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51.
Hara-Kaonga B Gao YA Havrda M Harrington A Bergquist I Liaw L 《Transgenic research》2006,15(1):101-106
Cre recombinase has become a ubiquitous tool in transgenic strategies for regulation of transgene expression in a tissue-specific
manner. We report analysis of two SM22αCre lines and their ability to mediate genomic recombination in five independent Cre-responsive transgenic lines. One of the
SM22αCre lines developed was a tet-on system based on the reverse tetracycline transactivator. Our goal was to use this strategy
to inhibit the Notch signaling pathway specifically in smooth muscle cells. Our responder transgenes contained a constitutively
expressed marker gene (chloramphenicol acetyltransferase, CAT), flanked by loxP sites in direct orientation, upstream of Notch-related transgenes. We developed two dominant negative Notch transgenic responder
lines activated by Cre-mediated DNA recombination. The first is the extracellular domain of human Jagged1, and the second
is the extracellular domain of the human Notch2 receptor. Despite high expression of the marker gene in all responder lines,
we found that Cre-mediated genomic recombination between these five lines was highly variable, ranging from 46 to 93% of individuals
using an SM22αCre activating strain, or 8–58% of individuals using an inducible SM22αrtTACre. In all cases examined, detection of recombination by PCR correlated with expression of the transgene as determined
by Western blot analysis. Our studies reflect the variability in recombination success based on the responder strain, presumably
due to inaccessibility of the locus of integration of the responder allele. 相似文献
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Bergquist AJ Clair JM Collins DF 《Journal of applied physiology (Bethesda, Md. : 1985)》2011,110(3):627-637
Neuromuscular electrical stimulation (NMES) can be delivered over a nerve trunk or muscle belly and can generate contractions by activating motor (peripheral pathway) and sensory (central pathway) axons. In the present experiments, we compared the peripheral and central contributions to plantar flexion contractions evoked by stimulation over the tibial nerve vs. the triceps surae muscles. Generating contractions through central pathways follows Henneman's size principle, whereby low-threshold motor units are activated first, and this may have advantages for rehabilitation. Statistical analyses were performed on data from trials in which NMES was delivered to evoke 10-30% maximum voluntary torque 2-3 s into the stimulation (Time(1)). Two patterns of stimulation were delivered: 1) 20 Hz for 8 s; and 2) 20-100-20 Hz for 3-2-3 s. Torque and soleus electromyography were quantified at the beginning (Time(1)) and end (Time(2); 6-7 s into the stimulation) of each stimulation train. H reflexes (central pathway) and M waves (peripheral pathway) were quantified. Motor unit activity that was not time-locked to each stimulation pulse as an M wave or H reflex ("asynchronous" activity) was also quantified as a second measure of central recruitment. Torque was not different for stimulation over the nerve or the muscle. In contrast, M waves were approximately five to six times smaller, and H reflexes were approximately two to three times larger during NMES over the nerve vs. the muscle. Asynchronous activity increased by 50% over time, regardless of the stimulation location or pattern, and was largest during NMES over the muscle belly. Compared with NMES over the triceps surae muscles, NMES over the tibial nerve produced contractions with a relatively greater central contribution, and this may help reduce muscle atrophy and fatigue when NMES is used for rehabilitation. 相似文献
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Sigrid N?ss Benedicte A. Lie Espen Melum Marita Olsson Johannes R. Hov Peter J. P. Croucher Jochen Hampe Erik Thorsby Annika Bergquist James A. Traherne Erik Schrumpf Kirsten Muri Boberg Stefan Schreiber Andre Franke Tom H. Karlsen 《PloS one》2014,9(12)
Background
Genetic variants within the major histocompatibility complex (MHC) represent the strongest genetic susceptibility factors for primary sclerosing cholangitis (PSC). Identifying the causal variants within this genetic complex represents a major challenge due to strong linkage disequilibrium and an overall high physical density of candidate variants. We aimed to refine the MHC association in a geographically restricted PSC patient panel.Methodology/Principal Findings
A total of 365 PSC cases and 368 healthy controls of Scandinavian ancestry were included in the study. We incorporated data from HLA typing (HLA-A, -B, -C, -DRB3, -DRB1, -DQB1) and single nucleotide polymorphisms across the MHC (n = 18,644; genotyped and imputed) alongside previously suggested PSC risk determinants in the MHC, i.e. amino acid variation of DRβ, a MICA microsatellite polymorphism and HLA-C and HLA-B according to their ligand properties for killer immunoglobulin-like receptors. Breakdowns of the association signal by unconditional and conditional logistic regression analyses demarcated multiple PSC associated MHC haplotypes, and for eight of these classical HLA class I and II alleles represented the strongest association. A novel independent risk locus was detected near NOTCH4 in the HLA class III region, tagged by rs116212904 (odds ratio [95% confidence interval] = 2.32 [1.80, 3.00], P = 1.35×10−11).Conclusions/Significance
Our study shows that classical HLA class I and II alleles, predominantly at HLA-B and HLA-DRB1, are the main risk factors for PSC in the MHC. In addition, the present assessments demonstrated for the first time an association near NOTCH4 in the HLA class III region. 相似文献56.
G. Thorsn J. Bergquist 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2000,745(2)
A method is presented for the chiral analysis of amino acids in biological fluids using micellar electrokinetic chromatography (MEKC) and laser-induced fluorescence (LIF). The amino acids are derivatized with the chiral reagent (+/−)-1-(9-anthryl)-2-propyl chloroformate (APOC) and separated using a mixed micellar separation system. No tedious pre-purification of samples is required. The excellent separation efficiency and good detection capabilities of the MEKC-LIF system are exemplified in the analysis of urine and cerebrospinal fluid. This is the first time MEKC has been reported for chiral analysis of amino acids in biological fluids. The amino acids
-alanine,
-glutamine, and
-aspartic acid have been observed in cerebrospinal fluid, and
-alanine and
-glutamic acid in urine. To the best of our knowledge no measurements of either
-alanine in cerebrospinal fluid or
-glutamic acid in urine have been presented in the literature before. 相似文献
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59.
Effects on water chemistry after drainage of a bog for forestry 总被引:1,自引:1,他引:0
Drainage for forestry has received increasing interest during recent decades. Generally, drainage concerns wet mineral soils while the utilization of peatlands is a matter of controversy. The peatlands mainly involved are fens, while forestry on bogs is an insignificant activity. Consequently, hydrology of bogs and effects of drainage on their hydrochemistry are little known.The investigation performed aimed at elucidating the parent conditions and the drainage impact on the hydrology and hydrochemistry of an ombrotrophic bog. Two bogs were first compared during a calibration period of two years and then, after drainage of one of them, during a period of three years. The second bog was kept virgin as a control.Considerable influences on runoff and stream water quality were found from the surrounding mineral soil uplands of the bog. Significant differences occurred between the chemical composition of the groundwater in the mineral soil and in the bog peat.Effects on runoff water from drainage of the bog deviate from drainage of minerotrophic peatlands with respect to decreased concentrations and losses of organic carbon and nitrogen. From two small bog catchments within the drained bog, there generally were greater losses of nutrients than from the catchment as a whole. Furthermore, the runoff from the drained bog decreased in comparison with the undrained condition. However, there were also similarities to drainage of other peatlands as regards increased pH, alkalinity and concentrations of sulphate. Also, concentrations of total-phosphorus increased in spite of a decreased phosphate (MRP) concentration. 相似文献
60.