Cytomorphological and Genetic Characterization of Troglobitic Leptolyngbya Strains Isolated from Roman Hypogea▿

Six Leptolyngbya strains, isolated from the archaeological surfaces of hypogean sites, were phenotypically and genetically characterized by light and electron microscopy and 16S rRNA gene and 16S-23S internally transcribed spacer (ITS) sequencing. Three phycoerythrin-rich (red) and three phycocyanin-rich (green) isolates were assigned to different operational taxonomic units (OTUs). Among the green isolates, one strain showed an OTU intraspecific variation due to differences in the ITS sequences and genomic polymorphism. Within the ITS sequence, variable regions, conserved domains and tRNAIle and tRNAAla genes showed high sequence identity among the phylotypes. Together, these data indicated a relatedness of the six strains to other Leptolyngbya from subaerophytic and geothermal environments and allowed the definition of novel Leptolyngbya OTUs

Epidemiology of cytomegalovirus infection in pregnant women living in the Greater Romagna Area, Italy

Background. Aim of this study was to assess the incidence of Cytomegalovirus (CMV) infection in pregnant women living in Romagna area, in North East Italy to implement the best management of this infection. Materials and Methods. In 2012, 23,727 serological tests for CMV IgG and IgM antibodies were performed in the Microbiology Unit, the Hub Laboratory of the Greater Romagna Area: 6931 were pregnant women. Results and Conclusions. 179 subjects were positive for CMV IgM anti- bodies: 82 were not pregnant; 97 were IgM positive during pregnancy or in the course of a pre-conception evaluation. The detected incidence of the CMV infection in pregnancy (calculated at 1.40%) actually validates the literature data. This study\u2019s findings clearly underline the usefulness of testing the CMV specific immune response in the pre-conception period or as early as possible during pregnancy

A Method for DNA Extraction from the Desert Cyanobacterium Chroococcidiopsis and Its Application to Identification of ftsZ

A method was developed for extraction of DNA from Chroococcidiopsis that overcomes obstacles posed by bacterial contamination and the presence of a thick envelope surrounding the cyanobacterial cells. The method is based on the resistance of Chroococcidiopsis to lysozyme and consists of a lysozyme treatment followed by osmotic shock that reduces the bacterial contamination by 3 orders of magnitude. Then DNase treatment is performed to eliminate DNA from the bacterial lysate. Lysis of Chroococcidiopsis cells is achieved by grinding with glass beads in the presence of hot phenol. Extracted DNA is further purified by cesium-chloride density gradient ultracentrifugation. This method permitted the first molecular approach to the study of Chroococcidiopsis, and a 570-bp fragment of the gene ftsZ was cloned and sequenced

Pneumatic pyloric dilatation for the treatment of gastric outlet obstruction in a child

Acquired causes of gastric outlet obstruction (GOO) are rarely encountered in infancy, having an approximate incidence of 1 per 100,000 live births. Reports of short-term exposure to non-steroidal anti-inflammatory drugs having adverse events are few. We present the case of a previously healthy 3-year-old boy who developed severe chronic gastric outlet obstruction and antral stenosis after a short-term ingestion of liquid ibuprofen at a dosage not thought to be associated with unfavorable effects. Even though the optimal management of these cases remains to be determined, we report on a prompt and successful endoscopic treatment for this condition

The seroprevalence of anti-hepatitis E IgG antibodies in blood donors in the Greater Romagna area, Italy

Objective: Data about Hepatitis E virus (HEV) prevalence are variable in Europe and also limited inItaly.WeaimedtodescribetheseroprevalenceofimmunoglobulinG(IgG)antibodiesagainst HEV infection among blood donors (BDs) in the Greater Romagna area, North-Eastern Italy. Materials and Methods: All serum samples were at first screened for anti-HEV IgG antibodies by a commercial enzyme-linked immunosorbent assay (ELISA) (DIA.PRO Diagnostic Bioprobes, Sesto San Giovanni (MI), Italy), used in our diagnostic routine and so considered as the reference test. Then, all serum samples were re-tested for anti-HEV IgG antibodies by the other two commercial ELISA (Wantai, Biologic Pharmacy Enterprise, Beijing, China; Euroimmun, L\ufcbeck, Germany). The seroprevalence rate was calculated based on the immunoblotting (IB) assay confirmation. Results: A total of 500 BDs samples were tested by three commercial ELISA tests; positive and borderline results were confirmed by an IB assay. The overall anti-HEV IgG prevalence was 3.8% (CI: 2.303 \u2013 5.871), 1.6% (CI: 0.693 \u2013 3.128) and 2.8% (CI: 1.539 \u2013 4.653) when samples were tested with DIA.PRO, Euroimmun and Wantai methods, respectively. The seroprevalence of IgG antibodies against HEV infection was found to be similar or lower than the previous reports in Italy. The results obtained by the different ELISA tests were similar. Conclusion: The data obtained in this study underline the need for achieving a harmonised testing algorithm that is necessary in order to be able to compare data from different studies. We suggest that the identification of potential HEV human infection should be added to the standard laboratory work-up for viral hepatitis

