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Roberto Verlato,Francesco Zanon,Emanuele Bertaglia,Pietro Turrini,Maria Stella Baccillieri,Enrico Baracca,Maria Grazia Bongiorni,Aldo Zampiero,Pietro Zonzin,Pietro Pascotto,Diego Venturini,Giorgio Corbucci OBJECTIVES: To evaluate the prevalence of severe right atrial conduction delay in patients with sinus node dysfunction (SND) and atrial fibrillation (AF) and the effects of pacing in the right atrial appendage (RAA) and in the inter-atrial septum (IAS). METHODS: Forty-two patients (15 male, 72 +/- ... ( view more )7 years) underwent electrophysiologic study to measure the difference between the conduction time from RAA to coronary sinus ostium during stimulation at 600 ms and after extrastimulus (DeltaCTos). Patients were classified as group A if DeltaCTos > 60 ms and group B if < 60 ms. Each Group was randomized to RAA/IAS pacing and algorithms ON/OFF. RESULTS: Fifteen patients (36%, group A) had DeltaCTos = 76 +/- 11 ms and 27 patients (64%, group B) had DeltaCTos = 36 +/- 20 ms. Twenty-two patients were paced at the RAA and 20 at the IAS. During the study, no AF recurrences were reported in 11 of 42 (26%) patients, independently of RAA or IAS pacing. Patients from group A and RAA pacing had 0.79 +/- 0.81 episodes of AF/day during DDD, which increased to 1.52 +/- 1.41 episodes of AF/day during DDDR + Alg (P = 0.046). Those with IAS pacing had 0.5 +/- 0.24 episodes of AF/day during DDD, which decreased to 0.06 +/- 0.08 episodes of AF/day during DDDR + Alg (P = 0.06). In group B, no differences were reported between pacing sites and pacing modes. CONCLUSIONS: Severe right atrial conduction delay is present in one-third of patients with SND and AF: continuous pacing at the IAS is superior to RAA for AF recurrences. In patients without severe conduction delay, no differences between pacing site or mode were observed. ( view less ) Cinzia Di Pietro,Marco Ragusa,Laura Duro,Maria Rosa Guglielmino,Davide Barbagallo,Alisia Carnemolla,Alessandro Laganà,Pietro Buffa,Rosario Angelica,Antonella Rinaldi,Maria Stella Calafato,Ionella Milicia,Cinzia Caserta,Rosalba Giugno,Alfredo Pulvirenti,Veronica Giunta,Antonella Rapisarda,Valentina Di Pietro,Agata Grillo,Angelo Messina,Alfredo Ferro,Karl Heinz Grzeschik,Michele Purrello TBPL2 is the most recently discovered and less characterized member of the TATA box binding protein (TBP) family that also comprises TBP, TATA box binding protein-like 1 (TBPL1), and Drosophila melanogaster TBP related factor (TRF). In this paper we report our in silico and in vitro data on (i) the... ( view more ) genomics of the TBPL2 gene in Homo sapiens, Pan troglodytes, Mus musculus, Rattus norvegicus, Gallus gallus, Xenopus tropicalis, and Takifugu rubripes; (ii) its evolution and phylogenetic relationship with TBP, TBPL1, and TRF; (iii) the structure of the TBPL2 proteins that belong to the recently identified group of the intrinsically unstructured proteins (IUPs); and (iv) TBPL2 expression in different organs and cell types of Homo sapiens and Rattus norvegicus. Similar to TBP, both the TBPL2 gene and protein are bimodular. The 3' region of the gene encoding the DNA binding domain (DBD) was well conserved during evolution. Its high homology to vertebrate TBP suggests that TBPL2 also should bind to the TATA box and interact with the proteins binding to TBP carboxy-terminal domain, such as the TBP associated factors (TAFs). As already demonstrated for TBP, TBPL2 amino-terminal segment is intrinsically unstructured and, even though variable among vertebrates, comprises a highly conserved motif not found in any other known protein. Absence of TBPL2 from the genome of invertebrates and plants demonstrates its specific origin within the subphylum of vertebrates. Our RT-PCR analysis of human and rat RNA shows that, similar to TBP, TBPL2 is ubiquitously synthesized even though at variable levels that are at least two orders of magnitude lower. Higher expression of TBPL2 in the gonads than in other organs suggests that it could perform important functions in gametogenesis. Our genomic and expression data should contribute to clarify why TBP has a general master role within the transcription apparatus (TA), whereas both TBPL1 and TBPL2 perform tissue-specific functions. ( view less ) G Divita,R S Goody,D C Gautheron,A Di Pietro The intrinsic tryptophan fluorescence of Schizosaccharomyces pombe mitochondrial F1 is a very sensitive probe to differentiate nucleotide binding to catalytic and noncatalytic sites (Divita, G., Di Pietro, A., Roux, B., and Gautheron, D. C. (1992) Biochemistry 31, 5791-5798), the catalytic site sat... ( view more )uration producing quenching of Trp-257 fluorescence (Divita, G., Jault, J.