ABSTRACT
AIMS: A molecular method for a rapid detection of viable Legionella pneumophila of all serogroups in tap water samples was developed as an alternative to the reference method (ISO). Legionellae are responsible for Legionnaires' disease, a severe pneumonia in humans with high lethality. METHODS AND RESULTS: The developed method is based on a nutritional stimulation and detection of an increase in precursor 16S rRNA as an indicator for viability. For quantification, DNA was detected by qPCR. This method was compared to the ISO method using water samples obtained from public sports facilities in Switzerland. The sensitivity and specificity were 91 and 97%, respectively, when testing samples for compliance with a microbiological criterion of 1000 cell equivalents per l. CONCLUSION: The new method is sensitive and specific for Leg. pneumophila and allows results to be obtained within 8 h upon arrival, compared to one week or more by the ISO method. SIGNIFICANCE AND IMPACT OF THE STUDY: The method represents a useful tool for a rapid detection of viable Leg. pneumophila of all serogroups in water by molecular biology. It can be used as an alternative to the ISO method for official water analysis for legionellae and particularly when a short test time is required.
Subject(s)
Fresh Water/microbiology , Legionella pneumophila/isolation & purification , Real-Time Polymerase Chain Reaction/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , DNA, Bacterial/genetics , Humans , Legionella pneumophila/classification , Legionella pneumophila/genetics , Legionella pneumophila/growth & development , Legionnaires' Disease/microbiology , Microbial Viability , RNA, Ribosomal, 16S/genetics , Sensitivity and Specificity , Water MicrobiologyABSTRACT
There are sex differences in free-running rhythms, activity level and activity distribution that are attributed, in part, to the action of gonadal hormones. We tested the hypothesis that non-classical estrogenic signaling pathways at estrogen receptor subtype 1 (ESR1) modify the amplitude and phase of activity. We used ESR1 knock-out mice (ERKO) and non-classical estrogen receptor knock-in mice (NERKI). ERKO animals are unable to respond to estrogen at the ESR1 and NERKI animals lack the ability to respond to estrogens via the estrogen response element-mediated pathway, but can still respond via non-classical mechanisms. We compared intact male and female ERKO, NERKI and wildtype (WT) mice with respect to total wheel-running activity, activity distribution across the 24-h day, phase angle of activity onset and free-running period (τ) and the duration of activity in constant conditions. WT females had significantly greater activity than WT males, and this activity was more consolidated to the dark phase of the light:dark cycle. These sex differences were absent in the NERKI and ERKO animals. Among females, NERKI and ERKO animals had greater activity during the light phase than WT counterparts. Additionally, we have identified a novel contribution of non-classical estrogen signaling pathways on the distribution of activity. Our data suggest that total activity is ESR1-dependent and daily activity patterns depend on both classical and non-classical actions of estrogens. These data will aid in identifying the mechanisms underlying sex differences in sleep-wake cycles and the influence of steroid hormones on circadian patterns.
Subject(s)
Circadian Rhythm/genetics , Estrogen Receptor alpha/genetics , Activity Cycles/genetics , Animals , Female , Male , Mice , Mice, Transgenic , Motor Activity/genetics , Sex CharacteristicsABSTRACT
A system for the creation of computer-generated sound patterns of two-dimensional line graphs is described. The objectives of the system are to provide the blind with a means of understanding line graphs in the holistic manner used by those with sight. A continuously varying pitch is used to represent motion in the x direction. To test the feasibility of using sound to represent graphs, a prototype system was developed and human factors experimenters were performed. Fourteen subjects were used to compare the tactile-graph methods normally used by the blind to these new sound graphs. It was discovered that mathematical concepts such as symmetry, monotonicity, and the slopes of lines could be determined quickly using sound. Even better performance may be expected with additional training. The flexibility, speed, cost-effectiveness, and greater measure of independence provided the blind or sight-impaired using these methods was demonstrated.