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Rocco Barazzoni,Annamaria Bernardi,Franco Biasia,Annamaria Semolic,Alessandra Bosutti,Mariapia Mucci,Franca Dore,Michela Zanetti,Gianfranco Guarnieri In spite of association between high plasma adiponectin and high metabolic and cardiovascular (CV) risk, highest adiponectin increments retain CV and metabolic protective effects in advanced chronic kidney disease (CKD). Passive accumulation can favor CKD-associated hyperadiponectinemia but potenti... ( view more )al additional regulation by adipose tissue remains undefined. Oxidative stress (OS) is associated with metabolic and CV disease and with CKD [increasing from conservative treatment (CT) to maintenance hemodialysis (MHD)], and OS can reduce adiponectin expression in experimental models. OS (in the form of plasma thiobarbituric acid-reactive substances: TBARS), subcutaneous adipose adiponectin mRNA, and plasma adiponectin were studied in CKD patients (stages 4 and 5) on CT (n = 7) or MHD (n = 11). Compared with CT and controls (C: n = 6) MHD had highest TBARS and lowest adiponectin mRNA (P < 0.05) with lower adipose adiponectin protein (P < 0.05 vs. CT). MHD also had lower plasma adiponectin than CT, although both had higher adiponectin than C (P < 0.05). In renal transplant recipients (RT: CKD stage 3; n = 5) normal TBARS were, in turn, associated with normal adiponectin mRNA (P < 0.05 vs. MHD). In all CKD (n = 23), adiponectin mRNA was associated positively with adiponectin plasma concentration (P < 0.01). In all subjects (n = 29), adiponectin mRNA was related (P < 0.05) negatively with TBARS after adjusting for plasma C-reactive protein (CRP) or CRP and creatinine. Thus altered OS, adiponectin expression, and plasma concentration represent a novel cluster of metabolic and CV risk factors in MHD that are normalized in RT. The data suggest novel roles of 1) MHD-associated OS in modulating adiponectin expression and 2) adipose tissue in contributing to circulating adiponectin in advanced CKD. ( view less ) Annamaria De Bellis,Annamaria Colao,Rosario Pivonello,Antonella Savoia,Marina Battaglia,Giuseppe Ruocco,Gilda Tirelli,Gaetano Lombardi,Antonio Bellastella,Antonio Bizzarro Hyperprolactinemia is often observed in lymphocytic hypophysitis (LYH). To clarify the possible autoimmune pituitary involvement in patients with apparently idiopathic hyperprolactinemia we investigated the presence of antipituitary antibodies (APA) in hyperprolactinemic patients with idiopathic hy... ( view more )perprolactinemia and in those with prolactinoma. Sixty-six hyperprolactinemic patients (52 F, 14 M age range 28-42 years, group 1) were studied. Of them, 34 out of 66 showed clinical features of hyperprolactinemia and subsequently underwent cabergoline therapy; the 32 out of 66 patients without symptoms of hyperprolactinemia did not receive cabergoline therapy. Moreover, 32 patients (24 F/8M, age range 23-44 years) with hyperprolactinemia due to microprolactinoma (group 2) were also studied. APA, by immunofluorescence method, and anterior pituitary function were evaluated in both groups of patients. APA were present in 17 out of 66 (25.7%) patients in group 1 with titers ranging from 1/16 to 1/64. All patients of group 2 were considered APA negative because these antibodies were found at low titer ( ( view less ) Annamaria Municinó,Annamaria Nicolino,Manlio Milanese,Edoardo Gronda,Bruno Andreuzzi,Fabrizio Oliva,Francesco Chiarella,Cardio-HKT Study Group  BACKGROUND: In-water exercise, hydrotherapy, may offer an attractive alternative to conventional training in markedly compromised patients with advanced HF. This Pilot Study evaluates the safety and efficacy of Cardio-Hydrokinesitherapy (Cardio-HKT) in patients with advanced HF on optimal medical t... ( view more )herapy. Cardio-HKT is a novel rehabilitation program that includes training sessions in warm water (31 degrees C), integrated by educational and psycho-behavioural sessions to promote healthy life style modifications. METHODS: We studied 18 adult patients with advanced HF, LVEF < 35%, NYHA functional class > II and peak oxygen uptake (peak VO2) < 18 ml/kg/min. Cardio-HKT consisted of a 3 weeks daily in-water training, combined to educational and psycho-behavioural sessions. Patients underwent a six-minute-walking-test (6mWT), a cardiopulmonary exercise test at baseline and after 3 weeks of Cardio-HKT. Quality of life was assessed with the Minnesota Living with Heart Failure Questionnaire (MLHF). RESULTS: All patients completed the Cardio-HKT rehabilitation program without complications. The 6mWT improved from 453 +/- 172 m to 571 +/- 120 m (p < 0.01), peak VO2 from 13.0 +/- 3.1 to 14.5 +/- 2.9 ml/kg/min (p = 0.03), whereas VE/ CO2 slope declined from 37 +/- 10 to 33 +/- 9 (p = 0.01). MLHF markedly improved from 56 (68-27) to 18 (40-7) (p < 0.01). CONCLUSIONS: Our results support the safety and efficacy of the innovative Cardio-HKT rehabilitation program in patients with advanced HF. ( view less ) Davide Tagliapietra,Michele Cornello,Annamaria Volpi Ghiradini,Annamaria Volpi GhirardiniIn this study, the use of reduced assemblages of benthic invertebrate