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    Olga Glinskii

    Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they... more
    Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they interact with tumor cells. Here we demonstrate that, while FAK/Src/RhoA/ROCK/MKK3/6/p38 pathway is fully activated downstream of α3β1 in EC, endothelial ERK1/2 is progressively dephosphorylated despite time‐dependent MEK1/2 activation. We further demonstrate that ERK1/2 dephosphorylation results from a crosstalk between p38 and ERK1/2 pathways whereby p38‐dependent increase in the expression of dual specificity phosphatase MKP‐3 causes progressive ERK1/2 deactivation. The resulting outcome of this complex crosstalk between the two major endothelial MAP kinases is p38‐dependent activation of MT1‐MMP, increase in MMP‐9 expression, and progressive release of MMP‐2 from ECs. We conclude that upon interaction with ECs, metastatic cells highjack EC MMP system via α3β1‐induced crosstalk between major MAPK pathways to facilitate tumor cell extravasation.Grant Funding Source: Supported by the VA BLR&D Service Award 1I01BX000609 and NIH Grant R01CA160461 to VV Glinsky
    Detection, segmentation, and quantification of microvascular structures are the main steps towards studying microvascular remodeling. Combined with appropriate staining, confocal microscopy imaging enables exploration of the full 3D... more
    Detection, segmentation, and quantification of microvascular structures are the main steps towards studying microvascular remodeling. Combined with appropriate staining, confocal microscopy imaging enables exploration of the full 3D anatomical characteristics of microvascular systems. Segmentation of confocal microscopy images is a challenging task due to complexity of anatomical structures, staining and imaging issues, and lack of annotated training data. In this paper, we propose a deep learning system for robust segmentation of cranial vasculature of mice in confocal microscopy images. The proposed system is an ensemble of two deep-learning cascades consisting of two coarse-to-fine subnetworks with skip connections in between. One cascade aims to improve sensitivity, while the other aims to improve precision of the segmentation results. Our experiments on mice cranial vasculature showed promising results achieving segmentation accuracy of 92.02% and dice score of 81.45% despite being trained on very limited confocal microscopy data.
    Glycobioinformatics is a rapidly developing field providing a vital support for MS‐based glycoproteomics research. Recent advances in MS greatly increased technological capabilities for high throughput glycopeptide analysis. However,... more
    Glycobioinformatics is a rapidly developing field providing a vital support for MS‐based glycoproteomics research. Recent advances in MS greatly increased technological capabilities for high throughput glycopeptide analysis. However, interpreting MS output, in terms of identifying glycan structures, attachment sites and glycosylation linkages still presents multiple challenges. Here, we discuss current strategies used in MS‐based glycoproteomics and bioinformatics tools available for MS‐based glycopeptide and glycan analysis. We also provide a brief overview of recent efforts in glycobioinformatics such as the new initiative UniCarbKB directed toward developing more comprehensive and unified glycobioinformatics platforms. With regards to glycobioinformatics tools and applications, we do not express our personal preferences or biases, but rather focus on providing a concise description of main features and functionalities of each application with the goal of assisting readers in making their own choices and identifying and locating glycobioinformatics tools most suitable for achieving their experimental objectives.
    Glutamate Carboxypeptidase II (also known as Prostate Specific Membrane Antigen-PSMA) is an important marker in the diagnosis of prostate cancer, however, relatively little is known about its biochemical and structure-function... more
    Glutamate Carboxypeptidase II (also known as Prostate Specific Membrane Antigen-PSMA) is an important marker in the diagnosis of prostate cancer, however, relatively little is known about its biochemical and structure-function characteristics. We have expressed mutant forms of PSMA and have started to address the roles of three putative domains of PSMA in its cellular localization and peptidase activity. Three mutants, a full-length recombinant PSMA (rPSMA-FL), one expressing only the proposed extracellular domain of PSMA (rPSMA-ECD) and one form omitting the proposed transmembrane domain (rPSMA-deltaTMD) have been produced in human cells via a mammalian expression vector system. We show that rPSMA-FL is associated with the cell surface membrane; so too is rPSMA-deltaTMD even though it lacks the proposed transmembrane domain, whereas rPSMA-ECD has a cytosolic localization. Only rPSMA-FL retains functional hydrolytic activity and is similarly glycosylated to PSMA found in the cultured prostate cancer cell line LNCaP.
