A molecular case-control study of the Merkel cell polyomavirus in colon cancer. - Campello, Cesare; Comar, Manola; D'Agaro, Pierlanfranco; Minicozzi, Anna; Rodella, Luca; Poli, Albino (J Med Virol, 2011)
To explore the putative role of the Merkel cell polyomavirus in human colon cancer, a prospective molecular case-control study was undertaken in patients and their relatives enrolled during a screening program. Fresh tissue samples from 64 cases of colon cancer (mean age 69.9 ± 11.0 years; 40 males) and fresh biopsies from 80 relatives (mean age 53.7 ± 8.6 years; 43 male; 55 son/daughter, 23 brother/sister, 2 parents) were analyzed by PCR and sequencing. Pre-cancerous lesions, namely adenomas and polyps, were detected in 15 (18.8%) and 9 (11.2%) of the controls, respectively. In addition, 144 blood samples were examined. Merkel cell polyomavirus DNA was detected in 6.3% of cases and 8.8% of controls. This difference was not statistically significant in the logistic regression analysis, after adjustment for age. Whereas blood samples from both cases and controls tested negative, the DNA Merkel cell polyomavirus was identified in 12.5% of adenoma/polyp tissues. No statistically significant difference was found when prevalence rates of Merkel cell polyomavirus in normal, pre-cancerous and cancer tissues were compared. Sequence analysis of the viral LT3 and VP1 regions showed high homology (>99%) with those of strains circulating worldwide, especially with genotypes detected in France. The findings of this survey are consistent with the hypothesis that the Merkel cell polyomavirus, in addition to other human polyomaviruses, can be recovered frequently from the gastrointestinal tract, because it is transmitted throughout the fecal-oral route. Moreover, the study does not indicate a role for Merkel cell polyomavirus in the genesis of colon cancer.
Keywords: Aged; Aged, 80 and over; Case-Control Studies; Cluster Analysis; Colonic Neoplasms; DNA, Viral; Female; Genotype; Humans; Italy; Male; Merkel Cells; Middle Aged; Molecular Sequence Data; Polymerase Chain Reaction; Polyomavirus; Polyomavirus Infections; Prevalence; Sequence Analysis, DNA; Tumor Virus Infections

Molecular mechanism of glucocorticoid resistance in inflammatory bowel disease. - De Iudicibus, Sara; Franca, Raffaella; Martelossi, Stefano; Ventura, Alessandro; Decorti, Giuliana (World J Gastroenterol, 2011)
Natural and synthetic glucocorticoids (GCs) are widely employed in a number of inflammatory, autoimmune and neoplastic diseases, and, despite the introduction of novel therapies, remain the first-line treatment for inducing remission in moderate to severe active Crohn's disease and ulcerative colitis. Despite their extensive therapeutic use and the proven effectiveness, considerable clinical evidence of wide inter-individual differences in GC efficacy among patients has been reported, in particular when these agents are used in inflammatory diseases. In recent years, a detailed knowledge of the GC mechanism of action and of the genetic variants affecting GC activity at the molecular level has arisen from several studies. GCs interact with their cytoplasmic receptor, and are able to repress inflammatory gene expression through several distinct mechanisms. The glucocorticoid receptor (GR) is therefore crucial for the effects of these agents: mutations in the GR gene (NR3C1, nuclear receptor subfamily 3, group C, member 1) are the primary cause of a rare, inherited form of GC resistance; in addition, several polymorphisms of this gene have been described and associated with GC response and toxicity. However, the GR is not self-standing in the cell and the receptor-mediated functions are the result of a complex interplay of GR and many other cellular partners. The latter comprise several chaperonins of the large cooperative hetero-oligomeric complex that binds the hormone-free GR in the cytosol, and several factors involved in the transcriptional machinery and chromatin remodeling, that are critical for the hormonal control of target genes transcription in the nucleus. Furthermore, variants in the principal effectors of GCs (e.g. cytokines and their regulators) have also to be taken into account for a comprehensive evaluation of the variability in GC response. Polymorphisms in genes involved in the transport and/or metabolism of these hormones have also been suggested as other possible candidates of interest that could play a role in the observed inter-individual differences in efficacy and toxicity. The best-characterized example is the drug efflux pump P-glycoprotein, a membrane transporter that extrudes GCs from cells, thereby lowering their intracellular concentration. This protein is encoded by the ABCB1/MDR1 gene; this gene presents different known polymorphic sites that can influence its expression and function. This editorial reviews the current knowledge on this topic and underlines the role of genetics in predicting GC clinical response. The ambitious goal of pharmacogenomic studies is to adapt therapies to a patient's specific genetic background, thus improving on efficacy and safety rates.
Keywords: Drug Resistance; Glucocorticoids; Humans; Inflammatory Bowel Diseases; P-Glycoproteins; Polymorphism, Genetic; Receptors, Glucocorticoid; Signal Transduction; Transcription, Genetic

Molecular targets for selective killing of TRAIL-resistant leukemic cells. - Zauli, Giorgio; Bosco, Raffaella; Secchiero, Paola (Expert Opin Ther Targets, 2011)
INTRODUCTION: TNF-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family of cytokines, and shows promising therapeutic activity against solid tumors and lymphomas, in a variety of Phase I and II clinical trials. In contrast, primary leukemias have shown poor susceptibility to TRAIL-mediated cytotoxicity, suggesting the need for sensitizing TRAIL-resistant leukemic cells, by combining soluble recombinant TRAIL either with chemotherapeutic drugs, or with targeted small molecules. AREAS COVERED: This review discusses potential therapeutic applications of combinations able to restore the sensitivity of leukemic cells to either recombinant TRAIL or anti-TRAIL-receptor agonistic antibodies for the treatment of hematological malignancies. EXPERT OPINION: Up-to-date knowledge of the most innovative anti-leukemic therapies including functional screening of specific-sensitizers, enhancing TRAIL-mediated cytotoxicity. Strategies aimed to enhance TRAIL-mediated apoptosis, include the combination of novel sensitizers, functionally identified from libraries of pharmaceutically active, synthetic or naturally derived compounds. Other approaches aim to employ the administration of stem cells engineered to express TRAIL, in the leukemic stem cell niche, and promise to be a successful treatment with reduced specific toxicity.
Keywords: Humans; Leukemia; TNF-Related Apoptosis-Inducing Ligand

A multicenter, case-control study on risk factors for antepartum stillbirth. - Facchinetti, Fabio; Alberico, Salvatore; Benedetto, Chiara; Cetin, Irene; Cozzolino, Sabrina; Di Renzo, Gian Carlo; Del Giovane, Cinzia; Ferrari, Francesca; Mecacci, Federico; Menato, Guido; Tranquilli, Andrea L; Baronciani, Dante (J Matern Fetal Neonatal Med, 2011)
OBJECTIVE: As the influence of socio-demographic variables, lifestyle and medical conditions on the epidemiology of stillbirth (SB) is modified by population features, we aimed at investigating the role played by these factors on the incidence of SB in a developed country. STUDY DESIGN: Multivariate logistic regression analysis (OR with 95% CI) was utilized in a prospective multicentre nested case-control study to compare in a 1:2 ratio stillborn of >22 weeks gestation with matched for gestational age live-born (LB) infants. Intrapartum SB were excluded. RESULTS: Two hundred fifty-four consecutive SBs and 497 LBs were enrolled. Socio-demographic variables were equally distributed. Fetal malformations (7.96, 2.69-23.55), severe intrauterine growth restriction (IUGR) (birthweight ≤ 5(th) %ile) (4.32, 2.27?8.24), BMI > 25 (2.87, 1.90-4.33), and preeclampsia (PE, 0.40, 0.21-0.77) were recognized as independent predictors for SB. At term, only BMI > 25 was associated with SB (7.70, 2.9-20.5). CONCLUSION: Fetal malformations, severe IUGR and maternal BMI > 25 were associated with a significant increase in the risk of SB; PE presented instead a protective role. Maternal BMI > 25 was the only risk factor for SB identified in term pregnancies.
Keywords: Adult; Case-Control Studies; Cause of Death; Congenital Abnormalities; Female; Fetal Death; Fetal Growth Retardation; Humans; Infant, Newborn; Male; Obstetric Labor Complications; Pre-Eclampsia; Pregnancy; Risk Factors; Stillbirth; Young Adult

A multicenter, randomized, placebo-controlled trial of levetiracetam in children and adolescents with newly diagnosed absence epilepsy. - Fattore, Cinzia; Boniver, Clementina; Capovilla, Giuseppe; Cerminara, Caterina; Citterio, Antonietta; Coppola, Giangennaro; Costa, Paola; Darra, Francesca; Vecchi, Marilena; Perucca, Emilio (Epilepsia, 2011)
PURPOSE: To evaluate the potential efficacy of levetiracetam as an antiabsence agent in children and adolescents with newly diagnosed childhood or juvenile absence epilepsy. METHODS: Patients were randomized in a 2:1 ratio to receive de novo monotherapy with levetiracetam (up to 30 mg/kg/day) or placebo for 2 weeks under double-blind conditions. Responder status (primary end point) was defined as freedom from clinical seizures on days 13 and 14 and from electroencephalographic (EEG) seizures during a standard EEG recording with hyperventilation and intermittent photic stimulation on day 14. The double-blind phase was followed by an open-label follow-up. KEY FINDINGS: Nine of 38 patients (23.7%) were responders in the levetiracetam group, compared with one of 21 (4.8%) in the placebo group (p = 0.08). Seven of 38 patients (18.4%) were free from clinical and EEG seizures during the last 4 days of the trial (including 24-h EEG monitoring on day 14) compared with none of the patients treated with placebo (p = 0.04). Seventeen patients remained seizure-free on levetiracetam after 1 year follow-up. Of the 41 patients who discontinued levetiracetam due to lack of efficacy (n = 39) or adverse events (n = 2), 34 became seizure-free on other treatments. SIGNIFICANCE: Although superiority to placebo just failed to reach statistical significance for the primary end point, the overall findings are consistent with levetiracetam having modest efficacy against absence seizures. Further controlled trials exploring larger doses and an active comparator are required to determine the role of levetiracetam in the treatment of absence epilepsy.
Keywords: Adolescent; Age Factors; Anticonvulsants; Child; Child, Preschool; Double-Blind Method; Drug Resistance; Epilepsy, Absence; Female; Humans; Male; Outcome Assessment (Health Care); Piracetam

Multiple loci are associated with white blood cell phenotypes. - Nalls, Michael A; Couper, David J; Tanaka, Toshiko; van Rooij, Frank J A; Chen, Ming-Huei; Smith, Albert V; Toniolo, Daniela; Zakai, Neil A; Yang, Qiong; Greinacher, Andreas; Wood, Andrew R; Garcia, Melissa; et al. (PLoS Genet, 2011)
White blood cell (WBC) count is a common clinical measure from complete blood count assays, and it varies widely among healthy individuals. Total WBC count and its constituent subtypes have been shown to be moderately heritable, with the heritability estimates varying across cell types. We studied 19,509 subjects from seven cohorts in a discovery analysis, and 11,823 subjects from ten cohorts for replication analyses, to determine genetic factors influencing variability within the normal hematological range for total WBC count and five WBC subtype measures. Cohort specific data was supplied by the CHARGE, HeamGen, and INGI consortia, as well as independent collaborative studies. We identified and replicated ten associations with total WBC count and five WBC subtypes at seven different genomic loci (total WBC count-6p21 in the HLA region, 17q21 near ORMDL3, and CSF3; neutrophil count-17q21; basophil count- 3p21 near RPN1 and C3orf27; lymphocyte count-6p21, 19p13 at EPS15L1; monocyte count-2q31 at ITGA4, 3q21, 8q24 an intergenic region, 9q31 near EDG2), including three previously reported associations and seven novel associations. To investigate functional relationships among variants contributing to variability in the six WBC traits, we utilized gene expression- and pathways-based analyses. We implemented gene-clustering algorithms to evaluate functional connectivity among implicated loci and showed functional relationships across cell types. Gene expression data from whole blood was utilized to show that significant biological consequences can be extracted from our genome-wide analyses, with effect estimates for significant loci from the meta-analyses being highly corellated with the proximal gene expression. In addition, collaborative efforts between the groups contributing to this study and related studies conducted by the COGENT and RIKEN groups allowed for the examination of effect homogeneity for genome-wide significant associations across populations of diverse ancestral backgrounds.
Keywords: Genetic Loci; Genome-Wide Association Study; Humans; Leukocyte Count; Leukocytes; Molecular Epidemiology; Multigene Family; Phenotype; Polymorphism, Single Nucleotide; Ubiquitin-Protein Ligases

Mutations in ANKRD26 are responsible for a frequent form of inherited thrombocytopenia: analysis of 78 patients from 21 families. - Noris, Patrizia; Perrotta, Silverio; Seri, Marco; Pecci, Alessandro; Gnan, Chiara; Loffredo, Giuseppe; Pujol-Moix, Núria; Zecca, Marco; Scognamiglio, Francesca; De Rocco, Daniela; Punzo, Francesca; Melazzini, Federica; et al. (Blood, 2011)
Until recently, thrombocytopenia 2 (THC2) was considered an exceedingly rare form of autosomal dominant thrombocytopenia and only 2 families were known. However, we recently identified mutations in the 5'-untranslated region of the ANKRD26 gene in 9 THC2 families. Here we report on 12 additional pedigrees with ANKRD26 mutations, 6 of which are new. Because THC2 affected 21 of the 210 families in our database, it has to be considered one of the less rare forms of inherited thrombocytopenia. Analysis of all 21 families with ANKRD26 mutations identified to date revealed that thrombocytopenia and bleeding tendency were usually mild. Nearly all patients had no platelet macrocytosis, and this characteristic distinguishes THC2 from most other forms of inherited thrombocytopenia. In the majority of cases, platelets were deficient in glycoprotein Ia and α-granules, whereas in vitro platelet aggregation was normal. Bone marrow examination and serum thrombopoietin levels suggested that thrombocytopenia was derived from dysmegakaryopoiesis. Unexplained high values of hemoglobin and leukocytes were observed in a few cases. An unexpected finding that warrants further investigation was a high incidence of acute leukemia. Given the scarcity of distinctive characteristics, the ANKRD26-related thrombocytopenia has to be taken into consideration in the differential diagnosis of isolated thrombocytopenias.
Keywords: Adolescent; Adult; Aged; Aged, 80 and over; Ankyrin Repeat; Child; Cohort Studies; Family; Female; Gene Frequency; Humans; Inheritance Patterns; Male; Middle Aged; Mutation; Pedigree; Thrombocytopenia; Transcription Factors; Young Adult

Mutations in the 5' UTR of ANKRD26, the ankirin repeat domain 26 gene, cause an autosomal-dominant form of inherited thrombocytopenia, THC2. - Pippucci, Tommaso; Savoia, Anna; Perrotta, Silverio; Pujol-Moix, Núria; Noris, Patrizia; Castegnaro, Giovanni; Pecci, Alessandro; Gnan, Chiara; Punzo, Francesca; Marconi, Caterina; Gherardi, Samuele; Loffredo, Giuseppe; et al. (Am J Hum Genet, 2011)
THC2, an autosomal-dominant thrombocytopenia described so far in only two families, has been ascribed to mutations in MASTL or ACBD5. Here, we show that ANKRD26, another gene within the THC2 locus, and neither MASTL nor ACBD5, is mutated in eight unrelated families. ANKRD26 was also found to be mutated in the family previously reported to have an ACBD5 mutation. We identified six different ANKRD26 mutations, which were clustered in a highly conserved 19 bp sequence located in the 5' untranslated region. Mutations were not detected in 500 controls and are absent from the 1000 Genomes database. Available data from an animal model and Dr. Watson's genome give evidence against haploinsufficiency as the pathogenetic mechanism for ANKRD26-mediated thrombocytopenia. The luciferase reporter assay suggests that these 5' UTR mutations might enhance ANKRD26 expression. ANKRD26 is the ancestor of a family of primate-specific genes termed POTE, which have been recently identified as a family of proapoptotic proteins. Dysregulation of apoptosis might therefore be the pathogenetic mechanism, as demonstrated for another thrombocytopenia, THC4. Further investigation is needed to provide evidence supporting this hypothesis.
Keywords: Ankyrin Repeat; Base Sequence; Chromosome Breakage; Chromosome Disorders; Conserved Sequence; Female; Genes, Dominant; Genetic Loci; Haploinsufficiency; Humans; Male; Molecular Sequence Data; Mutation; Pedigree; Thrombocytopenia

Mutations in TTC19 cause mitochondrial complex III deficiency and neurological impairment in humans and flies. - Ghezzi, Daniele; Arzuffi, Paola; Zordan, Mauro; Da Re, Caterina; Lamperti, Costanza; Benna, Clara; d'Adamo, Pio; Diodato, Daria; Costa, Rodolfo; Mariotti, Caterina; Uziel, Graziella; Smiderle, Cristina; et al. (Nat Genet, 2011)
Although mutations in CYTB (cytochrome b) or BCS1L have been reported in isolated defects of mitochondrial respiratory chain complex III (cIII), most cIII-defective individuals remain genetically undefined. We identified a homozygous nonsense mutation in the gene encoding tetratricopeptide 19 (TTC19) in individuals from two families affected by progressive encephalopathy associated with profound cIII deficiency and accumulation of cIII-specific assembly intermediates. We later found a second homozygous nonsense mutation in a fourth affected individual. We demonstrated that TTC19 is embedded in the inner mitochondrial membrane as part of two high-molecular-weight complexes, one of which coincides with cIII. We then showed a physical interaction between TTC19 and cIII by coimmunoprecipitation. We also investigated a Drosophila melanogaster knockout model for TTC19 that showed low fertility, adult-onset locomotor impairment and bang sensitivity, associated with cIII deficiency. TTC19 is a putative cIII assembly factor whose disruption is associated with severe neurological abnormalities in humans and flies.
Keywords: Adult; Animals; Brain; Codon, Nonsense; Drosophila melanogaster; Electron Transport Complex III; Female; Gene Knockdown Techniques; Humans; Male; Membrane Proteins; Mitochondria; Mitochondrial Proteins; Nervous System Diseases

Neonatal necrotizing tracheobronchitis. - Bua, Jenny; Trappan, Antonella; Demarini, Sergio; Grasso, Domenico; Schleef, Jurgen; Zennaro, Floriana (J Pediatr, 2011)
Keywords: Bronchitis; Bronchoscopy; High-Frequency Ventilation; Humans; Infant, Newborn; Male; Necrosis; Respiratory Insufficiency; Respiratory Mucosa; Tracheitis

Neuromotor deficits in developmental coordination disorder: evidence from a reach-to-grasp task. - Biancotto, Marina; Skabar, Aldo; Bulgheroni, Maria; Carrozzi, Marco; Zoia, Stefania (Res Dev Disabil, 2011)
Developmental coordination disorder (DCD) has been classified as a specific learning disability, nonetheless the underlying cognitive mechanisms are still a matter of discussion. After a summary of the main hypotheses on the principal neuromotor causes of DCD, this study applies a causal model framework to describe the possible coexistence of more than one deficit in this disorder. For this purpose, kinematic analysis was applied to an ecological task, the reach-to-grasp action, introducing the manipulation of three variables: vision, distance and object size. After a thorough neurological and neuropsychological evaluation, 9 children with DCD (7-9 years old) were selected and compared to 27 age-matched control children. The results suggest that children with DCD have a normal neurological characterization of the reaching and grasping movements, in terms of proximal to distal action, but their grasping aperture (MGA) was always wider with respect to controls, particularly when vision was not allowed. In addition, the performance of children with DCD was always slower, more dependent on vision and more variable than that of controls. The MGA of children with DCD could be explained by a deficit in the internal construction of movement for a forward model, while slowness could be related to a control problem in the neuronal firing of the muscles. The idea of a possible coexistence of these two deficits is discussed in accordance to a causal model framework and also addressed considering recent neurophysiologic evidences.
Keywords: Analysis of Variance; Biomechanical Phenomena; Child; Female; Humans; Male; Motor Skills Disorders; Psychometrics; Psychomotor Performance; Reaction Time; Space Perception

A new case of duplication of the MDS region identified by high-density SNP arrays and a review of the literature. - Faletra, Flavio; Devescovi, Raffaella; Pecile, Vanna; Fabretto, Antonella; Carrozzi, Marco; Gasparini, Paolo (J Appl Genet, 2011)
Keywords: 1-Alkyl-2-acetylglycerophosphocholine Esterase; Child; Female; Gene Duplication; Humans; Microtubule-Associated Proteins; Myelodysplastic Syndromes; Oligonucleotide Array Sequence Analysis; Polymorphism, Single Nucleotide; Prognosis

Non-invasive assessment of hemispheric language dominance by optical topography during a brief passive listening test: a pilot study. - Bembich, Stefano; Demarini, Sergio; Clarici, Andrea; Massaccesi, Stefano; Grasso, Domenico Loenardo (Med Sci Monit, 2011)
BACKGROUND: The Wada test is usually used for pre-surgical assessment of language lateralization. Considering its invasiveness and risk of complications, alternative methods have been proposed but they are not always applicable to non-cooperative patients. In this study we explored the possibility of using optical topography (OT)--a multichannel near-infrared system--for non-invasive assessment of hemispheric language dominance during passive listening. MATERIAL/METHODS: Cortical activity was monitored in a sample of healthy, adult Italian native speakers, all right-handed. We assessed changes in oxy-haemoglobin concentration in temporal, parietal and posterior frontal lobes during a passive listening of bi-syllabic words and vowel-consonant-vowel syllables lasting less then 3 minutes. Activated channels were identified by t tests. RESULTS: Left hemisphere showed significant activity only during the passive listening of bi-syllabic words. Specifically, the superior temporal gyrus, the supramarginal gyrus and the posterior inferior parietal lobe were activated. CONCLUSIONS: During passive listening of bi-syllabic words, right handed healthy adults showed a significant activation in areas already known to be involved in speech comprehension. Although more research is needed, OT proved to be a promising alternative to the Wada test for non-invasive assessment of hemispheric language lateralization, even if using a particularly brief trial, which has been designed for future applications with non-cooperative subjects.
Keywords: Acoustic Stimulation; Adult; Cerebrum; Diagnostic Techniques and Procedures; Dominance, Cerebral; Female; Humans; Language; Male; Middle Aged; Oxyhemoglobins; Pilot Projects; Spectroscopy, Near-Infrared

Osteoprotegerin induces morphological and functional alterations in mouse pancreatic islets. - Toffoli, Barbara; Bernardi, Stella; Candido, Riccardo; Sabato, Nicoletta; Carretta, Renzo; Corallini, Federica; Secchiero, Paola; Zauli, Giorgio; Fabris, Bruno (Mol Cell Endocrinol, 2011)
Although serum osteoprotegerin (OPG) is significantly increased in diabetic subjects, its potential role in beta cell dysfunction has not been investigated. This study aimed to assess the effect of full-length OPG administered in vivo in mice on pancreatic islet structure and function and its interaction with the renin-angiotensin system (RAS). OPG-treated mice showed increased islet monocyte/macrophage infiltration, fibrosis and apoptosis with reduction of islet function. The remodeling of islet architecture was associated with increased pancreatic expression of components of the RAS, growth factor genes (transforming growth factor β and connective tissue growth factor) and inflammatory molecules (monocyte chemotactic protein-1 and vascular adhesion molecule type 1). Prevention of these changes with improvement of insulin secretion was observed in ramipril treated animals. Our data suggest that OPG might play an important role in promoting beta cell dysfunction and that the upregulation of the local RAS represents one possible mechanism responsible for the OPG-induced beta cell dysfunction.
Keywords: Animals; Apoptosis; Blood Glucose; Blood Pressure; Body Weight; Cell Lineage; Cell Movement; Chemokine CCL2; Connective Tissue Growth Factor; Fibrosis; Gene Expression Regulation; Humans; Insulin; Islets of Langerhans; Macrophages; Mice; Monocytes; Organ Size; Osteoprotegerin; Peptidyl-Dipeptidase A; Receptor, Angiotensin, Type 1; Systole; Transforming Growth Factor beta; Vascular Cell Adhesion Molecule-1

Osteoprotegerin promotes vascular fibrosis via a TGF-β1 autocrine loop. - Toffoli, Barbara; Pickering, Raelene J; Tsorotes, Despina; Wang, Bo; Bernardi, Stella; Kantharidis, Phillip; Fabris, Bruno; Zauli, Giorgio; Secchiero, Paola; Thomas, Merlin C (Atherosclerosis, 2011)
BACKGROUND: This study was designed to evaluate the potential role of osteoprotegerin (OPG) in arterial fibrosis. METHODS: Aortic samples were analyzed after in vivo treatment of ApoE(-/-) mice with recombinant human OPG. Mouse vascular smooth muscle cells (VSMC) were exposed in vitro to recombinant OPG and analyzed for markers of inflammation and fibrosis, such as fibronectin, collagen I, III, IV and transforming growth factor-β1 (TGF-β1). Conversely, the potential modulation of endogenous OPG expression and release by VSMC was analyzed in response to different pro-atherosclerotic cytokines, TGF-β1, platelet derived growth factor (PDGF) and angiogensin II (Ang II). RESULTS: In vivo treatment with human OPG induced signs of fibrosis and up-regulated the arterial expression of TGF-β1. Consistently, in vitro treatment of VSMC with human OPG induced the expression of fibronectin, collagen type I, III, IV, metalloprotein-2 (MMP-2) and MMP-9, as well as of TGF-β1. On the other hand, exposure to recombinant TGF-β1 promoted the expression/release of endogenous OPG and mediated the increase of OPG release induced by PDGF and Ang II in VSMC. CONCLUSIONS: Taken together, these data support a pathogenic role for OPG in the development and progression of atherosclerotic lesions and suggest the existence of a vicious circle between TGF-β1 and OPG.
Keywords: Animals; Apolipoproteins E; Cell Proliferation; Collagen; Fibronectins; Fibrosis; Gene Expression Regulation; Humans; Mice; Mice, Inbred C57BL; Mice, Transgenic; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Osteoprotegerin; Platelet-Derived Growth Factor; Transforming Growth Factor beta1

Paediatric Behçet's disease presenting with recurrent papillitis and episcleritis: a case report. - Parentin, Fulvio; Lepore, Loredana; Rabach, Ingrid; Pensiero, Stefano (J Med Case Rep, 2011)
INTRODUCTION: Behçet's disease is a chronic multisystem vasculitis characterized by mucocutaneous, articular, neurological, gastrointestinal and ophthalmological lesions. Ocular involvement is mainly represented by recurrent uveitis, especially posterior uveitis; however, iridocyclitis, retinal and choroidal vasculitis, optic neuritis and retinal vascular occlusion can also occur. CASE PRESENTATION: A 12-year-old Caucasian boy with a history of recurrent buccal aphthosis and nonspecific gastrointestinal symptoms was admitted to our hospital with blurred vision associated with acute episcleritis and papillitis. The patient's pathergy test was positive, suggesting a diagnosis of Behçet's disease. Corticosteroid and cyclosporine therapy was started, but further episodes were noted in both eyes. The patient was then switched to intravenous infliximab, with complete resolution of the inflammation after the second infusion. CONCLUSION: Episcleritis and papillitis should be added to the list of uncommon manifestations of pediatric Behçet's disease. Infliximab is an effective, new therapeutic approach for Behçet's disease that is refractory to the conventional corticosteroid and immunosuppressive therapy.

