FLUKE 62 MAX PLUS EPUB
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An arbitrary waveform generator (Fluke, Everett, WA) was used to control the nm data using SlideWrite Plus 6 (Advanced Graphics Software, Inc., Encinitas CA). . Importantly, these results also show that optical pacing at 10% of maximum . which has also been reported by others in whole heart tissue samples. Feb 11, We measured the temperature next to the swabbed hibernating bat using a laser temperature thermometer (Fluke 62 MAX Plus Infrared. 6,  with the maximum-likelihood (ML) model as described previously . For the S. stercor- . Liver fluke, hookworm and S. stercoralis are the gastrointestinal helminths detected in our In HVR-IV the same 5 worms plus another 28 infective .. Epub /08/ yazik.info PMID.
For example, finding that hosts in disease-endemic areas are resistant to a disease would suggest that phylogenetically and ecologically similar hosts in invading regions might also persist by evolving resistance. However, few, if any, studies exist that have explicitly compared disease dynamics in introduced and endemic regions.
The mechanisms of host persistence with novel pathogens also determine, in large part, the intensity of transmission. Two mechanisms, reduced host density for pathogens in which transmission is density-dependent, and poor environmental conditions for pathogen survival or replication outside hosts, will result in lower pathogen transmission [ 7 , 12 ].
Fluke 62 MAX Infrared Thermometer, to °F ( to °C)
Similarly, if hosts in the introduced region are inherently resistant to the pathogen or evolve to become resistant, then this will limit transmission [ 9 , 13 ]. By contrast, if hosts are tolerant or evolve tolerance then transmission of the pathogen will be maintained at a much higher intensity, with correspondingly large impacts on intolerant, non-resistant species. White-nose syndrome WNS , a recently emerged disease of hibernating bats, was first detected in North America in [ 15 ] and has caused precipitous declines in temperate bat populations across eastern and midwestern North America [ 16 — 18 ].
The pathogen that causes WNS, Pseudogymnoascus destructans, is a cold growing fungus that infects bats' skin during their hibernation period [ 19 — 21 ], and can persist in the environment for long periods of time in the absence of bats [ 22 , 23 ]. The resulting infections lead to the disruption of homeostatic processes and ultimately mortality [ 24 , 25 ].
Pseudogymnoascus destructans has been documented widely across Europe and Asia on multiple species of bats [ 26 — 28 ]. European isolates of P. The widespread declines observed in North America have not been observed in Asia or Europe [ 29 ] and genetic data suggest that P.
By understanding how bats persist with P. Here, we present the first comparison of P.
Material and methods a Sample collection and testing We sampled bats and hibernacula caves and mines substrate for P. The North American sites were sampled during the first 3 years of pathogen invasion to each site where populations were experiencing severe declines [ 16 , 18 ].
We selected sites in Asia and North America at similar latitudes to control for climate and winter severity. The average daily above-ground temperature at the sites in Asia during winter when sampling was conducted 1 October—30 April was 0.
Samples were collected using identical methods and during the same time period March in the two regions to make the comparison as similar as possible. Past studies indicate that prevalence and infection intensities measured as the amount of fungal DNA detected in a sample and WNS lesions on bats throughout North America increased over the winter, and were the highest in late winter [ 19 , 21 ], making March an ideal sampling time to examine and compare infection prevalence and fungal growth.
In both regions, we estimated fungal prevalence and infection intensity based on the quantity of fungus on the surface of the skin by rubbing sterile swabs on bats' wings and muzzles as previously described [ 19 , 32 , 33 ]. We stored samples in RNAlater prior to testing.
All samples were run in duplicate, with quantification standards on each plate and 16 negative controls per plate. All quantification standards were within a consistent range and all negative controls had no fungal detection. IR optical pacing has been previously demonstrated as a robust tool for controlling heat rate in embryonic quails[ 16 , 17 ], rat neonatal cardiomyocytes[ 18 ] and adult rabbits[ 19 ].
However, it is unknown if IR optical pacing can be used to reliably control beating rate of a confluent hCM monolayer. Another barrier to using hCM in a high-throughput format is that the measurement of electrophysiological parameters e. For example, multielectrode arrays MEA are only able to estimate the timing of action potential depolarization and repolarization from the extracellular potential, which can be inaccurate when assessing drug response and when using high-pass filtering[ 20 ].
Fluorescent indicators may be better suited for this purpose[ 1 , 21 ] and can be used to measure a wide range of cellular parameters e. However, the small assay size and, thus, small fluorescent signal associated with a high throughput screening format e.
Furthermore, Fluovolt has recently been demonstrated to reflect repolarization in cardiac applications[ 20 , 26 ]. However, Fluovolt has not been systematically validated for safely assessing multiple electrophysiological parameters, including impulse conduction velocity, in a high throughput assay.
Thus, innovative methods are needed to foster investigation of human genetic disease mechanisms and cardiac safety testing that rely on hCM. Herein, a new assay is described that incorporates both optical pacing using IR light and high fidelity fluorescent mapping to create a fully contactless assay for controlling beating rate and for quantifying multiple cardiac electrophysiology parameters.
Material and methods Cell isolation and culture Human cardiac myocytes derived from induced pluripotent stem cells hCM were downloadd from Cellular Dynamics Inc. Cell pellets in the cryoprecipitate tube were thawed and cultured as monolayers according to the protocol provided by the manufacturer. Cells were plated onto fibronectin coated Biolite well plates Catalog , ThermoFisher Scientific, Waltham, Massachusetts prior to experimentation at 1.
Culture media was changed every 2 days, until day 14—20 when experiments were performed. A well plate sits atop an XYZ stage with a multi-well plate adapter. The CCD was configured for 10x10 pixel binning with additional 2x2 binning in software, resulting in 20x14 binned pixels.
No subsequent spatial or temporal filtering was utilized.If you've ever used an infrared thermometer, then you may have been fooled regarding the temperature reading.
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Many electrophysiological parameters that are mechanistically linked to arrhythmia, such as repolarization[ 6 , 7 ], impulse conduction velocity[ 8 ], and cardiac alternans[ 9 ], are highly sensitive to beating rate. However, it is unknown if IR optical pacing can be used to reliably control beating rate of a confluent hCM monolayer.
Accordingly, long reads from third generation sequencing and additional RNAseq data will be needed to improve gene predictions, as demonstrated for S. Using stage-specific gene expression data, we identified liver fluke proteins likely involved in host-parasite interactions, and using immunolocalization, we confirmed Neorickettsia in organs and tissues of the adult trematode.