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Spigaglia P.,Parasitic and Immune mediated Diseases | Barbanti F.,Parasitic and Immune mediated Diseases | Castagnola E.,Infectious Diseases Unit | Bandettini R.,Clinical Pathology Laboratory Unit
Anaerobe | Year: 2015

In this report, the first two cases of pediatric Clostridium difficile infection (CDI) due to the hypervirulent PCR-ribotype 027 in Italy are described as emblematic of the role of both the infecting C. difficile strain and patient status in the occurrence and clinical manifestation of CDI in children. © 2015 Elsevier Ltd. Source

Castagnola E.,Infectious Diseases Unit | Caviglia I.,Infectious Diseases Unit | Pescetto L.,Clinical Pathology Laboratory Unit | Bagnasco F.,Biostatistics and Committees Unit | And 2 more authors.
Future Microbiology | Year: 2015

Background: Monotherapy is recommended as the first choice for initial empirical therapy of febrile neutropenia, but local epidemiological and antibiotic susceptibility data are now considered pivotal to design a correct management strategy. Aim: To evaluate the proportion of Gram-negative rods isolated in bloodstream infections in children with cancer resistant to antibiotics recommended for this indication. Materials & methods: The in vitro susceptibility to ceftazidime, piperacillin-tazobactam, meropenem and amikacin of Gram-negatives isolated in bacteremic episodes in children with cancer followed at the Istituto "Giannina Gaslini", Genoa, Italy in the period of 2001-2013 was retrospectively analyzed using the definitions recommended by EUCAST in 2014. Data were analyzed for any single drug and to the combination of amikacin with each β-lactam. The combination was considered effective in absence of concomitant resistance to both drugs, and not evaluated by means of in vitro analysis of antibiotic combinations (e.g., checkerboard). Results: A total of 263 strains were evaluated: 27% were resistant to piperacillin-tazobactam, 23% to ceftazidime, 12% to meropenem and 13% to amikacin. Concomitant resistance to β-lactam and amikacin was detected in 6% of strains for piperacillin-tazobactam, 5% for ceftazidime and 5% for meropenem. During the study period there was a nonsignificant increase in the proportions of strains resistant to β-lactams indicated for monotherapy, and also increase in the resistance to combined therapies. Conclusion: in an era of increasing resistance to antibiotics guideline-recommended monotherapy could be not appropriate for initial empirical therapy of febrile neutropenia. Strict local survey on etiology and antibiotic susceptibility is mandatory for a correct management of this complication in cancer patients. © 2015 Future Medicine Ltd. Source

Barco S.,Clinical Pathology Laboratory Unit | Bandettini R.,Clinical Pathology Laboratory Unit | Maffia A.,Clinical Pathology Laboratory Unit | Tripodi G.,Clinical Pathology Laboratory Unit | And 2 more authors.
Journal of Chemotherapy | Year: 2015

Therapeutic drug monitoring is a cornerstone of antibacterial therapy, especially in an era of increasing antibacterial resistance in individualizing antimicrobial therapy. Liquid chromatography/tandem mass spectrometry (LC–MS/MS) assay was used for the simultaneous measurement of piperacillin, tazobactam, meropenem, ceftazidime, and linezolid in 50 ml plasma samples over a wide range. The overall turnaround time for the assay was 20 minutes. Intra-assay precision and accuracy for quality control samples ranged within 1.8–8.5 and 91.4–106.7%, respectively. Inter-assay precision and accuracy ranged within 1.3–14.4 and 95.8–104.6%, respectively. The lower limit of quantification was below 1.5 mg/ml for all the five antibiotics. No ion suppression due to matrix effects was found. A simple and rapid LC–MS/MS method which provides high specificity, precision and accuracy for the simultaneous quantification of piperacillin, tazobactam, meropenem, ceftazidime, and linezolid in human plasma has been developed and validated. The present method is suitable for therapeutic drug monitoring in paediatrics. © 2015 Edizioni Scientifiche per l’Informazione su Farmaci e Terapia. Source