Periodontal infection and preterm birth: successful periodontal therapy reduces the risk of preterm birth. - Di Mario, S; Spettoli, D; Alessandrini, C; Erenbourg, A; Ronfani, L; Basevi, V (BJOG, 2011)
Keywords: Female; Humans; Periodontal Diseases; Pregnancy; Pregnancy Complications, Infectious; Premature Birth; Randomized Controlled Trials as Topic; Risk Factors

A Phase II study on the safety and efficacy of a single dose of pegfilgrastim for mobilization and transplantation of autologous hematopoietic stem cells in pediatric oncohematology patients. - Cesaro, Simone; Zanazzo, Andrea Giulio; Frenos, Stefano; Luksch, Roberto; Pegoraro, Anna; Tridello, Gloria; Dallorso, Sandro (Transfusion, 2011)
BACKGROUND: Limited data are available on the use of pegfilgrastim in pediatric patients as a mobilizing agent in association with chemotherapy. STUDY DESIGN AND METHODS: This was a prospective, multicenter, Phase II study to evaluate the safety and efficacy of a single dose of 100 µg/kg pegfilgrastim in mobilizing peripheral blood stem cells (PBSCs) in pediatric patients. The primary endpoint of the study was the percentage of good mobilizers with pegfilgrastim (blood peak of CD34+ cells ≥ 20 × 10(6) /L). The results were compared with a historical control group. RESULTS: Thirty of 36 recruited patients were classified as good mobilizers (83%). The median value of circulating CD34+ at leukapheresis was 143 × 10(6) /L (range, 20 × 10(6) -1988 × 10(6) /L). No significant adverse effects were associated with the use of pegfilgrastim and no patient was withdrawn from using the drug. A blood peak of 20 × 10(6) /L or more CD34+ was observed in 33 of 36 control patients (92%) and the median CD34+ count at leukapheresis was 158 × 10(6) /kg (range, 28 × 10(6) -4529 × 10(6) /kg; p = 0.7). No significant differences were found between the two groups in terms of toxicity or other variables of mobilization. As at October 2008, 23 patients of the pegfilgrastim group and 32 patients of the filgrastim group underwent autologous transplant. No significant differences were found in terms of early toxicity, myeloid recovery, and Day 100 survival. CONCLUSION: A single dose of 100 µg/kg pegfilgrastim was safe and effective for PBSC collection in pediatric patients. We suggest that these results support the use of pegfilgrastim for pediatric patients.
Keywords: Adolescent; Adult; Child; Child, Preschool; Female; Granulocyte Colony-Stimulating Factor; Hematopoietic Stem Cell Mobilization; Humans; Infant; Male; Neoplasms; Peripheral Blood Stem Cell Transplantation; Prospective Studies; Recombinant Proteins; Transplantation, Autologous

Phospholipase C-β3 is a key modulator of IL-8 expression in cystic fibrosis bronchial epithelial cells. - Bezzerri, Valentino; d'Adamo, Pio; Rimessi, Alessandro; Lanzara, Carmen; Crovella, Sergio; Nicolis, Elena; Tamanini, Anna; Athanasakis, Emmanouil; Tebon, Maela; Bisoffi, Giulia; Drumm, Mitchell L; Knowles, Michael R; et al. (J Immunol, 2011)
Respiratory insufficiency is the major cause of morbidity and mortality in patients affected by cystic fibrosis (CF). An excessive neutrophilic inflammation, mainly orchestrated by the release of IL-8 from bronchial epithelial cells and amplified by chronic bacterial infection with Pseudomonas aeruginosa, leads to progressive tissue destruction. The anti-inflammatory drugs presently used in CF patients have several limitations, indicating the need for identifying novel molecular targets. To address this issue, we preliminarily studied the association of 721 single nucleotide polymorphisms from 135 genes potentially involved in signal transduction implicated in neutrophil recruitment in a cohort of F508del homozygous CF patients with either severe or mild progression of lung disease. The top ranking association was found for a nonsynonymous polymorphism of the phospholipase C-β3 (PLCB3) gene. Studies in bronchial epithelial cells exposed to P. aeruginosa revealed that PLCB3 is implicated in extracellular nucleotide-dependent intracellular calcium signaling, leading to activation of the protein kinase Cα and Cβ and of the nuclear transcription factor NF-κB p65. The proinflammatory pathway regulated by PLCB3 acts by potentiating the Toll-like Receptors' signaling cascade and represents an interesting molecular target to attenuate the excessive recruitment of neutrophils without completely abolishing the inflammatory response.
Keywords: Adenosine Triphosphate; Calcium; Cell Line, Transformed; Cystic Fibrosis; Enzyme Activation; Epithelial Cells; Gene Expression; Gene Frequency; Genotype; Green Fluorescent Proteins; Host-Pathogen Interactions; Humans; Interleukin-8; Isoenzymes; Lung Diseases; Microscopy, Fluorescence; Phospholipase C beta; Polymorphism, Single Nucleotide; Protein Kinase C; Protein Kinase C beta; Pseudomonas aeruginosa; RNA Interference; Toll-Like Receptors; Transcription Factor RelA

A polymorphism in PRF1 gene is associated with HIV-1 vertical transmission in Brazilian children. - Padovan, Lara; Segat, Ludovica; Crovella, Sergio (AIDS, 2011)
We investigated the possible association between PRF1 gene polymorphisms and HIV-1 vertical transmission in Brazilian children by analyzing PRF1 gene coding and untranslated regions in 173 perinatally infected children (HIV+), 51 exposed uninfected (HIV-), and 171 HIV-unexposed uninfected children. Seven single nucleotide polymorphisms were identified in our samples. The rs885822 C allele and CC genotype were significantly more frequent in HIV-negative than in HIV-positive patients and associated with a protective effect toward HIV vertical transmission.
Keywords: Brazil; Female; HIV Infections; HIV-1; Humans; Infant, Newborn; Infectious Disease Transmission, Vertical; Male; Polymorphism, Genetic; Pore Forming Cytotoxic Proteins; Pregnancy

A polymorphism in the 5' UTR of the DEFB1 gene is associated with the lung phenotype in F508del homozygous Italian cystic fibrosis patients. - Crovella, Sergio; Segat, Ludovica; Amato, Annalisa; Athanasakis, Emmanouil; Bezzerri, Valentino; Braggion, Cesare; Casciaro, Rosaria; Castaldo, Giuseppe; Colombo, Carla; Covone, Angela Elvira; De Rose, Virginia; Gagliardini, Rolando; et al. (Clin Chem Lab Med, 2011)
BACKGROUND: The identification of cystic fibrosis (CF) patients who are at greater risk of lung damage could be clinically valuable. Thus, we attempted to replicate previous findings and verify the possible association between three single nucleotide polymorphisms (SNPs c.-52G>A, c.-44C>G and c.-20G>A) in the 5' untranslated region (5' UTR) of the β defensin 1 (DEFB1) gene and the CF pulmonary phenotype. METHODS: Genomic DNA from 92 Italian CF patients enrolled in different regional CF centres was extracted from peripheral blood and genotyped for DEFB1 SNPs using TaqMan(®) allele specific probes. In order to avoid genetic confounding causes that can account for CF phenotype variability, all patients were homozygous for the F508del CFTR mutation, and were then classified on the basis of clinical and functional data as mild lung phenotype (Mp, n=50) or severe lung phenotype patients (Sp, n=42). RESULTS: For the c.-20G>A SNP, the frequency of the A allele, as well as the AA genotype, were significantly more frequent in Mp than in Sp patients, and thus this was associated with a protective effect against severe pulmonary disease (OR=0.48 and 0.28, respectively). The effect of the c.-20G>A A allele is consistent with a recessive model, and the protective effect against Sp is exerted only when it is present in homozygosis. For the other two SNPs, no differences were observed as allelic and genotypic frequency in the two subgroups of CF patients. CONCLUSIONS: Our results, although necessary to be confirmed in larger and multiethnic populations, reinforce DEFB1 as a candidate modifier gene of the CF pulmonary phenotype.
Keywords: 5' Untranslated Regions; Adult; beta-Defensins; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Genotype; Homozygote; Humans; Italy; Male; Mutation; Phenotype; Polymorphism, Genetic; Young Adult

Prevalence of anaemia in children with thromboembolism in IBD. - Bramuzzo, Matteo; Lazzerini, Marzia (Arch Dis Child, 2011)
Keywords: Anemia, Iron-Deficiency; Female; Humans; Male; Stroke

Procalcitonin in detecting neonatal nosocomial sepsis. - Auriti, Cinzia; Fiscarelli, Ersilia; Ronchetti, Maria Paola; Argentieri, Marta; Marrocco, Gabriella; Quondamcarlo, Anna; Seganti, Giulio; Bagnoli, Francesco; Buonocore, Giuseppe; Serra, Giovanni; Bacolla, Gianfranco; Mastropasqua, Savino; et al. (Arch Dis Child Fetal Neonatal Ed, 2011)
OBJECTIVE: To investigate the accuracy of procalcitonin (PCT) as a diagnostic marker of nosocomial sepsis (NS) and define the most accurate cut-off to distinguish infected from uninfected neonates. SETTING: Six neonatal intensive care units (NICUs). PATIENTS: 762 neonates admitted to six NICUs during a 28-month observational study for whom at least one serum sample was taken on admission. MAIN OUTCOME MEASURES: Positive and negative predictive values at different PCT cut-off levels. RESULTS: The overall probability of an NS was doubled or more if PCT was >0.5 ng/ml. In very-low-birth-weight (VLBW) infants, a cut-off of >2.4 ng/ml gave a positive predictive value of NS near to 50% with a probability of a false-positive diagnosis of NS in about 10% of the patients. CONCLUSIONS: In VLBW neonates, a serum PCT value >2.4 ng/ml prompts early empirical antibiotic therapy, while in normal-birth-weight infants, a PCT value ≤2.4 ng/ml carries a low risk of missing an NS.

Promoting mother’s milk use in very low birth weight infants: when nutritional hierarchy deals with the professional value system. - Davanzo, Riccardo (J Hum Lact, 2011)
Keywords: Attitude of Health Personnel; Breast Feeding; Health Promotion; Humans; Infant, Newborn; Infant, Very Low Birth Weight; Intensive Care Units, Neonatal

[Proteomic applications in gynecology-obstetrics]. - De Seta, F; Banco, R; Guaschino, S; De Santo, D; Turrisi, A; Piva, C (Minerva Ginecol, 2011)
Proteomics has recently emerged as a powerful approach both for discovering biomarkers as well as for understanding the physiopathology of unclear gynecological-obstetrical disorders. Currently, several biological fluids and fetal tissues were successfully tested, including maternal plasma, amniotic fluid, cervical-vaginal fluid, urine, saliva, placental trophoblast, amnio-chorionic membranes and cord blood. The potential of proteomics on the polycystic ovary syndrome (PCOS) involves biomarkers discovery for a more accurate diagnosis of the syndrome and identification, within the patients with PCOS, those who respond more easily to treatment and those who will be at increased risk for future metabolic complications. The proteomic approach applied to patients with endometriosis would allow not only a non-invasive early diagnosis, but also a staging of the disease and a prediction of infertility risk. Proteomics also involves oncological field, in order to discover biomarkers that allow early diagnosis and prognosis of female genital malignancies. In addition to this, proteomics could be used to understand and predict obstetrical complications such as recurrent spontaneous abortion, preterm birth and preeclampsia. However, further studies are needed on a larger cohort of patients to introduce these biomarkers in clinical practice.
Keywords: Endometriosis; Female; Genital Diseases, Female; Genital Neoplasms, Female; Humans; Polycystic Ovary Syndrome; Pregnancy; Pregnancy Complications; Proteomics

Quality of maternal and neonatal care in Albania, Turkmenistan and Kazakhstan: a systematic, standard-based, participatory assessment. - Tamburlini, Giorgio; Siupsinskas, Gelmius; Bacci, Alberta (PLoS One, 2011)
BACKGROUND: Progress in maternal and neonatal mortality has been slow in many countries despite increasing access to institutional births, suggesting deficiencies in the quality of care. We carried out a systematic assessment of the quality of maternal and newborn care in three CEE/CIS countries, using an innovative approach to identify priority issues and promote action. METHODS: A standard-based tool, covering over 400 items grouped in 13 main areas ranging from support services to case management, was used to assess a sample of ten maternity hospitals in Albania, Kazakhstan and Turkmenistan. Sources of information were visit to services, medical records, observation of cases, and interviews with staff and mothers. A score (range 0 to 3) was attributed to each item and area of care. The assessment was carried out by a multidisciplinary team of international and national professionals. Local managers and staff provided the necessary information and were involved in discussing the findings and the priority actions. RESULTS: Quality of care was found to be substandard in all 13 areas. The lowest scores (between one and two) were obtained by: management of normal labour, delivery, obstetric complications and sick babies; infection prevention; use of guidelines and audits; monitoring and follow-up. Neonatal care as a whole scored better than obstetric care. Interviewed mothers identified lack of information, insufficient support during labour and lack of companionship as main issues. Actions to improve quality of care were identified at facility as well as at central level and framed according to main health system functions. CONCLUSIONS: Quality of care is a key issue to improve maternal and neonatal outcomes, particularly in countries such as CEE/CIS where access to institutional births is nearly universal. Approaches that involve health professionals and managers in comprehensive, action-oriented assessments of quality of care are promising and should be further supported.
Keywords: Albania; Child Health Services; Female; Humans; Infant, Newborn; Kazakhstan; Maternal Health Services; Pregnancy; Quality of Health Care; Turkmenistan

Quantification of heteroplasmic mitochondrial DNA mutations for DNA samples in the low picogram range by nested real-time ARMS-qPCR. - Biffi, Stefania; Bortot, Barbara; Carrozzi, Marco; Severini, Giovanni Maria (Diagn Mol Pathol, 2011)
In many mitochondrial diseases, different clinical manifestations are related to tissue-specific distribution of mutated mitochondrial DNA (mtDNA). In this study, we describe an assay for the determination of mutated mtDNA copy number in small clinical samples, using standard polymerase chain reaction (PCR) followed by SYBR Green real-time allelic-specific PCR [amplification refractory mutation system-quantitative PCR (ARMS-qPCR)]. To assess the degree of heteroplasmy in a patient harboring 2 cosegregating mtDNA mutations (4415A>G and 9922A>C) starting from picogram amounts of DNA, we first amplified the mutated target sequence by standard PCR, and then analyzed it by real-time ARMS-qPCR. To validate this method, we analyzed by real-time ARMS-qPCR the PCR amplification products derived from different mixtures containing known proportions of mutant and wild-type cloned mtDNA fragments. The correlation coefficient of 0.994 between expected and observed values for the percentage of mutant A4415G confirms that the relative proportion of mutated and wild-type mtDNA was maintained after the first PCR amplification. This method allows the precise quantification of heteroplasmic mutations in DNA samples extracted from hairs, urine, small stomach biopsies, and, more importantly, single-muscle fiber, with a limit of detection close to 0.5%. This nested real-time ARMS-PCR represents a rapid, efficient, and less expensive method for the detection and quantification of heteroplasmic mutant mtDNA, even in very small clinical samples.
Keywords: Child; DNA, Mitochondrial; Humans; Mitochondrial Diseases; Mutation; Polymerase Chain Reaction; Sensitivity and Specificity

Recent advances in the therapeutic perspectives of Nutlin-3. - Secchiero, Paola; Bosco, Raffaella; Celeghini, Claudio; Zauli, Giorgio (Curr Pharm Des, 2011)
Nutlin-3 is a small molecule inhibitor of the MDM2/p53 interaction, which leads to the non-genotoxic p53 stabilization, activation of cell cycle arrest and apoptosis pathways. A series of recent studies have strengthened the concept that selective, non-genotoxic p53 activation by Nutlin-3 might represent an alternative to the current cytotoxic chemotherapy, in particular for pediatric tumors and for hematological malignancies, which retain a high percentage of p53(wild-type) status at diagnosis. Like most other drugs employed in cancer therapy, it will be unlikely that Nutlin-3 will be used as a monotherapy. In this respect, Nutlin-3 shows a synergistic cytotoxic effect when used in combination with innovative drugs, such as TRAIL or bortozemib. Although Nutlin-3 is currently in phase I clinical trial for the treatment of retinoblastoma, its effects on normal tissues and cell types remain largely to be determined and will require further investigation in the future years.
Keywords: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Drug Synergism; Genes, p53; Humans; Imidazoles; Neoplasms; Piperazines; Proto-Oncogene Proteins c-mdm2; Tumor Suppressor Protein p53

Recent advances in the understanding and management of MYH9-related inherited thrombocytopenias. - Balduini, Carlo L; Pecci, Alessandro; Savoia, Anna (Br J Haematol, 2011)
MYH9-related disease (MYH9-RD) is one of the most frequent forms of inherited thrombocytopenia. It is transmitted in an autosomal dominant fashion and derives from mutations of MYH9, the gene for the heavy chain of non-muscle myosin IIA. Patients present with congenital macrothrombocytopenia with mild bleeding tendency and may develop kidney dysfunction, deafness and cataracts later in life. The term MYH9-RD encompasses four autosomal-dominant thrombocytopenias that were previously described as distinct disorders, namely May-Hegglin Anomaly, Sebastian, Fechtner and Epstein syndromes. Thrombocytopenia is usually mild and derives from complex defects of megakaryocyte maturation and platelet formation. It is easily diagnosed, in that the presence of giant platelets in peripheral blood raises the suspicion of MYH9-RD and a simple immunofluorescence test on blood films confirms the diagnostic hypothesis. However, genotype/phenotype correlations have been recognized and mutation screening is therefore required to define the risk of acquiring extra-haematological defects. Results of a small clinical study suggested that a non-peptide thrombopoietin mimetic might greatly benefit both thrombocytopenia and bleeding tendency of MYH9-RD patients.
Keywords: Animals; Disease Models, Animal; Genetic Predisposition to Disease; Genotype; Humans; Mice; Molecular Motor Proteins; Mutation; Myosin Heavy Chains; Myosin Type II; Phenotype; Thrombocytopenia

Refractory iron-deficiency anaemia in a child with portal cavernoma. - Pastore, Serena; Londero, Margherita; Cont, Gabriele; Di Leo, Grazia; Ventura, Alessandro (Gut, 2011)
Keywords: Anemia, Iron-Deficiency; Antihypertensive Agents; Child; Hemangioma, Cavernous; Humans; Hypertension, Portal; Ileal Diseases; Male; Propranolol; Vascular Neoplasms

The resurgence of rheumatic fever in a developed country area: the role of echocardiography. - Pastore, Serena; De Cunto, Angela; Benettoni, Alessandra; Berton, Emanuela; Taddio, Andrea; Lepore, Loredana (Rheumatology (Oxford), 2011)
OBJECTIVES: The annual incidence of ARF ranges from 5 to 51/100, 000 population worldwide in the 5- to 15-year age group. In the past, there was a decline in the incidence of ARF; however, focal outbreaks have been reported. This study evaluated the incidence of ARF in 2007-08 in a region of a developed country compared with the previous decade. METHODS: A retrospective review of all admission records for ARF in Trieste between January 2007 and December 2008 was undertaken. The diagnosis of ARF was established by the Jones criteria according to the 1992 revision. RESULTS: Between January 2007 and December 2008: 13 cases of ARF were recorded, 11 females and 2 males. The estimated incidence was 23 and 27/100, 000 population new cases each year, respectively, in the 5- to 15-year age group. Migratory polyarthritis occurred in 6/13, chorea in 7/13 and clinical carditis in 5/13 cases. Five out of 13 patients had only echocardiographic abnormalities, with no clinical cardiac manifestations. Another two patients did not fulfil diagnostic criteria for ARF, presenting with only three minor criteria, but they revealed silent carditis at echocardiography evaluation. During the follow-up, in one case the carditis receded and in the other it significantly improved. CONCLUSIONS: Our experience underlines that ARF has not yet disappeared in industrialized countries. We observed a high incidence of chorea, always associated with mild carditis. Echocardiographic assessment should be routinely performed in all patients with suspected ARF in order to identify those subclinical cases of valvulitis that would otherwise pass undiagnosed without receiving proper prophylaxis.
Keywords: Adolescent; Child; Child, Preschool; Chorea; Developed Countries; Diagnosis, Differential; Echocardiography; Female; Humans; Italy; Male; Myocarditis; Retrospective Studies; Rheumatic Fever

Role of DC-SIGN and L-SIGN receptors in HIV-1 vertical transmission. - da Silva, Ronaldo Celerino; Segat, Ludovica; Crovella, Sergio (Hum Immunol, 2011)
The innate immune system acts in the first line of host defense against pathogens. One of the mechanisms used involves the early recognition and uptake of microbes by host professional phagocytes, through pattern recognition receptors (PRRs). These PRRs bind to conserved microbial ligands expressed by pathogens and initiate both innate and adaptative immune responses. Some PRRs located on the surface of dendritic cells (DCs) and other cells seem to play an important role in human immunodeficiency virus type 1 (HIV-1) transmission. Dendritic cell-specific intercellular adhesion molecule-3 grabbing non-integrin, CD209 (DC-SIGN) and its homolog, DC-SIGN-related (DC-SIGNR or L-SIGN) receptors are PPRs able to bind the HIV-1 gp120 envelope protein and, because alterations in their expression patterns also occur, they might play a role in both horizontal and vertical transmission as well as in disseminating the virus within the host. This review aims to explore the involvement of the DC-SIGN and L-SIGN receptors in HIV-1 transmission from mother to child.
Keywords: Cell Adhesion Molecules; HIV Infections; HIV-1; Humans; Immunity, Innate; Infectious Disease Transmission, Vertical; Lectins, C-Type; Polymorphism, Genetic; Receptors, Cell Surface

The role of mannose-binding lectin gene polymorphisms in susceptibility to HIV-1 infection in Southern Brazilian patients. - da Silva, Gabriela Kniphoff; Guimarães, Rafael; Mattevi, Vanessa Suñé; Lazzaretti, Rosmeri Kuhmmer; Sprinz, Eduardo; Kuhmmer, Regina; Brandão, Lucas; Crovella, Sergio; Chies, José Artur Bogo (AIDS, 2011)
OBJECTIVE: This study investigates the role of mannose-binding lectin (MBL) in the susceptibility to HIV-1 infection analyzing polymorphisms located at the MBL2 promoter and exon 1 regions. MATERIALS AND METHODS: The prevalence of MBL2 variant alleles was investigated in 410 HIV-1-infected patients from the South Brazilian HIV cohort and in 345 unexposed uninfected healthy individuals. The promoter variants were genotyped using polymerase chain reaction with sequence-specific primers (PCR-SSP) and exon 1 variants were analyzed by real-time PCR using a melting temperature assay and were confirmed by PCR-restriction fragment length polymorphism (RFLP). MBL2 genotypic and allelic frequencies were compared between HIV-1-infected patients and controls using the chi-squared tests. RESULTS: The analyses were performed subdividing the individuals according to their ethnic origin. Among Euro-derived individuals a higher frequency of the LX/LX genotype was observed in patients when compared to controls (P < 0.001). The haplotypic analysis also showed a higher frequency of the haplotypes associated with lower MBL levels among HIV-1-infected patients (P = 0.0001). Among Afro-derived individuals the frequencies of LY/LY and HY/HY genotypes were higher in patients when compared to controls (P = 0.009 and P = 0.02). CONCLUSIONS: An increased frequency of MBL2 genotypes associated with low MBL levels was observed in Euro-derived patients, suggesting a potential role for MBL in the susceptibility to HIV-1 infection in Euro-derived individuals.
Keywords: Adult; Aged; Brazil; Female; Genetic Predisposition to Disease; HIV Infections; HIV-1; Humans; Lectins, C-Type; Male; Mannose-Binding Lectin; Mannose-Binding Lectins; Middle Aged; Polymorphism, Genetic; Promoter Regions, Genetic; Receptors, Cell Surface; Young Adult

Rubella susceptibility profile in pregnant women with HIV. - Floridia, Marco; Pinnetti, Carmela; Ravizza, Marina; Tibaldi, Cecilia; Sansone, Matilde; Fiscon, Marta; Guaraldi, Giovanni; Guerra, Brunella; Alberico, Salvatore; Spinillo, Arsenio; Castelli, Paula; Dalzero, Serena; et al. (Clin Infect Dis, 2011)
Keywords: Female; HIV Infections; Humans; Pregnancy; Pregnancy Complications, Infectious; Rubella

Safety and efficacy of high-dose acarbose treatment for dumping syndrome. - De Cunto, Angela; Barbi, Egidio; Minen, Federico; Ventura, Alessandro (J Pediatr Gastroenterol Nutr, 2011)
Dumping syndrome (DS) is a complication of Nissen fundoplication. Dietary strategies can ameliorate symptoms, but this approach is not always foolproof. Limited evidence reports the efficacy of acarbose for children who are unresponsive to feeding manipulations. We report 8 patients with DS aged between 7 and 24 months. In 4 of 8 nutritional strategies failed, and acarbose treatment was started. The initial dose was 25 mg for meals, and increased until postprandial glucose was stable. In 3 of 4 children the final dose was higher than previously reported, without adverse effects. Acarbose is useful to treat DS in cases of failure of dietary strategies.
Keywords: Acarbose; Child, Preschool; Dumping Syndrome; Female; Humans; Hyperglycemia; Hypoglycemic Agents; Infant; Male; Postprandial Period; Treatment Outcome

Secondary lymphoid tissue as an important site for WU polyomavirus infection in immunocompetent children. - Comar, Manola; Zanotta, Nunzia; Rossi, Tatiana; Pelos, Giorgio; D'Agaro, Pierlanfranco (J Med Virol, 2011)
The polyomaviruses KI and WU (KIPyV and WUPyV) have been identified in respiratory specimens from children with acute respiratory infections, which suggests the respiratory tract as a possible site of infection. However, the persistence of infection in the lymphoid system is unknown. Fresh samples (n = 211) of tonsils, adenoids, and peripheral blood mononuclear cells (PBMCs) from 83 immunocompetent children (mean age 4.8 years) were tested for amplification of the KIPyV VP1 and WUPyV VP2 genes. The known BK and JC polyomaviruses and the lymphotropic human herpesvirus (HHV)-6 were also investigated by quantitative real-time PCR and direct sequencing. In addition, 98 nasopharyngeal swabs collected from children (mean age 6.2 years) affected by seasonal influenza-like illness were tested. Of the lymphoid tissues, 34.9% were positive for WUPyV, 4.8% for BK virus, and 33.8% for HHV-6. KIPyV and JC virus were not detected in these specimens. None of the polyomaviruses were detected in PBMCs. Among the nasopharyngeal samples, the prevalence of WUPyV was 27.5%, although 70% of the positive samples were co-infected with at least one of the following respiratory viruses: influenza virus, adenovirus, and respiratory syncytial virus. Phylogenetic analysis revealed high sequence homology (99%) between lymphoid- and nasopharynx-derived WUPyV strains. These results suggest that the tonsils and adenoids of immunocompetent children are a reservoir for WUPyV infection; probably due to the respiratory route of transmission. In addition, the prevalence of WUPyV was high among the children, and the virus was identified more frequently in older children than during the first years of life.
Keywords: Adenoids; Child; Child, Preschool; DNA, Viral; Female; Humans; Infant; Leukocytes, Mononuclear; Male; Nasopharynx; Palatine Tonsil; Phylogeny; Polymerase Chain Reaction; Polyomavirus; Polyomavirus Infections; Prevalence; Sequence Analysis, DNA; Sequence Homology

Serum anti-Müllerian hormone as a predictive marker of polycystic ovarian syndrome. - Parco, Sergio; Novelli, Caterina; Vascotto, Fulvia; Princi, Tanja (Int J Gen Med, 2011)
BACKGROUND: The anti-Müllerian hormone (AMH) is a dimeric protein secreted by the female ovaries and has two fundamental roles in follicle genesis. It delays the entrance of the primordial follicle into the pool of follicles in growth and diminishes the sensitivity of the ovarian follicle towards follicle-stimulating hormone (FSH). The purpose of this work was to study the AMH (nv 2.0-6.8 ng/mL) as a marker during assisted reproductive technology (ART), in order to identify cases of infertility due to polycystic ovarian syndrome (PCOS). This syndrome affects 10% of women with infertility problems, and a new biological marker could be useful to general practitioners of internal medicine to help generate the suspicion of PCOS so that they can refer the patient to the gynecologist for confirmation. METHODS: This study enrolled 236 patients aged 26-46 years undergoing assisted reproductive technology at the Institute for Maternal and Child Health, Trieste, Italy. On the third day of the ovarian cycle, the patients were given doses of AMH, FSH, and luteinizing hormone (LH, in cases of AMH < 2.0-6.8 ng/mL). A control pelvic ultrasound was also carried out. RESULTS: We identified 57 patients who were starting in vitro fertilization or embryo transfer with AMH values within the normal range (3.64 ± 1.51 ng/mL), 77 with values below normal (1.38 ± 0.32 ng/mL), and 96 cases with undetectable values of AMH. Six patients had very high AMH levels (10.0 ± 2.28 ng/mL) and, of these, five were found to have PCOS on pelvic ultrasound examination (P < 0.05). We also found inverse correlations between AMH levels and age (r = -0.52) and between AMH and FSH levels (r = -0.32). CONCLUSION: In clinical practice it is common to encounter patients who turn to medicine in search of a cure for female infertility. In our experience, AMH two or three times the normal amount (10 ± 2.28 ng/mL), is a good indication of PCOS and infertility.