taxa is proposed to describe similarity relationships between samples from transitional environments. A data set from four different studies, made up of 641 samples for a total 203 species, was analysed using permutation randomis... ( view more )ation tests in order to extract a pool of taxa able to approximate the full set of species. The identified "operational set", comprising 19 taxa, was capable of adequately reproduce similarity relationships between samples (Rho>0.90, p<0.001). All selected taxa were easily extractable from the samples and easy identifiable by non-specialised technicians; these characteristics were considered appropriate for the development of rapid and cost-effective monitoring procedures based on benthic invertebrate assemblages. ( view less ) Pasqualina Buono,Ottavia Barbieri,Andreina Alfieri,Annamaria Rosica,Simonetta Astigiano,Daniela Cantatore,Annamaria Mancini,Olimpia Fattoruso,Francesco SalvatoreAldolase C is selectively expressed in the hippocampus and Purkinje cells in adult mammalian brain. The gene promoter regions governing cell-specific aldolase C expression are obscure. We show that aldolase C messenger expression in the hippocampus is restricted to CA3 neurons. The human distal pro... ( view more )moter region (-200/-1200 bp) is essential for beta-galactosidase (beta-gal) expression in CA3 neurons and drives high stripe-like beta-gal expression in Purkinje cells. The 200 bp proximal promoter region is sufficient to drive low brain-specific and stripe-like beta-gal expression in Purkinje cells. Thus, the human aldolase C gene sequences studied drive endogenous-like expression in the brain. ( view less ) Rita Genesio,Genesio Rita,Daniele De Brasi,De Brasi Daniele,Anna Conti,Conti Anna,Annamaria Borghese,Borghese Annamaria,Pasqua Di Micco,Di Micco Pasqua,Pasquale Di Costanzo,Di Costanzo Pasquale,Dario Paladini,Paladini Dario,Paola Ungaro,Ungaro Paola,Lucio Nitsch,Nitsch Lucio We describe the cytogenetic and molecular characterization of an inverted duplication of chromosome 15q with evidence of a terminal deletion of the same rearranged chromosome. The proband was a multiple congenital malformed female with a prenatal diagnosis of trisomy 15q and an extremely severe cli... ( view more )nical course. The phenotype of the patient was characterized by marked intrauterine growth retardation, congenital heart defect, "horseshoe" kidney, hand contractures, and clubfeet. The exitus came at 20 days because of progressive cardio-respiratory impairment. Overall, the clinical phenotype appeared more severe than usual trisomy 15q syndrome. Postnatal cytogenetic and molecular studies unraveled a "de novo" inverted duplication of 15q (q21.3-->q26.3), associated with the deletion of the 15q telomere and part of the band 15q26.3. A single copy region spanning approximately 600 kb between the duplicated segments was present. Correlation between the clinical findings of the patient and the phenotype of trisomy 15q reported in literature is also provided. ( view less ) M Di Maio,F Perrone,C Gallo,R V Iaffaioli,L Manzione,F V Piantedosi,S Cigolari,A Illiano,S Barbera,S F Robbiati,E Piazza,G P Ianniello,L Frontini,E Veltri,F Castiglione,F Rosetti,E De Maio,P Maione,C Gridelli,Antonio Rossi,Emiddio Barletta,Maria Luisa Barzelloni,Giuseppe Signoriello,Domenico Bilancia,Angela Dinota,Gerardo Rosati,Domenico Germano,Alfredo Lamberti,Vittorio Pontillo,Luigi Brancacio,Carlo Crispino,Maria Esposito,Ciro Battiloro,Giovanni Tufano,Angela Cioffi,Vincenzo Guardasole,Valentina Angelini,Giovanna Guidetti,Santi Barbera,Francesco Renda,Francesco Romano,Antonio Volpintesta,Sergio Federico Robbiati,Mirella Sannicolò,Virginio Filipazzi,Gabriella Esani,Anna Gambaro,Sabrina Ferrario,Vincenza Tinessa,Maria Grazia Caprio,Sabrina Zonato,Mary Cabiddu,Alberto Raina,Enzo Veltri,Modesto D'Aprile,Giorgio Pistillucci,Gianfranco Porcile,Oliviero Ostellino,Orazio Vinante,Giuseppe Azzarello,Vittorio Gebbia,Nicola Borsellino,Antonio Testa,Giampietro Gasparini,Alessandra Morabito,Domenico Gattuso,Sante Romito,Francesco Carrozza,Sergio Fava,Anna Calcagno,Emanuela Grimi,Oscar Bertetto,Libero Ciuffreda,Giuseppe Parello,Luigi Maiorino,Antonio Santoro,Massimiliano Santoro,Giuseppe Failla,Rosa Anna Aiello,Alessandra Bearz,Roberto Sorio,Simona Scalone,Maurizia Clerici,Roberto Bollina,Paolo Belloni,Cosimo Sacco,Angela Sibau,Vincenzo Adamo,Giuseppe Altavilla,Antonino Scimone,Mario Spatafora,Vincenzo Bellia,Maria Raffealla Hopps,Silvio Monfardini,Adolfo Favaretto,Micaela Stefani,Giuliana Mara Corradini,Gianfranco Pavia,Giorgio Scagliotti,Silvia Novello,Giovanni Selvaggi,Maurizio Tonato,Samir Darwish,Giovanni Michetti,Maria Ori Belometti,Roberto Labianca,Antonello Quadri,Filippo De Marinis,Maria Rita Migliorino,Olga Martelli,Giuseppe Colucci,Dominico Galetta,Francesco Giotta,Luciano Isa,Paola Candido,Nestore Rossi,Antonio Calandriello,Francesco Ferraù,Emilia Malaponte,Sandro Barni,Marina Cazzaniga,Nicola Gebbia,Maria Rosaria Valerio,Mario Belli,Giuseppe Colantuoni,Matteo Antonio Capuano,Michele Angiolillo,Francesco Sollitto,Antonio Ardizzoia,Gino Luporini,Maria Cristina Locatelli,Franca Pari,Enrico Aitini,Tonino Pedicini,Antonio