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Cellular homeostasis of magnesium is still unclear. Several studies documented the occurrence of fluxes of magnesium across the plasmamembrane within minutes from the application of metabolic or hormonal stimuli. These fluxes, however, result in limited variation of free Mg2+ intracellular concentration and large changes in total Mg content. It has been reported that a stimulation with cyclic AMP caused a movement of total magnesium within 10 min after treatment in cardiomyocytes. In this study we tested this hypothesis in HL60 leukemic cells, not excitable but highly proliferating cell model. We evaluated Mg flux by DCHQ5, the phenyl-derivative of hydroxyquinoline fluorescent probe family. We observed a drastic decrease of intracellular total magnesium in the first 3 min. We also verified that at least 10% of the total intracellular amount of magnesium moved in the supernatant of stimulated cells.

Magnesium (Mg) is essential for biological processes, but its cellular homeostasis has not been thoroughly elucidated, mainly because of the inadequacy of the available techniques to map intracellular Mg distribution. Recently, particular interest has been raised by a new family of fluorescent probes, diaza-18-crown-hydroxyquinoline (DCHQ), that shows remarkably high affinity and specificity for Mg, thus permitting the detection of the total intracellular Mg. The data obtained by fluori- metric and cytofluorimetric assays performed with DCHQ5 are in good agreement with atomic absorption spectroscopy, confirming that DCHQ5 probe allows both qualitative and quantitative determination of total intracellular Mg.

A recent study showed the antitumor activity of a new indole-derivative – MM-67 – inducing mitochondria-mediated apoptosis and a decrease of intracellular magnesium (Mg) concentration in HT29 colon cancer cells. Aim of this work was to assess cellular Mg levels throughout MM-67-induced apoptosis from the early to the final stage of the process and to evaluate the correlation with mitochondrial membrane potential (ΔΨm) variations. All analysis were performed by flow cytometry: ΔΨm was assessed by using mitochondrial potential sensitive dye DiOC6, while free and total intracellular cation concentrations were assessed by using the commercial probe MagFluo4-AM (Kd=4.7 mM), and the new synthesized DCHQ5 (Kd=8.3 mM), respectively. Our results evidenced that the MM67 induced apoptosis is characterized by a direct correlation between ΔΨ and free intracellular Mg content variations.

Integrated Coastal Zone Management is an emerging research area. The aim is to provide a global view of different and heterogeneous aspects interacting in a geographical area. Decision Support Systems, integrating Computational Intelligence methods, can be successfully used to estimate useful anthropic and environmental indexes. Bayesian Networks have been widely used in the environmental science domain. In this paper a Bayesian model for estimating the Sustainable Coastal Index is presented. The designed Bayesian Network consists of 17 nodes, hierarchically organized in 4 layers. The first layer is initialized with the season and the physiographic region information. In the second layer, the first-order indexes, depending on raw data, of physiographic regions are computed. The third layer estimates the second-order indexes of the analyzed physiographic regions. In the fourth layer, the global Sustainable Coastal Index is inferred. Processed data refers to 13 physiographic regions in the Province of Trapani, western Sicily, Italy. Gathered data describes the environmental information, the agricultural, fisheries, and economical behaviors of the local population and land. The Bayesian Network was trained and tested using a real dataset acquired between 2000 and 2006. The developed system presents interesting results.

Nell’ambito delle azioni promosse dal CISIS-CPSG riguardanti lo sviluppo delle Infrastrutture di Dati Territoriali (IDT), è stato svolto uno studio per comporre un quadro di riferimento conoscitivo, propedeutico alla definizione delle linee guida e dei contenuti tecnici “minimi comuni” per la formazione di IDT regionali di tipo federato su base interregionale, in un’ottica d’interoperabilità e di riuso dei dati. The standing Committee on GIS (CPSG) of the Interregional Centre for ICT, Geographic and Statistical Systems (CISIS) has carried out a knowledge framework useful to define the guidelines for the creation of regional federated SDI on an interregional basis. The study has taken into consideration European and National initiatives concerning ICT and GI policies, the SDI concept shift in respect of the geospatial data use changing and the expertise of the Regions on this matter, already acquired within inter-institutional National and European projects. The article shows the main findings of the study and provides a synthetic description of the methodological approach in order to organize and plan the next actions.