-M., Gautheron, D. C., and Di Pietro, A. (1993) Biochemistry 32, 1017-1024). The present results indicate that two types of fluorescent nucleotide analogues, bearing either 2'(3')N-methylanthraniloyl (mant) or 2',3'-O-(2,4,6-trinitrophenyl) (TNP) group, exhibit high-affinity binding and behave similarly to the corresponding unmodified nucleotides. Selective binding of mant GDP to the catalytic site produces a marked quenching of intrinsic fluorescence which is due to resonance energy transfer between Trp-257 and the mant group. The high efficiency of the transfer allows the determination of a short distance, 10.5 A, indicating the close proximity of catalytic site and alpha-subunit Trp-257. Selective saturation of the noncatalytic site by TNP-ADP produces a marked quenching of the extrinsic fluorescence of mant GDP bound to the catalytic site, which is correlated to an important resonance energy transfer between the two fluorescent groups. A rather short distance of 17.5 A is calculated, indicating vicinity of catalytic and noncatalytic sites. ( view less ) Irene Aprile,Enrico Di Stasio,Federica Romitelli,Stefano Lancellotti,Pietro Caliandro,Pietro Tonali,Alessandro Gilardi,Luca Padua PRIMARY OBJECTIVE: To evaluate the effects of rehabilitation on disability and quality of life (QoL) in patients with chronic stroke. Research design: This is an observational, prospective and multidimensional study. METHODS AND PROCEDURES: Sixty-six consecutive inpatients were enrolled. A rehabili... ( view more )tation programme of 2 months consisting of six exercise periods per week was performed. An impairment of cognitive function was the exclusion criteria for patient enrolment. MAIN OUTCOMES AND RESULTS: European Stroke Scale, Barthel Index, Modified Rankin Scale, Deambulation Index and SF-36 were performed both before and after rehabilitation. A short-term follow-up (2 months after discharge) was performed by phone administration of SF-36. Significant improvement in clinical and disability measurements was observed after rehabilitation. For QoL, a significant improvement was observed both in physical function and in social function immediately after rehabilitation. At follow-up the social function improvement was maintained, while physical function improvement was lost. CONCLUSIONS: The data show that rehabilitation in patients with chronic stroke lessens disability and improves physical and social function and that repeated cycles of treatment are needed to maintain the level of improvement reached. ( view less ) Pietro Crispino,Giorgio De Toma,Antonio Ciardi,Antonino Bella,Margherita Rivera,Giuseppe Cavallaro,Andrea Polistena,Francesca Fornari,Hans Unim,Roberta Pica,Claudio Cassieri,Pietro Luigi Mingazzini,Paolo Paoluzi BACKGROUND: Colorectal cancer (CRC) metastasis is enhanced in patients with venous embolization increasing the risk of recurrence and therefore mortality rate. Several evidences indicate that stage II patients have an abrupt recurrence within five years from surgery. This fact, led us to investigat... ( view more )e the role played by different histological variables on CRC invasiveness. AIM: To demonstrate if quantitative and qualitative desmoplastic response and lymphocytic infiltration are prognostic factor involved in the recurrence of CRC within five years from surgery, considering possible clinical and therapeutical implications. METHODS: Thirty-four patients with CRC underwent colectomy and the UICC-TNM classification was applied for disease staging. Histological variables were semi-quantitatively evaluated. Qualitative evaluation of desmoplasia was obtained with the hematoxillin-eosin method. RESULTS: Survival rate arose 88% at stage II, at five years of follow-up, and the 12% not treated with adjuvant chemotherapy developed metastasis. Desmoplasia is strongly associated with venous neoplastic invasiveness (OR: 21.93; 95%CI: 1.012-475.26, p = 0.02), and therefore, with mortality rate (OR: 14.33; 95%CI: 0.67-304, p = 0.04). Moreover, mortality rate was significantly higher in patients with immature desmoplasia compare to mature stromal tissue (OR: 15.61, 95%CI: 0.69-343.38, p = 0.04). CONCLUSIONS: These observations should prompt a future evaluation of desmoplasia to extent more suitably the use of adjuvant chemotherapy in II stage patients. Further clinical trials are needed to determine if these findings will be able to reduce mortality rate, in stage II CRC patients. ( view less ) Pietro Caliandro,Graziano Grugni,Luca Padua,Yllka Kodra,Pietro Tonali,Luigi Gargantini,Letizia Ragusa,Antonino Crinò,Domenica Taruscio AIM: The goals of this study are to investigate the quality of life of Prader-Willi syndrome patients and to evaluate the relationship between quality of life and the clinical picture. METHODS: We performed a multicentric study on 40 consecutive patients with Prader-Willi syndrome. Quality of life ... ( view more )was evaluated through the Short Form-36 and the Child Health Questionnaire-Parent Form-50 according to the age of patients. RESULTS: In patients older than 14 years old, quality of life is intensely impaired both in mental and physical aspects. Weight at the moment of the observation, birthweight and facial features are the main variables that influence quality of life. In patients who are 14 years old or younger, the Family Activity and Physical scores are lower for those patients with characteristic facial features and in patients with decreased fetal movement or infantile lethargy. Self-esteem is lower in patients with a higher Mini Mental Score. CONCLUSIONS: Physical and mental aspects of quality of life are impaired in Prader-Willi patients, weight is the clinical finding which mainly influences negatively the physical aspects of quality of life. However, weight does not