Public Domain Image Analysis Software Research Articles

A Simple and Inexpensive Image Analysis Technique to Study the Effect of Disintegrants Concentration and Diluents Type on Disintegration

Tablets disintegration is often the result of a size expansion of the tablets. In this study, we quantified the extent and direction of size expansion of tablets during disintegration, using readily available techniques, that is, a digital camera and public domain image analysis software. After validating the method, the influence of disintegrants concentration and diluents type on kinetics and mechanisms of disintegration were studied. Tablets containing diluent, disintegrant (sodium starch glycolate, crospovidone, or croscarmellose sodium), and lubricant were prepared by direct compression. Projected area and aspect ratio of the tablets were monitored using image analysis techniques. The developed method could describe the kinetics and mechanisms of disintegration qualitatively and quantitatively. Sodium starch glycolate and crospovidone acted purely by swelling and shape recovery mechanisms. Instead, croscarmellose sodium worked by a combination of both mechanisms, the extent of which changed depending on its concentration and the diluent type. We anticipate that the method described here could provide a framework for the routine screening of tablets disintegration using readily available equipment.

AB290. SPR-17 Prosthesis insertion into segmented biomechanics simulation models

ObjectiveBiomechanical simulation requires accurate representation of the geometry of the structures to be studied. Specifically, when simulating the interaction of implanted prosthetics with surrounding tissues, the geometric and mechanical properties of the prosthesis and the surrounding tissues need to be adequately represented. The present work describes methods for inserting test prostheses into magnetic resonance imaging (MRI) derived geometric models of the pelvic floor structures, in order to create realistic simulation models.MethodsWe modified an existing public domain image analysis software tool to allow placement and segmentation of arbitrarily shaped 3D objects into the output segmented geometry of a pelvic MRI image dataset. The tool was applied to create composite segmented geometry and 3D models of the MRI derived pelvic floor structures with the inserted prostheses in the intended anatomic locations, suitable for biomechanical simulation model creation.ResultsSegmentations of the organs in the source pelvic MRI datasets were created, showing the segmented embedded prostheses in the planned location. Three dimensional reconstructions of the segmented datasets were generated, which were viewable from multiple angles, and the ability to turn on and off all tissue and prosthesis layers was demonstrated.ConclusionsWe created a software application for inserting prostheses into segmented MRI based datasets. The output segmentations were suitable for input into a soft-tissue simulation tool suite, which generated simulation results suitable for analysis. This tool has the potential to enable patient specific, iterative surgical planning of prolapse repair strategies.Funding Source(s)None

Grading of Neuroendocrine Tumors With Ki-67 Requires High-quality Assessment Practices

Gastrointestinal and pancreatic neuroendocrine tumors (NETs) arise from disseminated neuroendocrine cells, expressing general and specific neuroendocrine markers. The World Health Organization 2010 classification of NETs is based on grading them according to the proliferation index (PI), which is determined by immunohistochemical staining of the nuclear antigen Ki-67. The classification introduces Ki-67 as the most important criterion for tumor grading, influencing patients' prognoses and the choice of treatment. The aim of this study was to evaluate the assessment of PI value in NETs and its influence on tumor grading. The tumor material consisted of 51 NETs from the pancreas (n=31) and ileum (n=20). The slides were stained with the Ki-67 antibody and visualized using a polymer kit. PI was assessed visually by microscope oculars and using a public domain image analysis software, ImmunoRatio. The PI was measured from the most proliferative areas of the tumor. The PI values and tumor grade by ImmunoRatio were highly reproducible as compared with conventional assessment, which suffered from variation especially if ascertained by different observers. Computer-aided assessments had almost perfect correlation (r=0.985, r=0.987, and r=0.995) (P=0.000) and reproducibility (κ=0.886, κ=0.886, and κ=1.000) (P=0.000) in PI values and tumor grades, respectively. The PI values and tumor grade between conventional and ImmunoRatio assessments by a qualified observer were in good agreement. ImmunoRatio is a qualified diagnostic aid to more objectively analyze Ki-67 PI-based tumor grade in NETs.

Death of human tumor endothelial cells in vitro through a probable calcium-associated mechanism induced by bevacizumab and detected via a novel method

AbstractWe isolated three dimensional cell clusters from fresh human solid tumors and also isolated human neoplastic and normal lymphatic cells. Cells were cultured for 96 hours with and without bevacizumab and other agents. At concentrations of bevacizumab which completely removed VEGF from the culture medium, dead microvascular cells were detected through Fast Green/H&E staining as previously described. These peculiar staining characteristics suggested the involvement of calcium, and this was confirmed through staining with Alizarin red S. Using Alizarin staining as a marker for endothelial cell death permitted the use of public domain image analysis software which resulted in a sensitive and specific system for identifying active pharmaceuticals which target the tumor microvasculature at the same time direct antitumor cell effects are determined. Our results suggest an important role for calcium in endothelial cell death mediated by bevacizumab and other agents and further suggest that agents promoting calcium influx may potentiate the activity of antiangiogenic agents.