Febbraro,Cesira Zollo,Francesco Di Costanzo,Roberta Bartolucci,Silvia Gasperoni,Fernando Gaion,Giovanni Palazzolo,Enzo Galligioni,Orazio Caffo,Enrico Cortesi,Giuliana D'Auria,Carlo Curcio,Matteo Vasta,Cesare Bumma,Alfredo Celano,Sergio Bretti,Giuseppe Nettis,Annamaria Anselmo,Rodolfo Mattioli,Cecilia Nisticò,Annamaria Aschelter,Paola Foa The present study describes supportive care (SC) in patients with advanced non-small-cell lung cancer (NSCLC), evaluating whether it is affected by concomitant chemotherapy, patient's performance status (PS) and age. Data of patients enrolled in three randomised trials of first-line chemotherapy, c... ( view more )onducted between 1996 and 2001, were pooled. The analysis was limited to the first three cycles of treatment. Supportive care data were available for 1185 out of 1312 (90%) enrolled patients. Gastrointestinal drugs (45.7%), corticosteroids (33.4%) and analgesics (23.8%) were the most frequently observed categories. The mean number of drugs per patient was 2.43; 538 patients (45.4%) assumed three or more supportive drugs. Vinorelbine does not produce substantial variations in the SC pattern, while cisplatin-based treatment requires an overall higher number of supportive drugs, with higher use of antiemetics (41 vs 27%) and antianaemics (10 vs 4%). Patients with worse PS are more exposed to corticosteroids (42 vs 30%). Elderly patients require drugs against concomitant diseases significantly more than adults (20 vs 7%) and are less frequently exposed to antiemetics (12 vs 27%). In conclusion, polypharmacotherapy is a relevant issue in patients with advanced NSCLC. Chemotherapy does not remarkably affect the pattern of SC, except for some drugs against side effects. Elderly patients assume more drugs for concomitant diseases and receive less antiemetics than adults. ( view less ) Rosario Pivonello,Annamaria De Bellis,Antongiulio Faggiano,Francesco Di Salle,Mario Petretta,Carolina Di Somma,Silvia Perrino,Paolo Altucci,Antonio Bizzarro,Antonio Bellastella,Gaetano Lombardi,Annamaria Colao Central diabetes insipidus (CDI) is a rare hypothalamus-pituitary disease due to the deficiency of arginine vasopressin (AVP) synthesis from the hypothalamus and/or secretion from the neurohypophysis. The etiology of CDI is unknown in over one third of cases, classified as idiopathic CDI. The aim o... ( view more )f this study was 2-fold: 1) to evaluate the occurrence of circulating autoantibodies to AVP-secreting cells (AVPcAb), and 2) to correlate it to clinical (sex, age of disease onset, disease duration, and degree), immunological (clinical history of autoimmune diseases and presence of related organ-specific autoantibodies), and radiological features (neurohypophyseal bright spot, pituitary stalk thickening, and empty sella) in a large cohort of patients with apparently idiopathic CDI or CDI of known etiology. To this purpose, 150 patients with CDI were studied: 64 idiopathic, 6 familial, 12 associated to granulomatous diseases, and 68 secondary to cranial trauma, tumor, or surgery. AVPcAb were measured by an indirect immunofluorescence method. AVPcAb were found in 23.3% of CDI patients: 21 idiopathic (32.8%) and 14 nonidiopathic (16.3%; chi(2) = 13.1; P < 0.001). AVPcAb were independently associated with age less than 30 yr at disease onset (P = 0.001) in patients with idiopathic CDI and with history of autoimmune diseases (P = 0.006 and P = 0.02, respectively) and radiological evidence of pituitary stalk thickening (P = 0.02 and P = 0.003, respectively) in both idiopathic and nonidiopathic CDI. The likelihood of autoimmunity in one patient with apparently idiopathic CDI with age of disease onset less than 30 yr was 53%, it increased to 91% when history of autoimmune diseases was associated and to 99% when pituitary stalk thickening was further associated. In conclusion, autoimmunity is associated with one third of patients with apparently idiopathic CDI, which should therefore be classified as autoimmune CDI. Autoimmune CDI is highly likely in young patients with a clinical history of autoimmune diseases and radiological evidence of pituitary stalk thickening. Conversely, autoimmunity probably represents an epiphenomenon in patients with nonidiopathic CDI. ( view less ) Annamaria Innoceta,Luca Galluzzi,Annamaria Ruzzo,Francesca Andreoni,Laura Chiarantini,Mauro Magnani Human monoblastoid cells (U937) grown in the presence of therapeutically relevant dideoxycytidine concentrations (0.1 microM) become resistant to the drug thanks to an altered deoxycytidine kinase. In this paper we show that deoxycytidine kinase mRNA is significantly reduced in drug-resistant U937 ... ( view more )cells (U937-R) although the deoxycytidine kinase promoter is normal. Anumber of nucleotide deletions, insertions and substitutions was found in the coding region of deoxycytidine kinase gene. Several identified mutations result in truncated forms of the enzyme or in the introduction of stop codons: in one case a complete lack of exon 4 was found. Thus, the deoxycytidine kinase gene accumulates mutations at a very high rate, as already reported for other cytidine analogues (i.e. Ara C) suggesting that