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According to European Guidelines for Legionnaires' Disease prevention and control, travel-associated Legionnaires' disease (TALD) cases are managed differently if classified as sporadic or as part of a cluster and more stringent control measures are deployed after clusters are identified. In this study, we propose to modify the current cluster definition: 'two or more cases of Legionnaires' disease (LD) who stayed at, or visited, the same commercial accommodation site 2-10 days before onset of illness and whose onset is within the same 2-year period' with a new cluster definition, i.e. accommodation sites associated with multiple cases regardless of the time elapsed between them. TALD cases occurred in Italy and in the Balearic Islands between 2005 and 2015 were analysed applying the current European Legionnaires' Disease Surveillance Network (ELDSNet) cluster definition. In a sample of selected accommodation sites with multiple cases, a microbiological study was also conducted. Using the new definition, 63 additional sites (16.4% increase) and 225 additional linked cases (19.5% increase) were identified. Legionella pneumophila sg1 was isolated from 90.7% of the selected accommodation sites. The use of the here proposed TALD cluster definition would warrant a full investigation for each new identified case. This approach should therefore increase the number of sites that will require a risk assessment and, in the presence of an increased risk, the adoption of LD control measures to hopefully prevent additional cases.

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An unusually long-lasting community-acquired outbreak of Legionnaires' disease (LD) occurred in the inhabitants of a town in northern Italy from 2005 to 2008. Overall, 43 cases were diagnosed including five deaths. Hundreds of water samples were collected for Legionella isolation but only two clinical samples were obtained. Clinical strains were ST23 as were environmental isolates detected in most Legionella-positive patients' homes and those from a public fountain. Although no Legionella was found in the municipal water mains, a continuous chlorination was applied in 2008. This action resulted in a halving of cases, although incidence remained tenfold higher than the Italian average incidence until the end of 2013, when it dropped to the expected rate. Retrospective analyses of prevalent wind direction suggested that a hidden cooling tower could have been the main cause of this uncommon outbreak, highlighting the importance of implementation of cooling tower registers in supporting LD investigations.

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According to the Italian Surveillance System for Legionnaires' disease (LD), physicians must fill in a form for every case and send it through the Local Health Units to the National Institute of Health (Istituto Superiore di Sanità, ISS). Forms reported in the period from 2000 to 2011 were analysed and discussed. A total of 9,803 cases of LD were reported to ISS during the study period. The median age of cases was 63 years, with a ratio male/female of 2.6 and a case fatality rate of 11.8%. The number of cases has been steadily increasing from 192 cases in 2000 to 1,235 in 2010 and 1,008 cases in 2011. The reported cases showed a geographical gradient, with the highest number notified in the north and the lowest in the south. The majority of cases (73.0%) were community-acquired, followed by travel-associated (13.5%) and healthcare-associated cases (9.3%), cases acquired in long-term care facilities (2.1%), and other types of exposure (2.1%). Even though the increasing trend of LD in Italy indicates an improvement in the ability to detect and report cases, the geographical gradient highlights the existence of low reporting areas where the epidemiological surveillance of LD should be further strengthened.

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As of 7 July 2009, a total of 158 laboratory-confirmed cases of influenza A(H1N1)v were reported in Italy, from half of the 21 Italian regions. To date all cases have had symptoms consistent with seasonal influenza and no severe or fatal cases have been reported. An active surveillance of cases has been set up in Italy in order to undertake appropriate measures to slow down the spread of the new virus. This report describes the routine and enhanced surveillance currently ongoing in Italy.

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Two clusters of invasive meningococcal disease in the north of Italy both due to serogroup C/ST-11 clonal complex are here described. The objective of the investigation was to analyse the phenotype and the genotype of meningococci involved in the two clusters which were of national relevance due to the fatal outcome of the majority of cases (six of the total of 10 cases). All the strains were C:2a:P1.5 ST-11/ET-37 clonal complex. Two pulsed field gel electrophoresis (PFGE) and variable number tandem repeats (VNTR) profiles were identified, one for each cluster. VNTRs were different from those detected in Italy for C/ST-11 strains isolated from sporadic cases in the same period. This laboratory surveillance report highlights the importance and the crucial role of molecular characterisation to confirm the relatedness among meningococci responsible for clusters of cases.