cause mental problems. These are mainly due to the presence of characteristic facial features. Interestingly, a high birthweight is associated with less impairment of the mental aspects of quality of life. ( view less ) Clelia Miracco,Vasileios Mourmouras,Maurizio Biagioli,Pietro Rubegni,Susanna Mannucci,Irene Monciatti,Elena Cosci,Piero Tosi,Pietro Luzi Tumour-infiltrating lymphocytes (TILs) represent the local immune response to cancer, however, their correlation with tumour behaviour is not unanimously considered in the literature. Most studies have not characterized TILs, that are known to comprise distinct subsets, bearing different roles in t... ( view more )he complex tumour microenvironment. Characterization of patient lymphocytes has been mainly performed in peripheral blood, that is not always representative of the local immune status. Only few investigations have been performed at the tissue level in cancer, including melanoma. TILs encompass different populations of effector and regulatory T cells (Tregs), and the relevance of the latter in tumour progression is widely accepted. The transcription factor gene product FOXP3 is considered the most reliable marker of Tregs. However, it has not been extensively evaluated in primary cutaneous melanoma. We analyzed 66 vertical growth phase primary cutaneous melanomas, aiming at finding differences in TIL subsets between two groups of cases, that behaved differently in terms of local recurrence. In our study, the percentage of Tregs, as characterized by CD25 and FOXP3 expression, both among tumour cells, inside tumour parenchyma and at its periphery, and among TILs, at the tumour-stroma boundary, was significantly higher in cases that recurred than in those that did not (p=0.00065; p=0.00014; p<0.00001, respectively). TIL characterization by immunohistochemistry in melanoma diagnostic reports, could add further information. The analysis of a larger series of patients and correlation with other clinical parameters, such as distant metastases and/or patient survival, are mandatory for validating its use as a prognostic indicator. ( view less ) Francesco Angelucci,Valerio Ricci,Massimiliano Pomponi,Gianluigi Conte,Aleksander A Mathé,Pietro Attilio Tonali,Pietro Bria Chronic cocaine and heroin users display a variety of central nervous system (CNS) dysfunctions including impaired attention, learning, memory, reaction time, cognitive flexibility, impulse control and selective processing. These findings suggest that these drugs may alter normal brain functions an... ( view more )d possibly cause neurotoxicity. Neurotrophins are a class of proteins that serve as survival factors for CNS neurons. In particular, nerve growth factor (NGF) plays an important role in the survival and function of cholinergic neurons while brain-derived neurotrophic factor (BDNF) is involved in synaptic plasticity and in the maintenance of midbrain dopaminergic and cholinergic neurons. In the present study, we measured by enzyme-linked immunosorbent assay (ELISA) the NGF and BDNF levels in serum of three groups of subjects: heroin-dependent patients, cocaine-dependent patients and healthy volunteers. Our goal was to identify possible change in serum neurotrophins in heroin and cocaine users. BDNF was decreased in heroin users whereas NGF was decreased in both heroin and cocaine users. These findings indicate that NGF and BDNF may play a role in the neurotoxicity and addiction induced by these drugs. In view of the neurotrophin hypothesis of schizophrenia the data also suggest that reduced level of neurotrophins may increase the risk of developing psychosis in drug users. ( view less ) Marco Sacchi,Pietro D Legge,Pietro Picozzi,Francesco Papa,Capuano Loreto Giovanni,Luigi Greco Surgical management of rectal cancer has undergone a significant change during the past two decades. Low anterior resection (LAR) with total mesorectal excision (TME) is, at the moment, the "gold standard" for carcinoma of the mid or lower rectum. Because the most specific complication following re... ( view more )ctal resection with anastomosis is symptomatic leakage, which is associated with 18% mortality rate, routine formation of a temporary stoma is suitable after sphincter-saving resection for anastomoses situated at or less than 5cm from the anal verge. Actually the preferred modes of fecal diversion following LAR with TME are loop ileostomy or loop transverse colostomy. Low anastomosis, preoperative radiation or chemoradiation, presence of intraoperative adverse events and male gender are independent risk factors for symptomatic anastomotic leakage. A defunctioning loop ileostomy or the classical "protective" colostomy requires subsequent reconstructive surgery with a significant postoperative morbidity. For these reasons we use an alternative to protect a high risk anastomosis with fashioning a proximal intraabdominal closed loop ileostomy called "virtual ileostomy". In a seven-year period from 1999 to 2005 a total of 107 patients underwent elective anterior resection of the rectum for carcinoma, in all cases was fashioned a virtual ileostomy. The incidence of symptomatic clinically evident anastomotic leakage was 13%; in all the cases (14 pts) the closed loop ileostomy was opened with a