the design of new antiviral or anticancer drugs of the cytidine family should take into account the potential development of cell resistance as a critical factor in drug failure. ( view less ) Gabor Nemeth,Szabolcs Felszeghy,Annamaria Kenyeres,Nora Szentmary,Andras Berta,Ildiko Suveges,Laszlo Modis The aim of the present study was to investigate the expression pattern of different cell adhesion molecules in corneal stromal dystrophies. Fifteen corneal buttons from patients diagnosed with three different types of stromal corneal dystrophies and healthy corneas were investigated. Paraffin embed... ( view more )ded sections were stained immunohistochemically with monoclonal antibodies against human intercellular adhesion molecule-1 (ICAM-1), endothelial selectin (E-selectin) and endothelial cadherin (E-cadherin) using the avidin-biotin-peroxidase-complex technique. The sections were compared to normal eye bank controls. In corneas from granular dystrophy patients ICAM-1 was expressed focally in epithelial cells and in keratocytes, and expressed diffusely in endothelial cells. In corneas from macular dystrophy patients diffuse epithelial staining was observed and the stromal and endothelial expression was found to be similar to that of granular dystrophy. In lattice dystrophy, only the epithelial cells and endothelium were intensively positive for ICAM-1. E-selectin was not present on any layer of the corneal specimens. E-cadherin was observed only in the epithelium of all three types of corneal dystrophies. Normal corneas did not express any of the investigated adhesion molecules. We found different expression patterns of adhesion molecules in corneas from stromal dystrophies. Our results suggest that adhesion molecules may be involved in the pathogenesis of corneal stromal dystrophies. ( view less ) Luca Olivotti,Shahram Moshiri,Gennaro Santoro,Annamaria Nicolino,Francesco Chiarella Coronary artery fistulae may sometimes be asymptomatic and may spontaneously close, but they may also cause angina, myocardial infarction, endocarditis, atrial fibrillation and heart failure. Therefore, in patients with big or symptomatic fistulae, intervention is mandatory. In the literature, most... ( view more ) of the patients who underwent transcatheter coil embolization of coronary artery fistulae were children; this is because of the early onset of symptoms in the most severe cases, but also because many adult patients are usually referred to surgery. We describe the case of an adult patient presenting with heart failure and a giant tortuous coronary fistula, with high shunt flow from right coronary artery to right atrium, which was successfully treated with transcatheter closure with free embolization coils. The most challenging technical aspect of transcatheter coil embolization is the precise release of the spirals. ( view less ) Annamaria Pratelli Canine coronavirus (CCoV) is responsible for mild or moderate enteritis in puppies. The virus is highly contagious and avoiding contact with infected dogs and their excretions is the only way to ensure disease prevention. Since no studies have yet focused on the sensitivity of CCoV to chemical bioc... ( view more )ides the present investigation examined the efficiency of physical and chemical methods of viral inactivation. CCoV infectivity was stable at +56 degrees C for up to 30min, but tended to decrease rapidly at +65 degrees C and +75 degrees C. Germicidal ultra-violet (UV-C) light exposure demonstrated no significant effects on virus inactivation for up to 3 days. CCoV was observed to be more stable at pH 6.0-6.5 while extreme acidic conditions inactivated the virus. Two tested aldehydes inactivated the virus but their action was temperature- and time-dependent. The methods for CCoV inactivation could be applied as animal models to study human coronavirus infection, reducing the risk of accidental exposure of researchers to pathogens during routine laboratory procedures. ( view less ) Miranda D Grounds,Hannah G Radley,Gordon S Lynch,Kanneboyina Nagaraju,Annamaria De Luca This review discusses various issues to consider when developing standard operating procedures for pre-clinical studies in the mdx mouse model of Duchenne muscular dystrophy (DMD). The review describes and evaluates a wide range of techniques used to measure parameters of muscle pathology in mdx mi... ( view more )ce and identifies some basic techniques that might comprise standardised approaches for evaluation. While the central aim is to provide a basis for the development of standardised procedures to evaluate efficacy of a drug or a therapeutic strategy, a further aim is to gain insight into pathophysiological mechanisms in order to identify other therapeutic targets. The desired outcome is to enable easier and more rigorous comparison of pre-clinical data from different laboratories around the world, in order to accelerate identification of the best pre-clinical therapies in the mdx mouse that will fast-track translation into effective clinical treatments for DMD. ( view less ) Massimo Breccia,Fabiana Gentilini,Laura Cannella,Roberto Latagliata,Ida Carmosino,Annamaria Frustaci,Giuliana