    <p>This poster is related to the following paper:</p> <p>V. B. S. Prasath, R. Pelapur, O. Glinskii, V. Glinskii, V. Huxley, K. Palaniappan. <strong>Multiscale Tensor Anisotropic Filtering of Fluorescence Microscopy... more
    <p>This poster is related to the following paper:</p> <p>V. B. S. Prasath, R. Pelapur, O. Glinskii, V. Glinskii, V. Huxley, K. Palaniappan. <strong>Multiscale Tensor Anisotropic Filtering of Fluorescence Microscopy for Denoising Microvasculature</strong>. IEEE International Symposium on Biomedical Imaging (ISBI), New York, USA.</p> <p> </p
    Little is known about mechanical properties of human bone marrow endothelial cells (HBMEC) as culture conditions change. Here, we used atomic force microscopy (AFM) to test the hypothesis that the degree of confluency modifies HBMEC... more
    Little is known about mechanical properties of human bone marrow endothelial cells (HBMEC) as culture conditions change. Here, we used atomic force microscopy (AFM) to test the hypothesis that the degree of confluency modifies HBMEC stiffness. Individual cells were compared in non-confluent and confluent phases. Two HBMEC lines HBME-1 and HBMEC-60 immortalized by different methods were cultured until reaching a desired growth phase. Primary HBMEC were used as a reference. Cell stiffness was calculated as Young’s Moduli using a modified Hertz model from AFM cell-surface force curves. Actin expression was analyzed by immunofluorescence and western blotting (WB). Young’s Moduli were significantly increased in confluent vs non-confluent HBME-1 (6.79±2.58 vs 4.73±1.78 kPa, p<0.01) and HBMEC-60 (4.25±1.14 vs 2.03±1.09 kPa, p<0.01) cells. Non-confluent HBME-1 exhibited similar stiffness to the primary HBME (5.49±2.06 kPa). Topographical images revealed denser networks of actin stress fibers in confluent cells. F...
    <p>Poster from:</p> <p>V. B. S. Prasath, O. Haddad, F. Bunyak, R. K. Singh, O. Glinskii, V. Glinskii, V. Huxley, K. Palaniappan. <em>Computerized Dura Mater Laminae Analysis of Fluorescence Microscopy... more
    <p>Poster from:</p> <p>V. B. S. Prasath, O. Haddad, F. Bunyak, R. K. Singh, O. Glinskii, V. Glinskii, V. Huxley, K. Palaniappan. <em>Computerized Dura Mater Laminae Analysis of Fluorescence Microscopy Images</em>. Missouri Life Sciences Week, April 2013.</p> <p> </p> <p>This poster is related to the following paper:</p> <p>V. B. S. Prasath, O. Haddad, F. Bunyak, O. Glinskii, V. Glinskii, V. Huxley, K. Palaniappan. <strong>Robust Robust Filtering Based Segmentation and Analysis of Dura Mater Vasculature using Epifluorescence Microscopy</strong>. 35th Annual International Conference EMBS (IEEE EMBS/EMBC 2013), Osaka, Japan.</p> <p> </p
    Segmentation and quantification of microvasculature structures are the main steps toward studying microvasculature remodeling. The proposed patch based semantic architecture enables accurate segmentation for the challenging... more
    Segmentation and quantification of microvasculature structures are the main steps toward studying microvasculature remodeling. The proposed patch based semantic architecture enables accurate segmentation for the challenging epifluorescence microscopy images. Our pixel-based fast semantic network trained on random patches from different epifluorescence images to learn how to discriminate between vessels versus non-vessels pixels. The proposed semantic vessel network (SVNet) relies on understanding the morphological structure of the thin vessels in the patches rather than considering the whole image as input to speed up the training process and to maintain the clarity of thin structures. Experimental results on our ovariectomized - ovary removed (OVX) - mice dura mater epifluorescence microscopy images shows promising results in both arteriole and venule part. We compared our results with different segmentation methods such as local, global thresholding, matched based filter approache...