Expression of phospholipase C beta family isoenzymes in C2C12 myoblasts during terminal differentiation

In the present work, we have analyzed the expression and subcellular localization of all the members of inositide-specific phospholipase C (PLCβ) family in muscle differentiation, given that nuclear PLCβ1 has been shown to be related to the differentiative process. Cell cultures of C2C12 myoblasts were induced to differentiate towards the phenotype of myotubes, which are also indicated as differentiated C2C12 cells. By means of immunochemical and immunocytochemical analysis, the expression and subcellular localization of PLCβ1, β2, β3, β4 have been assessed. As further characterization, we investigated the localization of PLCβ isoenzymes in C2C12 cells by fusing their cDNA to enhanced green fluorescent protein (GFP). In myoblast culture, PLCβ4 was the most expressed isoform in the cytoplasm, whereas PLCβ1 and β3 exhibited a lesser expression in this cell compartment. In nuclei of differentiated myotube culture, PLCβ1, isoform was expressed at the highest extent. A marked decrease of PLCβ4 expression in the cytoplasm of differentiated C2C12 cells was detected as compared to myoblasts. No relevant differences were evidenced as regards the expression of PLCβ3 at both cytoplasmatic and nuclear level, whilst PLCβ2 expression was almost undetactable. Therefore, we propose that the different subcellular expression of these PLC isoforms, namely the increase of nuclear PLCβ1 and the decrease of cytoplasmatic PLCβ4, during the establishment of myotube differentiation, is related to a spatial-temporal signaling event, involved in myogenic differentiation. Once again the subcellular localization appears to be a key step for the diverse signaling activity of PLCβs. © 2004 Wiley-Liss, Inc

Epigenetic regulation of nuclear inositide signalling in high-risk MDS patients

Inositide signalling pathways are involved in cell growth, diff erentiation and apoptosis and play a role in the progression of myelodysplastic syndromes (MDS) towards acute myeloid leukemia (AML). The combination of the DNA methyltransferase inhibitor azacitidine (AZA) and the HDAC inhibitor valproic acid (VPA) has been demonstrated to be active and associated with a high response rate in patients with high-risk MDS and unfavourable prognosis. In the last few years, our group demonstrated not only that phosphoinositide-phospholipase C beta1 (PI-PLCbeta1) promoter gene is hyper-methylated in higher-risk MDS, but also that it is aff ected by epigenetic therapy. Indeed, AZA, alone or in combination with VPA, was able to induce PI-PLCbeta1 demethylation and expression. In this study we further investigated the role of lipid signalling pathways during epigenetic therapy, focusing on the functional eff ect of AZA and VPA on PI-PLCbeta1 promoter in high-risk MDS patients. The study included 20 high-risk MDS patients (IPSS risk: intermediate-2 or high): 8 of them were treated with AZA alone (75 mg/m2/day SC for 7 days/28 days), whereas 6 of them received the combination of AZA with VPA (600-1500 mg/ daily orally) and the remaining 6 were treated only with best supportive care. For each patient we analyzed the eff ect of epigenetic therapy in correlation to PI-PLCbeta1 signalling, by analyzing the binding affi nity of transcription factors correlated to hematopoietic stem cell diff erentiation and proliferation. 8/20 (40%) of our MDS patients showed a favourable hematologic response to the epigenetic therapy and an increase in PI-PLCbeta1 expression, as compared with the pre-treatment period, thus confi rming the involvement of this molecule in response to demethylating agents. Moreover, MDS patients responding to the epigenetic treatment seem to involve the recruitment of specifi c transcription factors to PI-PLCbeta1 promoter during the regulation of methylation processes. Taken together, our data are therefore consistent with the hypothesis of a correlation between the epigenetic treatment and PI-PLCbeta1 signalling, thus hinting at a role for PI-PLCbeta1 in monitoring the effi cacy of epigenetic therapy and possibly paving the way for the development of innovative therapeutic strategies in high-risk MDS