AlimenaImatinib mesylate is a selective inhibitor of the bcr/abl, c-kit and PDGF receptor tyrosine kinases. Its ocular toxicity is little known with mild periorbital oedema being the most commonly reported side effect. We here describe our experience on ocular complications in imatinib treated Ph+ CML pat... ( view more )ients, which consisted of a wide spectrum of adverse effects ranging from periobital oedema to serious adverse events such as glaucoma. ( view less ) Matteo Landriscina,Settimia Anna Altamura,Leonarda Roca,Margherita Gigante,Annamaria Piscazzi,Elisabetta Cavalcanti,Eleonora Costantino,Carlo Barone,Mauro Cignarelli,Loreto Gesualdo,Elena Ranieri Reverse transcriptase (RT) inhibitors are emerging as a novel class of anticancer differentiating agents, active in several human tumor cell models, such as melanoma and prostate, thyroid and colon carcinoma. Indeed, much evidence suggests that they may act by inhibiting endogenous RT, a gene highl... ( view more )y expressed in undifferentiated and transformed cells. We therefore evaluated whether endogenous RT may represent a new molecular target in the treatment of human renal clear-cell carcinoma, a neoplasm with very low sensitivity to standard pharmacological therapies. Efavirenz and nevirapine, 2 non-nucleosidic RT inhibitors commonly used in HIV patients, either induced a reversible downregulation of cell proliferation or enhanced cell differentiation in primary cultures of human renal carcinoma cells characterized by high levels of endogenous RT activity. Both agents upregulated the expression of the vitamin D receptor and calbindin 28k genes, which are constitutively expressed in renal tubular cells, and induced vitamin D signaling by enhancing the ability of tumor cells to upregulate the vitamin D-dependent gene, CYP24. Furthermore, efavirenz- and nevirapine-differentiated tumor cells exhibited an immunogenic phenotype with an increased expression of HLA-I and CD40 antigens and an enhanced ability to elicit a specific T-cell response in mixed lymphocyte/tumor-cell cultures. Indeed, renal carcinoma cells exposed to efavirenz induced a CD8(+)CCR7-CD45RA(-) effector memory T-cell phenotype, whereas untreated RCC cells induced a CD8(+)CCR7(+)CD45RA(-) central memory T-cell phenotype. These data suggest that RT inhibitors may be a novel tool in the treatment of human renal clear-cell carcinoma, potentially able to enhance the immunogenic potential of tumor cell. ( view less ) Max Gassmann,Annamaria Manini,Thomas Stallmach,Bianca Saam,Gisela Kuhn,Beat Grenacher,Anna Y Bogdanova,Johannes Vogel We postulate that repeated pregnancy loss, intrauterine growth restriction, and preeclampsia are caused by impaired elevation of uterine blood flow due to disturbed arteriogenesis of the uterine arcade. This hypothesis is based on the observation that pregnant human erythropoietin-overexpressing (p... ( view more )lasma levels elevated 12-fold) mice (termed tg6 mice) suffering from excessive erythrocytosis generally abort at midgestation unless their hematocrit of 0.85 is drastically lowered. Transgenic mice show placental malformations that parallel those observed in pregnant women suffering from impaired uterine perfusion. Shear stress, a key factor inducing arteriogenesis, was 5-fold lower in tg6 mice compared with wildtype (WT) littermates. Consequently, uterine artery growth was reduced, and dramatically fewer viable pups (1.63 +/- 2.20 vs. 8.10 +/- 0.74 in WT) of lower weight (1.29 +/- 0.07 g vs. 1.62 +/- 0.12 g in WT) were delivered in first pregnancies. Only in subsequent pregnancies did tg6 deliver approximately the expected number of pups. Birth weights of tg6 offspring, however, remained reduced. As the spleen is a major site of extramedullary erythropoiesis in tg6 animals, splenectomy reduced the hematocrit to 0.6-0.7. In turn, shear stress increased to normal values, and splenectomized primiparous tg6 showed normal uterine artery growth and delivery of pups similar in number and weight compared with WT. We conclude that poor arteriogenesis is a previously unappreciated cause for clinically important pregnancy complications. ( view less ) Francesca Viparelli,Nunzianna Doti,Annamaria Sandomenico,Daniela Marasco,Nina A Dathan,Claudia Miele,Francesco Beguinot,Simona M Monti,Menotti Ruvo PLD's (Phospholipases D) are ubiquitously expressed proteins involved in many transphosphatidylation reactions. They have a bi-lobed structure composed by two similar domains which at their interface reconstitute the catalytic site through the association of the two conserved HxKx(4)Dx(6)GSxN motif... ( view more )s. PLD1 interacts with the small phosphoprotein PED-PEA15 by an unknown mechanism that, by enhancing PLD1 stability, apparently increases its enzymatic activity; the minimum interacting region of PLD1 was previously identified as spanning residues 712-1074 (D4 region). Since the D4/PED-PEA15 interaction has been claimed to be one of the multiple molecular events that can trigger type 2 diabetes, we purified the two recombinant proteins to study in vitro this binding by both ELISA and SPR techniques. Whilst PED-PEA15 was easily expressed and purified, expression