Setting research priorities to reduce global mortality from childhood pneumonia by 2015. - Rudan, Igor; El Arifeen, Shams; Bhutta, Zulfiqar A; Black, Robert E; Brooks, Abdullah; Chan, Kit Yee; Chopra, Mickey; Duke, Trevor; Marsh, David; Pio, Antonio; Simoes, Eric A F; Tamburlini, Giorgio; et al. (PLoS Med, 2011)
Keywords: Biomedical Research; Child, Preschool; Humans; Infant; Infant, Newborn; Pneumonia

Shwachman-Diamond syndrome and type 1 diabetes mellitus: more than a chance association? - Gana, S; Sainati, L; Frau, M R; Monciotti, C; Poli, F; Cannioto, Z; Comelli, M; Danesino, C; Minelli, A (Exp Clin Endocrinol Diabetes, 2011)
Shwachman-Diamond syndrome is a rare clinical condition consisting of exocrine pancreatic dysfunction, various degree of pancytopenia, and metaphyseal dysplasia. The majority of Shwachman-Diamond syndrome cases result from mutations in the Shwachman-Bodian-Diamond Syndrome gene. To date, type 1 diabetes mellitus has only been reported in 4 independent cases presenting with Shwachman-Diamond syndrome, 3 of them with molecular confirmation of the diagnosis. We describe 2 unrelated patients with clinical and molecular features typical of Shwachman-Diamond syndrome and type 1 diabetes mellitus. In addition, we report the occurrence rate of type 1 diabetes mellitus in the Italian registry for Shwachman-Diamond syndrome, which is low (3.23%) but increased at least 30-fold over the type 1 diabetes mellitus occurrence rate in the general population. No evidence of a direct correlation between Shwachman-Diamond syndrome and type 1 diabetes mellitus have been reported, therefore the presence of both diseases in the same patient might be a chance association, however we suggest that the defects in immune regulation of Shwachman-Diamond syndrome might play a role in the development of type 1 diabetes mellitus.
Keywords: Bone Marrow Diseases; CD4-CD8 Ratio; Diabetes Mellitus, Type 1; Exocrine Pancreatic Insufficiency; Female; Heterozygote; Humans; Immune System; Infant; Italy; Lipomatosis; Male; Mutation; Prevalence; Proteins; Registries

The significance of mannose-binding lectin gene polymorphisms on the risk of BK virus coinfection in women with human papillomavirus-positive cervical lesions. - Comar, Manola; Segat, Ludovica; Crovella, Sergio; Bovenzi, Massimo; Cortini, Enzo; Tognon, Mauro (Hum Immunol, 2011)
The simultaneous detection of oncogenic human papillomavirus (HPV) and BK virus (BKV) has been recently reported in cervical cancers, suggesting that these viruses may act together in the process of cell transformation; host genetic polymorphisms may also influence virus persistence/reactivation. To disclose a possible role of the gene encoding for the mannose-binding lectin, MBL2, in susceptibility to BKV infection, we analyzed functional polymorphisms in the first exon of MBL2 in women stratified for the presence/absence of BKV and affected by different grades of HPV-induced cervical precancerous lesions. All BKV-positive samples were also HPV positive (HPV 16), and all presented with high-grade squamous intraepithelial lesions. The MBL2 A allele was significantly more frequent in BKV-negative patients than in BKV-positive patients. These data indicate a possible role for the A allele in conferring protection to BKV infection in high-risk HPV-positive women (odds ratio 0.40, 95% confidence interval 0.20-0.85, p = 0.01).
Keywords: Adult; Aged; Alleles; BK Virus; Cervical Intraepithelial Neoplasia; Cervix Uteri; DNA Fingerprinting; DNA, Viral; Female; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Genotype; Human papillomavirus 16; Humans; Italy; Mannose-Binding Lectin; Odds Ratio; Papillomavirus Infections; Polymorphism, Genetic; Polyomavirus Infections; Risk; Viral Load

A singular case of congenital self-healing histiocytosis with skin, liver and atypical eye involvement. - Parentin, Fulvio; Ventura, Giovanna; Pastore, Serena; Kiren, Valentina; Bibalo, Chiara; Pensiero, Stefano; Lepore, Loredana (Ocul Immunol Inflamm, 2011)
PURPOSE: To describe a rare case of congenital self-healing Langerhans cell histiocytosis (CSHLCH) presenting with atypical eye involvement. DESIGN: Case report. METHODS: A female newborn presented with purpuric lesions over the trunk, limbs, and face. Liver ultrasonography revealed hypoechogenic lesions with blurred borders. Biomicroscopy showed right posterior synechiae with fibrinoid deposits on the lens. At 7 months she presented with right acute glaucoma. RESULTS: Biomicroscopy showed the presence of inflammatory pseudo-membrane covering the anterior surface of the lens, iris, and iridocorneal angle. Ab externo trabeculotomy was performed; access to the anterior chamber with capsulorrhexis forceps permitted a peeling of the pseudo-membrane with normalization of the intraocular pressure. Histologic examination of the membrane revealed an inflammatory tissue with CD1a and S-100 positive histiocytic cells. CONCLUSIONS: This is the first case of CSHLCH describing acute glaucoma secondary to a pseudo-inflammatory membrane with typical histiocytic cells, occluding the iridocorneal angle.
Keywords: Antigens, CD1; Convalescence; Female; Glaucoma; Histiocytosis, Langerhans-Cell; Humans; Infant; Intraocular Pressure; Iridocorneal Endothelial Syndrome; Liver Diseases; S100 Proteins; Skin Diseases; Trabeculectomy

Somatic mosaic IDH1 and IDH2 mutations are associated with enchondroma and spindle cell hemangioma in Ollier disease and Maffucci syndrome. - Pansuriya, Twinkal C; van Eijk, Ronald; d'Adamo, Pio; van Ruler, Maayke A J H; Kuijjer, Marieke L; Oosting, Jan; Cleton-Jansen, Anne-Marie; van Oosterwijk, Jolieke G; Verbeke, Sofie L J; Meijer, Daniëlle; van Wezel, Tom; Nord, Karolin H; et al. (Nat Genet, 2011)
Ollier disease and Maffucci syndrome are non-hereditary skeletal disorders characterized by multiple enchondromas (Ollier disease) combined with spindle cell hemangiomas (Maffucci syndrome). We report somatic heterozygous mutations in IDH1 (c.394C>T encoding an R132C substitution and c.395G>A encoding an R132H substitution) or IDH2 (c.516G>C encoding R172S) in 87% of enchondromas (benign cartilage tumors) and in 70% of spindle cell hemangiomas (benign vascular lesions). In total, 35 of 43 (81%) subjects with Ollier disease and 10 of 13 (77%) with Maffucci syndrome carried IDH1 (98%) or IDH2 (2%) mutations in their tumors. Fourteen of 16 subjects had identical mutations in separate lesions. Immunohistochemistry to detect mutant IDH1 R132H protein suggested intraneoplastic and somatic mosaicism. IDH1 mutations in cartilage tumors were associated with hypermethylation and downregulated expression of several genes. Mutations were also found in 40% of solitary central cartilaginous tumors and in four chondrosarcoma cell lines, which will enable functional studies to assess the role of IDH1 and IDH2 mutations in tumor formation.
Keywords: Adult; Case-Control Studies; Cell Line, Tumor; DNA Methylation; Enchondromatosis; Female; Gene Expression Profiling; Gene Expression Regulation; Genome-Wide Association Study; Humans; Isocitrate Dehydrogenase; Male; Middle Aged; Mosaicism; Mutation, Missense; Sequence Analysis, DNA; Transcription, Genetic; Young Adult

Splicing mutations in glycogen-storage disease type II: evaluation of the full spectrum of mutations and their relation to patients' phenotypes. - Zampieri, Stefania; Buratti, Emanuele; Dominissini, Silvia; Montalvo, Anna Lisa; Pittis, Maria Gabriela; Bembi, Bruno; Dardis, Andrea (Eur J Hum Genet, 2011)
Glycogen-storage disease type II is an autosomal recessive-inherited disorder due to the deficiency of acid α-glucosidase. A large number of mutations in the acid α-glucosidase gene have been described to date. Among them, ~15% are variations that may affect mRNA splicing process. In this study, we have for the first time comprehensively reviewed the available information on splicing mutations of the acid α-glucosidase gene and we have evaluated their possible impact on the splicing process using different in silico approaches. Out of the 39 different GAA-sequence variations described, an in silico analysis using seven different programs showed that 97% of them are predicted to have an impact on the splicing process. Moreover, this analysis showed a quite good correlation between the impact of the mutation on the splicing process and the clinical phenotype. In addition, we have performed the functional characterization of three novel sequence variants found in Italian patients and still uncharacterized. Using a minigene system, we have confirmed their pathogenic nature. In conclusion, this study has shown that in silico analysis represents a useful tool to select mutations that affect the splicing process of the acid α-glucosidase gene and provides an updated picture of all this kind of mutations reported till now.
Keywords: alpha-Glucosidases; Cell Line; Computational Biology; DNA Mutational Analysis; Exons; Glycogen Storage Disease Type II; Humans; Introns; Mutagenesis, Site-Directed; Mutation; Phenotype; RNA Splicing

Successful treatment of acne with isotretinoin in chronic granulomatous disease. - Barbi, Egidio; Berti, Irene; Minute, Marta; Zennaro, Floriana (Eur J Dermatol, 2011)
Keywords: Acne Vulgaris; Adult; Anti-Infective Agents; Comorbidity; Dermatologic Agents; Granulomatous Disease, Chronic; Humans; Isotretinoin; Male; Trimethoprim-Sulfamethoxazole Combination

Tag-single nucleotide polymorphism-based human leukocyte antigen genotyping in celiac disease patients from northeastern Italy. - Vatta, Serena; Fabris, Annalisa; Segat, Ludovica; Not, Tarcisio; Crovella, Sergio (Hum Immunol, 2011)
We genotyped celiac disease (CD)-associated haplotypes DQ2.5, DQ8, DQ2.2, and DQ7 in 1005 CD patients from North Eastern Italy using a Tag-single nucleotide polymorphism (SNPs) approach and real time PCR platform, checking the accuracy and reliability of the method and comparing it to traditional PCR-SSP. Only 14 of 2010 chromosomes analyzed (0.7%) showed discrepancies between the Tag-SNPs real-time polymerase chain reaction (PCR) method and the PCR-single-strand polymorphism (SSP) technique, indicating a high sensitivity and specificity (ranging from 0.987 to 1 and from 0.998 to 0.999, respectively) for tagging with respect to corresponding human leukocyte antigen (HLA) alleles identified by PCR-SSP. Moreover, the overall cost of the Tag-SNPs HLA typing method was low (3 to 4 €/sample instead of 35 to 70 €/sample with commercial kits), making it suitable for mass screenings. Hence, we believe that the Tag-SNPs HLA typing could be used to complement or replace classic HLA typing in at high-risk groups, for research purposes and eventually in population screening programs.
Keywords: Adolescent; Adult; Aged; Celiac Disease; Child; Child, Preschool; Female; Humans; Infant; Italy; Male; Mass Screening; Middle Aged; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity

Therapeutic approaches in the treatment of juvenile dermatomyositis in patients with recent-onset disease and in those experiencing disease flare: an international multicenter PRINTO study. - Hasija, Rachana; Pistorio, Angela; Ravelli, Angelo; Demirkaya, Erkan; Khubchandani, Raju; Guseinova, Dinara; Malattia, Clara; Canhao, Helena; Harel, Liora; Foell, Dirk; Wouters, Carine; De Cunto, Carmen; et al. (Arthritis Rheum, 2011)
OBJECTIVE: To evaluate response to therapy over a 24-month period in a large prospective international cohort of patients with juvenile dermatomyositis (DM). METHODS: The study included 145 patients with recent-onset juvenile DM and 130 juvenile DM patients experiencing disease flare, all of whom were <18 years old. Disease activity parameters and therapeutic approaches in 4 geographic areas were analyzed at baseline and at 6, 12, and 24 months. Response was assessed according to the Pediatric Rheumatology International Trials Organization (PRINTO) juvenile DM response criteria, and data were reported "as observed" and in the intent-to-treat (ITT) population. RESULTS: Patients with recent-onset juvenile DM at baseline had higher baseline disease activity and greater improvement over 24 months when compared to juvenile DM patients experiencing disease flare at baseline. Methotrexate (MTX) or high-dose corticosteroids were administered more frequently to patients with recent-onset juvenile DM, compared to juvenile DM patients experiencing disease flare, who were more likely to receive cyclosporine. Compared to patients from Western and Eastern Europe, a higher proportion of patients from South and Central America and North America received pulse steroids, and the average steroid dosage was higher in the North American and South and Central American patients. The use of MTX was similar in all 4 regions, while cyclosporin A was more frequently used in Western Europe. In the "as observed" analysis, 57.9% of the patients with recent-onset juvenile DM and 36.4% of the patients experiencing disease flare (P<0.001) reached at least a 70% response by PRINTO criteria at 6 months; these proportions had increased at month 24 to 78.4% and 51.2%, respectively (P<0.001). Corresponding results of the ITT analysis were much lower, with only one-third of the patients able to maintain the initial assigned therapy over 24 months. CONCLUSION: Patients with recent-onset juvenile DM are more likely to achieve significant clinical improvement over 24 months, when compared to patients experiencing flares of juvenile DM. Internationally, various therapeutic approaches are used to treat this disease.
Keywords: Adolescent; Adrenal Cortex Hormones; Child; Dermatologic Agents; Dermatomyositis; Female; Humans; Longitudinal Studies; Male; Methotrexate; Prospective Studies; Treatment Outcome

Thromboembolism in pediatric inflammatory bowel disease: systematic review. - Lazzerini, Marzia; Bramuzzo, Matteo; Maschio, Massimo; Martelossi, Stefano; Ventura, Alessandro (Inflamm Bowel Dis, 2011)
BACKGROUND: Several studies suggest an increased risk of venous and arterial thromboembolism (TE) in adults with inflammatory bowel disease (IBD) compared to the general population. We performed a systematic review of studies on incidence and characteristic of TE in children with IBD. METHODS: We searched Medline, LILACS, EMBASE, POPLINE, CINHAL, and reference lists of identified articles, without language restrictions, in August 2010. RESULTS: Population studies suggest that there is an increased risk of TE in children with IBD compared to controls. TE occurred in children with IBD in all age ranges, mostly (82.8%) during active disease, and more frequently in children with ulcerative colitis (odds ratio [OR] 3.7, 95% confidence interval [CI] 1.8-7.6). At least one specific risk factor for TE was recognized in 50% of cases; two risk factors were present in 24%. Out of 92 published cases of TE in children with IBD, 54.3% occurred in cerebral site, 26% in the limbs, 13% in the abdominal vessels, and the remaining in the retina and lungs. After a first episode of TE, an early recurrence was observed in 11.4% of children, a late recurrence in 10%. A number of different therapeutic schemes were used. Overall mortality was 5.7% and was mostly associated with cerebral TE. CONCLUSIONS: Population studies are needed to clarify the risk of TE in children with IBD, the relative weight of other risk factors, the characteristics of the events, and to define guidelines of therapy and prophylaxis.
Keywords: Adult; Child; Humans; Inflammatory Bowel Diseases; Risk Factors; Thromboembolism

Trail down-regulates the release of osteoprotegerin (OPG) by primary stromal cells. - Corallini, Federica; Celeghini, Claudio; Rimondi, Erika; di Iasio, Maria Grazia; Gonelli, Arianna; Secchiero, Paola; Zauli, Giorgio (J Cell Physiol, 2011)
The soluble member of the TNF-R superfamily osteoprotegerin (OPG) is abundantly released under basal conditions by both mesenchymal stem cells (MSC) and fibroblasts and by endothelial cells upon stimulation with inflammatory cytokines. Since MSC, fibroblasts and endothelial cells represent key elements of the normal and tumor microenvironment and express detectable levels of surface TRAIL receptors, we investigated the effect of TRAIL on OPG release. Unexpectedly, recombinant TRAIL decreased the spontaneous OPG release in all cell types examined. Moreover, TRAIL decreased OPG release also in stromal cells co-cultured with lymphoma cells and counteracted the OPG induction by TN-alpha in HUVEC and MSC. Such down-regulation was not due to a masking effect in the ELISA quantification of the OPG released in the culture supernatants due to binding of OPG to its ligands (TRAIL and RANKL), as demonstrated by competition experiments with recombinant TRAIL and by the lack of RANKL release/induction. In addition, OPG down-regulation was not due to induction of cytotoxic effects by TRAIL, since the degree of apoptosis in response to TRAIL was negligible in all primary cell types. With regards to the possible molecular mechanism accounting for the down-regulation of OPG release by TRAIL, we found that treatment of MSC with TRAIL significantly decreased the phosphorylation levels of p38/MAPK. There is a suggestion that this pathway is involved in the stabilization of OPG mRNA. In this respect, the ability of TRAIL to decrease the release of OPG, in the absence of cell cytotoxicity, was mimicked by the p38/MAPK inhibitor SB203580.
Keywords: Bone Marrow Cells; Cell Death; Cells, Cultured; Coculture Techniques; Down-Regulation; Endothelial Cells; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Humans; MAP Kinase Signaling System; Mesenchymal Stromal Cells; Osteoprotegerin; p38 Mitogen-Activated Protein Kinases; Protein Binding; Recombinant Proteins; Stromal Cells; TNF-Related Apoptosis-Inducing Ligand; Tumor Necrosis Factor-alpha

TRAIL promotes a pro-survival signal in erythropoietin-deprived human erythroblasts through the activation of an NF-kB/IkBalpha pathway. - Sancilio, S; Di Giacomo, V; Quaglietta, A M; Iacone, A; Angelucci, D; Tatasciore, U; Rana, R A; Cataldi, A; Zauli, G; Di Pietro, R (J Biol Regul Homeost Agents, 2011)
The biological activity of TNF-related apoptosis inducing ligand (TRAIL) was analyzed in primary human erythroblasts derived from mononuclear cells of blood donors, kept in culture in the presence of 20 percent foetal calf serum, growth factors (EPO, SCF, IL-3) and glucocorticoids (10-6 M dexamethasone, 10-6 M oestradiol) or under growth factor and serum starvation. In the presence of growth factors and serum, primary erythroblasts showed a differential expression of TRAIL-Receptors (Rs) at various degrees of maturation and responded to TRAIL treatment with a mild cytotoxicity. On the other hand, in the absence of serum and growth factors, TRAIL treatment unexpectedly up-regulated TRAIL-R4 decoy receptor and promoted erythroblast survival. The concomitant activation of NF-kB/IkB survival pathway was detected with Western blotting and immunofluorescence procedures and confirmed by experiments performed with SN50, a pharmacological inhibitor of the NF-kB/IkB pathway. Our study indicates that TRAIL has a twofold activity on erythroid lineages: it induces a mild erythroid cell cytotoxicity in the presence of serum and growth factors, while it promotes erythroid cell survival through the activation of the NF-kB/IkB pathway under starvation conditions.
Keywords: Cell Survival; Erythroblasts; Erythropoietin; Gene Expression Regulation; Humans; I-kappa B Kinase; Jurkat Cells; NF-kappa B; Peptides; TNF-Related Apoptosis-Inducing Ligand; Tumor Necrosis Factor Decoy Receptors

Two lumens, one diagnosis. - Pastore, Serena; Gortani, Giulia; Maschio, Massimo; Di Leo, Grazia; Ventura, Alessandro (J Pediatr, 2011)
Keywords: Capsule Endoscopy; Child, Preschool; Gastrointestinal Hemorrhage; Humans; Male; Meckel Diverticulum; Recurrence

An unusual case of buccal obstruction: the antrochoanal polyp. - De Cunto, Angela; Minen, Federico (Arch Dis Child, 2011)
Keywords: Adolescent; Cheek; Female; Humans; Mouth Diseases; Nasal Obstruction; Nasal Polyps; Oropharynx

Use of zidovudine-sparing HAART in pregnant HIV-infected women in Europe: 2000-2009. - Tariq, Shema; Townsend, Claire L; Cortina-Borja, Mario; Duong, Trinh; Elford, Jonathan; Thorne, Claire; Tookey, Pat A (J Acquir Immune Defic Syndr, 2011)
BACKGROUND: Increasing numbers of women in resource-rich settings are prescribed zidovudine (ZDV)-sparing highly active antiretroviral therapy (HAART) in pregnancy. We compare ZDV-sparing with ZDV-containing HAART in relation to maternal viral load at delivery, mother-to-child transmission (MTCT) of HIV, and congenital abnormality. METHODS: This is an analysis of data from the National Study of HIV in Pregnancy and Childhood and the European Collaborative Study. Data on 7573 singleton births to diagnosed HIV-infected women between January 2000 and June 2009 were analyzed. Logistic regression models were fitted to estimate adjusted odds ratios (AORs). RESULTS: Overall, 15.8% (1199 of 7573) of women received ZDV-sparing HAART, with increasing use between 2000 and 2009 (P < 0.001). Nearly a fifth (18.4%) of women receiving ZDV-sparing HAART in pregnancy had a detectable viral load at delivery compared with 28.6% of women on ZDV-containing HAART [AOR 0.90; 95% confidence interval (CI): 0.72 to 1.14, P = 0.4]. MTCT rates were 0.8% and 0.9% in the ZDV-sparing and ZDV-containing groups, respectively (AOR 1.81; 95% CI: 0.77 to 4.26, P = 0.2). The congenital abnormality rate was the same in both groups (2.7%, AOR 0.98; 95% CI: 0.66 to 1.45, P = 0.9), with no significant difference between the groups in a subanalysis of pregnancies with first trimester HAART exposure (AOR 0.79; 95% CI: 0.48 to 1.30, P = 0.4). CONCLUSIONS: We found no difference in risk of detectable viral load at delivery, MTCT, or congenital abnormality when comparing ZDV-sparing with ZDV-containing HAART. With increasing use of ZDV-sparing HAART, continued monitoring of pregnancy outcomes and long-term consequences of in utero exposure to these drugs is required.
Keywords: Adult; Anti-HIV Agents; Antiretroviral Therapy, Highly Active; CD4 Lymphocyte Count; Europe; Female; HIV Infections; Humans; Infant; Infectious Disease Transmission, Vertical; Pregnancy; Pregnancy Complications, Infectious; Retrospective Studies; Time Factors; Viral Load; Zidovudine

Usefulness of wireless capsule endoscopy for detecting inflammatory bowel disease in children presenting with arthropathy. - Taddio, Andrea; Simonini, Gabriele; Lionetti, Paolo; Lepore, Loredana; Martelossi, Stefano; Ventura, Alessandro; Cimaz, Rolando (Eur J Pediatr, 2011)
Inflammatory bowel disease (IBD) is a cause of chronic intestinal inflammation in children. In a subset of patients affected by IBD, arthropathy may be the leading presenting sign. In the past years, remarkable advances in gastrointestinal endoscopy techniques have been achieved; recently, the development of capsule endoscopy (CE) provided a non-invasive method for the complete endoscopic evaluation, including small bowel assessment. We report three children suffering from IBD but presenting with articular complaints in whom CE was a useful tool for detecting gut inflammation. Patients were investigated with the wireless CE: PillCam SB2 (Given Imaging, Yoqneam, Israel) capsule, the second-generation capsule, was used in our paediatric patients. Three patients were initially evaluated for arthropathy. Enteropathic arthritis was suspected for gastrointestinal symptoms and/or persistence of inflammatory markers elevation. In one of these children, conventional endoscopy was refused by parents, while in the other two children, CE was proposed as first-line diagnostic tool. In all patients, CE revealed to be safe and provided information that led to diagnosis. Paediatric rheumatologists should consider CE as a valid, non-invasive tool, eventually first level diagnostic approach in order to evaluate the presence of IBD in children presenting with chronic articular complaints.
Keywords: Adolescent; Arthritis, Juvenile; Capsule Endoscopy; Child; Colitis, Ulcerative; Colon; Crohn Disease; Diagnosis, Differential; Follow-Up Studies; Humans; Inflammatory Bowel Diseases; Intestine, Small; Male; Predictive Value of Tests; Sensitivity and Specificity; Severity of Illness Index; Treatment Outcome