Objectifying the sand tray: An initial example of three-dimensional art image analysis for assessment

The purpose of this research was to explore placement and count of standardized sand tray figures based on a sample of adults ( n = 5) completing the Erica Method in an effort to address the need for higher accuracy in sand tray assessment ( Bowyer, 1970; Danielson, 1998; Sjolund, 1981). This study demonstrated image analysis capabilities and limitations in assessing both 3-dimensional object count and placement, which are important clinical sand tray assessment components ( Bolgar & Fischer, 1947; Buhler, 1951; Danielson, 1998). Public domain image analysis software (PDIAS, Mattson, 2009) objectively determined total sand tray figure count and total sand tray figure area used. Human raters ( n = 4) assessed the same criteria. Human-PDIAS inter-rater reliability was higher than human inter-rater reliability. As a result, computerized rating may hold potential for more efficient rating based on this study.

Multiple Conformers in Active Site of Human Dihydrofolate Reductase F31R/Q35E Double Mutant Suggest Structural Basis for Methotrexate Resistance

Methotrexate is a slow, tight-binding, competitive inhibitor of human dihydrofolate reductase (hDHFR), an enzyme that provides key metabolites for nucleotide biosynthesis. In an effort to better characterize ligand binding in drug resistance, we have previously engineered hDHFR variant F31R/Q35E. This variant displays a >650-fold decrease in methotrexate affinity, while maintaining catalytic activity comparable to the native enzyme. To elucidate the molecular basis of decreased methotrexate affinity in the doubly substituted variant, we determined kinetic and inhibitory parameters for the simple variants F31R and Q35E. This demonstrated that the important decrease of methotrexate affinity in variant F31R/Q35E is a result of synergistic effects of the combined substitutions. To better understand the structural cause of this synergy, we obtained the crystal structure of hDHFR variant F31R/Q35E complexed with methotrexate at 1.7-A resolution. The mutated residue Arg-31 was observed in multiple conformers. In addition, seven native active-site residues were observed in more than one conformation, which is not characteristic of the wild-type enzyme. This suggests that increased residue disorder underlies the observed methotrexate resistance. We observe a considerable loss of van der Waals and polar contacts with the p-aminobenzoic acid and glutamate moieties. The multiple conformers of Arg-31 further suggest that the amino acid substitutions may decrease the isomerization step required for tight binding of methotrexate. Molecular docking with folate corroborates this hypothesis.

SU‐FF‐T‐284: Accurate Determination of a Linac Radiation Isocenter Using Iterative Analysis of Digitized Star Shot Images

Purpose: To develop an accurate method to measure radiation isocenter for linac with sub‐millimeter isocenter radius. Method and Materials: Conventional star shot film is produced for gantry and collimator radiation isocenters. The pattern of gantry rotation and collimator rotation is produced on the same Kodak XV ready pack film by placing the film at vertical, and then horizontal positions at isocenter using a carefully placed “Iso‐Align” device. The beams can be differentiated easily because of the well define length of testing beams for collimator rotation, and penetrating beams of the gantry rotations. The film was digitized and the image was enhanced to show clearly the beam edges and center line of each narrow beam using a public domain image analysis software. Each beam can be described by a linear equation that was obtained through sampling and linear regression of the beam's central line. Using iterative search algorithms and analytical calculations, the location of the isocenter and its radius were obtained. Because gantry isocenter and collimator isocenter were determined on the same image, we were able to measure the lateral deviation between these two isocenters. The impacts of beam arrangements on isocenter determination were also investigated. Results: We applied this method on a Varian Trilogy machine. A 0.2 mm radius was measured for collimator radiation isocenter. The gantry radiation isocenter was measured to be of 0.3 mm radius. The lateral distance between two isocenters was 0.3 mm. The results are highly reproducible. Conclusion: We established a method to determine accurately the radii of the radiation isocenters of a LINAC. The precision of this method had not been achievable by the traditional analysis of a star‐shot film, but it is necessary for evaluating a linac for high precision radiotherapy.