Fanconi anemia (FA) is a recessive DNA repair disease characterized by a high predisposition to developing neoplasms. DNA tumor polyomavirus simian virus 40 (SV40) transforms FA fibroblasts at high efficiency suggesting that FA patients could be highly susceptible to SV40 infection. To test this hypothesis, the large tumor (LT) antigen of SV40, BKV, JCV and Merkel Cell (MC) polyomaviruses were tested in blood samples from 89 FA patients and from 82 of their parents. Two control groups consisting of 47 no-FA patients affected by other genetic bone marrow failure diseases and 91 healthy subjects were also evaluated. Although JCV, BKV and MC were not found in any of the FA samples, the prevalence and viral load of SV40 were higher in FA patients (25%; mean viral load: 1.1×10(2) copies/10(5)cells) as compared with healthy individuals (4.3%; mean viral load: 0.8×10(1) copies/10(5)cells) and genetic controls (0%) (p

Diamond Blackfan anemia typically presents in infants and is often associated with many kinds of malformations. Severity of anemia often needs transfusional support in the first months of life. We describe here a patient with Diamond Blackfan anemia related to a RPL5 mutation. The patient had no physical abnormalities and experienced a very late onset of transfusion dependency.

Evans Syndrome is a rare autoimmune disease consisting of hemolytic anemia, thrombocytopenia and/or neutropenia. It may be associated with other autoimmune or lymphoproliferative diseases. Its course can be extremely serious and, rarely, even life-threatening; thus it represents a excellent treatment challenge for the pediatric hematologist. First line treatment consists of steroids and/or immunoglobulin; further therapy with rituximab, vincristine, cyclophosphamide and other immunosuppressive drugs can be considered in unresponsive patients. We describe a baby with refractory Evans Syndrome that was cured by prolonged administration of mycophenolate mofetil and remained disease-free for 4 years after the discontinuation of treatment.

Research has recently focused on pasture species intake by ruminants due to their influence on animal product quality. A field-applicable method which investigates species intake and selection, was tested on two dairy cow grazing systems: continuous grazing on a highly-biodiverse pasture (C) and rotational grazing on a moderately-diverse sward (R). In addition to the grazed class method, which evaluates the percentage of grazed dry matter (DM) per species according to the residual height of the plant grazed, further measurements were introduced to quantify DM consumption and selection index per species. Six and four representative species were studied in the C and R systems respectively. We found an exponential regression between the presence of a species and its contribution to the cattle’s daily intake (P

Two cases of bladder exposure of TVT sling material at 23 and 12 months after the implantation are reported. In both cases the urinary problems developed immediately after the TVT procedure and only few millimetres of one arm of the sling were exposed into the bladder. It is assumed therefore that the graft was inserted into the bladder lumen or into the bladder wall during the procedure rather than rejected. The whole sling was removed and both patients were cured of dysuria but in one recurrence of stress incontinence was noted.

Les prairies permanentes occupent environ un tiers de la surface agricole utile et présentent un grand potentiel de diversité biologique. L’élevage a de ce fait un rôle majeur à jouer dans la préservation de la biodiversité sur le territoire français. L’objectif de cette synthèse est d’aborder la question de la diversité floristique des prairies et de l’élevage en s’interrogeant sur l’intérêt de cette diversité construite par les éleveurs, pour les ruminants et les produits animaux. Une seule composante de la diversité biologique des prairies est prise en compte : la diversité floristique. Il est réalisé dans cette synthèse un état des connaissances pluridisciplinaires des effets de cette diversité sur la valeur nutritive des fourrages, l’ingestion des animaux, les caractéristiques sensorielles et nutritionnelles des produits laitiers et carnés, enfin sur deux aspects de la santé animale, la lutte contre les infestations par les strongles digestifs et la prévention des processus de peroxydation. Dans chacune des thématiques abordées, des effets liés à la composition botanique des prairies ont été mis en évidence, mais sans qu’il puisse être toujours fait la part entre les effets liés à la présence d’un grand nombre d’espèces dans la parcelle, les effets liés à la présence de certaines espèces et les effets liés au stade phénologique. Le rôle probable important des dicotylédones et notamment des légumineuses a été souligné à plusieurs reprises, en particulier du fait de leur plus grande teneur en métabolites secondaires que les graminées. Enfin, l’intérêt de fournir aux animaux des sources d’alimentation diversifiées a également été évoqué en particulier sur les aspects liés à l’ingestion et à la santé animale.