Vertebral defects in patients with Peters plus syndrome and mutations in B3GALTL. - Faletra, Flavio; Athanasakis, Emmanouil; Minen, Federico; Fornasier, Federico; Marchetti, Federico; Gasparini, Paolo (Ophthalmic Genet, 2011)
Keywords: Abnormalities, Multiple; Alternative Splicing; Child, Preschool; Cleft Lip; Consanguinity; Cornea; Female; Galactosyltransferases; Glucosyltransferases; Growth Disorders; Humans; Limb Deformities, Congenital; Point Mutation; Spine

X-ray fluorescence elemental mapping and microscopy to follow hepatic disposition of a Gd-based magnetic resonance imaging contrast agent. - Delfino, Riccarda; Altissimo, Matteo; Menk, Ralf Hendrik; Alberti, Roberto; Klatka, Tomasz; Frizzi, Tommaso; Longoni, Antonio; Salomè, Murielle; Tromba, Giuliana; Arfelli, Fulvia; Clai, Milan; Vaccari, Lisa; et al. (Clin Exp Pharmacol Physiol, 2011)
1. Spatially resolved X-ray fluorescence (XRF) spectroscopy with synchrotron radiation is a technique that allows imaging and quantification of chemical elements in biological specimens with high sensitivity. In the present study, we applied XRF techniques at a macro and micro level to carry out drug distribution studies on ex vivo models to confirm the hepatobiliary disposition of the Gd-based magnetic resonance imaging contrast agent B22956/1. 2. Gd presence was selectively quantified allowing the determination of the time dependent disappearance of the drug from blood and its hepatic accumulation in mice after administration. Elemental mapping highlighted the drug distribution differences between healthy and diseased livers. XRF microanalyses showed that in CCl(4) -induced hepatitis, B22956/1 has greatly reduced hepatic accumulation, shown as a 20-fold reduction of Gd presence. Furthermore, a significant increase of Fe presence was found in steatotic compared with healthy livers, in line with the disease features. 3. The present results show that XRF might be useful in preclinical pharmacological studies with drugs containing exogenous elements. Furthermore, quantitative and high-sensitivity elemental mapping allows simultaneous detection of chemical variation, showing pathological conditions. This approach was useful in suggesting reduced B22956/1 accumulation in steatotic livers, thus opening possible new diagnostic perspectives for this drug.
Keywords: Animals; Contrast Media; Fatty Liver; Female; Gadolinium; Hepatitis; Iron; Liver; Magnetic Resonance Imaging; Mice; Mice, Inbred CBA; Organometallic Compounds; Spectrometry, X-Ray Emission

A 3'UTR SNP in NLRP3 gene is associated with susceptibility to HIV-1 infection. - Pontillo, Alessandra; Brandão, Lucas A; Guimarães, Rafael L; Segat, Ludovica; Athanasakis, Emmanouil; Crovella, Sergio (J Acquir Immune Defic Syndr, 2010)
OBJECTIVES: Innate immunity genes polymorphisms are known to be involved in the multifactorial susceptibility to HIV-1 infection. Recently it has been hypothesized that inflammasomes could play an important role in the host response to viruses. The aim of our study is to verify if single-nucleotide polymorphisms (SNPs) in genes encoding for NALPs-innate immune receptors that form molecular complexes leading to the production of IL-1beta and the activation of immune response-could influence the individual susceptibility to HIV-1. DESIGN: We performed an association study analyzing 2 NLRP1 and NLRP3 SNPs in HIV-1 vertically infected Brazilian children (n = 135), HIV-1-infected Brazilain adults (n = 192) and HIV-1-positive Italian seropositive subjects (n = 192). RESULTS: The 3'UTR NLRP3 rs10754558 SNP was associated with HIV-1 infection in all the studied groups. The frequency of rs10754558 G allele was differently distributed within seropositive subjects (HIV+) and controls, and in particular the GG genotype was less frequent in HIV+. CONCLUSIONS: susceptibility to HIV-1 infection is associated with a 3'UTR NLRP3 polymorphism. This is the first report linking SNPs in the NALPs with HIV-1 infection, and further epidemiologic and functional studies are needed to deeper investigate the role of inflammasome in the susceptibility to HIV-1 infection.
Keywords: Adult; Brazil; Carrier Proteins; Case-Control Studies; Child; Child, Preschool; Female; Genetic Predisposition to Disease; Genotype; HIV Infections; HIV-1; Humans; Infant; Infectious Disease Transmission, Vertical; Male; Middle Aged; Polymorphism, Single Nucleotide; Pregnancy; Young Adult

Abatacept improves health-related quality of life, pain, sleep quality, and daily participation in subjects with juvenile idiopathic arthritis. - Ruperto, Nicolino; Lovell, Daniel J; Li, Tracy; Sztajnbok, Flavio; Goldenstein-Schainberg, Claudia; Scheinberg, Morton; Penades, Inmaculada Calvo; Fischbach, Michael; Alcala, Javier Orozco; Hashkes, Philip J; Hom, Christine; Jung, Lawrence; et al. (Arthritis Care Res (Hoboken), 2010)
OBJECTIVE: To assess health-related quality of life (HRQOL) in abatacept-treated children/adolescents with juvenile idiopathic arthritis (JIA). METHODS: In this phase III, double-blind, placebo-controlled trial, subjects with active polyarticular course JIA and an inadequate response/intolerance to ≥1 disease-modifying antirheumatic drug (including biologics) received abatacept 10 mg/kg plus methotrexate (MTX) during the 4-month open-label period (period A). Subjects achieving the American College of Rheumatology Pediatric 30 criteria for improvement (defined "responders") were randomized to abatacept or placebo (plus MTX) in the 6-month double-blind withdrawal period (period B). HRQOL assessments included 15 Child Health Questionnaire (CHQ) health concepts plus the physical (PhS) and psychosocial summary scores (PsS), pain (100-mm visual analog scale), the Children's Sleep Habits Questionnaire, and a daily activity participation questionnaire. RESULTS: A total of 190 subjects from period A and 122 from period B were eligible for analysis. In period A, there were substantial improvements across all of the CHQ domains (greatest improvement was in pain/discomfort) and the PhS (8.3 units) and PsS (4.3 units) with abatacept. At the end of period B, abatacept-treated subjects had greater improvements versus placebo in all domains (except behavior) and both summary scores. Similar improvement patterns were seen with pain and sleep. For participation in daily activities, an additional 2.6 school days/month and 2.3 parents' usual activity days/month were gained in period A responders with abatacept, and further gains were made in period B (1.9 versus 0.9 [P = 0.033] and 0.2 versus -1.3 [P = 0.109] school days/month and parents' usual activity days/month, respectively, in abatacept- versus placebo-treated subjects). CONCLUSION: Improvements in HRQOL were observed with abatacept, providing real-life tangible benefits to children with JIA and their parents/caregivers.
Keywords: Adolescent; Arthritis, Juvenile; Child; Double-Blind Method; Female; Health Status; Humans; Immunoconjugates; Male; Pain; Quality of Life; Questionnaires; Sleep Stages

Acute respiratory failure in a child after talc inhalation. - Patarino, Federica; Norbedo, Stefania; Barbi, Egidio; Poli, Furio; Furlan, Stefano; Savron, Fabio (Respiration, 2010)
Keywords: Female; Humans; Infant; Inhalation Exposure; Respiratory Insufficiency; Talc

An alternative role of C1q in cell migration and tissue remodeling: contribution to trophoblast invasion and placental development. - Agostinis, Chiara; Bulla, Roberta; Tripodo, Claudio; Gismondi, Angela; Stabile, Helena; Bossi, Fleur; Guarnotta, Carla; Garlanda, Cecilia; De Seta, Francesco; Spessotto, Paola; Santoni, Angela; Ghebrehiwet, Berhane; et al. (J Immunol, 2010)
Fetal trophoblast cells invading the decidua in the early phase of pregnancy establish complex interaction with the maternal extracellular matrix. We discovered that C1q was widely distributed in human decidual stroma in the absence of C4 and C3 and was actively synthesized by migrating extravillous trophoblasts. The cells expressed the messages for the three chains of C1q and secreted this complement component that interacted with the proteins of the decidual extracellular matrix. Solid phase-bound C1q promoted trophoblast adhesion and migration, and cell binding to C1q resulted in activation of ERK1/2 MAPKs. Ab inhibition experiments showed that the receptors for the globular head of C1q/p33 and α(4)β(1) integrin were both involved in this process and were colocalized on the cell surface following binding of C1q to trophoblasts. We also found that C1q(-/-) mice manifested increased frequency of fetal resorption, reduced fetal weight, and smaller litter sizes compared with wild-type mice. C1q deficiency was associated with impaired labyrinth development and decidual vessel remodeling. Collectively, these data suggest that C1q plays an important role in promoting trophoblast invasion of decidua and that defective local production of C1q may be involved in pregnancy disorders, such as pre-eclampsia, characterized by poor trophoblast invasion.
Keywords: Animals; Cell Adhesion; Chemotaxis, Leukocyte; Complement C1q; Female; Humans; Immunoblotting; Immunohistochemistry; Immunoprecipitation; Mice; Mice, Inbred C57BL; Microscopy, Confocal; Placentation; Pre-Eclampsia; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; Trophoblasts

Analysis of DEFB1 regulatory SNPs in cystic fibrosis patients from North-Eastern Italy. - Segat, L; Morgutti, M; Athanasakis, E; Trevisiol, C; Amaddeo, A; Poli, F; Crovella, S (Int J Immunogenet, 2010)
Cystic fibrosis (CF) transmembrane regulator protein (CFTR) gene is undoubtedly the main genetic factor involved in the modulation of CF phenotype. However, other factors such as human defensins and the genes encoding for these antimicrobial peptides have been hypothesized as possible modifiers influencing airways infection in CF patients, but their role in the pathogenesis of lung disease is still debated. Since DEFB1 gene encoding for human beta-defensin 1 displays features such as antimicrobial or chemotactic activity playing a role in inflammation, it has been considered as a possible candidate CF modifier gene. We analysed three single nucleotide polymorphisms (SNPs) in the 5'-untranslated region of the DEFB1 gene (namely g-52G>A, g-44C>G and g-20G>A) in a group of 62 CF patients from North Eastern Italy, and in 130 healthy controls, with the aim of verifying the possible association of these functional SNPs with the pulmonary phenotype of CF patients. DEFB1 SNPs have been genotyped by using Taqman allele-specific fluorescent probes and a real-time PCR platform. No significant differences were found for allele, genotype and haplotype frequencies of DEFB1 g-52G>A, g-44C>G and g-20G>A SNPs in CF patients stratified for Pseudomonas aeruginosa infection, as well as in patients with a severe and mild clinical phenotype or in patients stratified for CFTR genotypes. DEFB1 allele, genotype and haplotype frequencies of CF patients globally considered were similar to those of healthy controls. Our findings are discordant with respect to another recent study performed on CF patients coming from Southern Italy, probably due to different ethnicity of the patients.
Keywords: 5' Untranslated Regions; Adolescent; Alleles; beta-Defensins; Case-Control Studies; Child; Child, Preschool; Chronic Disease; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Female; Gene Frequency; Genotype; Haplotypes; Humans; Immunity, Innate; Infant; Infant, Newborn; Italy; Male; Polymorphism, Single Nucleotide; Pseudomonas Infections

Anti transglutaminase antibodies cause ataxia in mice. - Boscolo, Sabrina; Lorenzon, Andrea; Sblattero, Daniele; Florian, Fiorella; Stebel, Marco; Marzari, Roberto; Not, Tarcisio; Aeschlimann, Daniel; Ventura, Alessandro; Hadjivassiliou, Marios; Tongiorgi, Enrico (PLoS One, 2010)
BACKGROUND: Celiac disease (CD) is an autoimmune gastrointestinal disorder characterized by the presence of anti-transglutaminase 2 (TG2) and anti-gliadin antibodies. Amongst the neurological dysfunctions associated with CD, ataxia represents the most common one. METHODS: We analyzed by immunohistochemistry, the anti-neural reactivity of the serum from 20 CD patients. To determine the role of anti-TG2 antibodies in ataxia, two anti-TG2 single chain variable fragments (scFv), isolated from a phage-display IgA antibody library, were characterized by immunohistochemistry and ELISA, and injected in mice to study their effects on motor coordination. We found that 75% of the CD patient population without evidence of neurological involvement, has circulating anti-neural IgA and/or IgG antibodies. Two anti-TG2 scFvs, cloned from one CD patient, stained blood vessels but only one reacted with neurons. This anti-TG2 antibody showed cross reactivity with the transglutaminase isozymes TG3 and TG6. Intraventricular injection of the anti-TG2 or the anti-TG2/3/6 cross-reactive scFv provoked transient, equally intensive ataxia in mice. CONCLUSION: The serum from CD patients contains anti-TG2, TG3 and TG6 antibodies that may potentially cause ataxia.
Keywords: Adult; Animals; Antibodies; Ataxia; Autoimmune Diseases; Brain; Celiac Disease; Female; Gliadin; Humans; Isoenzymes; Male; Mice; Mice, Inbred C57BL; Middle Aged; Motor Skills; Rats; Rats, Sprague-Dawley; Transglutaminases

Antimicrobial plant peptides isolated from native and crop species in Brazil: development of new drugs. - Crovella, Sergio (Curr Protein Pept Sci, 2010)
Keywords: Antimicrobial Cationic Peptides; Biodiversity; Brazil; Crops, Agricultural; Drug Discovery; Genomics; Plants

Beta defensin-1 gene (DEFB1) polymorphisms are not associated with atopic dermatitis in children and adolescents from northeast Brazil (Recife, Pernambuco). - Segat, Ludovica; Guimarães, Rafael L; Brandão, Lucas A C; Rocha, Cintia R C; Zanin, Valentina; Trevisiol, Chiara; de Lima Filho, José Luiz; Crovella, Sergio (Int J Dermatol, 2010)
BACKGROUND: Atopic dermatitis (AD) is a common inflammatory skin disease resulting from the interplay between environmental, immunological and genetic factors. In our study, we investigated the role of three single nucleotide polymorphisms (SNPs) at 5'-UTR of DEFB1 gene, encoding for the human beta defensin-1, on the susceptibility to develop AD in a group of Brazilian children and adolescents. METHODS: Three SNPs, -20 G/A (rs11362), -44 C/G (rs1800972), and -52 G/A (rs1799946) at 5'-UTR of DEFB1 gene were genotyped in two groups of children and adolescents, one affected by AD (96 subjects), the other healthy (191 individuals), from northeast Brazil. RESULTS: -44 C/G frequencies were comparable between the two groups. The -20 GG genotype was more frequent in AD subjects than in healthy controls; the -52 GG, conversely, was more frequent in healthy controls than in AD. However, both these differences did not reach statistical significance. Also, association between SNPs and AD severity has been shown. The analysis of DEFB1 haplotypes did not highlight any association of the three SNPs with AD development or disease severity. CONCLUSIONS: Our results seem to exclude a role for the -44 C/G DEFB1 SNPs on the pathogenesis and severity of AD, while for the -20 C/G and -52 G/A, even if not statistically significant, we evidenced a slight trend for susceptibility (-20 GG) and protection (-52 GG) for the development of AD. However, as controversial findings have been reported in the literature, the role of DEFB1 in the development of AD and in the severity of the phenotype deserves further investigation.
Keywords: Adolescent; beta-Defensins; Brazil; Child; Child, Preschool; Dermatitis, Atopic; Female; Gene Frequency; Genetic Predisposition to Disease; Haplotypes; Humans; Infant; Male; Polymorphism, Single Nucleotide; Risk Factors

A case of lymphedema-distichiasis syndrome carrying a new de novo frameshift FOXC2 mutation. - Fabretto, Antonella; Shardlow, Alison; Faletra, Flavio; Lepore, Loredana; Hladnik, Uros; Gasparini, Paolo (Ophthalmic Genet, 2010)
PURPOSE: Lymphedema-Distichiasis (LD, OMIM 153400) is an autosomal dominant disorder with variable expression. The mutated gene implicated is FOXC2, which encodes for a forkhead transcription factor involved in the development of the lymphatic and vascular system. LD is characterized by late childhood or pubertal onset lymphedema of the limbs and distichiasis. Other associations have been reported, including congenital heart disease, ptosis, scoliosis. CONCLUSIONS: Here we describe a case of LD carrying a de novo frameshift mutation of FOXC2 who presented a prepubertal onset of lower limbs lymphedema and mild distichiasis associated with other anomalies such as webbing neck and ptosis.
Keywords: Abnormalities, Multiple; Adolescent; Eye Abnormalities; Eyelashes; Face; Forkhead Transcription Factors; Frameshift Mutation; Humans; Lymphedema; Male; Syndrome

A child with pain after mild trauma. - Londero, Margherita; Pastore, Serena; Zanazzo, Giulio A; Bruno, Irene; Ventura, Alessandro (J Pediatr, 2010)
Keywords: Antigens, CD; Antigens, CD31; Antigens, CD34; Antigens, Differentiation, Myelomonocytic; Biopsy; Child; Factor VIII; Fingers; Hand Injuries; Hemangioendothelioma; Humans; Immunohistochemistry; Injury Severity Score; Male; Osteolysis; Pain; Pain Measurement; S100 Proteins; Vascular Neoplasms

Circulating levels of frizzled-related protein (FRZB) are increased in patients with early rheumatoid arthritis and decrease in response to disease-modifying antirheumatic drugs. - Corallini, Federica; Secchiero, Paola; Castellino, Gabriella; Montecucco, Maurizio; Trotta, Francesco; Zauli, Giorgio (Ann Rheum Dis, 2010)
Keywords: Antirheumatic Agents; Arthritis, Rheumatoid; Biological Markers; Glycoproteins; Humans

Corticosteroids do not cause harmful increase of viral load in severe H1N1 virus infection. - Confalonieri, Marco; D'Agaro, Pierlanfranco; Campello, Cesare (Intensive Care Med, 2010)
Keywords: Adult; Antiviral Agents; Bronchoalveolar Lavage Fluid; Extracorporeal Membrane Oxygenation; Glucocorticoids; Humans; Influenza A Virus, H1N1 Subtype; Influenza, Human; Male; Methylprednisolone; Oseltamivir; Respiration, Artificial; Severity of Illness Index; Viral Load; World Health Organization

Declining HCV seroprevalence in pregnant women with HIV. - Floridia, M; Tamburrini, E; Anzidei, G; Tibaldi, C; Muggiasca, M L; Guaraldi, G; Fiscon, M; Vimercati, A; Martinelli, P; Donisi, A; Dalzero, S; Ravizza, M (Epidemiol Infect, 2010)
We assessed recent trends in hepatitis C virus (HCV) prevalence in pregnant women with HIV using data from a large national study. Based on 1240 pregnancies, we observed a 3.4-fold decline in HCV seroprevalence in pregnant women with HIV between 2001 (29.3%) and 2008 (8.6%). This decline was the net result of two components: a progressively declining HCV seroprevalence in non-African women (from 35.7% in 2001 to 16.7% in 2008), sustained by a parallel reduction in history of injecting drug use (IDU) in this population, and a significantly growing presence (from 21.2% in 2001 to 48.6% in 2008) of women of African origin, at very low risk of being HCV-infected [average HCV prevalence 1%, adjusted odds ratio (aOR) for HCV 0.09, 95% CI 0.03-0.29]. Previous IDU was the stronger determinant of HCV co-infection in pregnant women with HIV (aOR 30.9, 95% CI 18.8-51.1). The observed trend is expected to translate into a reduced number of cases of vertical HCV transmission.
Keywords: Chi-Square Distribution; Female; Hepatitis C; HIV Infections; Humans; Italy; Logistic Models; Pregnancy; Risk Factors; Seroepidemiologic Studies

Decreased cholesterol levels reflect a consumption of anti-inflammatory isoprenoids associated with an impaired control of inflammation in a mouse model of mevalonate kinase deficiency. - Marcuzzi, Annalisa; Decorti, Giuliana; Pontillo, Alessandra; Ventura, Alessandro; Tommasini, Alberto (Inflamm Res, 2010)
OBJECTIVE: The aim of this study was to evaluate, in a mouse model of mevalonate kinase deficiency (MKD), the possible link between inflammatory symptoms and serum cholesterol levels. MATERIALS AND METHODS: Balb/c mice were treated with alendronate and bacterial muramyl dipeptide. Body temperature, interleukin-1 beta (IL-1 beta) secretion and serum cholesterol levels were measured. RESULTS: An increased production of the pro-inflammatory cytokine IL-1 beta (p < 0.05) and a rise in body temperature (p < 0.05) was observed, while, in parallel, serum cholesterol concentration significantly decreased (p < 0.05). These effects were completely reversed when animals were treated with exogenous isoprenoids. CONCLUSIONS: In the mouse model of MKD, the inflammatory response is associated with a reduction in cholesterol levels, and hence this parameter could be used as an indicator of isoprenoid consumption. In addition, plant derived isoprenoids could represent candidate treatments for this disease.
Keywords: Animals; Anti-Inflammatory Agents; Cholesterol; Disease Models, Animal; Inflammation; Interleukin-1beta; Male; Mevalonate Kinase Deficiency; Mice; Mice, Inbred BALB C; Random Allocation; Terpenes

Defective FOXP3 expression in patients with acute Kawasaki disease and restoration by intravenous immunoglobulin therapy. - Olivito, Biagio; Taddio, Andrea; Simonini, Gabriele; Massai, Cristina; Ciullini, Sara; Gambineri, Eleonora; de Martino, Maurizio; Azzari, Chiara; Cimaz, Rolando (Clin Exp Rheumatol, 2010)
OBJECTIVES: The aims of this study were: 1) to investigate forkhead box P3 (FOXP3) expression in patients with Kawasaki disease (KD), exploring possible differences during the acute phase and after defervescence; 2) to evaluate a possible association of the FOXP3 single nucleotide polymorphism (SNP) 543 (SNP ID: rs2232367) with KD. METHODS: FOXP3 expression was evaluated on 8 children with KD and 15 healthy children by flow cytometry and Real-Time polymerase chain reaction (RT-PCR). FOXP3 SNP 543 was genotyped by denaturing high-performance liquid chromatography (DHPLC) and sequencing on DNA samples from 58 additional children with KD and 114 healthy donors. RESULTS: The frequencies of CD4+CD25 +FOXP3+ regulatory T cells were significantly (p=0.0002) lower during the acute phase of KD than in sex- and age-matched healthy donors (median % + SD: 4.8+/-1.3 vs. 7.7+/-1.7) and a similar tendency was revealed for FOXP3 mRNA transcripts (p<0.0001). FOXP3 expression increased significantly, at both protein and mRNA levels, after intravenous immunoglobulin (IVIG) therapy treatment and achieving complete remission of disease (at least 48 hrs; median 9.5 days, range 2-30). Of the 58 patients screened, only one female subject (1.7%) carried the presence of 543 SNP in heterozygosis (C>T; for a total of 1 allele out of 88), with no difference between KD patients and controls (0.0%, 0/203 alleles). CONCLUSIONS: Our data reinforce the notion of an impaired immunoregulation in KD, suggesting also a role of IVIG treatment in modifying the Treg compartment. FOXP3 SNP 543 does not seem to be involved in susceptibility to KD in Italian children.
Keywords: Acute Disease; Child; Child, Preschool; Female; Flow Cytometry; Forkhead Transcription Factors; Genotype; Humans; Immunoglobulins, Intravenous; Infant; Italy; Male; Mucocutaneous Lymph Node Syndrome; Polymorphism, Single Nucleotide; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; T-Lymphocytes, Regulatory

Design and development of a computer program for the evaluation of the healthcare executive - biomed 2010. - Zotti, Daniel; Bava, Michele; Delendi, Mauro (Biomed Sci Instrum, 2010)
According to the Italian law which regulates executive healthcare contracts, the professional evaluation is mandatory. The goal of the periodic evaluation is to enhance and motivate the professional involved. In addition this process should 1. increase the sense of duty towards the patients, 2. become aware of ones own professional growth and aspirations and 3. enhance the awareness of the healthcare executive regarding the companys strategies. To satisfy these requirements a data sheet has been modeled for every evaluated subject, divided in two sections. In the first part, the chief executive officer (CEO) scores: 1. behavioral characteristics, 2. multidisciplinary collaboration and involvement, 3. organizational skills, 4. professional quality and training, 5. relationships with the citizens. The scores for these fields are decided by the CEO. In the second part the CEO evaluates: 1. quantitative job dimension, 2.technology innovation, 3. scientific and educational activities. The value scores of these fields are decided by the CEO together with the professional under evaluation. A previously established correction coefficient can be used for all the scores. This evaluation system model has been constructed according to the enhancement quality approaches (Deming cycle) and a web-based software has been developed on a Linux platform using LAMP technology and php programming techniques. The program replicates all the evaluation process creating different profiles of authentications and authorizations which can then give to the evaluator the possibility to make lists of the professionals to evaluate, to upload documents regarding their activities and goals, to receive individual documents in automatically generated folders, to change the correction coefficients, to obtain year by year the individual scores. The advantages of using this web-based software include easy data consultation and update, the implementation of IT security issues, the easy portability and scalability of the system itself in different contexts even beyond healthcare.

