Molecular epidemiology of non-viral sexually transmitted infections in the central Alpine province of Bolzano, northern Italy from April 2016 to March 2017

Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis and Mycoplasma genitalium are established or presumed as (??) STI pathogens. The present study aims at ng describing the one-year molecular epidemiology of these seven pathogens in the Province of Bolzano, Northern Italy. From April 2016 to March 2017, a total of 2,949 patients, mainly females, were enrolled and 3,427 urine, vaginal, endocervical and/or urethral samples were subjected to simultaneous analysis of the seven pathogens by means of Real Time Polymerase Chain Reaction (AnyplexTM II STI-7 Detection Kit Seegene, Seoul, Korea). At least one of the seven microorganisms was detected in 40.7% of patients, with an uneven distribution: 43.1% in females (F) and 29.8% (p<0.001) in males (M). The prevalence of microorganisms was as follows: 30.3% U. parvum (F: 35.6%, M: 8.3%), 6.9% U. urealyticum (F: 6.8%, M: 7.0%), 4.9% M. hominis (F: 5.4%, M: 2.3%), 4.9% C. trachomatis (F: 3.4%, M: 11.4%), 1.1% M. genitalium (F: 1.0%, M: 1.2%), 1.2% N. gonorrhoeae (F: 0.17%, M: 5.6%) and 0.40% T. vaginalis (F: 0.38%, M: 0.53%). Mixed infections were detected in 7.4% of patients. The highest prevalence was observed for U. parvum, followed by U. urealyticum and M. hominis and a significant presence of multi-pathogen infections was registered

Cell-Cycle Inhibition by Helicobacter pylori L-Asparaginase

Helicobacter pylori (H. pylori) is a major human pathogen causing chronic gastritis, peptic ulcer, gastric cancer, and mucosa-associated lymphoid tissue lymphoma. One of the mechanisms whereby it induces damage depends on its interference with proliferation of host tissues. We here describe the discovery of a novel bacterial factor able to inhibit the cell-cycle of exposed cells, both of gastric and non-gastric origin. An integrated approach was adopted to isolate and characterise the molecule from the bacterial culture filtrate produced in a protein-free medium: size-exclusion chromatography, non-reducing gel electrophoresis, mass spectrometry, mutant analysis, recombinant protein expression and enzymatic assays. L-asparaginase was identified as the factor responsible for cell-cycle inhibition of fibroblasts and gastric cell lines. Its effect on cell-cycle was confirmed by inhibitors, a knockout strain and the action of recombinant L-asparaginase on cell lines. Interference with cell-cycle in vitro depended on cell genotype and was related to the expression levels of the concurrent enzyme asparagine synthetase. Bacterial subcellular distribution of L-asparaginase was also analysed along with its immunogenicity. H. pylori L-asparaginase is a novel antigen that functions as a cell-cycle inhibitor of fibroblasts and gastric cell lines. We give evidence supporting a role in the pathogenesis of H. pylori-related diseases and discuss its potential diagnostic application

Letter to the editor regarding Shibayama et al.: Biochemical and pathophysiological characterization of Helicobacter pylori asparaginase.

Gli autori replicano ad alcune critiche da parte di Shibayama et al. al loro lavoro precedente

Onychomycosis by Paecilomyces lilacinus in a patient with leukonychia

Paecilomyces lilacinus, a filamentous fungus living saprophytic in the environment, very rarely causes infection in immunocompetent humans, mostly ocular or cutaneous and sub-cutaneous infections.The fungal isolates usually show low susceptibility to conventional antifungal drugs in vitro and variable susceptibility to novel triazoles. During a period of 2 years, 4 cultures of nail fragments were grown from a 41 years old female on modified Sabouraud/Dextrose agar (S.D.A.) (CAF bioMérieux, CAF+CEX Biolife). Cultures were continued in suspension and the hyphae stained with lactophenole blue analysed microscopically. 2 samples were tested with molecular methods, i.e. DNA extraction, amplification of internal transcribed spacer (ITS) region 1 and 2 and automated sequencing. The sequence results were evaluated using a validated reference database for humanpathogenic fungi. Additionally the BLAST algorithm on NIH database GenBank was used. A growth of a filamentous hyaline fungus with white colonies and lila shade was observed in all samples. The presence of Penicillium like conidiophores with divergent long terminal phialides was seen with little elliptical conidia. DNA sequencing determined the species as Paecilomyces lilacinus with 99% homology with GenBank isolate AY213667. The infection of the right hallux did not improve by systemic (Lamisil/terbinafine) and local treatment (Locetar/amorolfin nail lacquer).Also change to Sporanox/itraconazol only led to deterioriation, so finally the extraction of the nail had to be performed given the known difficulties with treatment described in the literature. This case highlights the usefulness of molecular techniques, especially use of the highly discriminative ITS region for fungal typing

Helicobacter pylori L-asparaginase: A promising chemotherapeutic agent

Bacterial L -asparaginases are amidohydrolases that catalyse the conversion of L -asparagine to L -aspartate and ammonia and are used as anti-cancer drugs. The current members of this class of drugs have several toxic side effects mainly due to their associated glutaminase activity. In the present study, we report the molecular cloning, biochemical characterisation and in vitro cytotoxicity of a novel L -asparaginase from the pathogenic strain Helicobacter pylori CCUG 17874. The recombinant enzyme showed a strong preference for L -asparagine over L -glutamine and, in contrast to most L -asparaginases, it exhibited a sigmoidal behaviour towards L -glutamine. The enzyme preserved full activity after 2 h incubation at 45 C. In vitro cytotoxicity assays revealed that different cell lines displayed a variable sensitivity towards the enzyme, AGS and MKN28 gastric epithelial cells being the most affected. These findings may be relevant both for the interpretation of the mechanisms underlying H. pylori associated diseases and for biomedical applications