    Segmenting microvascular structures is an important requirement in understanding angioadaptation by which vascular networks remodel their morphological structures. Accurate segmentation for separating microvasculature structures is... more
    Segmenting microvascular structures is an important requirement in understanding angioadaptation by which vascular networks remodel their morphological structures. Accurate segmentation for separating microvasculature structures is important in quantifying remodeling process. In this work, we utilize a deep convolutional neural network (CNN) framework for obtaining robust segmentations of microvasculature from epifluorescence microscopy imagery of mice dura mater. Due to the inhomogeneous staining of the microvasculature, different binding properties of vessels under fluorescence dye, uneven contrast and low texture content, traditional vessel segmentation approaches obtain sub-optimal accuracy. We consider a deep CNN for the purpose keeping small vessel segments and handle the challenges posed by epifluorescence microscopy imaging modality. Experimental results on ovariectomized — ovary removed (OVX) — mice dura mater epifluorescence microscopy images show that the proposed modified CNN framework obtains an highest accuracy of 99% and better than other vessel segmentation methods.
    Brain is a common site of breast cancer metastasis and is associated with significant neurologic morbidity, decreased quality of life, and greatly shortened survival. However, the molecular and cellular mechanisms underpinning brain... more
    Brain is a common site of breast cancer metastasis and is associated with significant neurologic morbidity, decreased quality of life, and greatly shortened survival. However, the molecular and cellular mechanisms underpinning brain colonization by breast carcinoma cells are poorly understood. In this study, we used 2D-DIGE (Difference in Gel Electrophoresis) proteomic analysis followed by LC-tandem mass spectrometry to identify the proteins differentially expressed in brain-targeting breast carcinoma cells (MB231-Br) compared with parental MDA-MB-231 cell line. Between the two cell lines, we identified 12 proteins consistently exhibiting greater than 2-fold (p This work was supported in part by Award Number 1I01BX000609 from the Biomedical Laboratory Research & Development Service of the VA Office of Research and Development to VVG and AHA National SDG 0830287N to OVG. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5124. doi:10.1158/1538-7445.AM2011-5124
    Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Metastatic cell arrest in distant organ microvessels constitutes an essential rate-limiting step in hematogenous cancer metastasis. Previously, in a series of works... more
    Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL Metastatic cell arrest in distant organ microvessels constitutes an essential rate-limiting step in hematogenous cancer metastasis. Previously, in a series of works we demonstrated the importance of interactions between cancer-associated Thomsen-Friedenreich antigen (TF-Ag) and endothelial galectin-3 (Gal-3) in facilitating the initial carbohydrate-mediated tumor cell adhesion to distant organ microvasculature. However, it was unclear which molecules stabilize TF-Ag/Gal-3 mediated adhesion and how these interactions effect subsequent events in a metastatic cascade. Here, the results of a pull down assay using TF-Ag coated gold nanoparticles followed by Western analysis identified endothelial integrin α3β1 being physically associated with TF-Ag/Gal-3 complexes. Blocking α3β1 did not affect the initial tumor cell adhesion to the endothelium, but reduced >10-fold shear force necessary to displace cancer cells stably attached to endothelial monolayers, indicating that α3β1 stabilizes tumor-endothelial cell adhesion initiated by TF-Ag/Gal-3 interactions. Subsequent Quadrupole Time-of-Flight mass spectrometry analysis revealed additional 7 proteins forming a macromolecular complexes with TF-Ag, Gal-3 and α3β1 such as filamin B, talin, IQ motif containing GTPase activating protein 1, vinculin, α actinin 4, zyxin, and T-plastin, all of which map onto focal adhesion pathway. Our data demonstrate an induction of complex signaling pathways downstream of this macromolecular complex facilitating endothelial cell retraction and metastatic cell transendothelial migration. Thus, endothelial α3β1 mobilization downstream of TF-Ag/Gal-3 interactions stabilizes tumor-endothelial cell adhesion, initiates focal adhesion formation, and induces signaling pathways in endothelial cells promoting cancer metastasis. This work was supported in part by Award Number 1I01BX000609 from the Biomedical Laboratory Research & Development Service of the VA Office of Research and Development to VVG and AHA National SDG 0830287N to OVG. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2436. doi:10.1158/1538-7445.AM2011-2436
    The cornea and cranial dura mater share sensory innervation. This link raises the possibility that pathological impulses mediated by corneal injury may be transmitted to the cranial dura, trigger dural perivascular/connective tissue... more
    The cornea and cranial dura mater share sensory innervation. This link raises the possibility that pathological impulses mediated by corneal injury may be transmitted to the cranial dura, trigger dural perivascular/connective tissue nociceptor responses, and induce vascular and stromal alterations affecting dura mater blood and lymphatic vessel functionality. In this study, using a mouse model, we demonstrate for the first time that two weeks after the initial insult, alkaline injury to the cornea leads to remote pathological changes within the coronal suture area of the dura mater. Specifically, we detected significant pro-fibrotic changes in the dural stroma, as well as vascular remodeling characterized by alterations in vascular smooth muscle cell (VSMC) morphology, reduced blood vessel VSMC coverage, endothelial cell expression of the fibroblast specific protein 1, and significant increase in the number of podoplanin-positive lymphatic sprouts. Intriguingly, the deficiency of a ...