of recombinant D4 was more problematic and only the fusion protein with Thioredoxin A and a six Histidine Tag (Trx-His(6)-D4) demonstrated sufficient stability for further characterization. We have found that Trx-His(6)-D4 is present as two different oligomeric forms, though only the monomeric variant is able to interact with PED-PEA15. All these findings may have important implications for both the mechanisms of phospholipase activity and PED-PEA15 regulative functions. ( view less ) Francesco Noè,Allan-Hermann Pool,Jari Nissinen,Marco Gobbi,Ross Bland,Massimo Rizzi,Claudia Balducci,Francesco Ferraguti,Gunther Sperk,Matthew J During,Asla Pitkänen,Annamaria Vezzani Temporal lobe epilepsy remains amongst the most common and drug refractory of neurological disorders. Gene therapy may provide a realistic therapeutic approach alternative to surgery for intractable focal epilepsies. To test this hypothesis, we applied here a gene therapy approach, using a recombin... ( view more )ant adeno-associated viral (rAAV) vector expressing the human neuropeptide Y (NPY) gene, to a progressive and spontaneous seizure model of temporal lobe epilepsy induced by electrical stimulation of the temporal pole of the hippocampus, which replicates many features of the human condition. rAAV-NPY or a control vector lacking the expression cassette (rAAV-Empty) was delivered into the epileptic rat hippocampi at an early progressive stage of the disease. Chronic epileptic rats were video-EEG monitored to establish pre-injection baseline recordings of spontaneous seizures and the effect of rAAV-NPY versus rAAV-Empty vector injection. Both non-injected stimulated controls and rAAV-empty injected rats showed a similar progressive increase of spontaneous seizure frequency consistent with epileptogenesis. The delivery of rAAV-NPY in epileptic rat brain leads to a remarkable decrease in the progression of seizures as compared to both control groups and this effect was correlated with the NPY over-expression in the hippocampus. Moreover, spontaneous seizure frequency was significantly reduced in 40% of treated animals as compared to their pre-injection baseline. Our data show that this gene therapy strategy decreases spontaneous seizures and suppresses their progression in chronic epileptic rats, thus representing a promising new therapeutic strategy. ( view less ) Oriana Simonetti,Oscar Cirioni,Roberto Ghiselli,Gaia Goteri,Alessandro Scalise,Fiorenza Orlando,Carmela Silvestri,Alessandra Riva,Vittorio Saba,Kiran D Madanahally,Annamaria Offidani,Naomi Balaban,Giorgio Scalise,Andrea Giacometti Quorum sensing is a mechanism through which a bacterial population receives input from neighboring cells and elicits an appropriate response to enable survival within the host. Inhibiting quorum sensing by RNAIII-inhibiting peptide (RIP) has been demonstrated as a very effective mode of prevention ... ( view more )and therapy for device-associated staphylococcal infections and was tested here for healing of wounds that are otherwise resistant to conventional antibiotics. Wounds, established through the panniculus carnosus of BALB/c mice, were inoculated with 5 x 10(7) CFU of methicillin-resistant Staphylococcus aureus. Mice were treated with Allevyn, RIP-soaked Allevyn (containing 20 microg RIP), daily intraperitoneal teicoplanin (7 mg/kg of body weight), Allevyn and teicoplanin, and RIP-soaked Allevyn and daily intraperitoneal teicoplanin. The main outcome measures were quantitative bacterial culture and histological examination with assessment of microvessel density and of vascular endothelial growth factor (VEGF) expression in tissue sections. Treatment with RIP-soaked Allevyn together with teicoplanin injection greatly reduced the bacterial load to 13 CFU/g (control untreated animals had 10(8) CFU/g bacteria). All other treatments were also significantly effective but only reduced the bacterial load to about 10(3) CFU/ml. Histological examination indicated that only treatment with RIP-soaked Allevyn with teicoplanin injection restored epithelial, granulation, and collagen scores, as well as microvessel density and VEGF expression, to the levels found with uninfected mice. In conclusion, we observed that RIP may be useful for the management of infected wounds and that it could represent an exciting and future alternative to the conventional antibiotics, at present considered the gold-standard treatments for methicillin-resistant S. aureus infections. ( view less ) Marco Guerrini,Daniela Beccati,Zachary Shriver,Annamaria Naggi,Karthik Viswanathan,Antonella Bisio,Ishan Capila,Jonathan C Lansing,Sara Guglieri,Blair Fraser,Ali Al-Hakim,Nur Sibel Gunay,Zhenqing Zhang,Luke Robinson,Lucinda Buhse,Moheb Nasr,Janet Woodcock,Robert Langer,Ganesh Venkataraman,Robert J Linhardt,Benito Casu,Giangiacomo Torri,Ram Sasisekharan Recently, certain lots of heparin have been associated with an acute, rapid onset of serious side effects indicative of an allergic-type reaction. To identify potential causes for this sudden rise in side effects, we examined lots of heparin that correlated with adverse events using orthogonal high... ( view more )-resolution analytical