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For several years, over 50% of the cases of travel-associated Legionnaires' disease (TALD) reported to the European Working Group for Legionella Infections (EWGLINET) have been among travellers to France, Italy, and Spain. We describe clusters of TALD cases reported in these countries during a four-year period. We analysed data from EWGLINET and from the individual countries. In all three countries, upon notification of a cluster, local health authorities are alerted by the national collaborator and immediately begin an environmental investigation at the accommodation site, which includes risk assessments and analysis of water samples. From July 1, 2002 to June 30, 2006, 2,101 accommodation sites were associated with TALD cases and reported by EWGLINET to Italian, Spanish and French collaborators. Of these, 252 sites (12%) were associated with clusters: 13.8% (96/697) in Italy, 13.2% (81/615) in Spain and 9.5% (75/789) in France. Overall, 641 cases were reported. Hotels, camping sites and ships and other sites represented respectively 83%, 10% and 7% of the total accommodation sites, with similar proportions in the three countries. In 99% of the sites, samples were collected; 62% of them were found to be positive for Legionella. The findings of this study highlight that disinfection and long-term preventive measures were correctly implemented by the large majority of sites. However, additional efforts must be made to further reduce the percentage of re-offending sites so as to reduce the number of accommodations that are contaminated by Legionella.

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The objective of this study was to evaluate the degree of underreporting to the Italian National Legionellosis Register (NLR). For the year 2002, all cases of Legionellosis notified to the NLR were compared with cases recorded in the hospital discharge record (HDR) database. The number of unreported cases and the total number of cases in the population were estimated using the capture-recapture method with two independent data sources. Seventeen out of 21 Italian regions participated in the study. Overall, underreporting was estimated to be 21.4% and was found to be significantly greater in the Centre-South (28.2%) than in the North (20.0%). However, even after taking into account the higher degree of underreporting, a significantly lower incidence of the disease is registered in central-southern Italy. The hypothesis, which needs to be verified, is that, in addition to underreporting, under-diagnosis of legionellosis is more widespread in this geographical area.

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The influence of intracaudate administration of N-methyl-D-aspartic acid (NMDA) and of the competitive NMDA receptor antagonist, DL-2-amino-5-phosphonovaleric acid (AP-5) was studied on thyrotropin-releasing hormone (TRH)-induced scratching in rabbits. NMDA (28 nmol) significantly increased the latency of TRH-induced scratching but did not modify the duration of this behaviour. Conversely, AP-5 (0.5 mumol) significantly potentiated scratching duration. Since TRH-induced scratching has been reported to be a dopamine-dependent behaviour, these results suggest that NMDA receptor ligands modulate dopaminergic neurotransmission.

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A method is described for the simultaneous determination of biogenic amines, adenosine and their metabolites in rat striatal tissue using high-performance liquid chromatography with ultraviolet spectrophotometric and electrochemical detection. Peaks in the chromatograms of striatal tissue extracts were identified by retention times and by on-line analysis of peak spectra for adenosine and its metabolites, and by comparing current ratios of the dual-electrode coulometric detector for monoamines and metabolites. The assay gives a linear response over the concentration range of 0.15-0.60 micrograms/ml for biogenic amines, 0.5-2.0 micrograms/ml for serotonin, 5-20 micrograms/ml for hypoxanthine, adenosine and N-methyladenosine, and 10-40 micrograms/ml for inosine. The limit of detection for striatal homogenates was 3.5 ng/g for monoamines, 9 ng/g for serotonine, 140 ng/g for hypoxanthine, 290 ng/g for inosine and 80 ng/g for adenosine. The recovery ranged from 88.5% for vanillylmandelic acid to 110.3% for dopamine. The method was used to measure biogenic amines, adenosine and related metabolites in rat striatal tissues.

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The influence of CGS 21680 (2-[p-(2-carboxyethyl)phenethylamino]-5'-N-ethylcarboxamido-adenos ine), an adenosine A2 receptor agonist, was tested in an animal model of Huntington's disease. Male Wistar rats received bilateral intrastriatal injections of quinolinic acid and then, 1 and 2 weeks later, they were treated with intrastriatal CGS 21680 (3 micrograms/2 microliters) or saline. While quinolinic acid-lesioned rats not treated with CGS 21680 showed the typical motor hyperresponsiveness to d-amphetamine (1 mg/kg i.p.), the intrastriatal injection of CGS 21680 completely prevented this effect.