Detection of SV40 in colon cancer: a molecular case-control study from northeast Italy. - Campello, Cesare; Comar, Manola; Zanotta, Nunzia; Minicozzi, Anna; Rodella, Luca; Poli, Albino (J Med Virol, 2010)
To explore the involvement of the simian polyomavirus SV40 in human colon cancer, a molecular case-control study was undertaken in patients and in their relatives living in an area where the spread of SV40 has already been documented. From 2006 to 2008, 94 colon cancer patients (age: 37-90 years) and 91 subjects (age: 32-70 years) relatives of each index case were enrolled. A blood sample and a specimen of cancer tissue or biopsy were collected, from each patient or control, respectively. Samples were analyzed twice for Polyomavirus (i.e., SV40, JCV, and BKV) by PCR and by quantitative real-time PCR (RT-qPCR) with reproducible results. No BKV/JCV was detected either in normal or pathological tissues. SV40 was not present in control subjects, either normal tissue or in biopsies from adenomas or polyps. All blood samples were negative. Conversely, six adenocarcinoma specimens were positive for SV40 sequences (overall prevalence 6.4%, P = 0.03 in comparison with controls). Nevertheless, the SV40-associated colon cancer risk proved statistically not significant (OR = 3.91; P = 0.115) when adjusted for age. Quantitation of SV40 DNA performed by RT-qPCR showed a low viral load ranging from 6.2 x 10(1) to 9 x 10(3) copies per reaction. This molecular case-control survey showed, for the first time in fresh samples and by RT-qPCR, that SV40 can be detected in colon cancer tissue. However, the finding was not statistically significant when compared with a well-structured community control group. Thus, the role of SV40 and other polyomavirus in colon cancer genesis deserves further investigation.
Keywords: Adenocarcinoma; Adult; Aged; Aged, 80 and over; BK Virus; Case-Control Studies; Colonic Neoplasms; DNA, Viral; Female; Humans; Italy; JC Virus; Male; Middle Aged; Polymerase Chain Reaction; Polyomavirus Infections; Simian virus 40; Tumor Virus Infections

Determinants of nosocomial infection in 6 neonatal intensive care units: an Italian multicenter prospective cohort study. - Auriti, Cinzia; Ronchetti, Maria Paola; Pezzotti, Patrizio; Marrocco, Gabriella; Quondamcarlo, Anna; Seganti, Giulio; Bagnoli, Francesco; De Felice, Claudio; Buonocore, Giuseppe; Arioni, Cesare; Serra, Giovanni; Bacolla, Gianfranco; et al. (Infect Control Hosp Epidemiol, 2010)
BACKGROUND: Nosocomial infections are still a major cause of morbidity and mortality among neonates admitted to neonatal intensive care units (NICUs). OBJECTIVE: To describe the epidemiology of nosocomial infections in NICUs and to assess the risk of nosocomial infection related to the therapeutic procedures performed and to the clinical characteristics of the neonates at birth and at admission to the NICU, taking into account the time between the exposure and the onset of infection. DESIGN: A multicenter, prospective cohort study. PATIENTS AND SETTING: A total of 1,692 neonates admitted to 6 NICUs in Italy were observed and monitored for the development of nosocomial infection during their hospital stay. METHODS: Data were collected on the clinical characteristics of the neonates admitted to the NICUs, their therapeutic interventions and treatments, their infections, and their mortality rate. The cumulative probability of having at least 1 infection and the cumulative probability of having at least 1 infection or dying were estimated. The hazard ratio (HR) for the first infection and the HR for the first infection or death were also estimated. RESULTS: A total of 255 episodes of nosocomial infection were diagnosed in 217 neonates, yielding an incidence density of 6.9 episodes per 1,000 patient-days. The risk factors related to nosocomial infection in very-low-birth-weight neonates were receipt of continuous positive airway pressure (HR, 3.8 [95% confidence interval CI, 1.7-8.1]), a Clinical Risk Index for Babies score of 4 or greater (HR, 2.2 [95% CI, 1.4-3.4]), and a gestational age of less than 28 weeks (HR, 2.1 [95% CI, 1.2-3.8]). Among heavier neonates, the risk factors for nosocomial infection were receipt of parenteral nutrition (HR, 8.1 [95% CI, 3.2-20.5]) and presence of malformations (HR, 2.3 [95% CI, 1.5-3.5]). CONCLUSIONS: Patterns of risk factors for nosocomial infection differ between very-low-birth-weight neonates and heavier neonates. Therapeutic procedures appear to be strong determinants of nosocomial infection in both groups of neonates, after controlling for clinical characteristics.
Keywords: Bacteremia; Birth Weight; Cross Infection; Gestational Age; Hospitals, University; Humans; Incidence; Infant, Newborn; Infant, Very Low Birth Weight; Intensive Care Units, Neonatal; Italy; Length of Stay; Proportional Hazards Models; Prospective Studies; Risk Factors; Sepsis; Time Factors

Development of paediatric quality of inpatient care indicators for low-income countries - A Delphi study. - Ntoburi, Stephen; Hutchings, Andrew; Sanderson, Colin; Carpenter, James; Weber, Martin; English, Mike (BMC Pediatr, 2010)
BACKGROUND: Indicators of quality of care for children in hospitals in low-income countries have been proposed, but information on their perceived validity and acceptability is lacking. METHODS: Potential indicators representing structural and process aspects of care for six common conditions were selected from existing, largely qualitative WHO assessment tools and guidelines. We employed the Delphi technique, which combines expert opinion and existing scientific information, to assess their perceived validity and acceptability. Panels of experts, one representing an international panel and one a national (Kenyan) panel, were asked to rate the indicators over 3 rounds and 2 rounds respectively according to a variety of attributes. RESULTS: Based on a pre-specified consensus criteria most of the indicators presented to the experts were accepted: 112/137(82%) and 94/133(71%) for the international and local panels respectively. For the other indicators there was no consensus; none were rejected. Most indicators were rated highly on link to outcomes, reliability, relevance, actionability and priority but rated more poorly on feasibility of data collection under routine conditions. There was moderate to substantial agreement between the two panels of experts. CONCLUSIONS: This Delphi study provided evidence for the perceived usefulness of most of a set of measures of quality of hospital care for children proposed for use in low-income countries. However, both international and local experts expressed concerns that data for many process-based indicators may not currently be available. The feasibility of widespread quality assessment and responsiveness of indicators to intervention should be examined as part of continued efforts to improve approaches to informative hospital quality assessment.
Keywords: Child; Child, Hospitalized; Delphi Technique; Developing Countries; Expert Testimony; Female; Humans; Inpatients; Male; Pediatrics; Quality Indicators, Health Care; World Health Organization

Does infant gastro-oesophageal reflux really deserve medical attention? - Declich, V; Badina, L; Ventura, A (Arch Dis Child, 2010)
Keywords: Beds; Gastroesophageal Reflux; Humans; Infant; Infant Care; Posture; Prognosis

Eltrombopag for the treatment of the inherited thrombocytopenia deriving from MYH9 mutations. - Pecci, Alessandro; Gresele, Paolo; Klersy, Catherine; Savoia, Anna; Noris, Patrizia; Fierro, Tiziana; Bozzi, Valeria; Mezzasoma, Anna Maria; Melazzini, Federica; Balduini, Carlo L (Blood, 2010)
Platelet transfusion is currently the primary medical treatment for reducing thrombocytopenia in patients with inherited thrombocytopenias. To evaluate whether stimulating megakaryopoiesis could increase platelet count in these conditions, we treated patients with a severe thrombocytopenia induced by MYH9 mutations (MYH9-related disease) with a nonpeptide thrombopoietin receptor agonist, eltrombopag. Twelve adult patients with MYH9-RD and platelet counts of less than 50 × 10(9)/L received 50 mg of eltrombopag orally per day for 3 weeks. Patients who achieved a platelet count higher than 150 × 10(9)/L stopped therapy, those with 100 to 150 platelets × 10(9)/L continued treatment at the same eltrombopag dose for 3 additional weeks, while those with less than 100 platelets × 10(9)/L increased the eltrombopag dose to 75 mg for 3 weeks. Major responses (platelet count of at least 100 × 10(9)/L or 3 times the baseline value) were obtained in 8 patients, minor responses (platelet counts at least twice the baseline value) in 3. One patient did not respond. Bleeding tendency disappeared in 8 of 10 patients with bleeding symptoms at baseline. Mild adverse events were reported in 2 patients. The availability of thrombopoietin mimetics opened new prospects in the treatment of inherited thrombocytopenias. This study is registered at www.clinicaltrials.gov as NCT01133860 (European Union Drug Regulating Authorities Clinical Trials number 2008-001903-42).
Keywords: Administration, Oral; Adolescent; Adult; Benzoates; Dose-Response Relationship, Drug; Female; Genetic Predisposition to Disease; Humans; Hydrazines; Male; Molecular Motor Proteins; Mutation; Myosin Heavy Chains; Platelet Aggregation; Platelet Count; Pyrazoles; Receptors, Thrombopoietin; Survival Rate; Thrombocytopenia; Treatment Outcome; Young Adult

Emx2 and Foxg1 inhibit gliogenesis and promote neuronogenesis. - Brancaccio, Marco; Pivetta, Chiara; Granzotto, Marilena; Filippis, Carol; Mallamaci, Antonello (Stem Cells, 2010)
Neural stem cells (NSCs) give rise to all cell types forming the cortex: neurons, astrocytes, and oligodendrocytes. The transition from the former to the latter ones takes place via lineage-restricted progenitors in a highly regulated way. This process is mastered by large sets of genes, among which some implicated in central nervous system pattern formation. The aim of this study was to disentangle the kinetic and histogenetic roles exerted by two of these genes, Emx2 and Foxg1, in cortico-cerebral precursors. For this purpose, we set up a new integrated in vitro assay design. Embryonic cortical progenitors were transduced with lentiviral vectors driving overexpression of Emx2 and Foxg1 in NSCs and neuronal progenitors. Cells belonging to different neuronogenic and gliogenic compartments were labeled by spectrally distinguishable fluoroproteins driven by cell type-specific promoters and by cell type-specific antibodies and were scored via multiplex cytofluorometry and immunocytofluorescence. A detailed picture of Emx2 and Foxg1 activities in cortico-cerebral histogenesis resulted from this study. Unexpectedly, we found that both genes inhibit gliogenesis and promote neuronogenesis, through distinct mechanisms, and Foxg1 also dramatically stimulates neurite outgrowth. Remarkably, such activities, alone or combined, may be exploited to ameliorate the neuronal output obtainable from neural cultures, for purposes of cell-based brain repair.
Keywords: Animals; Cell Differentiation; Cell Enlargement; Cells, Cultured; Female; Forkhead Transcription Factors; Homeodomain Proteins; Mice; Nerve Tissue Proteins; Neurogenesis; Neuroglia; Neurons; Stem Cells; Transcription Factors

Epidemiological and molecular assessment of a rubella outbreak in North-Eastern Italy. - D'Agaro, Pierlanfranco; Dal Molin, Gianna; Zamparo, Emanuela; Rossi, Tatiana; Micuzzo, Michele; Busetti, Marina; Santon, Daniela; Campello, Cesare (J Med Virol, 2010)
From January to June 2008, a rubella outbreak involving 111 laboratory confirmed cases occurred in the Friuli Venezia Giulia (FVG) region of North-Eastern Italy. The outbreak occurred initially in two residential homes for young adults disabled mentally and physically. Subsequently, the epidemic spread to the general population. Young adult cohorts were mostly affected and the mean age of the patients was 26.8 years; the majority of cases were male (73.8%), with a mean age of 26.6 years in males and 27.4 in females. Three pregnant women had a primary infection and two had their pregnancies terminated. Genotyping of 16 isolates showed the circulation of RUBV 2B, a genotype originating from Asia and South Africa and now present in Europe. In addition, molecular analysis revealed a well defined space-temporal spread of two viruses showing distinct sequences. A seroepidemiological survey carried out in a city within the same geographical area showed that the proportion of women of childbearing age still susceptible to rubella virus was 5.5%, fairly close to the figure (<5%) expected by 2010.
Keywords: Adolescent; Adult; Antibodies, Viral; Disease Outbreaks; Epidemics; Female; Genotype; Humans; Immunoglobulin G; Italy; Male; Pregnancy; Pregnancy Complications, Infectious; Rubella; Rubella virus; Young Adult

Ethnobotanical bioprospection of candidates for potential antimicrobial drugs from Brazilian plants: state of art and perspectives. - Benko-Iseppon, Ana Maria; Crovella, Sergio (Curr Protein Pept Sci, 2010)
Despite of the high biological diversity and traditional use of medicinal plants in Brazil, no comprehensive ethnobotanic review of plants with potential antimicrobial effects is available. In the present work own field information is aggregated with a literature review, identifying 433 Brazilian plant species potentially useful for identification of antimicrobial peptides. They included mainly woody species, distributed on 100 plant families (93 angiosperms and 7 pteridophytes) and 266 genera, covering all Brazilian regions and ecosystems. Main plant parts and indications for their use are presented and discussed, revealing the high potential that these plants present for the future planning strategies regarding the future development of antimicrobial drugs.
Keywords: Anti-Infective Agents; Biodiversity; Brazil; Ethnobotany; Plants

EULAR/PRINTO/PRES criteria for Henoch-Schönlein purpura, childhood polyarteritis nodosa, childhood Wegener granulomatosis and childhood Takayasu arteritis: Ankara 2008. Part I: Overall methodology and clinical characterisation. - Ruperto, Nicolino; Ozen, Seza; Pistorio, Angela; Dolezalova, Pavla; Brogan, Paul; Cabral, David A; Cuttica, Ruben; Khubchandani, Raju; Lovell, Daniel J; O'Neil, Kathleen M; Quartier, Pierre; Ravelli, Angelo; et al. (Ann Rheum Dis, 2010)
OBJECTIVES: To report methodology and overall clinical, laboratory and radiographic characteristics for Henoch-Schönlein purpura (HSP), childhood polyarteritis nodosa (c-PAN), c-Wegener granulomatosis (c-WG) and c-Takayasu arteritis (c-TA) classification criteria. METHODS: The preliminary Vienna 2005 consensus conference, which proposed preliminary criteria for paediatric vasculitides, was followed by a EULAR/PRINTO/PRES - supported validation project divided into three main steps. Step 1: retrospective/prospective web-data collection for HSP, c-PAN, c-WG and c-TA, with age at diagnosis <or=18 years. Step 2: blinded classification by consensus panel of a subgroup of 280 cases (128 difficult cases, 152 randomly selected) enabling expert diagnostic verification. Step 3: Ankara 2008 Consensus Conference and statistical evaluation (sensitivity, specificity, area under the curve, kappa-agreement) using as 'gold standard' the final consensus classification or original treating physician diagnosis. RESULTS: A total of 1183/1398 (85%) samples collected were available for analysis: 827 HSP, 150 c-PAN, 60 c-WG, 87 c-TA and 59 c-other. Prevalence, signs/symptoms, laboratory, biopsy and imaging reports were consistent with the clinical picture of the four c-vasculitides. A representative subgroup of 280 patients was blinded to the treating physician diagnosis and classified by a consensus panel, with a kappa-agreement of 0.96 for HSP (95% CI 0.84 to 1), 0.88 for c-WG (95% CI 0.76 to 0.99), 0.84 for c-TA (95% CI 0.73 to 0.96) and 0.73 for c-PAN (95% CI 0.62 to 0.84), with an overall kappa of 0.79 (95% CI 0.73 to 0.84). CONCLUSION: EULAR/PRINTO/PRES propose validated classification criteria for HSP, c-PAN, c-WG and c-TA, with substantial/almost perfect agreement with the final consensus classification or original treating physician diagnosis.
Keywords: Adolescent; Biopsy; Child; Delphi Technique; Granulomatosis with Polyangiitis; Humans; International Cooperation; Internet; Polyarteritis Nodosa; Purpura, Schoenlein-Henoch; Reproducibility of Results; Takayasu Arteritis

Exenatide in type 2 diabetes. - Verzegnassi, Federico; Chinello, Matteo (Lancet, 2010)
Keywords: Biological Markers; Diabetes Mellitus, Type 2; Energy Intake; Humans; Hypoglycemic Agents; Insulin; Insulin, Long-Acting; Peptides; Quality of Life; Venoms

Expression and association data strongly support JARID2 involvement in nonsyndromic cleft lip with or without cleft palate. - Scapoli, Luca; Martinelli, Marcella; Pezzetti, Furio; Palmieri, Annalisa; Girardi, Ambra; Savoia, Anna; Bianco, Anna Monica; Carinci, Francesco (Hum Mutat, 2010)
Nonsyndromic cleft lip with or without cleft palate (CL/P) affects approximately 1 in 1,000 births. Genetic studies have provided evidence for the role of several genes and candidate loci in clefting; however, conflicting results have frequently been obtained and much have to be done to unravel the complex genetics of CL/P. In the present investigation we have focused on the candidate region in 6p23, a region that have been found linked to CL/P in several investigations, in the attempt to find out the susceptibility gene provisionally named OFC1. Gene expression experiments in mice embryo of positional candidate genes revealed that JARID2 was highly and specifically expressed in epithelial cells in merging palatal shelves. A family-based linkage disequilibrium study confirmed the pivotal role of JARID2 in orofacial development and strongly supports a role for this gene in CL/P etiology (multiallelic haplotype test P=6 x 10(-5)). Understanding the molecular role of JARID2 within facial development may offer additional information to further unravel the complex genetics of CL/P.
Keywords: Animals; Cleft Lip; Cleft Palate; DNA Mutational Analysis; Family Health; Female; Gene Expression; Gene Expression Regulation, Developmental; Genetic Predisposition to Disease; Haplotypes; Humans; In Situ Hybridization; Linkage Disequilibrium; Male; Mice; Mice, Inbred C57BL; Mutation; Nerve Tissue Proteins; Palate; Polycomb Repressive Complex 2; Polymorphism, Single Nucleotide; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction

Eye movement impairment recovery in a Gaucher patient treated with miglustat. - Accardo, Agostino; Pensiero, Stefano; Ciana, Giovanni; Parentin, Fulvio; Bembi, Bruno (Neurol Res Int, 2010)
In Gaucher Disease (GD) the enzyme (imiglucerase) replacement therapy (ERT) is not able to stop the progression of the neurological involvement, while the substrate reduction therapy (SRT), performed by N-Butyldeoxynojirimycin (miglustat), is an alternative that should be evaluated. Two sisters, presenting the same genotype (R353G/R353G), were diagnosed as suffering from GD; one of them later developed neurological alterations identified by quantitative saccadic eye movements analysis. The aim of the study was to quantitatively measure the miglustat effects in this GD neurological patient. Eye movement analysis during subsequent controls was performed by estimating the characteristic parameters of saccadic main sequence. The study demonstrates that the SRT alone can be effective in GD3. Moreover, it confirms that quantitative eye movement analysis is able to precociously identify also slight neurological alterations, permitting more accurate GD classification.

Fasting increases tobramycin oral absorption in mice. - De Leo, Luigina; Di Toro, Nicola; Decorti, Giuliana; Malusà, Noelia; Ventura, Alessandro; Not, Tarcisio (Antimicrob Agents Chemother, 2010)
The pharmacokinetics of the aminoglycoside tobramycin was evaluated after oral administration to fed or fasting (15 h) mice. As expected, under normal feeding conditions, oral absorption was negligible; however, fasting induced a dramatic increase in tobramycin bioavailability. The dual-sugar test with lactulose and l-rhamnose confirmed increased small bowel permeability via the paracellular route in fasting animals. When experiments aimed at increasing the oral bioavailability of hydrophilic compounds are performed, timing of fasting should be extremely accurate.
Keywords: Administration, Oral; Animals; Anti-Bacterial Agents; Biological Availability; Fasting; Injections, Intramuscular; Injections, Intravenous; Intestinal Absorption; Lactulose; Male; Mice; Mice, Inbred BALB C; Rhamnose; Tobramycin

First case in Italy of acquired resistance to oseltamivir in an immunocompromised patient with influenza A/H1N1v infection. - Campanini, Giulia; Piralla, Antonio; Rovida, Francesca; Puzelli, Simona; Facchini, Marzia; Locatelli, Franco; Minoli, Lorenzo; Percivalle, Elena; Donatelli, Isabella; Baldanti, Fausto (J Clin Virol, 2010)
A pandemic influenza A/H1N1v strain with the neuraminidase H274Y mutation was detected in nasal secretions of a 2-year-old leukemic patient with influenza-like illness after 18 days of treatment with oseltamivir. At baseline, no drug-resistant virus was found, while 4 days after treatment initiation a mixture of wild-type and mutated virus was detected. After treatment interruption, the wild type influenza virus re-emerged and became prevalent in nasal secretions after a few days, suggesting the lower fitness of the mutated virus strain. The patient slowly improved concurrently with a decrease in virus load, which resulted negative 42 days after diagnosis. No other drug-resistant influenza A/H1N1v virus strains have been detected in Italy (up to the end of November 2009) since the first case of the novel A/H1N1v virus was identified in the country (May 2009).
Keywords: Amino Acid Substitution; Antiviral Agents; Bodily Secretions; Child, Preschool; Drug Resistance, Viral; Female; Humans; Immunocompromised Host; Influenza A Virus, H1N1 Subtype; Influenza, Human; Italy; Molecular Sequence Data; Mutation, Missense; Neuraminidase; Nose; Oseltamivir; RNA, Viral; Sequence Analysis, DNA; Treatment Outcome; Viral Load; Viral Proteins; Withholding Treatment

Five new OTOF gene mutations and auditory neuropathy. - Zadro, Cristina; Ciorba, Andrea; Fabris, Annalisa; Morgutti, Marcello; Trevisi, Patrizia; Gasparini, Paolo; Martini, Alessandro (Int J Pediatr Otorhinolaryngol, 2010)
OBJECTIVE: Purpose of this paper is to analyse OTOF gene in a series of subjects affected by auditory neuropathy. METHODS: Four children showing mild to profound prelingual deafness, confirmed by the absence of a clear and detectable responses at auditory brainstem responses (ABR), associated with the presence of bilateral OAE, were enrolled in the study. RESULTS AND CONCLUSIONS: Genetic analysis identified five new mutations (a nonsense, a small and a large deletion and two splicing site mutations), and one missense mutation (F1795C) previously described. These results further confirm the role of OTOF gene in auditory neuropathy. In the absence of a context of neurological syndrome, the combination of absent ABR and positive OAE responses should lead to an auditory neuropathy diagnosis and to a mutational screening in OTOF.
Keywords: Auditory Diseases, Central; Deafness; Evoked Potentials, Auditory, Brain Stem; Female; Humans; Male; Membrane Proteins; Mutation; Otoacoustic Emissions, Spontaneous; Polymerase Chain Reaction

Follow-up and quality of life of patients with cryopyrin-associated periodic syndromes treated with Anakinra. - Lepore, Loredana; Paloni, Giulia; Caorsi, Roberta; Alessio, Maria; Rigante, Donato; Ruperto, Nicola; Cattalini, Marco; Tommasini, Alberto; Zulian, Francesco; Ventura, Alessando; Martini, Alberto; Gattorno, Marco (J Pediatr, 2010)
OBJECTIVE: To evaluate the quality of life and long-term follow-up of patients enrolled in the Italian registry of cryopyrin-associated periodic syndromes (CAPS). STUDY DESIGN: Since 2004, 20 patients with CAPS were enrolled in a common registry from different Italian Centers of Pediatric Rheumatology; 14 patients were treated with Anakinra in an open fashion. Both treated and untreated patients were routinely followed according to standard of care. The Child Health Questionnaire (CHQ-PF 50) was used to assess the health-related quality of life. RESULTS: The mean duration of follow-up was 37.5 months. In all treated patients, a complete and persistent control of the inflammatory manifestations was observed with no further progression of the disease. At enrollment in the registry, patients showed a poorer health-related quality of life than healthy children in both physical and the psychosocial summary scores. Treatment was associated with a dramatic and sustained amelioration of a variety of measures of poor quality of life, particularly in those concerning the global health perception, bodily pain-discomfort, and other physical domains. CONCLUSIONS: Long-term IL-1 blockade produces a significant and persistent improvement in the clinical manifestations associated with the disease and on the overall quality of life.
Keywords: Adolescent; Adult; Case-Control Studies; Child; Child, Preschool; Cryopyrin-Associated Periodic Syndromes; Female; Follow-Up Studies; Humans; Inflammation; Interleukin 1 Receptor Antagonist Protein; Interleukin-1; Male; Phenotype; Quality of Health Care; Quality of Life; Questionnaires; Syndrome; Treatment Outcome

A G to C transversion at the last nucleotide of exon 25 of the MYH9 gene results in a missense mutation rather than in a splicing defect. - Vettore, Silvia; De Rocco, Daniela; Gerber, Bernhard; Scandellari, Raffaella; Bianco, Anna Monica; Balduini, Carlo L; Pecci, Alessandro; Fabris, Fabrizio; Savoia, Anna (Eur J Med Genet, 2010)
MYH9-related disease (MYH9-RD) is a rare autosomal dominant disorder caused by mutations in MYH9, the gene encoding the heavy chain of non-muscle myosin IIA. Patients present with congenital macrothrombocytopenia and inclusion bodies in neutrophils and might develop sensorineural deafness, presenile cataract, and/or progressive nephropathy leading to end-stage renal failure. In two families with macrothrombocytopenia we identified a novel c.3485G > C mutation in the last nucleotide of exon 25. Bioinformatic tools for splice site prediction and minigene functional test predicted splicing anomalies of exon 25. However, analysis of RNA purified from patient's peripheral blood did not allowed us to detect any anomalies, suggesting that RNA processing is correct at least in this tissue. Therefore, we concluded that c.3485G > C leads to a novel missense mutation (p.Arg1162Thr) of myosin-9, which resulted to be slightly degraded in patient platelets. A precise definition of the effect of mutations is fundamental to improve our knowledge into the pathogenetic mechanisms responsible for the disease.
Keywords: Adolescent; Adult; Blood Platelets; Computational Biology; Exons; Female; Humans; Inclusion Bodies; Kidney Failure, Chronic; Molecular Motor Proteins; Mutation, Missense; Myosin Heavy Chains; Neutrophils; Nonmuscle Myosin Type IIA; Nucleotides; RNA Splicing; Thrombocytopenia

Genetic polymorphism of inosine-triphosphate-pyrophosphatase influences mercaptopurine metabolism and toxicity during treatment of acute lymphoblastic leukemia individualized for thiopurine-S-methyl-transferase status. - Stocco, Gabriele; Crews, Kristine R; Evans, William E (Expert Opin Drug Saf, 2010)
IMPORTANCE OF THE FIELD: Although genetic polymorphisms in the gene encoding human thiopurine methyltransferase (TPMT) are known to have a marked effect on mercaptopurine metabolism and toxicity, there are many patients with wild-type TPMT who develop toxicity. Furthermore, when mercaptopurine dosages are adjusted in patients who are heterozygous at the TPMT locus, there are still some patients who develop toxicity for reasons that are not fully understood. Therefore, we recently studied the effects of a common polymorphism in another gene encoding an enzyme involved in mercaptopurine metabolism (SNP rs1127354 in inosine-triphospate-pyrophosphatase, ITPA), showing that genetic polymorphism of ITPA is a significant determinant of mercaptopurine metabolism and of febrile neutropenia following combination chemotherapy of acute lymphoblastic leukemia (ALL) in which mercaptopurine doses are individualized based on TPMT genotype. AREA COVERED IN THIS REVIEW: In this review, we summarize the knowledge available about the effect and clinical relevance of TPMT and ITPA on mercaptopurine pharmacogenomics, with a particular focus on the use of this medication in pediatric patients with ALL. WHAT THE READER WILL GAIN: Reader will gain insights into: i) the effects of pharmacogenomic traits on mercaptopurine toxicity and efficacy for the treatment of ALL and ii) individualization strategies that can be used to mitigate toxicity without compromising efficacy in pediatric patients with ALL. TAKE HOME MESSAGE: Mercaptopurine dose can be adjusted on the basis of TPMT genotype to mitigate toxicity in pediatric patients with ALL. As treatment is individualized in this way for the most relevant genetic determinant of drug response (i.e., for mercaptopurine, TPMT), the importance of other genetic polymorphisms emerges (e.g., ITPA).
Keywords: 6-Mercaptopurine; Antineoplastic Combined Chemotherapy Protocols; Child; Humans; Individualized Medicine; Methyltransferases; Models, Biological; Neutropenia; Nucleotide Transport Proteins; Polymorphism, Genetic; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Pyrophosphatases

