Occurrence of Legionella in showers at recreational facilities.

Critical environments, including water systems in recreational settings, represent an important source of Legionella pneumophila infection in humans. In order to assess the potential risk for legionellosis, we analyzed Legionella contamination of water distribution systems in 36 recreational facilities equipped with swimming pools. One hundred and sixty water samples were analyzed from shower heads or taps located in locker rooms or in bathrooms. By culture method and polymerase chain reaction, 41/160 samples were positive for Legionella from 12/36 recreational centers. Hotels (57.1%) and sports centers (41.2%) were the most contaminated. L. pneumophila serotypes 2–14 (25/41) were more frequently found than serotype 1 (10/41). Samples at temperature ≥30 °C were more frequently positive than samples at temperature <30 °C (n = 39 vs n = 2, p < 0.00001). The presence of L. pneumophila was investigated by comparison with heterotrophic plate count (HPC), an indicator of water quality. The presence of L. pneumophila was associated more frequently with high and intermediate HPC load at 37 °C, therefore should be considered a potential source when HPC at 37 °C is >10 CFU/mL. Maintenance, good hygiene practices, interventions on the hydraulic system and regular controls must be implemented to minimize exposure to L. pneumophila infection risk

Brain function and clinical characterization in the Boston adolescent neuroimaging of depression and anxiety study

We present a Human Connectome Project study tailored toward adolescent anxiety and depression. This study is one of the first studies of the Connectomes Related to Human Diseases initiative and is collecting structural, functional, and diffusion-weighted brain imaging data from up to 225 adolescents (ages 14-17 years), 150 of whom are expected to have a current diagnosis of an anxiety and/or depressive disorder. Comprehensive clinical and neuropsychological evaluations and longitudinal clinical data are also being collected. This article provides an overview of task functional magnetic resonance imaging (fMRI) protocols and preliminary findings (N = 140), as well as clinical and neuropsychological characterization of adolescents. Data collection is ongoing for an additional 85 adolescents, most of whom are expected to have a diagnosis of an anxiety and/or depressive disorder. Data from the first 140 adolescents are projected for public release through the National Institutes of Health Data Archive (NDA) with the timing of this manuscript. All other data will be made publicly-available through the NDA at regularly scheduled intervals. This article is intended to serve as an introduction to this project as well as a reference for those seeking to clinical, neurocognitive, and task fMRI data from this public resource.R01 MH101521 - NIMH NIH HHS; R01 EB021265 - NIBIB NIH HHS; S10 RR019307 - NCRR NIH HHS; R01 AT007257 - NCCIH NIH HHS; U01 MH108168 - NIMH NIH HHS; R37 MH068376 - NIMH NIH HHS; P41 EB019936 - NIBIB NIH HHS; U01 EB026996 - NIBIB NIH HHS; R01 EB020740 - NIBIB NIH HHS; F32 MH114525 - NIMH NIH HHS; S10 RR023043 - NCRR NIH HHS; R01 MH099021 - NIMH NIH HHS; R01 MH096987 - NIMH NIH HHS; S10 RR023401 - NCRR NIH HHSPublished versio

Discriminazione Rapida Tra Legionella Pneumophila E Altre Specie Di Legionella Con Metodi Molecolari

OBIETTIVI Obiettivi specifici - Da vari anni sono stati descritti diversi metodi molecolari per la ricerca di Legionella che spesso non permettono di discriminare Legionella pneumophila (Lpn) dalle altre specie di Legionella (Lspp). La Legionella è diffusa negli ambienti acquatici naturali ed artificiali, quali fiumi, stagni e laghi, piscine, torri di raffreddamento, acque termali e sistemi di distribuzione idrica ed è importante negli studi epidemiologici identificare i ceppi isolati. Studi recenti indicano che altre specie di Legionella, oltre Lpn, possono essere importanti nell’eziologia di polmoniti acquisite in comunità e che la loro prevalenza può essere sottostimata a causa degli attuali metodi diagnostici spesso inadeguati. L’identificazione delle specie isolate è fondamentale nelle indagini epidemiologiche per riconoscere casi con comune sorgente di infezione, vie di diffusione e distribuzione nelle reti idriche. A tale scopo abbiamo utilizzato una PCR (Polimerase Chain Reaction) seguita da una seminested-PCR per migliorare la ricerca di Legionella e, soprattutto, discriminare rapidamente Lpn dalle altre Lspp. MATERIALI Materiali e metodi – Per la messa a punto del test molecolare, si è utilizzato DNA estratto da ceppi di Lpn ( ceppo di riferimento ATCC 33152 L. pneunophila ssp pneumophila - Culti-Loops) e Lspp (Legionella micdadei) cresciuti su BCYE con L-cisteina e incubati a 37 °C in microaerofilia. L’estrazione del DNA è stata fatta con QIAmp DNA mini kit della QIAGEN. Il test consiste in una prima PCR in cui si utilizzano primer derivati dalla sequenza genica per l’rRNA 16S specifica per tutte le specie di Legionella, seguita da una seminested-PCR con uno specifico primer per Lpn o per Lspp. Per ottimizzare le seminested-PCR, sono state provate diverse concentrazioni di MgCl2 nel mix di reazione di PCR (da 3 a 4 mM) e diverse temperature di annealing per ciascuna coppia di primer (da 55 a 72 °C). RIASSUNTO Risultati – Dalle prove effettuate si è stabilito che le condizioni sperimentali che permettono di distinguere chiaramente Lpn da Lspp attraverso bande specifiche di DNA amplificato visibili su gel d’agarosio generate dalla seminested-PCR, sono la concentrazione di MgCl2 e la temperatura di annealing che devono essere comprese, rispettivamente, tra 3 e 3.5 mM e tra 65 e 70 °C. CONCLUSIONI Conclusioni – Dai risultati sperimentali preliminari, si può affermare che attraverso questo metodo è possibile discriminare facilmente e rapidamente Lpn da Lspp. Ulteriori prove saranno necessarie per stabilirne i vantaggi rispetto ad altri metodi in uso e l’effettiva applicabilità in indagini su campioni ambientali e clinici

The effect of lipophilic amines on the growth of hepatitis A virus in Frp/3 cells

The effect of lipophilic amines on hepatitis A virus infection in a monkey cell line (Frp/3 cells) was studied. Ammonium chloride, amantadine, methylamine and dansylcadaverine inhibited viral antigen synthesis when added to the cells at least one hour after the attachment step. Results obtained suggest that the HAV entry pathway in Frp/3 cells follows an endocytic route and that viral uncoating takes probably place in endosomes and/or lysosomes

Study of the chemical nature of Frp/3 cell recognition units for hepatitis A virus

Research has been carried out in order to clarify the chemical nature of cell receptors interacting with a fast growing strain of hepatitis A virus (HAV) producing a cytopathic effect on Frp/3 cells. Cell surface susceptibility to HAV attachment has been studied after treatment with enzymes acting on different chemical groupings. Results obtained showed a lowering of cell susceptibility to HAV infection following the action of phospholipase A2, phospholipase C, trypsin and beta-galactosidase. These data suggested that phospholipids, proteins and galactose participate to the cellular receptorial area for HAV