Accessible image analysis for art assessment

Until now, art therapists relied on manual ratings for instruments such as the Face Stimulus Assessment, which is a series of stimulus drawings used to garner projective information from clients [FSA, Betts, 2003; Hamilton, M. K. (2008). Developing a standardized rating system for the Face Stimulus Assessment ( FSA) using nine scales adapted from the Formal Elements Art Therapy Scale ( FEATS). Unpublished master's thesis, Avila University, Kansas City, MO]. The current study outlines several image analysis techniques intended to complement interpretive analysis through demonstrating public domain image analysis software (PDIAS) capabilities adapted for measuring formal elements [Gantt, L., & Tabone, C. (1998). The formal elements art therapy scale. Morgantown, WV: Gargoyle Press]. An FSA drawing completed by an individual formally diagnosed with schizophrenia serves as the model. This study also extends manual [Gantt, L., & Tabone, C. (1998). The formal elements art therapy scale. Morgantown, WV: Gargoyle Press] and computer-assisted ratings [Kim, S. I., Bae, J., & Lee, Y. (2007). A computer system to rate the color-related formal elements in art therapy assessments. The Arts in Psychotherapy, 34, 3] through this model by using readily available PDIAS. Case study results indicate that PDIAS is able to analyze accurately formal elements of an FSA drawing. Research implications include improved inter-rater reliability in the adapted Prominence of Color Scale [Gantt, L., & Tabone, C. (1998). The formal elements art therapy scale. Morgantown, WV: Gargoyle Press; Hamilton, M. K. (2008). Developing a standardized rating system for the Face Stimulus Assessment (FSA) using nine scales adapted from the Formal Elements Art Therapy Scale (FEATS). Unpublished master's thesis, Avila University, Kansas City, MO] and the development of highly accurate art therapy assessment rating scales for those without extensive computer analysis backgrounds.

Cell culture detection of microvascular cell death in clinical specimens of human neoplasms and peripheral blood

Angiogenesis studies are limited by the clinical relevance of laboratory model systems. We developed a new method for measuring dead microvascular (MV) cells in clinical tissue, fluid and blood specimens, and applied this system to make several potentially novel observations relating to cancer pharmacology. Dead MV cells tend to have a hyperchromatic, refractile quality, further enhanced during the process of staining with Fast Green and counterstaining with either haematoxylin-eosin or Wright-Giemsa. We used this system to quantify the relative degree of direct antitumour versus anti-MV effects of cisplatin, erlotinib, imatinib, sorafenib, sunitinib, gefitinib and bevacizumab. Bevacizumab had striking anti-MV effects and minimal antitumour effects; cisplatin had striking antitumour effects and minimal anti-MV effects. The ;nib' drugs had mixed antitumour and anti-MV effects. Anti-MV effects of erlotinib and gefitinib were equal to those of sunitinib and sorafenib. There was no detectable VEGF in culture medium without cells; tumour cells secreted copious VEGF, reduced to undetectable levels by bevacizumab, greatly reduced by cytotoxic levels of cisplatin + anguidine, and variably reduced by DMSO and/or ethanol. We observed anti-MV additivity between bevacizumab and other drugs on an individual patient basis. Peripheral blood specimens had numerous MV cells which were strikingly visualized for quantification with public domain image analysis software using bevacizumab essentially as an imaging reagent. This system could be adapted for simple, inexpensive and sensitive/specific detection of tissue and circulating MV cells in a variety of neoplastic and non-neoplastic conditions, and for drug development and individualized cancer treatment.

Quantitative imaging assay for NF-κB nuclear translocation in primary human macrophages

Quantitative measurement of NF-κB nuclear translocation is an important research tool in cellular immunology. Established methodologies have a number of limitations, such as poor sensitivity, high cost or dependence on cell lines. Novel imaging methods to measure nuclear translocation of transcriptionally active components of NF-κB are being used but are also partly limited by the need for specialist imaging equipment or image analysis software. Herein we present a method for quantitative detection of NF-κB rel A nuclear translocation, using immunofluorescence microscopy and the public domain image analysis software ImageJ that can be easily adopted for cellular immunology research without the need for specialist image analysis expertise and at low cost. The method presented here is validated by demonstrating the time course and dose response of NF-κB nuclear translocation in primary human macrophages stimulated with LPS, and by comparison with a commercial NF-κB activation reporter cell line.

3D morpho-topological analysis of asymmetric neuronal morphogenesis in developing zebrafish

3D morpho-topological analysis of asymmetric neuronal morphogenesis in developing zebrafish

Videomimicography: the standards of normal revised.