reduction of the originally planned number of ileostomies by over 80%. The procedure is easy to perform and well accepted by the patients. It avoids a second operation. ( view less ) Katia Marinelli,Patrizia Bagnarelli,Gianni Gaffi,Silvia Trappolini,Pietro Leoni,Alessandra Mataloni Paggi,Agnese Della Vittoria,Giorgio Scalise,Pietro Emanuele Varaldo,Stefano MenzoTesting for viral BKV-DNA in urine is a non-invasive early detection and monitoring tool in the diagnostic of BKV-related pathologies: quantitative analysis by Real-Time PCR can provide useful information in addition to cytologic analysis, although our study suggests that high BKV viruria is not ne... ( view more )cessarily associated with kidney or bladder damage. ( view less ) Pietro Cugini,Mario Curione,Claudia Castro,Angela Napoli,Pietro Francia,Antonietta Colatrella,Simonetta Di Bona,Camillo Cammarota,Gianbattista Cardarelli,Silvia Amato,Francesco Fallucca OBJECTIVE: The scope of this study is to detect whether or not the entropy (E) circadian rhythm (CR) is maintained preserved in sinusal R-R intervals (SRRI), its loss being the expression of a transition to an indeterministic chaos in heart rate variability (HRV). METHODS: The E of SRRI was estimat... ( view more )ed in 14 type I diabetic pregnant women (DPW) in the first trimester of an apparently uncomplicated gestation (7 patients - mean age = 30.3 +/- 4.1 y - without clinical and laboratory evidence of cardiac autonomic neuropathy, and 7 patients - mean age = 30.7 +/- 3.6 y - with positive tests for a cardiac dysautonomia). The E CR was studied via the single cosinor method, and summarized via the population-mean cosinor method. RESULTS: The E CR was found not to be preserved in both the investigated type I DPW, despite the occurrence of the SRRI CR. CONCLUSIONS: The loss of the E CR confirms that in type I DPW there is a transition to an indeterministic disorder in HRV due to the lack of an autocorrelated periodic chaos in cardiac pacing. Such an unphysiological neurovegetative regulation suggests a new silent cardiac dysautonomic syndrome, that we intend to call "minimum delirium cordis syndrome" (MDCS). Can the MDCS be regarded as a condition of cardiovascular risk? To answer this question, it seems justified to suggest that the study of the E CR should be added to the routine tests that are presently applied to clinical analysis of the Holter ECG, being the classic tests of linear analysis not methodologically suitable for detecting the indeterministic chaos of the MDCS. ( view less ) Carmelo Romeo,Teresa Arrigo,Pietro Impellizzeri,Agatino Manganaro,Pietro Antonuccio,Giuseppe Di Pasquale,Maria Francesca Messina,Lucia Marseglia,Isabella Formica,Biagio Zuccarello BACKGROUND/PURPOSE: Different studies have evaluated testicular hormonal dysfunction in adolescent varicocele but with variable results. Recently, inhibin B has been proposed as a marker for spermatogenesis and Sertoli cell function. The aim of the present study was to study in a homogeneous cohort... ( view more ) of adolescents inhibin B and other hormones to detect whether untreated varicocele may be associated with any modifications of these factors. METHODS: Sixteen adolescents (mean age, 14.5 +/- 1.0 years), at Tanner stages 4 to 5 with grade II or III left-sided varicocele, underwent hormonal evaluation of inhibin B, basal testosterone, and both baseline and GnRH stimulated FSH and LH levels. Thirteen unaffected adolescents of age-matched and pubertal development were used as controls. RESULTS: Patients with varicocele showed a significant reduction in the testicular volume of the affected side (13.3 +/- 4.1 vs 15.8 +/- 4.8 mL; P = .002) and significant reduced levels of inhibin B compared with controls (271.9 +/- 70.2 vs 327.1 +/- 34.9 pg/mL; P = .042). Inhibin B levels were significantly correlated with testes volume (r = 0.62; P = .0097). Other hormonal parameters were similar in both varicocele and control groups. CONCLUSION: Inhibin B is reduced in adolescents with untreated varicocele, and it is positively correlated with testicular volume. This could be an early marker for Sertoli cell damage, with possible implications for spermatogenesis and could represent a new indication for varicocele repair. ( view less ) Laura Settimi,Franca Davanzo,Pietro Carbone,Fabrizio Sesana,Carlo Locatelli,Maria Luisa Farina,Pietro Maiozzi,Paolo Roazzi,Fabio Maccari,Luigi Macchi,Antonio Fanuzzi Between 1 February and 31 March 2006, the Poison Control Centers (PPC) active in Lombardy collaborated with an integrated surveillance system carried out in Piedmont during the Olympic Games 2006. The collaborating PPC notified to the system 697 human cases of exposure occurred in Piedmont during t... ( view more )he observation period. Among these cases, 70% were exposed accidentally, 40% were 6 years old or younger, and 45% reported at least a clinical effect. The agents more frequently reported were: cleaning substances (household) (110 cases), fumes/gases/vapors (63 cases, comprising 38 cases accidentally exposed to carbon monoxide), and sedative/hypnotics/antipsychotics (53 cases). Although very limited, the available observations focused the attention on specific hazards and were able to highlight the potential of a toxic