    Research Interests:
    Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they... more
    Our recent results demonstrate that metastatic cell adhesive interactions with endothelial cells (EC) are stabilized by the endothelial integrin α3β1 leading to the activation of p38 and ERK1/2 pathways in endothelial cells when they interact with tumor cells. Here we demonstrate that, while FAK/Src/RhoA/ROCK/MKK3/6/p38 pathway is fully activated downstream of α3β1 in EC, endothelial ERK1/2 is progressively dephosphorylated despite time-dependent MEK1/2 activation. We further demonstrate that ERK1/2 dephosphorylation results from a crosstalk between p38 and ERK1/2 pathways whereby p38-dependent increase in the expression of dual specificity phosphatase MKP-3 causes progressive ERK1/2 deactivation. The resulting outcome of this complex crosstalk between the two major endothelial MAP kinases is p38-dependent activation of MT1-MMP, increase in MMP-9 expression, and progressive release of MMP-2 from ECs. We conclude that upon interaction with ECs, metastatic cells highjack EC MMP system via α3β1-induced cross...
    Little is known about mechanical properties of human bone marrow endothelial cells (HBMEC) as culture conditions change. Here, we used atomic force microscopy (AFM) to test the hypothesis that the degree of confluency modifies HBMEC... more
    Little is known about mechanical properties of human bone marrow endothelial cells (HBMEC) as culture conditions change. Here, we used atomic force microscopy (AFM) to test the hypothesis that the degree of confluency modifies HBMEC stiffness. Individual cells were compared in non-confluent and confluent phases. Two HBMEC lines HBME-1 and HBMEC-60 immortalized by different methods were cultured until reaching a desired growth phase. Primary HBMEC were used as a reference. Cell stiffness was calculated as Young’s Moduli using a modified Hertz model from AFM cell-surface force curves. Actin expression was analyzed by immunofluorescence and western blotting (WB). Young’s Moduli were significantly increased in confluent vs non-confluent HBME-1 (6.79±2.58 vs 4.73±1.78 kPa, p<0.01) and HBMEC-60 (4.25±1.14 vs 2.03±1.09 kPa, p<0.01) cells. Non-confluent HBME-1 exhibited similar stiffness to the primary HBME (5.49±2.06 kPa). Topographical images revealed denser networks of actin stress fibers in confluent cells. F...
    The pathogenesis of headaches is a matter of ongoing discussion of two major theories describing it either as a vascular phenomenon resulting from vasodilation or primarily as a neurogenic process accompanied by secondary vasodilation... more
    The pathogenesis of headaches is a matter of ongoing discussion of two major theories describing it either as a vascular phenomenon resulting from vasodilation or primarily as a neurogenic process accompanied by secondary vasodilation associated with sterile neurogenic inflammation. While summarizing current views on neurogenic and vascular origins of headache, this mini review adds new insights regarding how smooth muscle-free microvascular networks, discovered within dura mater connective tissue stroma (previously thought to be "avascular"), may become a site of initial insult generating the background for the development of headache. Deficiencies in estrogen-dependent control of microvascular integrity leading to plasma protein extravasation, potential activation of perivascular and connective tissue stroma nociceptive neurons, and triggering of inflammatory responses are described. Finally, possible avenues for controlling and preventing these pathophysiological change...