Gestational diabetes and fetal growth acceleration: induction of labour versus expectant management. - Alberico, S; Businelli, C; Wiesenfeld, U; Erenbourg, A; Maso, G; Piccoli, M; Ronfani, L (Minerva Ginecol, 2010)
AIM: The aim of the study was to compare elective induction of labour at 38 weeks versus expectant management in A1 and A2 gestational diabetes (GDM) pregnancies with fetal growth acceleration. Primary outcome of the study was C-section (CS) rate, while secondary outcomes were macrosomia incidence and adverse perinatal outcomes. METHODS: A retrospective cohort study was carried out. Data were collected between 1996 and 2006 and evaluated through patients' records analysis. Differences between the two study groups were investigated using non-parametric tests for continuous variables and χ2 test for categorical ones. RESULTS: There was no significant difference between induction and expectant management in terms of caesarean section rate. A trend favoring women in the induction group in terms of incidence of macrosomia and neonatal outcomes was identified, but results were not statistically significant. CONCLUSION: Labour induction at 38 weeks in GDM patients with fetal growth acceleration does not seem to determine an increased incidence of C-section in comparison to expectant management, particularly in case of maternal obesity.
Keywords: Adult; Body Mass Index; Cesarean Section; Diabetes, Gestational; Elective Surgical Procedures; Female; Fetal Development; Fetal Macrosomia; Gestational Age; Humans; Incidence; Infant, Newborn; Italy; Labor, Induced; Medical Records; Obesity; Pregnancy; Pregnancy Outcome; Retrospective Studies; Risk Factors; Statistics, Nonparametric; Watchful Waiting

Hepatic glycogenosis in an adolescent with diabetes. - Bua, Jenny; Marchetti, Federico; Faleschini, Elena; Ventura, Alessandro; Bussani, Rossana (J Pediatr, 2010)
Keywords: Adolescent; Diabetes Mellitus, Type 1; Female; Glycogen Storage Disease; Humans; Liver Diseases

High polymorphism of the MBL2 gene in patients with atopic dermatitis. - Carréra, Matilde Campos; Moura, Patrícia; Crovella, Sergio; de Souza, Paulo Roberto Eleutério; de Alencar, Luiz Cláudio Arraes; Sarinho, Emanuel (Ann Allergy Asthma Immunol, 2010)
BACKGROUND: Low serum levels of mannose-binding lectin (MBL) are determined mainly by variant alleles of the MBL2 gene and it has been suggested that MBL may play a role in the susceptibility to atopic dermatitis (AD). OBJECTIVE: The aim was to investigate the difference of the frequency of MBL2 variant alleles in AD patients and in a group of individuals without AD, and associate the MBL2 alleles with AD severity. METHODS: MBL2 variant allele's frequency was investigated in 131 children with AD and 165 healthy children/adolescents matched by convenience. The severity of disease was graded according to the SCORing Atopic Dermatitis (SCORAD) index. The first exon variants were called "O" and the wild type "A". The variants in the promoter were H/L at -550 and X/Y at -221, determined by Real Time PCR. RESULTS: Children with AD had higher frequency of allele O and the genotypes related to low or deficient levels of MBL, when compared to the healthy group (p = 0.0012 and p < 0.001, respectively), but not with AD severity. CONCLUSION: Low or deficient MBL serum levels determined genetically may contribute to the predisposition for AD, but not for disease severity.
Keywords: Adolescent; Alleles; Child; Child, Preschool; Dermatitis, Atopic; Disease Progression; Female; Genetic Predisposition to Disease; Genotype; Humans; Infant; Male; Mannose-Binding Lectin; Polymorphism, Genetic; Promoter Regions, Genetic

Histatins in non-human primates: gene variations and functional effects. - Padovan, Lara; Segat, Ludovica; Pontillo, Alessandra; Antcheva, Nikolinka; Tossi, Alessandro; Crovella, Sergio (Protein Pept Lett, 2010)
Human histatins are histidine-rich, low molecular weight salivary proteins that contribute to the immune system of the oral cavity. In this work, nucleotide sequences of the HIS1 (coding for histatin 1) and HIS2 (coding for histatin 3) genes, homologous to the human ones, have been sequenced and analysed in five primates species including Great Ape, Hylobatidae and Cercopithecidae. In HIS1, the region corresponding to the putative mature peptide shows a premature stop codon in Macaca and Cercopithecus, while HIS2 a six codon insertion in the Cercopithecidae. Histatin 5, a 24-residue peptide derived from histatin 3, is the most antimicrobially active among human histatins, thus macaque and nomascus orthologues of histatin 5 were selected for chemical synthesis and functional characterization, in comparison to the human peptide. All synthesized histatins are predicted to be poorly amphipathic, depending on the charged state of His residues and assume partially a-helical conformations only in lipophilic conditions. Antimicrobial assays against Candida and Criptococcus spp. indicate somewhat different spectra of in vitro activity against the tested fungi. We have described HIS1 and HIS2 gene variations in primates and have analysed their functional effects on selected Hst5 orthologues. The human antimicrobial peptide has been proposed to represent an important lead for new generation of antimicrobial compounds for the treatment of oral mycoses, thus the information from the non-human primates histatins studied may aid strategies for drugs design.
Keywords: Amino Acid Sequence; Animals; Anti-Infective Agents; Base Sequence; Candida; Catarrhini; Cell Proliferation; Computational Biology; Cryptococcus; Genetic Variation; Histatins; Humans; Molecular Sequence Data; Phylogeny; Sequence Alignment; Sequence Analysis, DNA

HLA-G*0105N allele is associated with augmented risk for HIV infection in white female patients. - Segat, Ludovica; Catamo, Eulalia; Fabris, Annalisa; Morgutti, Marcello; D'Agaro, Pierlanfranco; Campello, Cesare; Crovella, Sergio (AIDS, 2010)
We analyzed HLA-G 3777G > C, HLA-G 14 bp deletion/insertion and HLA-G*0105N polymorphisms in HIV-positive white adult participants, infected through horizontal heterosexual transmission, and unexposed uninfected individuals, all from north eastern Italy. We report a new association between the HLA-G*0105N allele and HIV infection in adult white female participants, being HLA-G*0105N null allele correlated with an augmented risk (odds ratio = 4.35, 95% confidence interval = 1.38-18.07, P = 0.005) for HIV infection.
Keywords: Adolescent; Adult; Aged; Female; Genetic Predisposition to Disease; Histocompatibility Antigens Class I; HIV Infections; HIV-1; Humans; Middle Aged; Polymorphism, Genetic; Young Adult

Hot water and preparation of infant formula: how hot does it have to be to be safe? - Davanzo, Riccardo; Giurici, Nagua; Demarini, Sergio (J Pediatr Gastroenterol Nutr, 2010)
Keywords: Guidelines as Topic; Hot Temperature; Humans; Infant; Infant Formula; Water; World Health Organization

[Human papillomavirus cervical infection: viral genotyping and risk factors for high-grade squamous intraepithelial lesion and cervix cancer]. - de Mendonça, Vilma Guimarães; Guimarães, Maria José Bezerra; de Lima Filho, José Luiz; de Mendonça, Carolina Guimarães; Martins, Danyelly Bruneska Gondim; Crovella, Sergio; de Alencar, Luiz Cláudio Arraes (Rev Bras Ginecol Obstet, 2010)
PURPOSE: to analyze the characteristics of viral infection and the risk factors for high-grade squamous intraepithelial lesion and cervical carcinoma in women with cervical HPV infection. METHODS: a case-control study was conducted on women with cervical HPV at a Gynecology reference service enrolled at the Public Health System, located in Recife, Northeastern Brazil. The groups of cases (72 women with high-grade squamous intraepithelial lesion or cervical cancer) and controls (176 women with normal Pap smear or benign alterations) were investigated for six viral genotypes (HPV 16, 18, 31, 33, 6, 11) in ecto- and endocervical material using MY09/MY11 primers. The independent variables were ranked in three levels of determination: distal (sociodemographic), intermediate (behavioral) and proximal (previous Pap smear). The homogeneity of proportions was tested (χ2), unadjusted Odds Ratios (OR) were obtained and hierarchical logistic regression was applied to the final model, with adjustment of the effect of each variable to the outcome based on the variables in the same and previous levels of causality. RESULTS: the viral genotype of cervical infection was identified in 76.6% of the 248 women participating in the study. High-risk HPV genotypes (83.4% of cases and 67.1% of controls) were predominant, especially HPV 16 and 31. The distal risk factors identified were: living in a rural area (OR=2.71, 95%CI: 1.18-6.23), less than three years of study (OR=3.97, 95%CI: 2.09-7.54) and family income below two minimum wages (OR=3.30, 95%CI: 1.04-10.51); intermediate: four or more pregnancies (OR=2.00, 95%CI: 1.06-3.76); and proximal: absence of a previous Pap smear (OR=9.74, 95%CI: 2.48-38.28). CONCLUSIONS: genotypes 16 and 31 of cervical HPV infection are predominant among women assisted by the Public Health System in Northeastern Brazil. Socioeconomic and reproductive factors, as well as the absence of cytological screening, represent risk factors for the progression of infection to high-grade squamous intraepithelial lesion and cervical cancer.
Keywords: Adult; Carcinoma in Situ; Case-Control Studies; Female; Genotype; Humans; Papillomaviridae; Papillomavirus Infections; Risk Factors; Socioeconomic Factors; Uterine Cervical Neoplasms

Incidence of bacteremias and invasive mycoses in children with acute non-lymphoblastic leukemia: results from a multi-center Italian study. - Castagnola, Elio; Rossi, Mario R; Cesaro, Simone; Livadiotti, Susanna; Giacchino, Mareva; Zanazzo, Giulio; Fioredda, Francesca; Beretta, Chiara; Ciocchello, Francesca; Carli, Modesto; Putti, Maria Caterina; Pansini, Valeria; et al. (Pediatr Blood Cancer, 2010)
BACKGROUND: Data on the epidemiology of bacteremias and invasive fungal diseases (IFD) in children with acute myeloid leukemia (AML) are scarce. DESIGN AND METHODS: In a multi-center, retrospective study, we analyzed proportion, rate per 1,000 person-days at risk, and cumulative risk of bacteremias and IFD in children with AML. RESULTS: Between January 1998 and December 2005, 240 children were treated for AML at 8 Italian Centers, for a total of 521 treatment courses and 63,232 person-days at risk. Bacteremia was observed in 32% of treatment courses and IFD was seen in 10% (P < 0.0001), with rates of 2.62 and 0.84, respectively (P < 0.001). There was a significantly higher frequency of IFD during relapse treatment: proportion 15% versus 9% (P = 0.05), rate 2.10 versus 0.64 (P = 0.008) and cumulative risk 32% versus 12% (P = 0.007), while there were no differences in the proportion, rate and cumulative risk of bacteremia during front-line or relapse treatment. The epidemiology of bacteremias and IFD was different during front-line therapy for M3 as compared to other types of AML, but the differences were not statistically significant. CONCLUSIONS: Severe infectious complications are frequent during the treatment of pediatric AML, especially during relapse treatment, and bacteremias are more frequent than IFD.
Keywords: Antineoplastic Combined Chemotherapy Protocols; Bacteremia; Child; Female; Follow-Up Studies; Humans; Incidence; Italy; Leukemia, Myeloid, Acute; Male; Mycoses; Neoplasm Recurrence, Local; Retrospective Studies

Individual differences in prefrontal cortex activity during perception of bitter taste using fNIRS methodology. - Bembich, Stefano; Lanzara, Carmela; Clarici, Andrea; Demarini, Sergio; Tepper, Beverly J; Gasparini, Paolo; Grasso, Domenico L (Chem Senses, 2010)
Although bitter taste has a crucial role in nutrition by preventing the ingestion of toxic foods, there are few studies on bitter taste neuroimaging. To identify cortical areas involved in bitter taste perception and to determine if individual differences in taste sensitivity to 6-n-propylthiouracil (PROP) are represented in the brain by different cortical activation patterns, we examined 48 healthy volunteers using functional near-infrared spectroscopy. Participants rated the perceived intensity of filter paper disks impregnated with PROP and NaCl during the imaging procedure and were then classified as PROP tasters and nontasters. We monitored cortical activity in both the anterior and posterior regions of the dorsolateral prefrontal cortex (DLPFC) and in the ventrolateral prefrontal cortex (VLPFC). No activity was detected in the anterior DLPFC in any of the participants. However, during the administration of PROP, significant cortical activation was detected in the more posterior regions of the left DLPFC and in the left and right VLPFC but only in PROP tasters. PROP nontasters showed no cortical activity in these areas. These data suggest that the prefrontal cortex is involved in the conscious perception of the bitter taste of PROP and that the pattern of activity is consistent with individual differences in the ability to taste this compound. Thus, the PROP phenotype is associated with fundamental differences in cortical taste processing.
Keywords: Adult; Eating; Female; Food Preferences; Humans; Individuality; Male; Perception; Phenotype; Prefrontal Cortex; Propylthiouracil; Sodium Chloride; Spectroscopy, Near-Infrared; Taste; Taste Threshold; Young Adult

Insights into the binding of Phenyltiocarbamide (PTC) agonist to its target human TAS2R38 bitter receptor. - Biarnés, Xevi; Marchiori, Alessandro; Giorgetti, Alejandro; Lanzara, Carmela; Gasparini, Paolo; Carloni, Paolo; Born, Stephan; Brockhoff, Anne; Behrens, Maik; Meyerhof, Wolfgang (PLoS One, 2010)
Humans' bitter taste perception is mediated by the hTAS2R subfamily of the G protein-coupled membrane receptors (GPCRs). Structural information on these receptors is currently limited. Here we identify residues involved in the binding of phenylthiocarbamide (PTC) and in receptor activation in one of the most widely studied hTAS2Rs (hTAS2R38) by means of structural bioinformatics and molecular docking. The predictions are validated by site-directed mutagenesis experiments that involve specific residues located in the putative binding site and trans-membrane (TM) helices 6 and 7 putatively involved in receptor activation. Based on our measurements, we suggest that (i) residue N103 participates actively in PTC binding, in line with previous computational studies. (ii) W99, M100 and S259 contribute to define the size and shape of the binding cavity. (iii) W99 and M100, along with F255 and V296, play a key role for receptor activation, providing insights on bitter taste receptor activation not emerging from the previously reported computational models.
Keywords: Amino Acid Sequence; Calcium; Cell Line; Computational Biology; Dose-Response Relationship, Drug; Humans; Intracellular Space; Ligands; Models, Molecular; Molecular Sequence Data; Mutagenesis, Site-Directed; Phenylthiourea; Protein Binding; Protein Structure, Secondary; Receptors, G-Protein-Coupled

JCV/BKV and SV40 viral load in lymphoid tissues of young immunocompetent children from an area of north-east Italy. - Comar, Manola; Zanotta, Nunzia; Bovenzi, Massimo; Campello, Cesare (J Med Virol, 2010)
Polyomavirus infection occurring during childhood is followed by a lifelong latency in immunocompetent subjects. The major site of polyomavirus persistence are the uroepithelial cells which leads to oral transmission. It has recently been hypothesized that tonsils could be a possible reservoir. The role of tonsil, adenoid, and peripheral blood mononuclear cells (PBMCs) as possible sites of JCV, BKV, and SV40 latency in young healthy children was assessed. Two hundred fifteen fresh specimens, including 57 tonsil, 80 adenoid, and 78 PBMC samples from 80 immunocompetent children (mean age 4.8 years) were analyzed to determine the viral load by quantitative real-time PCR. The human herpes virus 6 (HHV-6) was tested as a lymphotropic reference virus. Polyomavirus was detected in 5/80 (6.2%) children while HHV-6 infection affected 27/80 children (33.7%) (P < 0.001). SV40 was detected in one adenoid sample, while footprints of BKV were found in one adenoid and three tonsil samples. JCV was never found in all samples. Polyomavirus sequences were not detected in the 78 blood samples. One adenoid and two tonsils from three children (1.4%) were positive for both polyomavirus and HHV-6. Infections were characterized by low replication rates ranging typically from 1 x 10e(2)/5.5 x 10e(4) to 6.8 x 10e(3)/8.5 x 10e(4) viral copies/number of cells. In conclusion, tonsils and adenoids of children could effectively harbor BKV and SV40, although only very few cells proved to be infected. Nevertheless, the low prevalence of polyomavirus, in comparison with the lymphotropic HHV-6, suggests that these tissues are unlikely to be the preferred site of polyomavirus latency, at least in younger children.
Keywords: Adenoids; BK Virus; Carrier State; Child; Child, Preschool; Female; Herpesvirus 6, Human; Humans; Immunocompetence; Italy; JC Virus; Male; Neutrophils; Palatine Tonsil; Polyomavirus Infections; Simian virus 40; Viral Load; Virus Latency

Leukocyte peroxidase and leptin: an associated link of glycemic tolerance and bronchial asthma? - Parco, Sergio (Diabetes Metab Syndr Obes, 2010)
Recent observations suggest the presence of an interaction between leptin and the inflammatory system during bronchial asthma. Although there is evidence of a positive association between asthma and obesity in adults and children, little is yet known about the role of serum leptin, as a potential mediator for bronchial epithelial homeostasis, and intraleukocyte myeloperoxidase (MPO), a hemoprotein with a molecular weight of 140 kDa, expression of the inflammatory system, in asthmatic children. Glycemic tolerance is an important pathogenetic element in developing type 2 mellitus diabetes and a confirmed predictor of incident asthma-like symptoms in adults. This work is aimed at assessing a possible correlation between basal leukocyte myeloperoxidase levels, basal leptin and insulin-glycemic tolerance in obese children. Thirty obese children aged between 7 and 15 years were examined. The analyzed data showed a normal response to the insulinemic stimulus in children of both sexes whose basal leptin and MPO values, expressed as MPO intracellular index, were within the normal range.

Long-term observational, non-randomized study of enzyme replacement therapy in late-onset glycogenosis type II. - Bembi, Bruno; Pisa, Federica Edith; Confalonieri, Marco; Ciana, Giovanni; Fiumara, Agata; Parini, Rossella; Rigoldi, Miriam; Moglia, Arrigo; Costa, Alfredo; Carlucci, Annalisa; Danesino, Cesare; Pittis, Maria Gabriela; et al. (J Inherit Metab Dis, 2010)
OBJECTIVES: Type II glycogenosis (GSDII) is a lysosomal storage disorder due to acid alpha-glucosidase (GAA) deficiency. Enzyme replacement therapy (ERT) with human recombinant alpha-glucosidase (rhGAA) has been demonstrated to be effective in the treatment of infantile forms of GSDII, but little information is available concerning late-onset phenotypes. Long-term follow-up studies are not available at present. The aim of this study was to evaluate the ERT long-term effects in late-onset GSDII. METHODS: Twenty-four patients, including 7 juveniles and 17 adults, received bi-weekly infusion of rhGAA (20 mg/kg) for at least 36 months. Clinical conditions, muscular function (6-min walking test, 6MWT; Walton scale, WS), respiratory function (vital capacity, VC; forced expiratory volume, FEV1; arterial pCO(2)), and muscle enzymes were assessed every 6 months. RESULTS: The 6MWT improved in both juvenile and adult patients (p = 0.01, p = 0.0002, respectively), as well as in patients with moderate to severe muscle function impairment (WS >3.5; p = 0.002). An overall improvement in WS was also observed (p = 0.0003). VC and FEV1 remained unchanged, while pCO(2) decreased (p = 0.017). Muscle enzymes decreased significantly (p < 0.0001). Two patients (8%) showed transient secondary events during ERT. CONCLUSIONS: Long-term ERT with rhGAA was shown to be safe, well tolerated, and effective in improving motor function and in stabilizing respiratory function in late-onset GSDII. The response pattern showed a progressive clinical improvement during the follow-up period in juvenile patients, while in adults it reached and maintained a plateau after the first year of treatment.
Keywords: Adolescent; Adult; Age of Onset; Aged; alpha-Glucosidases; Child; Enzyme Replacement Therapy; Female; Follow-Up Studies; Glycogen Storage Disease Type II; Humans; Male; Middle Aged; Observation; Time Factors; Treatment Outcome; Young Adult

Mannose-binding lectin is produced by vaginal epithelial cells and its level in the vaginal fluid is influenced by progesterone. - Bulla, R; De Seta, F; Radillo, O; Agostinis, C; Durigutto, P; Pellis, V; De Santo, D; Crovella, S; Tedesco, F (Mol Immunol, 2010)
Mannose-binding lectin (MBL) is a recognition molecule of the complement (C) system and binds to carbohydrate ligands present on a wide range of pathogenic bacteria, viruses, fungi, and parasites. MBL has been detected in the cervico-vaginal cavity where it can provide a first-line defence against infectious agents colonizing the lower tract of the reproductive system. Analysis of the cervico-vaginal lavage (CVL) obtained from 11 normal cycling women at different phases of the menstrual cycle revealed increased levels of MBL in the secretive phase. Part of this MBL derives from the circulation as indicated by the presence of transferrin in CVL tested as a marker of vascular and tissue permeability. The local synthesis of MBL is suggested by the finding that its level is substantially higher than that of transferrin in the secretive phase. The contribution of endometrium is negligible since the MBL level did not change before and after hysterectomy. RT-PCR and in situ RT-PCR analysis showed that the vaginal tissue, and in particular the basal layer of the epithelium, is a source of MBL which binds to the basal membrane and to cells of the outer layers of the epithelium. In conclusion, we have shown that MBL detected in CVL derives both from plasma as result of transudation and from local synthesis and its level is progesterone dependent increasing in the secretive phase of the menstrual cycle.
Keywords: Adolescent; Adult; Body Fluids; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Female; Humans; Immunohistochemistry; Mannose-Binding Lectin; Menstrual Cycle; Progesterone; Reverse Transcriptase Polymerase Chain Reaction; Vagina; Young Adult

Methotrexate withdrawal at 6 vs 12 months in juvenile idiopathic arthritis in remission: a randomized clinical trial. - Foell, Dirk; Wulffraat, Nico; Wedderburn, Lucy R; Wittkowski, Helmut; Frosch, Michael; Gerss, Joachim; Stanevicha, Valda; Mihaylova, Dimitrina; Ferriani, Virginia; Tsakalidou, Florence Kanakoudi; Foeldvari, Ivan; Cuttica, Ruben; et al. (JAMA, 2010)
CONTEXT: Novel therapies have improved the remission rate in chronic inflammatory disorders including juvenile idiopathic arthritis (JIA). Therefore, strategies of tapering therapy and reliable parameters for detecting subclinical inflammation have now become challenging questions. OBJECTIVES: To analyze whether longer methotrexate treatment during remission of JIA prevents flares after withdrawal of medication and whether specific biomarkers identify patients at risk for flares. DESIGN, SETTING, AND PATIENTS: Prospective, open, multicenter, medication-withdrawal randomized clinical trial including 364 patients (median age, 11.0 years) with JIA recruited in 61 centers from 29 countries between February 2005 and June 2006. Patients were included at first confirmation of clinical remission while continuing medication. At the time of therapy withdrawal, levels of the phagocyte activation marker myeloid-related proteins 8 and 14 heterocomplex (MRP8/14) were determined. INTERVENTION: Patients were randomly assigned to continue with methotrexate therapy for either 6 months (group 1 [n = 183]) or 12 months (group 2 [n = 181]) after induction of disease remission. MAIN OUTCOME MEASURES: Primary outcome was relapse rate in the 2 treatment groups; secondary outcome was time to relapse. In a prespecified cohort analysis, the prognostic accuracy of MRP8/14 concentrations for the risk of flares was assessed. RESULTS: Intention-to-treat analysis of the primary outcome revealed relapse within 24 months after the inclusion into the study in 98 of 183 patients (relapse rate, 56.7%) in group 1 and 94 of 181 (55.6%) in group 2. The odds ratio for group 1 vs group 2 was 1.02 (95% CI, 0.82-1.27; P = .86). The median relapse-free interval after inclusion was 21.0 months in group 1 and 23.0 months in group 2. The hazard ratio for group 1 vs group 2 was 1.07 (95% CI, 0.82-1.41; P = .61). Median follow-up duration after inclusion was 34.2 and 34.3 months in groups 1 and 2, respectively. Levels of MRP8/14 during remission were significantly higher in patients who subsequently developed flares (median, 715 [IQR, 320-1 110] ng/mL) compared with patients maintaining stable remission (400 [IQR, 220-800] ng/mL; P = .003). Low MRP8/14 levels indicated a low risk of flares within the next 3 months following the biomarker test (area under the receiver operating characteristic curve, 0.76; 95% CI, 0.62-0.90). CONCLUSIONS: In patients with JIA in remission, a 12-month vs 6-month withdrawal of methotrexate did not reduce the relapse rate. Higher MRP8/14 concentrations were associated with risk of relapse after discontinuing methotrexate. TRIAL REGISTRATION: isrctn.org Identifier: ISRCTN18186313.
Keywords: Adolescent; Antirheumatic Agents; Arthritis, Juvenile; ATP-Binding Cassette Transporters; Calgranulin B; Child; Child, Preschool; Female; Humans; Infant; Male; Methotrexate; Predictive Value of Tests; Prospective Studies; Recurrence; Remission Induction

Mode of delivery in HIV-infected pregnant women and prevention of mother-to-child transmission: changing practices in Western Europe. - Boer, K; England, K; Godfried, M H; Thorne, C (HIV Med, 2010)
OBJECTIVES: The aim of the study was to examine temporal and geographical patterns of mode of delivery in the European Collaborative Study (ECS), identify factors associated with elective caesarean section (CS) delivery in the highly active antiretroviral therapy (HAART) era and explore associations between mode of delivery and mother-to-child transmission (MTCT). METHODS: The ECS is a cohort study in which HIV-infected pregnant women are enrolled and their infants prospectively followed. Data on 5238 mother-child pairs (MCPs) enrolled in Western European ECS sites between 1985 and 2007 were analysed. RESULTS: The elective CS rate increased from 16% in 1985-1993 to 67% in 1999-2001, declining to 51% by 2005-2007. In 2002-2004, 10% of infants were delivered vaginally, increasing to 34% by 2005-2007. During the HAART era, women in Belgium, the United Kingdom and the Netherlands were less likely to deliver by elective CS than those in Italy and Spain [adjusted odds ratio (AOR) 0.07; 95% confidence interval (CI) 0.04-0.12]. The MTCT rate in 2005-2007 was 1%. Among MCPs with maternal HIV RNA<400 HIV-1 RNA copies/mL (n=960), elective CS was associated with 80% decreased MTCT risk (AOR 0.20; 95% CI 0.05-0.65) adjusting for HAART and prematurity. Two infants born to 559 women with viral loads <50 copies/mL were infected, one of whom was delivered by elective CS (MTCT rate 0.4%; 95% CI 0.04-1.29). CONCLUSIONS: Our findings suggest that elective CS prevents MTCT even at low maternal viral loads, but the study was insufficiently powered to enable a conclusion to be drawn as to whether this applies for viral loads <50 copies/mL. Diverging mode of delivery patterns in Europe reflect uncertainties regarding the risk-benefit balance of elective CS for women on successful HAART.
Keywords: Adult; Antiretroviral Therapy, Highly Active; Cesarean Section; Delivery, Obstetric; Epidemiologic Methods; Europe; Female; HIV Infections; Humans; Infant; Infant, Newborn; Infectious Disease Transmission, Vertical; Pregnancy; Pregnancy Complications, Infectious; Premature Birth; Prenatal Care; Reverse Transcriptase Inhibitors; Substance Abuse, Intravenous; Viral Load; Young Adult; Zidovudine