exposure surveillance system based on the information reported by the Italian PPC. ( view less ) Laura Spizzichino,Giovanna Pedone,Pietro Gattari,Anna Maria Luzi,Pietro Gallo,Rudi Valli,Giovanni RezzaWomen account for nearly half the people living with HIV worldwide. This situation makes it necessary to improve prevention actions targeting women: the female condom is a good option. The study was conducted, the first in Italy, in a public AIDS Center on a sample of 162 participants (66.7% female... ( view more ), 33.3% men) who requested the HIV test. The objectives were: assess the current knowledge of the female condom; collect information on opinions, impressions and willingness to use the female condom. Participants were administered a Lickert-scale questionnaire after post-test counselling. The results are in line with international studies and show an early positive response, characterized by interest and openness to innovation, followed by resisting to use the female condom. ( view less ) Pietro Delva,Maurizio Degan,Paola Vallerio,Enrico Arosio,Pietro Minuz,Gabriella Amen,Marzia Di Chio,Alessandro Lechi OBJECTIVE(S): The eventual role of blood pressure on the endothelial progenitor cell (EPC) has rarely been evaluated and data collected so far relate to patients with co-existing coronary heart disease. METHODS: We have studied the number and functional activity of EPC as well as the number of EPC ... ( view more )endothelial colony-forming units (CFU) in a carefully selected group of 36 patients with essential hypertension and 24 normotensive control subjects. RESULTS: In patients with essential hypertension, the EPC number was not statistically different from that found in control subjects (mean +/- SD, essential hypertension 58 +/- 29, controls 53 +/- 20; EPC/high power field). CFU per well were not statistically different in patients with essential hypertension compared with normotensive controls (mean +/- SD, patients with essential hypertension 2.4 +/- 2.6, normotensive controls 3 +/- 3.3 CFU/well). In essential hypertension patients, the EPC number was inversely correlated with both total (R=0.635, P < 0.0001) and low-density lipoprotein (LDL)-cholesterol (R=0.486, P < 0.05). Neither the EPC number nor the EPC CFU were correlated with age, systolic blood pressure, diastolic blood pressure, body mass index, lipoprotein(a), high-sensitivity C-reactive protein or homocysteine. CONCLUSIONS: The present study shows that essential hypertension is not characterized by the altered number or functional activity of EPC. Plasma total and LDL-cholesterol are independent predictors of reduced numbers of circulating EPC in essential hypertension patients. The absence of any correlation between the characteristics of EPC and several markers predictive of cardiovascular damage merits further investigation. ( view less ) Rosalia Patti,Piero Luigi Almasio,Almasio Piero Luigi,Matteo Arcara,Arcara Matteo,Sergio Sammartano,Sammartano Sergio,Pietro Romano,Romano Pietro,Calogero Fede,Fede Calogero,Gaetano Di Vita,Di Vita Gaetano PURPOSE: The maximum resting pressure in the anal canal is greatly raised after hemorrhoidectomy. This increase is likely to be the cause of postoperative pain, which is still the most troublesome early problem after hemorrhoidectomy. This study was designed to compare, after hemorrhoidectomy, the ... ( view more )effects of intrasphincter injection of botulinum toxin vs. application of glyceryl trinitrate ointment in improving wound healing and reducing postoperative pain at rest or during defecation. METHODS: Thirty patients with hemorrhoids of third and fourth degree were included in the study and randomized in two groups. Anorectal manometry was performed preoperatively and 5 and 40 days after hemorrhoidectomy. One group received one injection containing 20 IU of botulinum toxin, whereas the other an application of 300 mg of 0.2 percent glyceryl trinitrate ointment three times daily for 30 days. RESULTS: Five days after hemorrhoidectomy, maximum resting pressure was significantly reduced compared with baseline values in both groups (85 +/- 15 vs. 68 +/- 11 mmHg for the group treated with botulinum toxin, 87 +/- 11 vs. 78 +/- 11 mmHg for the group treated with glyceryl trinitrate ointment). Overall analysis of postoperative pain at rest showed a significant reduction in the botulinum toxin group vs. glyceryl trinitrate group, whereas pain during defecation and time of healing were similar. Adverse effects, such as headaches, were observed only in the glyceryl trinitrate group. Forty days after hemorrhoidectomy in the glyceryl trinitrate group, maximum resting pressure values were similar to preoperative ones, whereas the values were still reduced in the botulinum toxin group. CONCLUSIONS: A single intrasphincter injection of botulinum toxin was more effective and safer than repeated applications of glyceryl trinitrate in reducing early postoperative pain at rest but not during defecation. ( view less ) Angela Spanu,Orazio Schillaci,Pietro Pirina,Antonio Arru,Giordano Madeddu,Francesca Chessa,Pietro Marongiu,Maria Elisabetta Solinas,Giuseppe Madeddu A correct differential diagnosis between benign and malignant lesions is mandatory in patients with solitary pulmonary nodule (SPN). The aim of the present study was to investigate whether 99mTc-tetrofosmin SPECT may play a role in SPN evaluation. A consecutive series of 111 patients with an uncalc... ( view more )ified =3 cm (range: 0.8-3 cm) SPN, without definite benign findings and indeterminate at CT, were studied. Within 1 week of CT scan, following 740 MBq of 99mTc-tetrofosmin i.v. injection, all patients underwent chest SPECT using a rectangular dual head gamma camera with HR collimators. The images were analysed both qualitatively and semiquantitatively by calculating tumor/normal tissue ratio (T/N). All nodules were referred to a definitive diagnosis after scintigraphy: 84/111 nodules resulted malignant (primary lung carcinomas in 59 cases and metastases in 25), whereas 27/111 were benign. SPECT was true positive in 77/84 malignant nodules (overall sensitivity: 91.7%), detecting 55/59 carcinomas (93.2%) and 22/25 metastases (88%), whereas it was false negative in 4 carcinomas (3 adenocarcinomas and 1 squamous cell carcinoma, the latter with necrotic areas; range size: 1.5-2.4 cm) and in 3 metastases (range size: 1.0-1.2 cm). SPECT was true negative in 24/27 benign lesions (specificity: 88.9%) and false positive in 2 hamartomas and in 1 aspecific inflammation (range size: 0.8-2 cm), each with a T/N value =1.4. Accuracy, positive predictive value and negative predictive value were 91, 96.2 and 77.4%, respectively. Mean T/N value was significantly higher in malignant than in benign nodules (2.1+/-0.6 vs. 1.3+/-0.1, P<0.05), whereas no significant differences were observed between primary lung carcinomas and metastases (2.1+/-0.6 vs. 1.9+/-0.6) or in the different histologic types of carcinomas. 99mTc-tetrofosmin SPECT proved a highly sensitive imaging method in both primary and secondary malignant =3 cm SPNs detection, with a high accuracy value in discriminating malignant from benign lesions, also by adding semiquantitative analysis. A larger clinical application of this non-invasive, simple and widely available procedure is thus suggested in SPN management, especially when FDG-PET is not available. ( view less ) Vincenzo Di Lazzaro,Fabio Pilato,Michele Dileone,Federico Ranieri,Valerio Ricci,Paolo Profice,Pietro Bria,Pietro A Tonali,Ulf Ziemann Inhibition is of fundamental importance to regulate activity in cortical circuits. Inhibition is mediated through a diversity of different interneurones and gamma-aminobutyric acid A receptor (GABA(A)R) subtypes. Here we employed paired-pulse transcranial magnetic stimulation (TMS) to measure short... ( view more ) interval intracortical inhibition (SICI), a GABA(A)R-mediated inhibition in human motor cortex, to address the question of which GABA(A)R subtype is responsible for this form of inhibition. It has been shown that classical benzodiazepines (diazepam and lorazepam) have a non-selective affinity profile at different alpha-subunit-bearing subtypes of the GABA(A)R while zolpidem has a 10-fold greater affinity to the alpha1-subunit-bearing GABA(A)R compared with those bearing the alpha2- or alpha3-subunit. We found that, in seven healthy subjects, a single oral dose of 20 mg of diazepam or 2.5 mg of lorazepam significantly increased SICI, whereas 10 mg of zolpidem did not change SICI. This dissociation occurred despite equal sedation by all three drugs, an alpha1-subunit GABA(A)R-mediated effect. The findings strongly suggest that SICI is not mediated by the alpha1-subunit-bearing subtype of the GABA(A)R but by those bearing either the alpha2- or alpha3-subunit. This study represents an attempt by means of TMS to identify GABA(A)R subtype-specific action at the systems level of human cortex, a highly relevant issue because the different alpha-subunit-bearing subtypes of the GABA(A)R are differently involved in benzodiazepine-mediated effects such as sedation, amnesia or anxiolysis, in developmental cortical plasticity, and in neurological disorders such as epilepsy. ( view less ) Giuseppe Inama,Claudio Pedrinazzi,Ornella Durin,Pietro Agricola,Giulio Romagnoli,Pietro Gazzaniga Atrial flutter is a common arrhythmia that may cause significant symptoms, including palpitations, dyspnoea, chest pain and even syncope. Frequently, it is possible to diagnose atrial flutter with a 12-lead surface electrocardiogram (ECG), looking for distinctive waves in leads II, III, aVF, aVL, V... ( view more )1 and V2. Puech and Waldo developed the first classification of atrial flutter in the 1970s. These authors divided the dysrhythmia into types I and II. Therefore, in 2001, the European Society of Cardiology and the North American Society of Pacing and Electrophysiology developed a new classification of atrial flutter based not only on the ECG, but also on the electrophysiological mechanism. More recently, Scheinman and colleagues have provided an updated classification and nomenclature. Terms such as common, uncommon, typical, reverse typical or atypical flutter are abandoned, because they may generate confusion. The authors worked out a new terminology, which differentiates atrial flutter only on the basis of electrophysiological mechanism. ( view less ) Francesca Crippa,Chris Panzeri,Andrea Martinuzzi,Alessia Arnoldi,Francesca Redaelli,Alessandra Tonelli,Cinzia Baschirotto,Giovanni