A red line not to cross: Evaluating the limitation and properness of gel image tuning procedures

Currently, results of gel electrophoresis are commonly documented in digital formats by image acquisition instruments. In this study, gel images tuned by a common image processing software package, Photoshop, were assessed to understand the transforming algorithms and their impacts on quantitative analysis. TotalLab 100, an electrophoresis gel image analysis software package, was applied for image quantitation and evaluation. The three most frequently used image tuning functions—adjustments of the brightness, contrast, and grayscale span (level) of images—were investigated using both data generated from a standard grayscale tablet and an actual electrophoresis gel image. The influences of these procedures were analyzed for the grayscale transformation between the input and output images. Although all three procedures differentially improved the visualization of the input image, adjusting the contrast of images disrupted the quantitative information because of its nonlinear transforming algorithm. Under certain conditions, adjusting the brightness or the level of images could preserve the quantitative information because of the linear transforming algorithms. It was found that when the minimum and maximum grayscales of a gel image were recognized, using a commercial software package to maximally stretch the level may significantly improve the quality of a gel image without jeopardizing quantitative analysis.     

Dynamics and Distribution of Klothoβ (KLB) and fibroblast growth factor receptor-1 (FGFR1) in living cells reveal the fibroblast growth factor-21 (FGF21)-induced receptor complex

FGF21 stimulates FGFR1c activity in cells that co-express Klothoβ (KLB); however, relatively little is known about the interaction of these receptors at the plasma membrane. We measured the dynamics and distribution of fluorescent protein-tagged KLB and FGFR1c in living cells using fluorescence recovery after photobleaching and number and brightness analysis. We confirmed that fluorescent protein-tagged KLB translocates to the plasma membrane and is active when co-expressed with FGFR1c. FGF21-induced signaling was enhanced in cells treated with lactose, a competitive inhibitor of the galectin lattice, suggesting that lattice-binding modulates KLB and/or FGFR1c activity. Fluorescence recovery after photobleaching analysis consistently revealed that lactose treatment increased KLB mobility at the plasma membrane, but did not affect the mobility of FGFR1c. The association of endogenous KLB with the galectin lattice was also confirmed by co-immunoprecipitation with galectin-3. KLB mobility increased when co-expressed with FGFR1c, suggesting that the two receptors form a heterocomplex independent of the galectin lattice. Number and brightness analysis revealed that KLB and FGFR1c behave as monomers and dimers at the plasma membrane, respectively. Co-expression resulted in monomeric expression of KLB and FGFR1c consistent with formation of a 1:1 heterocomplex. Subsequent addition of FGF21 induced FGFR1 dimerization without changing KLB aggregate size, suggesting formation of a 1:2 KLB-FGFR1c signaling complex. Overall, these data suggest that KLB and FGFR1 form a 1:1 heterocomplex independent of the galectin lattice that transitions to a 1:2 complex upon the addition of FGF21.     

Impact of hexenuronic acids on xylanase-aided bio-bleaching of chemical pulps

Hexenuronic acids (HexA) of hemicellulosic heteroxylan were shown to play important role in brightness development of chemical pulps during xylanase-aided bio-bleaching. Industrial wood (eucalypt) kraft pulp and a few non-wood (giant reed) organosolv pulps were pre-treated with commercial xylanase preparations (endo-1,4-β-xylanase activity; EC 3.2.1.8) and bleached by simplified bleaching sequence. The HexA performance was examined and compared with control (enzyme-free) samples. The xylanase-assisted direct brightening effect, noted immediately after an enzymatic stage, was proved to be caused by exclusive HexA removal with solubilized HexA-carrying xylooligosaccharide fractions. Aldohexa- and aldopentahexenuronic acids (Xyl₅-HexA and Xyl₄-HexA) were found as predominant oligosaccharides, accounting for up to 65% of total acidic oligomers in enzymatic hydrolyzates. A strong positive correlation (R² = 0.91) was established between further brightness improvement of xylanase-treated pulps during subsequent chemical bleaching (bleach boosting) and the content of HexA. This underlined the role of HexA as one of the key factors in definition of final brightness of bio-bleached pulps, determining to a large extent the bleaching efficiency of xylanase application as a whole.     

Sensory response patterns and the evolution of visual signal design in anoline lizards

 The evolutionary relationship between visual system response and visual signal design was investigated in four species of anoline lizards which occupy distinctly different habitats. Anoles display with motion patterns of a colorful throat fan called the dewlap. We assessed signal visibility by recording evoked potentials from the optic tectum in response to a moving stimulus flag (a dewlap-like stimulus), and, in one species, by testing behavioral response. The motion pattern, intensity and spectral quality of the stimulus flag, and the background against which it was viewed, were independently manipulated. In all cases, high-velocity motion patterns with a high percentage of brightness contrast between stimulus and background produced the greatest response. Differences in spectral quality between stimulus and background (color contrast) had no effect on tectal responses, but did influence the behaviorally measured detection probability. Using habitat light data we estimated the visibility of the dewlap of each species in different natural habitats. Each species' dewlap was highly visible in its own habitat, but some were much less visible in the habitats of some other species. Habitat light conditions and visual system response properties appear to have constrained the evolution of dewlap design, in at least some of the species. Accepted: May 1998     

Is tissue harmonic ultrasound imaging (THI) of the prostatic urethra and rectum superior to brightness (B) mode imaging? An observer study

Quality ultrasound images are an essential part of prostate brachytherapy procedure. The authors have previously reported that tissue harmonic ultrasound images (THI) are superior to brightness (B) mode for the prostate. The objective of the current study was to compare both imaging modes for visualization of the prostatic urethra and rectum.B and THI mode transrectal ultrasound images were acquired for ten patients. The prostatic urethra and rectal wall were contoured by a radiation oncologist (RO) and five observers on randomly presented images. The contours on one patient were repeated four additional times by four observers. All the images were qualitatively scored using a five-level Likert scale.The values of the Pearson product-moment correlation coefficients showed that the observers were in close agreement with the RO. Two sample paired student t-test showed that the rectum volumes with THI were significantly smaller than B-mode, but no significant difference for urethra. Two-factor analysis of variances showed significant observer variability in defining the rectum and urethra in both imaging modes. Observer consistency of the rectum volumes, estimated by standard deviations as percentages of means was significantly improved for THI. The Likert scale based qualitative assessment supported quantitative observations.The significant improvement in image quality of the prostate (reported previously) and rectum with THI may offer better-quality treatment plans for prostate brachytherapy and potential improvement in local control.