Modeling the effect of 3 missense AGXT mutations on dimerization of the AGT enzyme in primary hyperoxaluria type 1. - Robbiano, Angela; Frecer, Vladimir; Miertus, Jan; Zadro, Cristina; Ulivi, Sheila; Bevilacqua, Elena; Mandrile, Giorgia; De Marchi, Mario; Miertus, Stanislav; Amoroso, Antonio (J Nephrol, 2010)
INTRODUCTION: Mutations of the AGXT gene encoding the alanine:glyoxylate aminotransferase liver enzyme (AGT) cause primary hyperoxaluria type 1 (PH1). Here we report a molecular modeling study of selected missense AGXT mutations: the common Gly170Arg and the recently described Gly47Arg and Ser81Leu variants, predicted to be pathogenic using standard criteria. METHODS: Taking advantage of the refined 3D structure of AGT, we computed the dimerization energy of the wild-type and mutated proteins. RESULTS: Molecular modeling predicted that Gly47Arg affects dimerization with a similar effect to that shown previously for Gly170Arg through classical biochemical approaches. In contrast, no effect on dimerization was predicted for Ser81Leu. Therefore, this probably demonstrates pathogenic properties via a different mechanism, similar to that described for the adjacent Gly82Glu mutation that affects pyridoxine binding. CONCLUSION: This study shows that the molecular modeling approach can contribute to evaluating the pathogenicity of some missense variants that affect dimerization. However, in silico studies--aimed to assess the relationship between structural change and biological effects--require the integrated use of more than 1 tool.
Keywords: Amino Acid Sequence; Female; Humans; Male; Models, Molecular; Molecular Sequence Data; Mutation, Missense; Protein Multimerization; Transaminases

Molecular surveillance of pandemic influenza A(H1N1) viruses circulating in Italy from May 2009 to February 2010: association between haemagglutinin mutations and clinical outcome. - Puzelli, S; Facchini, M; De Marco, M A; Palmieri, A; Spagnolo, D; Boros, S; Corcioli, F; Trotta, D; Bagnarelli, P; Azzi, A; Cassone, A; Rezza, G; et al. (Euro Surveill, 2010)
Haemagglutinin sequences of pandemic influenza A(H1N1) viruses circulating in Italy were examined, focusing on amino acid changes at position 222 because of its suggested pathogenic relevance. Among 169 patients, the D222G substitution was detected in three of 52 (5.8%) severe cases and in one of 117 (0.9%) mild cases, whereas the D222E mutation was more frequent and evenly distributed in mild (31.6%) and severe cases (38.4%). A cluster of D222E viruses among school children confirms reported human-to-human transmission of viruses mutated at amino acid position 222.
Keywords: Adolescent; Adult; Age Distribution; Aged; Amino Acid Substitution; Child; Child, Preschool; Female; Hemagglutinins; Humans; Infant; Influenza A Virus, H1N1 Subtype; Influenza, Human; Italy; Male; Middle Aged; Mutation; Pandemics; Population Surveillance; Reverse Transcriptase Polymerase Chain Reaction; Severity of Illness Index; Sex Distribution; Young Adult

Morbidity of pandemic H1N1 influenza in children with cancer. - Caselli, Désirée; Carraro, Francesca; Castagnola, Elio; Ziino, Ottavio; Frenos, Stefano; Milano, Giuseppe Maria; Livadiotti, Susanna; Cesaro, Simone; Marra, Nicoletta; Zanazzo, Giulio; Meazza, Cristina; Cellini, Monica; et al. (Pediatr Blood Cancer, 2010)
BACKGROUND: To define the mortality and the current impact of the H1N1 pandemic in pediatric hematology-oncology centers, we performed a specific survey. PROCEDURE: Pharyngeal swabs from patients with fevers of unknown origin, flu-like symptoms or bronchopneumonia were screened for H1N1 using PCR. RESULTS: Sixty-two patients with documented H1N1 infection were reported: 16 had recently stopped therapy, 2 were at the diagnosis stage, and 44 were receiving therapy. The clinical course was severe (requiring ICU admission) in only 1 patient, moderate (requiring hospital admission) in 38, and mild in the remaining 23 (37%), treated as outpatients. While none of the patients died of H1N1-related complications, two patients died of progressive cancer; in all of the remaining cases, symptoms resolved within 11 days. The clinical course was complicated by respiratory distress or bronchopneumonia in 10 cases. Oseltamivir was given to 82% of patients. Chemotherapy was temporarily withdrawn in 54% of cases for a median time of 21 days (range, 4-43 days). CONCLUSION: H1N1 infection in children with cancer was not reported as the cause of death in any case but resulted in reduced intensity of anti-cancer therapy.
Keywords: Adolescent; Antineoplastic Agents; Cause of Death; Child; Child, Preschool; Data Collection; Disease Outbreaks; Humans; Influenza A Virus, H1N1 Subtype; Influenza, Human; Leukemia; Lymphoma, Non-Hodgkin; Morbidity; Neoplasms; Treatment Outcome; Young Adult

Multidrug resistant Pseudomonas aeruginosa infection in children undergoing chemotherapy and hematopoietic stem cell transplantation. - Caselli, Désirée; Cesaro, Simone; Ziino, Ottavio; Zanazzo, Giulio; Manicone, Rosaria; Livadiotti, Susanna; Cellini, Monica; Frenos, Stefano; Milano, Giuseppe M; Cappelli, Barbara; Licciardello, Maria; Beretta, Chiara; et al. (Haematologica, 2010)
Pseudomonas aeruginosa is one leading gram-negative organism associated with nosocomial infections. Bacteremia is life-threatening in the immunocompromised host. Increasing frequency of multi-drug-resistant (MDRPA) strains is concerning. We started a retrospective survey in the pediatric hematology oncology Italian network. Between 2000 and 2008, 127 patients with Pseudomonas aeruginosa bacteremia were reported from 12 centers; 31.4% of isolates were MDRPA. Death within 30 days of a positive blood culture occurred in 19.6% (25/127) of total patients; in patients with MDRPA infection it occurred in 35.8% (14/39). In the multivariate analysis, only MDRPA had significant association with infection-related death. This is the largest series of Pseudomonas aeruginosa bacteremia cases from pediatric hematology oncology centers. Monitoring local bacterial isolates epidemiology is mandatory and will allow empiric antibiotic therapy to be tailored to reduce fatalities.
Keywords: Adolescent; Antineoplastic Agents; Bacteremia; Child; Child, Preschool; Drug Resistance, Multiple; Female; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Infant; Infant, Newborn; Italy; Male; Pseudomonas aeruginosa; Pseudomonas Infections; Retrospective Studies; Young Adult

Multiple segmental absence of intestinal musculature presenting as spontaneous isolated perforation in an extremely low-birth-weight infant. - Oretti, Chiara; Bussani, Rossana; Janes, Augusta; Demarini, Sergio (J Pediatr Surg, 2010)
Defect of the intestinal musculature is a rare condition. It may cause intestinal perforation or obstruction. It manifests itself mainly in the neonatal period and usually affects preterm infants. We describe one such case, which was first diagnosed as a spontaneous isolated intestinal perforation. Emergency laparotomy was performed and showed multiple perforations, with accompanying peritonitis and ascites. Pathologic examination showed partial or complete absence of the musculature, particularly of the inner circular layer, with fibrous tissue in the regions of missing muscle, and abnormal vasculature. The myenteric plexus was absent in areas of muscle loss but present in other sites. These findings suggest that the absence of muscle may not represent a congenital malformation but may be secondary to ischemic injury.
Keywords: Diseases in Twins; Female; Humans; Ileum; Infant, Extremely Low Birth Weight; Infant, Newborn; Infant, Premature; Intestinal Atresia; Intestinal Perforation; Laparotomy; Muscle, Smooth; Myenteric Plexus; Rare Diseases

MYH9-related disease: Report on five German families and description of a novel mutation. - Savoia, Anna; Germeshausen, Manuela; De Rocco, Daniela; Henschel, Bettina; Kratz, Christian P; Kuhlen, Michaela; Rath, Bettina; Steuhl, Klaus-Peter; Wermes, Cornelia; Ballmaier, Matthias (Ann Hematol, 2010)
Keywords: Adult; Aged; Child; Chromosome Disorders; DNA Mutational Analysis; Germany; Humans; Infant; Molecular Motor Proteins; Mutation; Myosin Heavy Chains; Young Adult

Natural isoprenoids inhibit LPS-induced-production of cytokines and nitric oxide in aminobisphosphonate-treated monocytes. - Marcuzzi, Annalisa; Tommasini, Alberto; Crovella, Sergio; Pontillo, Alessandra (Int Immunopharmacol, 2010)
The inhibition of mevalonate pathway through genetic defects (mevalonate kinase deficiency, MKD) or pharmacologic drugs (aminobisphosphonates) causes a shortage of intermediate compounds and, in particular, of geranylgeranyl-pyrophosphate (GGPP) associated to the activation of caspase-1 and IL-1beta release. Geraniol (GOH), farnesol (FOH), geranylgeraniol (GGOH) and menthol (MOH), due to their isoprenoid structure, are supposed to enter the mevalonate pathway and to by-pass the biochemical block, reconstituting the pathway. Considering the already known side effects of aminobisphosphonates, and the lack of a specific treatment for MKD, we evaluated the impact of these natural isoprenoids compounds in a RAW cell lines chemically treated with the aminobisphosphonate alendronate, and in monocytes isolated from 2 patients affected by MKD. GOH, FOH, GGOH and MOH were all capable to diminish inflammatory marker levels induced by LPS. These natural isoprenoids could be proposed as novel therapeutic approach for the still orphan drug MKD, but also considered for the evaluation of possible inflammatory side effects of aminobisphosphonates.
Keywords: Animals; Cell Line; Cells, Cultured; Child; Child, Preschool; Cytokines; Diphosphonates; Humans; Inflammation; Interleukin-1beta; Lipopolysaccharides; Male; Mevalonate Kinase Deficiency; Mice; Monocytes; Nitric Oxide; Terpenes

Neonatal jaundice and breastfeeding reputation. - Bramuzzo, Matteo; Davanzo, Riccardo (J Hum Lact, 2010)
Keywords: Bottle Feeding; Breast Feeding; Female; Humans; Infant, Newborn; Jaundice, Neonatal; Lactation Disorders; Milk, Human; Risk Factors

Oxidative stress-based cytotoxicity of delphinidin and cyanidin in colon cancer cells. - Cvorovic, Jovana; Tramer, Federica; Granzotto, Marilena; Candussio, Luigi; Decorti, Giuliana; Passamonti, Sabina (Arch Biochem Biophys, 2010)
Colorectal cancer is the second most frequent cause of cancer death in the western world. Although the prognosis has improved after the introduction of newer anticancer drugs, the treatment of metastatic colorectal cancer still remains a challenge due to a high percentage of drug-resistant tumor forms. We aimed at testing whether anthocyanidins exerted cytotoxicity in primary (Caco-2) and metastatic (LoVo and LoVo/ADR) colorectal cancer cell lines. Both cyanidin and delphinidin, though neither pelargonidin nor malvidin, were cytotoxic in metastatic cells only. The cell line most sensitive to anthocyanidins was the drug-resistant LoVo/ADR. There, cellular ROS accumulation, inhibition of glutathione reductase, and depletion of glutathione could be observed. This suggests that anthocyanidins may be used as sensitizing agents in metastatic colorectal cancer therapy.
Keywords: Anthocyanins; Antioxidants; Apoptosis; Caco-2 Cells; Camptothecin; Cell Line, Tumor; Colonic Neoplasms; Drug Resistance, Neoplasm; Glutathione; Glutathione Reductase; Humans; Oxidants; Oxidative Stress

Pharmacogenomics in pediatric leukemia. - Paugh, Steven W; Stocco, Gabriele; Evans, William E (Curr Opin Pediatr, 2010)
PURPOSE OF REVIEW: The therapeutic index of many medications, especially in children, is very narrow with substantial risk for toxicity at doses required for therapeutic effects. This is particularly relevant to cancer chemotherapy, when the risk of toxicity must be balanced against potential suboptimal (low) systemic exposure that can be less effective in patients with higher rates of drug clearance. The purpose of this review is to discuss genetic factors that lead to interpatient differences in the pharmacokinetics and pharmacodynamics of these medications. RECENT FINDINGS: Genome-wide agonistic studies of pediatric patient populations are revealing genome variations that may affect susceptibility to specific diseases and that influence the pharmacokinetic and pharmacodynamic characteristics of medications. Several genetic factors with relatively small effect may be combined in the determination of a pharmacogenomic phenotype and considering these polygenic models may be mandatory in order to predict the related drug response phenotypes. These findings have potential to yield new insights into disease pathogenesis, and lead to molecular diagnostics that can be used to optimize the treatment of childhood cancers. SUMMARY: Advances in genome technology, and their comprehensive and systematic deployment to elucidate the genomic basis of interpatient differences in drug response and disease risk, hold great promise to ultimately enhance the efficacy and reduce the toxicity of drug therapy in children.
Keywords: Antineoplastic Agents; Child; Genetic Predisposition to Disease; Genetic Testing; Humans; Individualized Medicine; Pharmacogenetics; Phenotype; Precursor Cell Lymphoblastic Leukemia-Lymphoma

Predictors of poor response to methotrexate in polyarticular-course juvenile idiopathic arthritis: analysis of the PRINTO methotrexate trial. - Vilca, Iris; Munitis, Pablo Garcia; Pistorio, Angela; Ravelli, Angelo; Buoncompagni, Antonella; Bica, Blanca; Campos, Lucia; Häfner, Renate; Hofer, Michael; Ozen, Seza; Huemer, Christian; Bae, Sang Cheol; et al. (Ann Rheum Dis, 2010)
OBJECTIVES: To determine whether baseline demographic, clinical, articular and laboratory variables predict methotrexate (MTX) poor response in polyarticular-course juvenile idiopathic arthritis. METHODS: Patients newly treated for 6 months with MTX enrolled in the Paediatric Rheumatology International Trials Organization (PRINTO) MTX trial. Bivariate and logistic regression analyses were used to identify baseline predictors of poor response according to the American College of Rheumatology pediatric (ACR-ped) 30 and 70 criteria. RESULTS: In all, 405/563 (71.9%) of patients were women; median age at onset and disease duration were 4.3 and 1.4 years, respectively, with anti-nuclear antibody (ANA) detected in 259/537 (48.2%) patients. With multivariate logistic regression analysis, the most important determinants of ACR-ped 70 non-responders were: disease duration > 1.3 years (OR 1.93), ANA negativity (OR 1.77), Childhood Health Assessment Questionnaire (CHAQ) disability index > 1.125 (OR 1.65) and the presence of right and left wrist activity (OR 1.55). Predictors of ACR-ped 30 non-responders were: ANA negativity (OR 1.92), CHAQ disability index > 1.14 (OR 2.18) and a parent's evaluation of child's overall well-being < or = 4.69 (OR 2.2). CONCLUSION: The subgroup of patients with longer disease duration, ANA negativity, higher disability and presence of wrist activity were significantly associated with a poorer response to a 6-month MTX course.
Keywords: Adolescent; Antibodies, Antinuclear; Antirheumatic Agents; Arthritis, Juvenile; Child; Child, Preschool; Disability Evaluation; Female; Follow-Up Studies; Humans; Immunosuppressive Agents; Male; Methotrexate; Prognosis; Treatment Outcome

Prepregnancy BMI influences maternal and fetal outcomes in women with isolated gestational hyperglycaemia: a multicentre study. - Lapolla, A; Bonomo, M; Dalfrà, M G; Parretti, E; Mannino, D; Mello, G; Di Cianni, G (Diabetes Metab, 2010)
AIM: This multicentre study analyzed the maternal and fetal outcomes of women who had one elevated 3-h oral glucose tolerance test (isolated gestational hyperglycaemia [IGH]). METHODS: From 1999 to 2003, data were collected for 606 IGH women from 31 Italian obstetric or diabetic centres, including time and mode of delivery, gestational hypertension, preeclampsia, eclampsia, congenital malformations, and neonatal mortality and morbidity, to compare them with the general pregnant Italian population. A prognostic model for the outcome of pregnancy was constructed, and the concurrence of certain specified conditions was considered a positive outcome, whereas pregnancies that failed to meet one or more of the stated conditions were classified as "complicated". RESULTS: Macrosomia was significantly more frequent in women with IGH than in the normal pregnant population (10.7 vs 7.4%, respectively; P=0.003). Stillbirth and neonatal mortality rates did not differ from those in normal pregnancies, while a slight rise in the frequency of major malformations was not statistically significant (1.48 vs 0.89%, respectively; P<0.11). Multivariate logistic analyses confirmed that the prepregnancy body mass index (BMI) was an independent predictor of a complicated pregnancy. As for fetal growth, multivariate logistic analyses according to BMI showed that being overweight or obese were strong predictors of macrosomia. CONCLUSION: These findings in a large cohort of Italian women with IGH confirm the detrimental effect of even minimally altered glucose tolerance on fetal outcome. Also, prepregnancy obesity plays an important role in raising the risk of adverse perinatal outcomes in such patients.
Keywords: Adult; Blood Glucose; Body Mass Index; Female; Fetal Macrosomia; Glucose Tolerance Test; Humans; Hyperglycemia; Italy; Models, Statistical; Obesity; Pregnancy; Pregnancy Complications; Pregnancy Outcome; Prognosis; Retrospective Studies; Risk Factors; Time Factors

Progress towards the achievement of MDG4 in the Commonwealth of Independent States: uncertain data, clear priorities. - Cattaneo, Adriano; Gafurov, Ilkhom; Bomestar, Tamara; Bacci, Marianna; Kumar, Sanjiv; Popovic, Dragoslav; Tamburlini, Giorgio (Health Res Policy Syst, 2010)
Data on under five mortality in the twelve countries of the Commonwealth of Independent States show important fluctuations over time due to variations in quality of data, definitions of neonatal deaths and methods of mortality estimation. Despite the uncertainties regarding mortality trends, the analysis of health and social information from different sources offers clues to identify priority areas and key strategic directions for accelerating the achievement of the 4th Millennium Development Goal. Neonatal deaths represent from 40% to over 50% of under five deaths in all these countries. Maternal mortality was above 50 per 100,000 in 2005, despite the good coverage with antenatal care and births assisted by skilled birth attendants. The scanty information on quality of perinatal care indicates widespread substandard care at all levels. Stunting in children under five is above 10% in ten out of twelve countries and coexists with emerging overweight. Exclusivity and duration of breastfeeding fall short of what is recommended. There are important inequalities in child and maternal mortality, malnutrition and access and use of health services within countries. Taken as a whole, the available information clearly indicates that priority should be given to improvement of the health of women in reproductive age and of the quality of perinatal care, including the establishment of reliable data collection systems. To achieve this, action will need to focus on strengthening the capacity of the health system to improve the technical content of service provision, and on improving access and appropriate use of services by the most disadvantaged groups. The involvement of other sectors will be necessary to improve reproductive health and nutrition at community level and to tackle inequity. Comparisons between countries with similar socioeconomic background but different health policies seem to indicate that gradual progression towards universal coverage with essential health care through a national health insurance system is associated with larger reduction of child mortality than troubled transition towards a privatized and unregulated health system.

Risk of preterm delivery in relation to maternal use of psychotropic medications during pregnancy: methodological issues. - Erenbourg, Anna; Wiesenfeld, Uri; Ronfani, Luca (Am J Obstet Gynecol, 2010)
Keywords: Benzodiazepines; Confounding Factors (Epidemiology); Data Interpretation, Statistical; Female; Humans; Multivariate Analysis; Pregnancy; Premature Birth; Psychotropic Drugs

Rotavirus vaccine efficacy in African and Asian countries. - Tamburlini, Giorgio; Cattaneo, Adriano; Monasta, Lorenzo (Lancet, 2010)
Keywords: Africa; Child; Dysentery; Gastroenteritis; Humans; Rotavirus Infections; Rotavirus Vaccines; Treatment Outcome

Screening for GJB2 and GJB6 gene mutations in patients from Campania region with sensorineural hearing loss. - Chinetti, Viviana; Iossa, Sandra; Auletta, Gennaro; Laria, Carla; De Luca, Maria; Di Leva, Francesca; Riccardi, Pasquale; Giannini, Pasquale; Gasparini, Paolo; Ciccodicola, Alfredo; Marciano, Elio; Franzè, Annamaria (Int J Audiol, 2010)
The aim of this study was to screen 349 patients affected by sensorineural hearing loss (SNHL), mostly from the Campania region (southern Italy), for GJB2 gene mutations and for two deletions of the GJB6 gene (del GJB6 -D13S1830 and del GJB6 -D13S1854). We identified pathogenetic GJB2 mutations in 51 cases (15% of patients). No GJB6 mutation was found. We also examined the audiologic features of the patients for whom we had an etiologic diagnosis, in order to identify correlations between the severity of hearing loss and the type of mutation.
Keywords: Acoustic Stimulation; Adolescent; Adult; Audiometry; Auditory Perception; Child; Child, Preschool; Connexins; Genetic Predisposition to Disease; Genetic Testing; Hearing Loss, Sensorineural; Heterozygote; Homozygote; Humans; Italy; Mass Screening; Middle Aged; Mutation; Phenotype; Risk Factors; Severity of Illness Index; Young Adult

Severe X-linked mitochondrial encephalomyopathy associated with a mutation in apoptosis-inducing factor. - Ghezzi, Daniele; Sevrioukova, Irina; Invernizzi, Federica; Lamperti, Costanza; Mora, Marina; d'Adamo, Pio; Novara, Francesca; Zuffardi, Orsetta; Uziel, Graziella; Zeviani, Massimo (Am J Hum Genet, 2010)
We investigated two male infant patients who were given a diagnosis of progressive mitochondrial encephalomyopathy on the basis of clinical, biochemical, and morphological features. These patients were born from monozygotic twin sisters and unrelated fathers, suggesting an X-linked trait. Fibroblasts from both showed reduction of respiratory chain (RC) cIII and cIV, but not of cI activities. We found a disease-segregating mutation in the X-linked AIFM1 gene, encoding the Apoptosis-Inducing Factor (AIF) mitochondrion-associated 1 precursor that deletes arginine 201 (R201 del). Under normal conditions, mature AIF is a FAD-dependent NADH oxidase of unknown function and is targeted to the mitochondrial intermembrane space (this form is called AIF(mit)). Upon apoptogenic stimuli, a soluble form (AIF(sol)) is released by proteolytic cleavage and migrates to the nucleus, where it induces "parthanatos," i.e., caspase-independent fragmentation of chromosomal DNA. In vitro, the AIF(R201 del) mutation decreases stability of both AIF(mit) and AIF(sol) and increases the AIF(sol) DNA binding affinity, a prerequisite for nuclear apoptosis. In AIF(R201 del) fibroblasts, staurosporine-induced parthanatos was markedly increased, whereas re-expression of AIF(wt) induced recovery of RC activities. Numerous TUNEL-positive, caspase 3-negative nuclei were visualized in patient #1's muscle, again indicating markedly increased parthanatos in the AIF(R201 del) critical tissues. We conclude that AIF(R201 del) is an unstable mutant variant associated with increased parthanatos-linked cell death. Our data suggest a role for AIF in RC integrity and mtDNA maintenance, at least in some tissues. Interestingly, riboflavin supplementation was associated with prolonged improvement of patient #1's neurological conditions, as well as correction of RC defects in mutant fibroblasts, suggesting that stabilization of the FAD binding in AIF(mit) is beneficial.
Keywords: Apoptosis; Apoptosis Inducing Factor; Caspase 3; Computer Simulation; Dietary Supplements; DNA Primers; DNA, Mitochondrial; Electron Transport; Female; Fibroblasts; Flavin-Adenine Dinucleotide; Genes, X-Linked; Humans; In Situ Nick-End Labeling; Infant, Newborn; Magnetic Resonance Imaging; Male; Mitochondrial Encephalomyopathies; Muscle, Skeletal; Mutation; Nervous System Diseases; Pedigree; Poly(ADP-ribose) Polymerases; Protein Conformation; Riboflavin; Staurosporine; Twins, Monozygotic

Splenomegaly as presentation of a wandering spleen. - Maschio, Massimo; Cozzi, Giorgio; Sanabor, Daniela; Zennaro, Floriana; Gloria, Pelizzo; Barbi, Egidio (J Pediatr, 2010)
Keywords: Child; Female; Humans; Splenomegaly; Wandering Spleen

State of the art and recommendations. Kangaroo mother care: application in a high-tech environment. - Nyqvist, K H; Anderson, G C; Bergman, N; Cattaneo, A; Charpak, N; Davanzo, R; Ewald, U; Ludington-Hoe, S; Mendoza, S; Pallás-Allonso, C; Peláez, J G; Sizun, J; et al. (Breastfeed Rev, 2010)
UNLABELLED: Since Kangaroo Mother Care (KMC) was developed in Colombia in the 1970s, two trends in clinical application emerged. In low-income settings, the original KMC modelis implemented. This consists of continuous (24 h/day; 7 days/week) and prolonged mother/parent-infant skin-to-skin contact; early discharge with the infant in the kangaroo position; (ideally) exclusive breastfeeding and, adequate follow up. In affluent settings, intermittent KMC with sessions of one or a few hours skin-to-skin contact for a limited period is common. As a result of the increasing evidence of the benefits of KMC for both infants and families in all intensive care settings, KMC in a high-tech environment was chosen as the topic for the first European Conference on KMC, and the clinical implementation of the KMC modelin all types of settings was discussed at the 7th International Workshop on KMC Kangaroo Mother Care protocols in high-tech Neonatal Intensive Care Units (NICU) should specify criteria for initiation, kangaroo position, transfer to/from KMC, transport in kangaroo position, kangaroo nutrition, parents'role, modification of the NICU environment, performance of care in KMC, and KMCin case of infant instability. CONCLUSION: Implementation of the original KMC method, with continuous skin-to-skin contact whenever possible, is recommended for application in high-tech environments, although scientific evaluation should continue.

