Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

This paper introduces a disposable battery-driven heating system for loop-mediated isothermal DNA amplification (LAMP) inside a centrifugally-driven DNA purification platform (LabTube). We demonstrate LabTube-based fully automated DNA purification of as low as 100 cell-equivalents of verotoxin-producing Escherichia coli (VTEC) in water, milk and apple juice in a laboratory centrifuge, followed by integrated and automated LAMP amplification with a reduction of hands-on time from 45 to 1 min. The heating system consists of two parallel SMD thick film resistors and a NTC as heating and temperature sensing elements. They are driven by a 3 V battery and controlled by a microcontroller. The LAMP reagents are stored in the elution chamber and the amplification starts immediately after the eluate is purged into the chamber. The LabTube, including a microcontroller-based heating system, demonstrates contamination-free and automated sample-to-answer nucleic acid testing within a laboratory centrifuge. The heating system can be easily parallelized within one LabTube and it is deployable for a variety of heating and electrical applications

X-Ray Phase-Contrast Tomography of Renal Ischemia-Reperfusion Damage

Purpose: The aim of the study was to investigate microstructural changes occurring in unilateral renal ischemia-reperfusion injury in a murine animal model using synchrotron radiation. Material and Methods: The effects of renal ischemia-reperfusion were investigated in a murine animal model of unilateral ischemia. Kidney samples were harvested on day 18. Grating-Based Phase-Contrast Imaging (GB-PCI) of the paraffin-embedded kidney samples was performed at a Synchrotron Radiation Facility (beam energy of 19 keV). To obtain phase information, a two-grating Talbot interferometer was used applying the phase stepping technique. The imaging system provided an effective pixel size of 7.5 mu m. The resulting attenuation and differential phase projections were tomographically reconstructed using filtered back-projection. Semi-automated segmentation and volumetry and correlation to histopathology were performed. Results: GB-PCI provided good discrimination of the cortex, outer and inner medulla in non-ischemic control kidneys. Post-ischemic kidneys showed a reduced compartmental differentiation, particularly of the outer stripe of the outer medulla, which could not be differentiated from the inner stripe. Compared to the contralateral kidney, after ischemia a volume loss was detected, while the inner medulla mainly retained its volume (ratio 0.94). Post-ischemic kidneys exhibited severe tissue damage as evidenced by tubular atrophy and dilatation, moderate inflammatory infiltration, loss of brush borders and tubular protein cylinders. Conclusion: In conclusion GB-PCI with synchrotron radiation allows for non-destructive microstructural assessment of parenchymal kidney disease and vessel architecture. If translation to lab-based approaches generates sufficient density resolution, and with a time-optimized image analysis protocol, GB-PCI may ultimately serve as a non-invasive, non-enhanced alternative for imaging of pathological changes of the kidney

Factors Associated with Deviant Behavior at Intermountain School

The types and frequency of deviance. and the relationship between deviance and factors in the pre-Intermountain. Intermountain. and post- Intermountain experiences were studied using the male population of the 1964 graduating class at Intermountain School, Brigham City, Utah. Among the important findings are the following. Of 233 total deviant acts. 8\u274 were time schedule violations and 66 were drinking and drinking-related violations. The quantitative pattern of deviance was about the same during the sophomore and junior years and then decreased during the senior year. There was an inverse relationship between family size and deviance. The lowest average deviance rate occurred among those students whose parents were living together. The students who started school at ages six to seven had the lowest average deviance rate. There\u27 was a negative correlation between grade point average and degree-of deviance. Deviants had higher average scores than non-deviants on ten of twenty characteristics evaluated by counselors and teachers. The former Intermountain students with the highest\u27 post-high school productive activity scores also had the highest high school average deviance rate

Epizootiological, Clinical and Immunological Features of an Experimentally Induced Strangles Epizootic

Die komplette Dissertation im pdf-Format : diss.pdfIn the present study serum and mucosal antibody responses to the M protein of Streptococcus equi subsp. equi were studied in a group of young horses following intranasal application of an avirulent strain of subsp. equi and during a subsequent experimentally induced strangles epizootic in an attempt to identify humoral immune responses that could be correlated with protection. Convalescent protective immunity was verified by intranasal inoculation of virulent subsp. equi 5 weeks after recovery. Additionally, epizootiological and clinical data were collected during the strangles epizootic to gain further insight into the epizootiology and dynamics of subsp. equi infection within the group. Serum IgG and mucosal IgA and IgG levels were measured by ELISA with electrophoretically purified acid extracted fragments of the M protein of subsp. equi. ELISA values were set in relation to total IgA and IgG antibody to determine M protein specific local IgA and IgG activities in nasal wash samples. Serum bactericidal responses were measured by counts of viable organisms after opsonization and incubation with horse neutrophils for 3 hours. Intranasal inoculation of 12 horses with an avirulent strain of subsp. equi did not elicit protective immune reponses. Following commingling exposure to 2 experimentally infected horses all 20 horses (12 inoculated and 8 controls) developed strangles of varying severity with a mean incubation period of 12 days. The horses were not medicated during the epizootic. Thirteen horses challenged by intranasal inoculation 5 weeks after recovery were solidly immune. At this time these horses exhibited strong serum bactericidal activity and high levels of M protein specific serum IgG. Additionally, nine horses showed high levels of M protein specific mucosal IgA at the time of challenge, whereas local IgG had already returned to preinfection values in most horses. Intranasal challenge did not produce a local or systemic booster response, indicating that protective antibodies may have prevented immune stimulation by the antigens presented at this time. A striking feature of the study was the great qualitative and quantitative heterogeneity of serum and mucosal antibody response among individual horses. Systemic and local antibody responses were not synchronized, suggesting independence of these responses. A substantial portion of M protein reactive serum IgG showed crossreactivity with S. equi subsp. zooepidemicus, suggesting extensive sharing of epitopes by the two subspecies. Local IgA concentrations were about 10 times higher than local IgG concentrations. IgG in nasal wash samples exhibited high M protein specific activity, suggesting local synthesis of this antibody. Nasal shedding of subsp. equi as studied intensively in 9 horses started after a latent period of 2-9 days and ceased after a mean of 14 days. Shedding by 2 horses continued for 61 and 106 days after clinical signs were first noticed. Both horses showed intermittent purulent nasal discharge until about 3 weeks before the last isolation of subsp. equi. Cessation of shedding was associated in most horses with appearance of high levels of M protein specific local IgA. Barn repopulation with a highly susceptible group of yearlings 8 months after the strangles epizootic was uneventful, despite the lack of preceding decontamination measures such as barn disinfection

Device and Method for Producing a Spatially Uniformly Intense Source of X-Rays

An x-ray source for producing a uniformly intense area x-ray beam. The x-ray source includes a vacuum chamber. An area electron emitter is disposed at a first end of the vacuum chamber. A target material is disposed at a second end of the vacuum chamber and spaced apart from the area electron emitter. The area electron emitter and the target material are correspondingly shaped and/or correspondingly curved. The x-ray source also includes at least one high voltage power source. The area electron emitter is electrically connected to a negative pole of one of the at least one high voltage power source and the target electrically connected to a positive pole of one of the at least one high voltage power source.Sponsorship: Illinois Institute of TechnologyUnited States Paten

Genome Sequencer System Rapid and Accurate Pyrosequencing of Serial Analysis of Gene Expression Ditags

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