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The influence of bilateral intrastriatal injection of quinolinic acid (QA, 300 nmol) was studied in male Wistar rats. Behavioral and electrophysiological experiments were conducted in 15 lesioned plus 15 vehicle-injected (control) animals. With respect to control animals, QA-lesioned rats showed marked, statistically significant alterations from both the behavioral (greater motor activation in response to d-amphetamine, place-learning deficit in the Morris water maze), and the electroencephalographic (reduced voltage amplitude and EEG power at the level of frontal cortex) points of view. In addition, a significant loss in body weight and a marked striatal gliosis (GFAP staining) were observed in lesioned rats. Conversely, QA-lesioned rats did not show modifications in posttetanic potentiation (P.T.P.) or long-term potentiation (L.T.P.) in CA1 hippocampal area. The present results confirm that QA lesions of rat striatum may be regarded as a suitable model of Huntington's disease (HD).

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The present article studies the influence of A2 (CGS 21680) and A1 (CPA) adenosine receptor agonists on TRH-induced behavioural excitation in rabbits. The results show that CGS 21680 was the most effective drug when administered into the nucleus caudate, while its intracerebroventricular (i.c.v.) administration did not influence the effects induced by TRH. Conversely, CPA exerted the same, slight effects after either intracaudate or i.c.v. administration. Since TRH-induced excitatory effects can be regarded as dopamine-related behaviours, the present results further confirm that the adenosine system inhibits the behavioural effects due to dopaminergic activation. The major involvement of striatal A2 receptors in this effect of adenosine is also discussed.

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Muscarinic cholinoceptor subtypes (M1 and M2) were studied in membrane particles of the rat frontoparietal cortex 72 h and 1, 2, 3, and 4 weeks after ipsilateral lesioning of the nucleus basalis magnocellularis (NBM). The affinity of the ligand used to characterize muscarinic cholinoceptors, 3H-quinuclidinyl benzilate did not significantly change in lesioned compared with sham-operated rats as well as the density of high affinity (M1) sites. Low affinity muscarinic cholinoceptors (M2 sites) were significantly decreased in NBM-lesioned rats 72 h and 1 week after lesioning. The density of M2 sites did not significantly differ in lesioned rats 2 or 3 weeks after NBM lesioning, but increased, in comparison with sham-operation 4 weeks after NBM lesioning. These findings suggest that frontoparietal M2 muscarinic cholinoceptors, which probably have a presynaptic localization, are sensitive to NBM lesions. Their changes at different times after NBM lesioning suggest the occurrence of loss, compensation and upregulation of cholinergic projections arising to the neocortex from the NBM.

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The probability of hippocampal long-term potentiation induction in the mossy fiber CA3 and commissural/associational CA3 responses and the cortical levels of choline acetyltransferase (ChAT) activity were compared in right nucleus basalis magnocellularis (NBM)-lesioned rats. A 50% reduction in the right cortical ChAT activity was demonstrated 4 weeks after an ibotenic acid lesion of the NBM. No significative differences were found in the probability of LTP induction of right hippocampal slices in sham-operated rats from 10 to 40 days after the injection into the right NBM. On the contrary, a progressive and significative increase in the probability of LTP induction was shown in right hippocampal slices of NBM-lesioned rats from 10 to 40 days after the injection of ibotenic acid into the right NBM. The results demonstrated the appearance of a paradoxical increase of hippocampal synaptic plasticity when the cortical cholinergic biochemical alterations are still present. This finding might be responsible for a behavioural recovery, in NBM-lesioned rats.

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A pattern of perioral dyskinesia was induced in adult male rabbits by concomitant stimulation of dopamine D1 receptors (SKF 38393) and blockade of dopamine D2 receptors (sulpiride). Rabbits treated with sulpiride (6 and 12.5 mg/kg i.v.) then, 90 min thereafter, with SKF 38393 (0.1, 1 and 10 mg/kg i.v.) showed a pattern of perioral dyskinesia characterized by compulsive and repetitive sniffing, licking and vacuous chewing. These effects were completely prevented by the administration of N-ethylcarboxamide adenosine (NECA), an A2 > A1 adenosine receptor agonist. The present results confirm that perioral dyskinesia is dependent on the activation of dopamine D1 receptors. They also show that, in order to induce perioral dyskinesia in rabbits, a concomitant blockade of dopamine D2 receptors is required. Finally, the antagonistic effect of NECA on the appearance of perioral movements confirms that adenosine receptors play a key role in the control of dopamine-mediated effects.