techniques. Through detailed structural analysis, the contaminant was found to contain a disaccharide repeat unit of glucuronic acid linked beta1-->3 to a beta-N-acetylgalactosamine. The disaccharide unit has an unusual sulfation pattern and is sulfated at the 2-O and 3-O positions of the glucuronic acid as well as at the 4-O and 6-O positions of the galactosamine. Given the nature of this contaminant, traditional screening tests cannot differentiate between affected and unaffected lots. Our analysis suggests effective screening methods that can be used to determine whether or not heparin lots contain the contaminant reported here. ( view less ) Annamaria Colao,Josef Marek,Miklos I Goth,Philippe Caron,Jean Marc Kuhn,Francesco M Minuto,Neil J Weissman Context: Excess GH and IGF-I in acromegaly are associated with reduced life expectancy due to cardiovascular complications. Objective: The objective of the study was to investigate the prevalence, incidence, and severity of cardiac valve regurgitation before and after somatostatin-analog treatment ... ( view more )in acromegaly. Design: This was a prospective, observer-blinded, multicenter, 12-month study. Setting: The study was conducted at 33 specialist centers. Patients: The study population consisted of 225 adult patients with acromegaly without significant cardiac valve abnormalities or prior valve-replacement surgery, matched for age, sex, and center/country/study. Interventions: Interventions included initiation/continuation of lanreotide (n = 107) or octreotide treatment (n = 118), tailored for optimal disease control. Main Outcome Measures: Relative risk of new/worsening regurgitation in any valve at 12 months compared with baseline, was measured. Results: At baseline, almost 80% of patients had some degree of cardiac valve regurgitation, although none was severe. The risk of developing new/worsening regurgitation in any valve at 12 months was nonsignificant and similar for the cohorts [adjusted odds ratio 0.86; 95% confidence interval (CI) 0.41-1.82; P = 0.694; relative risk 1.04; 95% CI 0.67-1.60; risk difference 0.01; 95% CI -0.13 to 0.16]. For 54% of patients, the severity of regurgitation stayed the same during the study. At baseline, significant valve regurgitation occurred in 18% of patients (lanreotide cohort) and 13% (octreotide cohort) and at 12 months in 18% of each cohort. Conclusions: The incidence of valve regurgitation did not change over 12 months of treatment with somatostatin analogs, and most cases were physiologic or mild in severity. There was no significant difference between somatostatin analogs in the risk of developing new/worsening valve regurgitation or significant regurgitation after 1 yr. ( view less ) Carlo Calabrese,Annamaria Pisi,Giulio Di Febo,Giuseppina Liguori,Gianfranco Filippini,Maurizio Cervellera,Valeria Righi,Patrizia Lucchi,Adele Mucci,Luisa Schenetti,Valeria Tonini,Maria Raffaella Tosi,Vitaliano Tugnoli Background and Aims: The metabolic profile and morphologic aspects of normal and pathologic human gastric mucosa were studied. The aim of the present research was the application of ex vivo high-resolution magic angle spinning magnetic resonance spectroscopy (HR-MAS MRS) to the human gastric tissue... ( view more ) to get information on the molecular steps involved in gastric carcinogenesis and the identification of biochemical markers useful for the development of in vivo MRS methodologies to diagnose gastric pathologies in clinical situations. METHODS: Twelve normal subjects, five with autoimmune atrophic gastritis, five with Helicobacter pylori infection, and five with adenocarcinoma were examined. Ten biopsies were taken during endoscopy from each patient. Specimens from carcinoma were also obtained during gastrectomy. Of the 10 biopsies, 4 were used for histologic evaluation, 4 were fixed in glutaraldehyde and processed for transmission and scanning electron microscopy, and 2 were immersed in liquid nitrogen and stored at -85 degrees C for monodimensional and bidimensional ex vivo HR-MAS MRS analysis. RESULTS: Ex vivo HR-MAS MRS identified glycine, alanine, free choline, and triglycerides as possible molecular markers related to the human gastric mucosa differentiation toward preneoplastic and neoplastic conditions. Ultrastructural studies of autoimmune atrophic gastritis and gastric adenocarcinoma revealed lipid accumulations intracellularly and extracellularly associated with a severe prenecrotic hypoxia and mitochondria degeneration. CONCLUSIONS: This is the first report of synergic applications of ex vivo HR-MAS MRS and electron microscopy in studying the human gastric mucosa differentiation. This research provides useful information about some molecular steps involved in gastric carcinogenesis. The biochemical data obtained on gastric pathologic tissue could represent the basis for clinical applications of in vivo MRS. (Cancer Epidemiol Biomarkers Prev 2008;17(6):1386-95). ( view less ) Miguel Saps,Licia Pensabene,Lucio Di Martino,Annamaria Staiano,Joshua Wechsler,Xiaotian Zheng,Carlo Di Lorenzo OBJECTIVE: To investigate the development of functional gastrointestinal disorders (FGIDs) after an episode of acute bacterial gastroenteritis (AGE) in children. STUDY DESIGN: A cohort study of children 3 to 19 years old with a positive result on a bacterial stool culture. 