Vazza,Maria Luisa Mostacciuolo,Andrea Daga,Genny Orso,Paolo Profice,Antonio Trabacca,Maria Grazia D'Angelo,Giacomo Pietro Comi,Sara Galbiati,Costanza Lamperti,Sara Bonato,Massimo Pandolfo,Giovanni Meola,Olimpia Musumeci,Antonio Toscano,Carlo Pietro Trevisan,Nereo Bresolin,Maria Teresa Bassi BACKGROUND: Hereditary spastic paraplegia (HSP) is a group of genetically heterogeneous disorders characterized by progressive spasticity of the lower limbs. Mutations in the SPG4 gene, which encodes spastin protein, are responsible for up to 45% of autosomal dominant cases. OBJECTIVE: To search fo... ( view more )r disease-causing mutations in a large series of Italian patients with HSP. DESIGN: Samples of DNA were analyzed by direct sequencing of all exons in SPG4. Samples from a subset of patients were also analyzed by direct sequencing of all exons in SPG3A, SPG6, SPG10, and SPG13. SETTING: Molecular testing facility in Italy. PATIENTS: Sixty unrelated Italian patients with pure (n = 50) and complicated (n = 10) HSP. MAIN OUTCOME MEASURES: Mutations in SPG4, SPG3A, SPG6, SPG10, and SPG13. RESULTS: We identified 12 different mutations, 8 of which were novel, in 13 patients. No mutations of any of the other HSP genes tested were found in 15 patients with sporadic pure HSP who did not have mutations in the SPG4 gene. CONCLUSIONS: The overall rate of mutation in the SPG4 gene within our sample was 22%, rising to 26% when only patients with pure HSP were considered. The negative result obtained in 15 patients without mutations in SPG4 in whom 4 other genes were analyzed (SPG3A, SPG6, SPG10, and SPG13) indicate that these genes are not frequently mutated in sporadic pure HSP. ( view less ) Massimiliano Veroux,Carmelo Madia,Pietro Fiamingo,Pietro Caglià,Maurizio Valastro,Corrado Amodeo,Pierfrancesco Veroux,Massimiliano Gagliano,Stefano Basso,Davide Francesco D'Amico BACKGROUND AND OBJECTIVES: This retrospective study was undertaken to evaluate if high resectability rate could improve the long-term outcome of patients with proximal bile duct cancer. METHODS: Between 1985 and 2001, 50 patients (34 male and 16 female) with proximal bile duct cancer were treated. ... ( view more )Thirty-six patients (72%) were considered suitable for surgery, while 14 underwent nonsurgical palliative procedures. Twenty patients had bile duct resection only. Ten patients had Roux-en-Y cholangiojejunostomy with two or three divided segmental hepatic ducts; in 10 patients, the cholangiojejunostomy was performed with four or five divided segmental hepatic ducts. Three patients were treated by palliative transtumoral intubation with Kehr tube. Thirteen patients had bile duct resection plus hepatectomy. Despite the curative intention of the operation, only in 19 (52.7%) patients did the histopathological examination reveal tumor-free margins. RESULTS: There was no operative mortality. Postoperative morbidity was 25%. Overall 1-, 3-, and 5-year survival of the entire surgical group was 61%, 22.5%, and 9%, respectively. In the 19 patients treated with curative intent the survival at 1, 3, and 5 years was 63.1%, 31.5%, and 15.8%, respectively, while in the group that had palliative treatment it was 45%, 15%, and 0%, respectively. CONCLUSIONS: Only margins free from tumor can guarantee an improvement in long-term outcome. Increasing resectability improves survival and could offer a chance of better long-term survival. ( view less ) Emanuele Bertaglia,Giuseppe Stabile,Gaetano Senatore,Pietro Turco,Giovanni Donnici,Antonio De Simone,Massimo Fazzari,Francesca Zerbo,Pietro Pascotto OBJECTIVES: To investigate the clinical outcome of right and left atrial radiofrequency ablation after the first 12 months in patients with drug-refractory persistent atrial fibrillation (AF), and to identify predictors of long-term success. METHODS: We analyzed the clinical outcome of 74 consecuti... ( view more )ve patients with a follow-up >12 months who underwent right and left atrial ablation for persistent AF. Patients who did not present symptomatic or asymptomatic atrial tachyarrhythmias (AT) lasting >30 seconds after the first 3 months of follow-up were defined responders to pulmonary veins ablation. RESULTS: After a mean follow-up of 20.2 +/- 6.3 months (12-36), 52/74 (70%) patients were deemed responders. AT relapsed within the first 12 months in 19/74 (26%) patients (17 AF and 2 left atrial flutter). Among those patients who did not relapse within the first 12 months, only 3 patients (5%) presented AF after the first year of follow-up. At the multivariate analysis presence of early AT relapse and history of AF >7 years inversely correlated with a successful long-term clinical outcome. CONCLUSION: Right and left atrial ablation, alone or in association with antiarrhythmic drugs, prevented AT relapses in 70% of patients with drug-refractory persistent AF also after the first 12 months. Presence of AT relapse within the first 3 months and history of AF >7 years identified patients with a lower probability of successful long-term clinical outcome. ( view less ) Clelia Miracco,Maria Caterina De Nisi,Felice Arcuri,Elena Cosci,Lorenzo Pacenti,Marzia Toscano,Anna Vittoria Lalinga,Maurizio Biagioli,Pietro Rubegni,Rosella Vatti,Emilia Maellaro,Barbara Del Bello,Daniela