuclear and intracytoplasmic viral particles. The MCPyV DNA in tumor cells was demonstrated with polymerase chain reaction analysis.

Pasteurella spp is the most frequently pathogen isolated from infected wound inflicted by domestic animal bites. Here we present a case of isolation of Pasteurella pneumotropica like organism from knee mega prosthesis infection occurred in 63 years old female.

International audience; IntroductionLouse-borne relapsing fever (LBRF) is an acute febrile infection that is typically characterized by one to three fairly regular waves of bacteremia [1,2]. It is caused by Borrelia recurrentis, a motile spirochete that measures 5 to 40 μm in length. The microorganism is transmitted from person to person by the human body louse (Pediculus humanus humanus). Disruptions in sanitation during wartime and mass migrations of people provide conditions that favor the propagation of body lice and thus the occurrence of outbreaks of the disease [1,3]. LBRF is endemic in East Africa (e.g., Ethiopia, Eritrea, Somalia, and Sudan) with the highest number of cases observed in Ethiopia, where it is the seventh most common cause of hospital admission and the fifth most common cause of death [4,5]. We report here the first case of imported LBRF observed in Lombardy (northern Italy) in a Somali refugee.

The extracellular matrix (ECM) from perilesional and colorectal carcinoma (CRC), but not healthy colon, sustains proliferation and invasion of tumor cells. We investigated the biochemical and physical diversity of ECM in pair-wised comparisons of healthy, perilesional and CRC specimens. Progressive linearization and degree of organization of fibrils was observed from healthy to perilesional and CRC ECM and was associated with a steady increase of stiffness and collagen crosslinking. In the perilesional ECM these modifications coincided with increased vascularization, whereas in the neoplastic ECM they were associated with altered modulation of matrisome proteins, increased content of hydroxylated lysine and lysyl oxidase. This study identifies the increased stiffness and crosslinking of the perilesional ECM predisposing an environment suitable for CRC invasion as a phenomenon associated with vascularization. The increased stiffness of colon areas may represent a new predictive marker of desmoplastic region predisposing to invasion, thus offering new potential application for monitoring adenoma with invasive potential.

PARV4 is a recently discovered human parvovirus widely distributed in injecting drug users in the USA and Europe, particularly in those co-infected with human immunodeficiency virus (HIV). Like parvovirus B19, PARV4 persists in previously exposed individuals. In bone marrow and lymphoid tissue, PARV4 sequences were detected in two sub-Saharan African study subjects with AIDS but without a reported history of parenteral exposure and who were uninfected with hepatitis C virus. PARV4 variants infecting these subjects were phylogenetically distinct from genotypes 1 and 2 (formerly PARV5) that were reported previously. Analysis of near-complete genome sequences demonstrated that they should be classified as a third (equidistant) PARV4 genotype. The availability of a further near-complete genome sequence of this novel genotype facilitated identification of conserved novel open reading frames embedded in the ORF2 coding sequence; one encoded a putative protein with identifiable homology to SAT proteins of members of the genus Parvovirus.

Post-kala-azar dermal leishmaniasis (PKDL) is a complication of visceral leishmaniasis (VL) observed mainly in Sudan and India where it follows treated VL in 50% and 10% of cases, respectively. We report a 46-year-old patient with acquired immune deficiency syndrome who, 7 months after diagnosis of VL, developed PKDL and uveal leishmaniasis following HAART-induced immune recovery. In southern Europe PKDL seems to be an emerging clinical presentation among human immunodeficiency virus (HIV)-infected patients experiencing HAART-induced immune recovery after a previous diagnosis of VL. The best treatment among HIV-infected patients remains to be determined.

Recent reports document resolution of human parvovirus B19-related pure red blood cell aplasia (PB19-PRCA) in HIV-infected patients upon commencement of highly active antiretroviral therapy (HAART). This article describes a patient with PB19-PRCA who, despite fully suppressive HAART, required cyclic administration of intravenous human immunoglobulin over a period of 17 months before PB19 seroconversion and subsequent resolution of relapsing severe anemia. All reports in the English literature describing PB19-related hematologic abnormalities in the post-HAART era are also described herein.

Highly active antiretroviral therapy (HAART) started shortly after birth resulted in reversion of human immunodeficiency virus (HIV) plasma viremia, proviral DNA in PBMC, viral culture, and serum HIV antibodies to negative. Discontinuation of HAART 2 years after apparent HIV eradication, however, was followed by virus replication, CD4 decline, and destruction of HIV-specific lymphocytes, epitomizing the impossibility of HIV eradication.

AimImmunohistochemical and molecular studies have suggested an oncogenic role for JCV in gastrointestinal carcinomas, but at least in colorectal cancers, the data are far from being unambiguous.MethodsTwo large series of formalin-fixed paraffin-embedded gastric and colorectal cancers were analysed for the expression of JCV large T Antigen (T-Ag) with a panel of five antibodies, and for the presence of T-Ag DNA sequences using two PCR systems.ResultsIntense nuclear staining was observed in 54/116 (46%) colorectal, and in 92/234 (39%) gastric cancers, using the PAb416 monoclonal antibody against large T-Ag. In colorectal cancers, PAb416-positivity was directly related to the presence of chromosomal instability, lymph node metastases and a more advanced tumour stage, and inversely related to proximal tumour site and the presence of microsatellite instability (MSI). In gastric cancers, the glandular histotype, the presence of lymph node metastases, a low frequency of MSI and EBV infection, and a worse prognosis were significantly associated with PAb416 immunoreactivity. Moreover, at both these sites, PAb416 expression was significantly associated with p53 nuclear accumulation. No positivity was obtained with all the other four anti-T-Ag-antibodies, and molecular analysis failed to demonstrate the presence of JCV DNA sequences in tested cases.ConclusionsOur immunohistochemical and molecular results do not support the idea that JCV T-Ag has a role in gastrointestinal carcinogenesis. It is possible that PAb416, besides binding the viral protein, may cross-react with a hitherto undefined protein whose expression is associated with a distinct pathological profile and, at least in gastric cancers, with worse prognosis.

Human parvovirus 4 (PARV4) is a recently discovered member of the Parvoviridae. We investigated the presence of this virus in bone-marrow aspirates of 35 Italian patients with AIDS. Viral DNA was detected by polymerase chain reaction in over 40% of patients (16/35). The infection was most prevalent in injection drug users (IDU; 12/18; 66.7%) as opposed to non-IDU (4/17; 23.5%). PARV4 infection is widespread in Italian patients with AIDS.

It is currently unknown if the use of a real-time polymerase chain reaction (PCR) adds value to the diagnosis and follow-up prognosis of patients affected by leishmaniasis. We performed a study using a real-time PCR directed against the alpha-polymerase gene and a semiquantitative PCR that target the SSU ribosomal RNA (rRNA) gene as control for the diagnosis and quantification of parasites in patients with visceral (VL) and cutaneous (CL) leishmaniasis. Our single copy real-time PCR missed one diagnosis of VL compared with the conventional PCR, whereas both PCR methods were able to detect Leishmania parasites in CL. Under anti-leishmania treatment the kinetics of parasitemia were comparable with the two methods. The real-time PCR directed against alpha-polymerase of Leishmania despite being able to make a more accurate quantification of parasites does not add to the decision-making management compared with a semiquantitative PCR, and it is comparatively expensive.