Cangemi G.,Clinical Pathology Laboratory Unit | Storti S.,CNR Institute of Neuroscience | Cantinotti M.,Clinical Pathology Laboratory Unit | Cantinotti M.,CNR Institute of Neuroscience | And 6 more authors.
Clinical Chemistry and Laboratory Medicine | Year: 2013

Background: Neutrophil gelatinase-associated lipocalin (NGAL) has been suggested as the most promising biomarker of acute kidney injury. However, there are no reliable data on analytical sensitivity and reference limits of urinary NGAL (uNGAL) assay in pediatric age. The aim of the present study is to evaluate the analytical sensitivity and the reference range of uNGAL measured in urine specimens of pediatric age with the fully automated platform ARCHITECT ® i1000. Methods: A total of 333 urine samples were collected from 25 healthy newborns (16 males and 9 females; age 1 - 4 days) and 308 children (150 males and 147 females; mean age 80.7 months, range 0.63 - 248 months) and assayed for uNGAL by two different Italian centers (Department of Laboratory Medicine of the Fondazione Toscana G. Monasterio of Pisa and Massa and the Clinical Pathology Laboratory Unit of Istituto Giannina Gaslini of Genova). Results: The calculated limits of blank (LOB) and detection (LOD) values were 0.5 ng/mL and 0.95 ng/mL, respectively. The distribution of uNGAL values approximated a lognormal distribution (median 5.2 ng/mL, interquartile range 2.5-12.8 ng/mL, 99th percentile 117.6 ng/mL). uNGAL values of the 25 neonates were significantly higher than those of 308 children (neonates: mean 44.2 ng/mL, median 30.3 ng/mL, range 5.2 - 137.4 ng/mL; children: mean 10.2 ng/mL, median 4.6 ng/mL, range 0.2 - 146.7 ng/mL; p < 0.0001). Conclusions: uNGAL assay shows a good analytical sensitivity and imprecision, which allows the measurement of uNGAL values around the cut-off value (i.e., 117.6 ng/mL) with an imprecision < 5 CV%. The distribution of uNGAL values in pediatric age approximates a log-normal distribution, with values which are higher in neonates compared to children. Source

Scapolla C.,University of Genoa | Cangemi G.,Clinical Pathology Laboratory Unit | Barco S.,Clinical Pathology Laboratory Unit | Barbagallo L.,Clinical Pathology Laboratory Unit | And 6 more authors.
Journal of Mass Spectrometry | Year: 2012

The levels of urinary catecholamine metabolites, such as homovanillic acid (HVA) and vanillylmandelic acid, are routinely used as a clinical tool in the diagnosis and follow-up of neuroblastoma (NB) patients. Recently, in the Clinical Pathology Laboratory Unit of G. Gaslini Children Hospital, a commercial method that employs liquid chromatography coupled to electrochemical detection (LC-EC) has been introduced for the measurement of these metabolites in the routine laboratory practice. Using this LC-EC method, an unknown peak could be observed only in samples derived from NB patients. To investigate the nature of this peak, we used a combination of liquid chromatography-time-of-flight mass spectrometry (LC-TOF-MS) and liquid chromatography-ion trap tandem mass spectrometry (LC-IT-MS). The first approach was used to obtain the elemental composition of the ions present in this new signal. To get additional structural information useful for the elucidation of unknown compounds, the ion trap analyzer was exploited. We were able to identify not just one, but three unknown signals in urine samples from NB patients which corresponded to three conjugated products of HVA: HVA sulfate and two glucuronoconjugate isomers. The enzymatic hydrolysis with β-glucuronidase confirmed the proposed structures, while the selective alkaline hydrolysis allowed us to distinguish the difference between phenol- and acyl-glucuronide of HVA. The latter was the unknown peak observed in LC-EC separations of urine samples from NB patients. Copyright © 2012 John Wiley & Sons, Ltd. Source

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