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Abstract Introduction Vincristine is an antineoplastic drug with a well known efficacy for the treatment of acute lymphoblastic leukemia and many solid tumors. No more than 20 pediatric patients with vincristine-induced vocal cord palsy have been reported, and to the best of our knowledge this is the first case where glutamic acid was administered with the aim of preventing a relapse of laryngeal dysfunction. Case presentation The larynx paralysis presented with hoarseness and stridor in a Caucasian 18-month-old girl and spontaneously resolved in about a month. In order to administer a subsequent full dose of vincristine, our patient received oral glutamic acid whose efficacy against vincristine neurological side effects has been previously reported. Conclusions Since in our patient the amino acid proved to be ineffective in the prevention of laryngeal paralysis relapse, we suggest that a dose reduction of vincristine should be preferred by oncologists as an initial approach after a case of drug-induced vocal cord palsy.

Abstract Castleman's disease (CD) is a rare, localized or generalized, lymphoproliferative disorder with a frequent mediastinal location, but possible in any lymph node or extra nodal site. It usually appears in young adults whilst it rarely occurs in childhood. There are only about 100 pediatric cases published, five of them in Italy. We report 3 cases of localized Castleman's disease, investigated in our Department in a 3 years period and reviewed the literature.

Abstract Hepatic hemangioma, adenoma and focal nodular hyperplasia are the most frequent benign lesions of the liver, but they are all infrequent among pediatric population. The reports of focal nodular hyperplasia in children have recently increased in number, with many cases associated to drug intake, particularly to chemotherapy. We here describe, to our knowledge, the first case of focal nodular hyperplasia in association with diabetes mellitus in childhood.

Abstract Background The clinical efficacy of camptothecin (CPT), a drug specifically targeting topoisomerase I (TopoI), is under evaluation for the treatment of malignant gliomas. Due to the high unresponsiveness of these tumours to chemotherapy, it would be very important to study the signalling network that drives camptothecin outcome in this type of cancer cells. To address this issue, we had previously compared the expression profile of human U87-MG glioblastoma cells with that of a CPT-resistant counterpart, giving evidence that the development of a robust inflammatory response was the main transcriptional effect associated with CPT resistance. Here we report time-related changes and cell line specific patterns of gene expression after CPT treatment by using two p53 wild-type glioblastoma cell lines, U87-MG and DBTRG-05, with different sensitivities to TopoI inhibition. Results First, we demonstrated that CPT treatment brings the two cell lines to completely different outcomes: accelerated senescence in U87-MG and apoptosis in DBTRG-05 cells. Then, to understand the different susceptibility to CPT, we used oligo-microarray to identify the genes whose expression was regulated during a time-course treatment, ranging from 2 h to 72 h. The statistical analysis of microarray data by MAANOVA (MicroArray ANalysis Of VAriance) showed much less modulated genes in apoptotic DBTRG-05 cells (155) with respect to the senescent U87-MG cells (3168), where the number of down-regulated genes largely exceeded that of the up-regulated ones (80% vs. 20%). Despite this great difference, the two data-sets showed a large overlapping (60% circa) mainly due to the expression of early stress responsive genes. The use of High-Throughput GoMINER and EASE tools, for functional analysis of significantly enriched GO terms, highlighted common cellular processes and showed that U87-MG and DBTRG-05 cells shared many GO terms, which are related to the down-regulation of cell cycle and mitosis and to the up-regulation of cell growth inhibition and DNA damage. Furthermore, the down-regulation of MYC and DP1 genes, which act as key transcription factors in cell growth control, together with the inhibition of BUB1, BUB3 and MAD2 mRNAs, which are known to be involved in the spindle checkpoint pathway, were specifically associated with the execution of senescence in U87-MG cells and addressed as critical factors that could drive the choice between different CPT-inducible effectors programs. In U87-MG cells we also found inflammation response and IL1-beta induction, as late transcriptional effects of Topo I treatment but these changes were only partially involved in the senescence development, as shown by IL1-beta gene silencing. Conclusion By comparing the transcription profile of two glioblastoma cell lines treated with camptothecin, we were able to identify the common cellular pathways activated upon Topo I inhibition. Moreover, our results helped in identifying some key genes whose expression seemed to be associated with the execution of senescence or apoptosis in U87-MG and DBTRG-05 cells, respectively.