Massi,Pietro Luzi,Piero Tosi Macrophage migration inhibitory factor (MIF) is a widely expressed cytokine involved in various biological processes. Although MIF's functions in cancer have not been completely elucidated, its expression has usually been correlated with tumour progression and aggressiveness, and it is currently di... ( view more )scussed as a new promising target for novel therapies. Recent studies seem to confirm its active role in melanoma pathobiology; however, its expression has not yet been extensively studied in melanocytic tumours. We evaluated MIF protein expression in 126 skin lesions, including benign and atypical nevi, melanoma and melanoma metastases. In 55 cases, we also assessed MIF mRNA expression by real-time RT-PCR. Benign nevi were subdivided into nevocytic and Spitz/blue types; and melanomas into the radial, and vertical growth phase. A strong cytoplasmic MIF positivity was found in most samples, although it was more heterogeneous in malignant tumours; MIF nuclear expression characterized Spitz/blue nevi, atypical nevi, melanomas and metastases. All samples expressed MIF mRNA but it was significantly lower in benign nevi vs atypical nevi, melanomas and metastases (p=0.001; p<0.0001; p=0.002, respectively). Our study shows a widespread distribution of MIF among melanocytic tumours. Whereas we observed a trend towards higher expression levels of mRNA in atypical and malignant tumours, MIF protein was highly expressed in all lesions, although limited to the cytoplasm in most benign nevi. These observations suggest differences in MIF protein storage, subcellular location and properties in most benign nevi vs atypical and malignant tumours that should be confirmed by further investigation and correlation with clinical outcome. ( view less ) Giuseppe Stabile,Emanuele Bertaglia,Gaetano Senatore,Antonio De Simone,Franco Zoppo,Giovanni Donnici,Pietro Turco,Pietro Pascotto,Massimo Fazzari,Dino Franco Vitale AIMS: We conducted a multi-centre, prospective, controlled, randomized trial to investigate the adjunctive role of ablation therapy to antiarrhythmic drug therapy in preventing atrial fibrillation (AF) relapses in patients with paroxysmal or persistent AF in whom antiarrhythmic drug therapy had alr... ( view more )eady failed. METHODS AND RESULTS: One hundred and thirty seven patients were randomized to ablation and antiarrhythmic drug therapy (ablation group) or antiarrhythmic drug therapy alone (control group). In the ablation group, patients underwent cavo-tricuspid and left inferior pulmonary vein (PV)-mitral isthmus ablation plus circumferential PV ablation. The primary end-point of the study was the absence of any recurrence of atrial arrhythmia lasting >30 s in the 1-year follow-up period, after 1-month blanking period. Three (4.4%) major complications were related to ablation: one patient had a stroke during left atrium ablation, another suffered transient phrenic paralysis, and the third had a pericardial effusion which required pericardiocentesis. After 12 months of follow-up, 63/69 (91.3%) control group patients had at least one AF recurrence, whereas 30/68 (44.1%) (P<0.001) ablation group patients had atrial arrhythmia recurrence (four patients had atrial flutter, 26 patients AF). CONCLUSION: Ablation therapy combined with antiarrhythmic drug therapy is superior to antiarrhythmic drug therapy alone in preventing atrial arrhythmia recurrences in patients with paroxysmal or persistent AF in whom antiarrhythmic drug therapy has already failed. ( view less ) C Di Pietro,S Piro,G Tabbì,M Ragusa,V Di Pietro,V Zimmitti,F Cuda,M Anello,U Consoli,E T Salinaro,M Caruso,C Vancheri,N Crimi,M G Sabini,G A P Cirrone,L Raffaele,G Privitera,A Pulvirenti,R Giugno,A Ferro,G Cuttone,S Lo Nigro,R Purrello,F Purrello,M Purrello Due to their ballistic precision, apoptosis induction by protons could be a strategy to specifically eliminate neoplastic cells. To characterize the cellular and molecular effects of these hadrons, we performed dose-response and time-course experiments by exposing different cell lines (PC3, Ca301D,... ( view more ) MCF7) to increasing doses of protons and examining them with FACS, RT-PCR, and electron spin resonance (ESR). Irradiation with a dose of 10 Gy of a 26,7 Mev proton beam altered cell structures such as membranes, caused DNA double strand breaks, and significantly increased intracellular levels of hydroxyl ions, are active oxygen species (ROS). This modified the transcriptome of irradiated cells, activated the mitochondrial (intrinsic) pathway of apoptosis, and resulted in cycle arrest at the G2/M boundary. The number of necrotic cells within the irradiated cell population did not significantly increase with respect to the controls. The effects of irradiation with 20 Gy were qualitatively as well as quantitatively similar, but exposure to 40 Gy caused massive necrosis. Similar experiments with photons demonstrated that they induce apoptosis in a significantly lower number of cells and in a temporally delayed manner. These data advance our knowledge on the cellular and molecular effects of proton irradiation and could be useful for improving current hadrontherapy protocols. ( view less )
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