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We have recently demonstrated that monolateral lesions of the Nucleus Basalis Magnocellularis (NBM), which is a nucleus sending cholinergic projections to the fronto-parietal cortex, cause a loss in the intensity of Timm staining in the intrahippocampal pathway of mossy fibres (MF). Moreover, these lesions induce ultrastructural changes consistent with the occurrence of degeneration of presynaptic buttons of MF. The present study was designed to quantify the effects of NBM lesioning on the morphology of the presynaptic buttons of MF. Moreover the effects of 4-week choline alfoscerate (alphaGFC) treatment on the density of Timm staining and on the ultrastructure of presynaptic buttons of MF were assessed, alphaGFC, which was given at an oral daily dose of 100 mg/kg, is a precursor in the biosynthesis of several brain phospholipids which increases the availability of choline in the nervous tissue. Monolateral lesions of NBM cause, 4 weeks after lesioning, a significant decrease in the intensity of Timm staining in the MF area accompanied by a loss of about 23% of presynaptic buttons of MF. Moreover about 40% of presynaptic buttons of MF show an impaired morphology. alphaGFC administration restored the intensity of Timm staining in the MF area. In alphaGFC-treated rats, the loss of presynaptic buttons and the number of impaired buttons were reduced to about 12% and 27%, respectively in comparison with non-treated animals. These results confirm and extend our previous observations indicative of the occurrence of transneuronal degenerations in the MF of the hippocampus after monolateral NBM lesioning. Moreover these findings show that alphaGFC treatment is able to counter in part these degenerative changes.

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The sulfide staining technique, known also as neo-Timm staining, predominantly stains associational fibres arising from cortical interneurons located primarily in the neuropil of layers I-III of the rat cerebral cortex. The density of these fibres, considered to have a possible role in cognitive and mnemonic processes, has been demonstrated to be related to the density of zinc-containing presynaptic buttons in the cerebral cortex. The unilateral injection of ibotenic acid into the nucleus basalis magnocellularis (NBM) resulted in no changes in the density of sulfide staining in the first 3 weeks after neurotoxin injection and in a significant loss of sulfide staining in the neuropil of cortical layers I-III 4 weeks after NBM lesion in the fronto-parietal cortex ipsilaterally to the lesion. These data suggest that unilateral lesioning of the NMB may cause changes of chemo-specific zinc-containing intracortical pathways. Moreover, they indicate that, in a manner similar to that described in the brain of Alzheimer's disease patients, NBM-lesioned rats show a decrease of zinc tissue stores in the fronto-parietal cortex.

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The interactions between dopamine D1 and D2 receptors in the model of thyrotropin-releasing hormone (TRH)-induced behavioural excitation were assessed in adult male rabbits. As we had previously observed with the dopamine D1 receptor antagonist SCH 23390 (0.01 mg/kg i.v.), sulpiride (12 mg/kg i.v.), a dopamine D2 receptor antagonist, also significantly antagonized the scratching behaviour elicited by TRH (100 micrograms/10 microliters i.c.v.). Sulpiride (6 and 12 mg/kg i.v.) also induced marked grooming. SKF 38393 (10 mg/kg i.v.), a dopamine D1 receptor agonist, did not modify the TRH-induced scratching. However, LY 17155 (0.5 mg/kg i.v.), a dopamine D2 receptor agonist, significantly increased it. The potentiating effects of LY 171555 were completely antagonized by SCH 23390. These results demonstrate that a concomitant activation of both dopamine D1 and D2 receptors is required for the expression of TRH-induced scratching. They also suggest that scratching behaviour may be a D2-dependent, D1-enabled response. The involvement of dopamine D1 and D2 receptors on grooming behaviour is also discussed.

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The administration of reserpine with alpha-methyl-p-tyrosine (2.5 and 200 mg kg-1 i.p., 24 and 3 h before the test, respectively) induced a marked akinesia in mice. This effect was significantly and dose-dependently reversed by the methylxanthine, caffeine. The anti-akinetic effect of caffeine within a pattern of catecholamine depletion has been interpreted as a dopamine mimetic activity of this drug. The possible involvement of the adenosine system in this effect of caffeine is discussed.

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