State of the art and recommendations. Kangaroo mother care: application in a high-tech environment. - Nyqvist, K H; Anderson, G C; Bergman, N; Cattaneo, A; Charpak, N; Davanzo, R; Ewald, U; Ludington-Hoe, S; Mendoza, S; Pallás-Allonso, C; Peláez, J G; Sizun, J; et al. (Acta Paediatr, 2010)
UNLABELLED: Since Kangaroo Mother Care (KMC) was developed in Colombia in the 1970s, two trends in clinical application emerged. In low income settings, the original KMC model is implemented. This consists of continuous (24 h/day, 7 days/week) and prolonged mother/parent-infant skin-to-skin contact; early discharge with the infant in the kangaroo position; (ideally) exclusive breastfeeding; and, adequate follow-up. In affluent settings, intermittent KMC with sessions of one or a few hours skin-to-skin contact for a limited period is common. As a result of the increasing evidence of the benefits of KMC for both infants and families in all intensive care settings, KMC in a high-tech environment was chosen as the topic for the first European Conference on KMC, and the clinical implementation of the KMC model in all types of settings was discussed at the 7th International Workshop on KMC. Kangaroo Mother Care protocols in high-tech Neonatal Intensive Care Units (NICU) should specify criteria for initiation, kangaroo position, transfer to/from KMC, transport in kangaroo position, kangaroo nutrition, parents' role, modification of the NICU environment, performance of care in KMC, and KMC in case of infant instability. CONCLUSION: Implementation of the original KMC method, with continuous skin-to-skin contact whenever possible, is recommended for application in high-tech environments, although scientific evaluation should continue.
Keywords: Attitude of Health Personnel; Female; Humans; Infant Care; Infant, Newborn; Intensive Care Units, Neonatal; Male; Parent-Child Relations; Practice Guidelines as Topic; Professional-Patient Relations; Role; Skin; Visitors to Patients

Storage of human milk: accepting certain uncertainties. - Davanzo, Riccardo; Travan, Laura; Demarini, Sergio (J Hum Lact, 2010)
Keywords: Food Handling; Humans; Milk, Human; Refrigeration

[Strategies for cardiovascular prevention in children]. - Spinelli, Angela; Nardone, Paola; Lamberti, Anna; Baglio, Giovanni (G Ital Cardiol (Rome), 2010)
Keywords: Body Mass Index; Cardiovascular Diseases; Child; Diet, Mediterranean; Food Habits; Humans; Life Style; Motor Activity; Obesity; Overweight; Patient Education as Topic; Population Surveillance; Questionnaires; Risk Factors

Structural aspects of plant antimicrobial peptides. - Padovan, Lara; Scocchi, Marco; Tossi, Alessandro (Curr Protein Pept Sci, 2010)
Antimicrobial peptides exert an important role in plant defence and their structure/activity relationship against pathogens is widely described. Although the most striking feature of these antimicrobial peptides is their molecular diversity, they share some common features, such as a relatively low molecular weight, and the presence of a variable number of cysteines residues that contribute to stabilize conserved scaffolds through disulphide bond formation, and can be assigned to different structural classes. Peptides from different classes in some cases act synergistically against pathogens when produced by the same tissue, and contribute to extending defence to a wider range of microbes. In this review we briefly describe the structure of some of the main plant antimicrobial peptide classes: thionins, defensins, lipid transfer proteins, cyclotides and snakins, and how they are reported to contribute to the plant protection. In many cases these antimicrobial peptides show a wider activity spectrum than that suggested by their name, exerting an action also against predatory insects and revealing useful antiviral activities. This extends their interest from defense of important food crops also to the design of novel anti-infective compounds for both pharmaceutical and agricultural applications.
Keywords: Amino Acid Sequence; Antimicrobial Cationic Peptides; Molecular Sequence Data; Plant Proteins; Plants; Sequence Alignment

Targeting farnesyl-transferase as a novel therapeutic strategy for mevalonate kinase deficiency: in vitro and in vivo approaches. - De Leo, Luigina; Marcuzzi, Annalisa; Decorti, Giuliana; Tommasini, Alberto; Crovella, Sergio; Pontillo, Alessandra (Pharmacol Res, 2010)
Mevalonate kinase deficiency (MKD) is a rare inborn auto-inflammatory disease due to the impairment of the pathway for the biosynthesis of cholesterol and non-sterol isoprenoids. The shortage of isoprenoids compounds and in particular of geranylgeranylpyrophosphate (GGPP) was recently associated to the MKD characteristic inflammatory attacks. The aim of this study is to demonstrate that the normalization of the mevalonate pathway intermediates levels and in particular of GGPP, through the specific inhibition of farnesyl-transferase (FT) with Manumycin A could ameliorate the inflammatory phenotype of MKD patients. The effect of Manumycin A was first evaluated in MKD mouse and cellular models, chemically obtained using the aminobisphosphonate alendronate (ALD), and then in monocytes isolated from 2 MKD patients. Our findings were compared to those obtained by using natural exogenous isoprenoids (NEIs). Manumycin A was able to significantly reduce the inflammatory marker serum amyloid A in ALD-treated Balb/c mice, as well as IL-1 beta secretion in ALD-monocytes and in MKD patients. These results clearly showed that, through the inhibition of FT, an increased number of mevalonate pathway intermediates could be redirected towards the synthesis of GGPP diminishing the inflammatory response. The importance in limiting the shortage of GGPP was emphasized by the anti-inflammatory effect of NEIs that, due to their biochemical structure, can enter the MKD pathway. In conclusion, manumycin A, as well as NEIs, showed anti-inflammatory effect in MKD models and especially in MKD-monocytes, suggesting novel approaches in the treatment of MKD, an orphan disease without any efficacious treatment currently available.
Keywords: Adult; Animals; Anti-Inflammatory Agents; Cells, Cultured; Child, Preschool; Enzyme Inhibitors; Farnesyltranstransferase; Female; Humans; Male; Mevalonate Kinase Deficiency; Mevalonic Acid; Mice; Mice, Inbred BALB C; Monocytes; Polyenes; Polyisoprenyl Phosphates; Polyunsaturated Alkamides; Young Adult

Therapeutic strategy in p47-phox deficient chronic granulomatous disease presenting as inflammatory bowel disease. - Freudenberg, Folke; Wintergerst, Uwe; Roesen-Wolff, Angela; Albert, Michael H; Prell, Christine; Strahm, Brigitte; Koletzko, Sibylle; Ehl, Stephan; Roos, Dirk; Tommasini, Alberto; Ventura, Alessandro; Belohradsky, Bernd H; et al. (J Allergy Clin Immunol, 2010)
Keywords: Age of Onset; Anti-Bacterial Agents; Antibodies, Monoclonal; Child; Drug Therapy, Combination; Gene Deletion; Granulomatous Disease, Chronic; Humans; Inflammatory Bowel Diseases; Male; NADPH Oxidase; Steroids; Treatment Outcome; Vidarabine

Thirty new loci for age at menarche identified by a meta-analysis of genome-wide association studies. - Elks, Cathy E; Perry, John R B; Sulem, Patrick; Chasman, Daniel I; Franceschini, Nora; He, Chunyan; Lunetta, Kathryn L; Visser, Jenny A; Byrne, Enda M; Cousminer, Diana L; Gudbjartsson, Daniel F; Esko, Tõnu; et al. (Nat Genet, 2010)
To identify loci for age at menarche, we performed a meta-analysis of 32 genome-wide association studies in 87,802 women of European descent, with replication in up to 14,731 women. In addition to the known loci at LIN28B (P = 5.4 × 10⁻⁶⁰) and 9q31.2 (P = 2.2 × 10⁻³³), we identified 30 new menarche loci (all P < 5 × 10⁻⁸) and found suggestive evidence for a further 10 loci (P < 1.9 × 10⁻⁶). The new loci included four previously associated with body mass index (in or near FTO, SEC16B, TRA2B and TMEM18), three in or near other genes implicated in energy homeostasis (BSX, CRTC1 and MCHR2) and three in or near genes implicated in hormonal regulation (INHBA, PCSK2 and RXRG). Ingenuity and gene-set enrichment pathway analyses identified coenzyme A and fatty acid biosynthesis as biological processes related to menarche timing.
Keywords: Adolescent; Aging; Body Height; Body Size; Child; DNA Copy Number Variations; Female; Genetic Loci; Genetic Predisposition to Disease; Genome-Wide Association Study; Humans; Inheritance Patterns; Menarche; Obesity; Polymorphism, Single Nucleotide; Quantitative Trait Loci; Reproducibility of Results; Time Factors

Ticks and Lyme borreliosis in an alpine area in northeast Italy. - Nazzi, F; Martinelli, E; Del Fabbro, S; Bernardinelli, I; Milani, N; Iob, A; Pischiutti, P; Campello, C; D'Agaro, P (Med Vet Entomol, 2010)
A 2-year study was conducted in a mountainous area of northeast Italy to evaluate the occurrence and distribution of ticks, as well as to assess the prevalence of the spirochaete Borrelia burgdorferi sensu lato. All ticks collected were Ixodes ricinus L. (Parasitiformes: Ixodidae). In general, most nymphs and adult ticks were collected from April to July. Tick density was highly variable among sites; however, two areas with different infestation levels were recognized. Prevalences of B. burgdorferi s.l. in nymphal stages were rather variable between sites; overall the prevalence of infected nymphs in the whole area was slightly higher than 20%. The prevalence of B. burgdorferi s.l. in nymphs does not seem to be correlated with nymph density. The correlation between the incidence of Lyme borreliosis (reported human cases/1000 inhabitants/year) and Borrelia prevalence in nymphs was not significant, although a significant correlation was found between borreliosis incidence and nymph density.
Keywords: Animals; Borrelia burgdorferi; Climate; Ecology; Humans; Incidence; Italy; Ixodes; Lyme Disease; Nymph; Population Density; Prevalence; Seasons; Ticks

Toward a consensus on guiding principles for health systems strengthening. - Swanson, Robert C; Bongiovanni, Annette; Bradley, Elizabeth; Murugan, Varnee; Sundewall, Jesper; Betigeri, Arvind; Nyonator, Frank; Cattaneo, Adriano; Harless, Brandi; Ostrovsky, Andrey; Labonté, Ronald (PLoS Med, 2010)
Keywords: Global Health; Humans; Public Health

Towards universal Kangaroo Mother Care: recommendations and report from the First European conference and Seventh International Workshop on Kangaroo Mother Care. - Nyqvist, K H; Anderson, G C; Bergman, N; Cattaneo, A; Charpak, N; Davanzo, R; Ewald, U; Ibe, O; Ludington-Hoe, S; Mendoza, S; Pallás-Allonso, C; Ruiz Peláez, J G; et al. (Acta Paediatr, 2010)
UNLABELLED: The hallmark of Kangaroo Mother Care (KMC) is the kangaroo position: the infant is cared for skin-to-skin vertically between the mother's breasts and below her clothes, 24 h/day, with father/substitute(s) participating as KMC providers. Intermittent KMC (for short periods once or a few times per day, for a variable number of days) is commonly employed in high-tech neonatal intensive care units. These two modalities should be regarded as a progressive adaptation of the mother-infant dyad, ideally towards continuous KMC, starting gradually and progressively with intermittent KMC. The other components in KMC are exclusive breastfeeding (ideally) and early discharge in kangaroo position with strict follow-up. Current evidence allows the following general statements about KMC in affluent and low-income settings: KMC enhances bonding and attachment; reduces maternal postpartum depression symptoms; enhances infant physiologic stability and reduces pain, increases parental sensitivity to infant cues; contributes to the establishment and longer duration of breastfeeding and has positive effects on infant development and infant/parent interaction. Therefore, intrapartum and postnatal care in all types of settings should adhere to a paradigm of nonseparation of infants and their mothers/families. Preterm/low-birth-weight infants should be regarded as extero-gestational foetuses needing skin-to-skin contact to promote maturation. CONCLUSION: Kangaroo Mother Care should begin as soon as possible after birth, be applied as continuous skin-to-skin contact to the extent that this is possible and appropriate and continue for as long as appropriate.
Keywords: Congresses as Topic; Female; Global Health; Humans; Infant Care; Infant, Newborn; Male; Parent-Child Relations; Practice Guidelines as Topic; Randomized Controlled Trials as Topic; Skin

Transmission of hemagglutinin D222G mutant strain of pandemic (H1N1) 2009 virus. - Puzelli, Simona; Facchini, Marzia; Spagnolo, Domenico; De Marco, Maria A; Calzoletti, Laura; Zanetti, Alessandro; Fumagalli, Roberto; Tanzi, Maria L; Cassone, Antonio; Rezza, Giovanni; Donatelli, Isabella (Emerg Infect Dis, 2010)
A pandemic (H1N1) 2009 virus strain carrying the D222G mutation was identified in a severely ill man and was transmitted to a household contact. Only mild illness developed in the contact, despite his obesity and diabetes. The isolated virus reacted fully with an antiserum against the pandemic vaccine strain.
Keywords: Adult; Amino Acid Substitution; Disease Outbreaks; Hemagglutinin Glycoproteins, Influenza Virus; Humans; Influenza A Virus, H1N1 Subtype; Influenza, Human; Male; Middle Aged; Mutation, Missense; Phylogeny; Retrospective Studies; RNA, Viral; Sequence Analysis, RNA; Severity of Illness Index

Treatment with recombinant tumor necrosis factor-related apoptosis-inducing ligand alleviates the severity of streptozotocin-induced diabetes. - Zauli, Giorgio; Toffoli, Barbara; di Iasio, Maria Grazia; Celeghini, Claudio; Fabris, Bruno; Secchiero, Paola (Diabetes, 2010)
OBJECTIVE: To evaluate the potential therapeutic effect of recombinant human tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) treatment in a model of type 1 diabetes. RESEARCH DESIGN AND METHODS: Recombinant TRAIL was added in vitro to primary human and mouse peripheral blood mononuclear cells (PBMCs) and isolated human islets to evaluate the expression of the immunoregulatory gene SOCS1. Diabetes was induced by five consecutive daily injections of low-concentration (50 mg/kg) streptozotocin (STZ) in C57 black mice (n = 24). A group of these mice (n = 12) was co-injected with recombinant TRAIL (20 microg/day) for 5 days, and the diabetic status (glycemia and body weight) was followed over time. After 6 weeks, circulating levels of insulin, TNF-alpha, and osteoprotegerin (OPG) were measured, and animals were killed to perform the histological analysis of the pancreas. RESULTS: The in vitro exposure of both PBMCs and human islets to recombinant TRAIL significantly upregulated the expression of SOCS1. With respect to STZ-treated animals, mice co-injected with STZ+TRAIL were characterized by 1) lower levels of hyperglycemia, 2) higher levels of body weight and insulinemia, 3) a partial preservation of pancreatic islets with normal morphology, and 4) a lower expression of both systemic (TNF-alpha and OPG) and pancreatic (vascular cell adhesion molecule [VCAM]-1) inflammatory markers. CONCLUSIONS: Overall, these data demonstrate that the administration of recombinant TRAIL ameliorates the severity of STZ-induced type 1 diabetes, and this effect was accompanied by the upregulation of SOCS1 expression.
Keywords: Animals; Cells, Cultured; Diabetes Mellitus, Experimental; Gene Expression; Humans; Islets of Langerhans; Leukocytes, Mononuclear; Mice; Recombinant Proteins; Suppressor of Cytokine Signaling Proteins; TNF-Related Apoptosis-Inducing Ligand

Two cases of Noonan syndrome with severe respiratory and gastroenteral involvement and the SOS1 mutation F623I. - Fabretto, Antonella; Kutsche, Kerstin; Harmsen, May-Britt; Demarini, Sergio; Gasparini, Paolo; Fertz, Maria Cristina; Zenker, Martin (Eur J Med Genet, 2010)
Noonan syndrome (NS) is an autosomal dominant, inherited disorder characterized by facial dysmorphism, congenital heart defects, and reduced postnatal growth. Dysregulated RAS-MAPK signalling is the common molecular basis for NS, a genetically heterogeneous disease. Germline mutations in genes encoding small GTPases of the RAS family (KRAS and NRAS), modulators of RAS function (PTPN11, SOS1 and SHOC2) or downstream signal transducers (RAF1) are causative for NS. SOS1 is the second major gene for NS after PTPN11. Compared to patients with mutations in other genes, SOS1 mutation-positive individuals in general tend to have a more favorable outcome, with less short stature and cognitive impairment. We describe two unrelated patients with NS carrying the same heterozygous SOS1 missense mutation (c.1867T > A/p.F623I). The phenotype of both patients is remarkable as they show uncommon clinical features such as pulmonary lymphangiectasis, congenital pleural effusions, severe feeding problems, and laryngomalacia. These findings may be related to the specific mutation present in our two patients, or be part of the SOS1 phenotype. Detailed clinical assessment of large cohorts of patients with NS and SOS1 mutation is required to clarify this initial observation.
Keywords: Child, Preschool; Genes, ras; Germ-Line Mutation; Heterozygote; Humans; Infant; Infant, Newborn; Male; Mutation, Missense; Noonan Syndrome; Phenotype; SOS1 Protein

The universe of immune deficiencies in Crohn's disease: a new viewpoint for an old disease? - Tommasini, Alberto; Pirrone, Angela; Palla, Gabriella; Taddio, Andrea; Martelossi, Stefano; Crovella, Sergio; Ventura, Alessandro (Scand J Gastroenterol, 2010)
Crohn's disease (CD) is generally considered a multifactorial disorder, since different genetic and environmental factors are thought to play a role in its pathogenesis. Recently, genome wide linkage studies allowed to identify the association of several loci with the increased risk of CD, although it is still unclear how they interact with environmental factors in causing the disease. The fact that many CD-risk-related genes are involved in the function of phagocytes seems in agreement with the well known role of these cells in CD histopathology. Functional defects in cytokine production or in clearance of bacteria in CD patients have recently been reported. Growing evidence that CD could arise from primary phagocyte immunodeficiency is also coming from the study of cases with early onset in infancy. We review such evidences starting from selected cases and discuss the clinical implications of these findings.
Keywords: Biological Markers; Crohn Disease; Cytokines; Evidence-Based Medicine; Granulomatous Disease, Chronic; Hematopoietic Stem Cell Transplantation; Humans; Immunosuppressive Agents; Phagocytes; Treatment Outcome; Wiskott-Aldrich Syndrome

Use of ketamine continuous infusion for pediatric sedation in septic shock. - Barbi, Egidio; Rizzello, Elisa; Taddio, Andrea (Pediatr Emerg Care, 2010)
Keywords: Anesthetics, Dissociative; Child; Conscious Sedation; Humans; Infusions, Intravenous; Ketamine; Male; Shock, Septic

Usefulness of the measurement of azathioprine metabolites in the assessment of non-adherence. - Stocco, Gabriele; Londero, Margherita; Campanozzi, Angelo; Martelossi, Stefano; Marino, Sara; Malusà, Noelia; Bartoli, Fiora; Decorti, Giuliana; Ventura, Alessandro (J Crohns Colitis, 2010)
Azathioprine is a thiopurine immunosuppressive antimetabolite used to chronically treat inflammatory bowel disease and autoimmune hepatitis. Azathioprine treatment is a long-term therapy and therefore it is at risk for non-adherence, which is considered an important determinant of treatment inefficacy. Measurement of 6-thioguanine and 6-methylmercaptopurine nucleotides has been recently suggested as a screener for non-adherence detection. We describe four young patients in which non-adherence to azathioprine therapy was detected only through the measurement of drug metabolite concentrations, and the criterion for non-adherence was undetectable metabolite levels. After the identification of non-adherence, patients and their families were approached and the importance of a correct drug administration was thoroughly enlightened and discussed; this allowed obtaining a full remission in all subjects. Our observations support the use of undetectable metabolite levels as indicators of non-adherence to therapy in azathioprine treated patients. The additional level of medical supervision given by this assay allows getting a better adherence to medical treatment, which results in an improvement in the response to therapy; these benefits may justify the costs associated with the assay.
Keywords: 6-Mercaptopurine; Adolescent; Azathioprine; Child; Child, Preschool; Female; Guanine Nucleotides; Hepatitis, Autoimmune; Humans; Immunosuppressive Agents; Inflammatory Bowel Diseases; Male; Medication Adherence; Thionucleotides; Young Adult

A web-based system for total parenteral nutrition prescription in a pediatric hospital - biomed 2010. - Bava, Michele; Bradashia, Fulvio; Rovere, Francesca; Maestro, Alessandra; Vecchi Brumatti, Liz; Accardo, Agostino; Paparazzo, Rossella; Zanon, Davide (Biomed Sci Instrum, 2010)
otal Parenteral Nutrition (TPN) is defined as feeding a patient by infusing nutrients intravenously, bypassing the usual process of eating and digestion. There are two kinds of TPN: short-term TPN may be used when a patient's digestive system is temporarily nonfunctional because of an interruption in its continuity; long-term TPN is used to treat patients with an impairment or a lack in nutrient absorption. TPN has extended the life of children born with nonexistent or severely deformed digestive organs and is a vital support for these patients. In Burlos Pediatric Department, pediatricians fill in a pharmacy request form in which nutritional needs are included for each patient. Subsequently, clinical pharmacists evaluate the patients individual data and decide which TPN formula to prepare. To enhance the TPN prescription process, an intranet web-based system has been developed to replicate the original paper-based forms. The software, developed in PHP and based on open source tools and services, has been constructed according to pharmacists requirements. These professionals, together with the Hospital Information System technicians, thanks to the availability of affordable instruments, perceive the advantages that such a system would have in improving clinical practice and quality of care. The system was devised with the goal to avoid common reading errors, to improve the correct text comprehension, to ensure prescription preparation, administration and tracking. According to a process of total quality control, the system reduces clinical risks regarding issues such as the correct and rapid availability of medical prescriptions and the incorrect identification of the patients. In comparison with paper-based TPN prescriptions, electronic-based forms have reduced the incidence of errors, the possible lack of patient data and reading misunderstandings. Regarding future improvements, IT technicians are defining the procedures to implement digital signature and medical aspects of the electronic TPN medical prescriptions.

Wired to be social: the ontogeny of human interaction. - Castiello, Umberto; Becchio, Cristina; Zoia, Stefania; Nelini, Cristian; Sartori, Luisa; Blason, Laura; D'Ottavio, Giuseppina; Bulgheroni, Maria; Gallese, Vittorio (PLoS One, 2010)
BACKGROUND: Newborns come into the world wired to socially interact. Is a propensity to socially oriented action already present before birth? Twin pregnancies provide a unique opportunity to investigate the social pre-wiring hypothesis. Although various types of inter-twins contact have been demonstrated starting from the 11(th) week of gestation, no study has so far investigated the critical question whether intra-pair contact is the result of motor planning rather then the accidental outcome of spatial proximity. METHODOLOGY/PRINCIPAL FINDINGS: Kinematic profiles of movements in five pairs of twin foetuses were studied by using four-dimensional ultrasonography during two separate recording sessions carried out at the 14(th) and 18(th) week of gestation. We demonstrate that by the 14th week of gestation twin foetuses do not only display movements directed towards the uterine wall and self-directed movements, but also movements specifically aimed at the co-twin, the proportion of which increases between the 14(th) and 18(th) gestational week. Kinematic analysis revealed that movement duration was longer and deceleration time was prolonged for other-directed movements compared to movements directed towards the uterine wall. Similar kinematic profiles were observed for movements directed towards the co-twin and self-directed movements aimed at the eye-region, i.e. the most delicate region of the body. CONCLUSIONS/SIGNIFICANCE: We conclude that performance of movements towards the co-twin is not accidental: already starting from the 14th week of gestation twin foetuses execute movements specifically aimed at the co-twin.
Keywords: Female; Fetus; Humans; Pregnancy; Social Behavior; Ultrasonography, Prenatal

Atomic models for the polypeptide backbones of myohemerythrin and hemerythrin. - Hendrickson, W A; Ward, K B (Biochem Biophys Res Commun, 1975)
Keywords: Animals; Cnidaria; Computers; Hemerythrin; Metalloproteins; Models, Molecular; Muscle Proteins; Protein Conformation; Species Specificity

Comparison between procaine and isocarboxazid metabolism in vitro by a liver microsomal amidase-esterase. - Moroi, K; Sato, T (Biochem Pharmacol, 1975)
Keywords: Amidohydrolases; Animals; Esterases; Hydrogen-Ion Concentration; In Vitro Techniques; Isocarboxazid; Kinetics; Male; Metals; Microsomes, Liver; Phospholipids; Procaine; Proteins; Rats; Subcellular Fractions; Temperature

Delineation of the intimate details of the backbone conformation of pyridine nucleotide coenzymes in aqueous solution. - Bose, K S; Sarma, R H (Biochem Biophys Res Commun, 1975)
Keywords: Fourier Analysis; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Conformation; NAD; NADP; Structure-Activity Relationship; Temperature

Effect of chloroquine on cultured fibroblasts: release of lysosomal hydrolases and inhibition of their uptake. - Wiesmann, U N; DiDonato, S; Herschkowitz, N N (Biochem Biophys Res Commun, 1975)
Keywords: Biological Transport; Cells, Cultured; Cerebroside-Sulfatase; Chloroquine; Dextrans; Fibroblasts; Glucuronidase; Humans; Leukodystrophy, Metachromatic; Lysosomes; Pinocytosis; Skin; Sulfatases

Formate assay in body fluids: application in methanol poisoning. - Makar, A B; McMartin, K E; Palese, M; Tephly, T R (Biochem Med, 1975)
Keywords: Aldehyde Oxidoreductases; Animals; Body Fluids; Carbon Dioxide; Formates; Haplorhini; Humans; Hydrogen-Ion Concentration; Kinetics; Methanol; Methods; Pseudomonas

Maturation of the adrenal medulla--IV. Effects of morphine. - Anderson, T R; Slotkin, T A (Biochem Pharmacol, 1975)
Keywords: Adrenal Medulla; Aging; Animals; Animals, Newborn; Body Weight; Catecholamines; Dopamine beta-Hydroxylase; Epinephrine; Female; Humans; In Vitro Techniques; Maternal-Fetal Exchange; Metaraminol; Morphine; Morphine Dependence; Pregnancy; Rats; Tyrosine 3-Monooxygenase

Metal substitutions incarbonic anhydrase: a halide ion probe study. - Smith, R J; Bryant, R G (Biochem Biophys Res Commun, 1975)
Keywords: Animals; Binding Sites; Cadmium; Carbonic Anhydrases; Cattle; Humans; Hydrogen-Ion Concentration; Magnetic Resonance Spectroscopy; Mercury; Protein Binding; Protein Conformation; Zinc

Radiochemical assay of glutathione S-epoxide transferase and its enhancement by phenobarbital in rat liver in vivo. - Marniemi, J; Parkki, M G (Biochem Pharmacol, 1975)
Keywords: Animals; Carrier Proteins; Epoxy Compounds; Glutathione; Glutathione Transferase; Hydrogen-Ion Concentration; Liver; Male; Methylcholanthrene; Phenobarbital; Rats; Stimulation, Chemical; Styrenes

Studies of oxygen binding energy to hemoglobin molecule. - Chow, Y W; Pietranico, R; Mukerji, A (Biochem Biophys Res Commun, 1975)
Keywords: Binding Sites; Cobalt; Hemoglobins; Humans; Hydrogen-Ion Concentration; Iron; Ligands; Mathematics; Oxygen; Oxyhemoglobins; Protein Binding; Spectrum Analysis