Studies aiming to objectively evaluate facial movements have focused on the technique of measurement, whereas the most pertinent measurements of basic facial movements have not been well characterized. To determine the best normal measures of 5 basic facial movements in healthy patients. In 5 healthy subjects, 11 facial landmarks were placed on the face, and 5 movements (forehead lift, eye closure, nose wrinkling, lip puckering, and smiling) with maximal contraction force were requested. Each subject repeated each movement 3 times, and the entire session was repeated on 4 different days. No specific immobilization of the head was performed. The session was filmed with a digital camera, and the frames with maximal movement were selected. Measurements were performed with Osiris public domain image analysis software. For each measure, the change from rest was computed. Intersubject and intrasubject variability were determined by a multivariate analysis of variance. In all movements, surface changes (mean +/- SD) were higher than distance changes. For forehead lifting and eye closure, the best measure was the eye surface changes of 13% +/- 5% and -32% +/- 9%, respectively. For nasal wrinkling, lip puckering, and smiling, the best measures were the paranasal area (change, -28% +/- 9%), upper lip area (change, -23% +/- 8%), and mouth area (change, 63% +/- 21%), respectively. Most distance changes were below 10%. Same-day repeatability variation was below 15%, and day-to-day repeatability variation was below 7%. In healthy subjects, more than 80% of the total variation was accounted for by the intersubject variability. Videomimicography is a simple and objective linear measurement system based on facial surface changes. The measures exhibit good reliability.

Porosity measurement of fragile agglomerates

Agglomeration processes in the food and pharmaceutical industries frequently produce porous, brittle agglomerates intended for redispersion in liquids (a.k.a. “instant powders”). Typical products have a porosity ranging from ε=0.4 to ε=0.8 and a particle size between 0.2 and 2 mm. The agglomerate porosity is an important product property, as it has a major influence on the wetting behaviour. Due to their high porosity and comparatively weak internal bondings, instant agglomerates are fragile. Compared to the wide range of currently available particle sizing methods, few porosity-measuring methods exist, most of which are not applicable to instant agglomerates. For controlling the porosity of instant agglomerates produced in our laboratory, we used individual particle weighing and sizing. A balance with 1 μg of precision allowed reliable weighing of individual agglomerates as small as 300 μm diameter. The volume of the particles was determined using a long-distance microscope with CCD camera attached to a computer running public domain image analysis software. Porosity measurements of agglomerated skim and whole milk powder, agglomerated pectin, agglomerated flour/maltodextrin mixture, and agglomerated rice starch/maltodextrin mixtures were carried out. The results of our measurements show, for the majority of the samples, an approximately constant porosity over a wide particle size range, with a tendency towards lower porosity for smaller particles.

A Syndrome of Joint Laxity and Impaired Tendon Integrity in Lumican- and Fibromodulin-deficient Mice

Lumican and fibromodulin regulate the assembly of collagens into higher order fibrils in connective tissues. Here, we show that mice deficient in both of these proteoglycans manifest several clinical features of Ehlers-Danlos syndrome. The Lum(-/-)Fmod(-/-) mice are smaller than their wild type littermates and display gait abnormality, joint laxity, and age-dependent osteoarthritis. Misaligned knee patella, severe knee dysmorphogenesis, and extreme tendon weakness are the likely causes for joint laxity in the double-nulls. Fibromodulin deficiency alone leads to significant reduction in tendon stiffness in the Lum(+/+)Fmod(-/-) mice, with further loss in stiffness in a Lum gene dose-dependent way. At the protein level, we show marked increase of lumican in Fmod(-/-) tendons, which may partially rescue the tendon phenotype in this genotype. These results establish fibromodulin as a key regulator and lumican as a modulator of tendon strength. A disproportionate increase in small diameter immature collagen fibrils and a lack of progression to mature, large diameter fibrils in the Fmod(-/-) background may constitute the underlying cause of tendon weakness and suggest that fibromodulin aids fibril maturation. This study demonstrates that the collagen fibril-modifying proteoglycans, lumican and fibromodulin, are candidate genes and key players in the pathogenesis of certain types of Ehlers-Danlos syndrome and other connective tissue disorders.

Quick and easy molecular weight determination with Macintosh computers and public domain image analysis software.

The program "molecular weights" allows a fast and easy estimation of molecular weights (M(r)), isoelectric point (pI) values and band intensities directly from scanned, polyacrylamide gels, two-dimensional protein patterns and DNA gel images. The image coordinates of M(r) and pI reference standards enable the program to calculate M(r) and pI values in a real time manner for any cursor position. The program requires NIH-Image for Macintosh computers and includes automatic band detection coupled with a densitometric evaluation.