44 patients in each arm (... ( view more )unidirectional alpha of 0.05, power of 0.80). Children presenting at two pediatric hospitals (United States and Italy) for AGE who tested positive for bacteria on stool culture (2001-2005) were contacted at least 6 months after the episode. Exposed children were matched with control subjects of similar age and sex consulting to the same hospitals for trauma or well-child visit within 4 weeks of the index case. Symptoms were evaluated with a validated questionnaire for FGIDs assessing pain, diarrhea, and disability. RESULTS: 88 patients (46 boys; mean age, 8.1 years; age range, 3-19 years) were recruited. Bacteria included Salmonella (54%), Campylobacter (32%), and Shigella (14%). 36% of exposed patients and 11% of control subjects complained of abdominal pain (P < .01). 87% had irritable bowel syndrome and 24% had dyspepsia. 56% reported onset of pain following the AGE. CONCLUSION: There is a significant increase in cases of FGIDs after bacterial infections in children. ( view less ) Francesco Orio,Francesco Giallauria,Stefano Palomba,Francesco Manguso,Marcello Orio,Domenico Tafuri,Gaetano Lombardi,Enrico Carmina,Annamaria Colao,Carlo Vigorito OBJECTIVE: The aim of the present study was to determine if the favourable cardiopulmonary and metabolic benefits induced by exercise training (ET) programme are maintained after its cessation. PATIENTS: Thirty-two young overweight polycystic ovary syndrome (PCOS) women matched for age and body mas... ( view more )s index (BMI) with other 32 PCOS patients was enrolled. The first group [PCOS-T (trained)] underwent 24-week ET programme, whereas the second [PCOS-DT (detrained)] underwent 12-week ET programme followed by 12-week detraining period. METHODS: At baseline, after 12- and 24-week follow-up, all PCOS women were studied for their hormonal (ovarian and adrenal androgens), metabolic (glucose and insulin) and lipid profile, and underwent cardiopulmonary exercise test. RESULTS: After the initial 12-week ET programme, both PCOS-T and PCOS-DT groups, without differences between groups, showed a similar significant (P < 0.05) improvement in BMI, fasting insulin, areas under curve insulin (AUC(INS)), glucose and insulin AUC (AUC(GLU/INS)), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C) and maximal oxygen consumption at cardiopulmonary exercise test (VO2max). At 24-week follow-up, PCOS-T group showed a significant (P < 0.05) improvement in BMI, fasting insulin, AUC(INS), AUC(GLU/INS), LDL-C, HDL-C and VO2max, in comparison to baseline and 12-week follow-up. At same follow-up visit, the all parameters resulted significantly (P < 0.05) worsened in PCOS-DT group in comparison to 12-week follow-up and PCOS-T group. In PCOS-DT group, no parameter assessed at 24-week follow-up was significantly different in comparison with baseline. CONCLUSION: In young PCOS women, 12-week detraining resulted in a complete loss of the favourable adaptations obtained after ET. ( view less ) Teresa Zotta,Annamaria Ricciardi,Felicia Ciocia,Rocco Rossano,Eugenio Parente Exponential and stationary phase cells of 56 strains of the dairy Streptococcus species S. thermophilus, S. macedonicus and S. salivarius, were exposed to acid, osmotic, oxidative and heat stresses to investigate the diversity of their responses. Three strains of dairy related streptococci, Lactoco... ( view more )ccus lactis ATCC11454, Enterococcus faecium DSM20477 and Enterococcus faecalis DSM20478, were included for comparison purposes. Acid and heat adaptation and cross-protection to stress were studied in ten strains with different stress response patterns. Cell death and the changes in protein expression were evaluated by plate counts and Sodium Dodecyl Sulfate Polyacrilamide Gel Electrophoresis, respectively. All strains of all species were highly tolerant of osmotic stress. With a few exceptions, acid and oxidative treatments reduced the number of viable cells by >5 log units but responses to heat stress were more variable. For some, but not all strains, stationary phase cells were more resistant to some or all stresses. Matrix cluster analysis was used to group strains on the basis of their pattern of stress response in seven clusters. Significant associations between the sources of strains and stress resistance were found for acid and oxidative stresses. Adaptation to stress during the exponential phase enhanced the survival of acid and heat stressed cells from 1 to 60,000-folds, but a detrimental effect of adaptation on cell viability was evident for oxidative and osmotic stresses for three strains. Adaptation and entry into the stationary phase resulted in significant changes of protein bands whose estimated molecular masses corresponded with those of proteins (DnaK, GprE, GroEL, and GroES) involved in the general stress response but no statistically significant correlation between stress response and band intensity was evident. ( view less )
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