    In this paper, we consider confocal microscopy based vessel segmentation with optimized features and random forest classification. By utilizing multi-scale vessel-specific features tuned to capture curvilinear structures such as Frobenius... more
    In this paper, we consider confocal microscopy based vessel segmentation with optimized features and random forest classification. By utilizing multi-scale vessel-specific features tuned to capture curvilinear structures such as Frobenius norm of the Hessian eigenvalues, Laplacian of Gaussians (LoG), oriented second derivative, line detector and intensity masked with LoG scale map. we obtain better segmentation results in challenging imaging conditions. We obtain binary segmentations using random forest classifier trained on physiologists marked ground-truth. Experimental results on mice dura mater confocal microscopy vessel segmentations indicate that we obtain better results compared to global segmentation approaches.
    The tumor-associated Thomsen-Friedenreich glycoantigen (TF-Ag) plays an important role in hematogenous metastasis of multiple cancers. The LTQ Orbitrap LC-MS/MS mass spectrometry analysis of cell surface TF-Ag proteome of metastatic... more
    The tumor-associated Thomsen-Friedenreich glycoantigen (TF-Ag) plays an important role in hematogenous metastasis of multiple cancers. The LTQ Orbitrap LC-MS/MS mass spectrometry analysis of cell surface TF-Ag proteome of metastatic prostate cancer cells reveals that several cell surface glycoproteins expressing this carbohydrate antigen in prostate cancer (CD44, α2 integrin, β1 integrin, CD49f, CD133, CD59, EphA2, CD138, transferrin receptor, profilin) are either known as stem cell markers or control important cancer stem-like cell functions. This outcome points to a potential link between TF-Ag expression and prostate cancer stem-like phenotype. Indeed, selecting prostate cancer cells for TF-Ag expression resulted in the enrichment of cells with stem-like properties such as enhanced clonogenic survival and growth, prostasphere formation under non-differentiating and differentiating conditions, and elevated expression of stem cell markers such as CD44 and CD133. Further, the analys...
    JAA-F11 is a highly specific mouse monoclonal to the Thomsen-Friedenreich Antigen (TF-Ag) which is an alpha-O-linked disaccharide antigen on the surface of ~80% of human carcinomas, including breast, lung, colon, bladder, ovarian, and... more
    JAA-F11 is a highly specific mouse monoclonal to the Thomsen-Friedenreich Antigen (TF-Ag) which is an alpha-O-linked disaccharide antigen on the surface of ~80% of human carcinomas, including breast, lung, colon, bladder, ovarian, and prostate cancers, and is cryptic on normal cells. JAA-F11 has potential, when humanized, for cancer immunotherapy for multiple cancer types. Humanization of JAA-F11, was performed utilizing complementarity determining regions grafting on a homology framework. The objective herein is to test the specificity, affinity and biology efficacy of the humanized JAA-F11 (hJAA-F11). Using a 609 target glycan array, 2 hJAA-F11 constructs were shown to have excellent chemical specificity, binding only to TF-Ag alpha-linked structures and not to TF-Ag beta-linked structures. The relative affinity of these hJAA-F11 constructs for TF-Ag was improved over the mouse antibody, while T20 scoring predicted low clinical immunogenicity. The hJAA-F11 constructs produced anti...
    ABSTRACT
    Interactions of metastatic cancer cells with vasculatory endothelium are critical during early stages of cancer metastasis. Understanding the molecular underpinnings of these interactions is essential for the development of new... more
    Interactions of metastatic cancer cells with vasculatory endothelium are critical during early stages of cancer metastasis. Understanding the molecular underpinnings of these interactions is essential for the development of new efficacious cancer therapies. Here we demonstrate that cancer-associated carbohydrate T antigen plays a leading role in docking breast and prostate cancer cells onto endothelium by specifically interacting with endothelium-expressed beta-galactoside-binding protein, galectin-3. Importantly, T antigen-bearing glycoproteins are also capable of mobilizing galectin-3 to the surface of endothelial cells, thus priming them for harboring metastatic cancer cells. The T antigen-mediated, tumor-endothelial cell interactions could be efficiently disrupted using synthetic compounds either mimicking or masking this carbohydrate structure. High efficiency of T antigen-mimicking and T antigen-masking inhibitors of tumor cell adhesion warrants their further development into ...
    Glutamate Carboxypeptidase II (also known as Prostate Specific Membrane Antigen—PSMA) is an important marker in the diagnosis of prostate cancer, however, relatively little is known about its biochemical and structure-function... more
    Glutamate Carboxypeptidase II (also known as Prostate Specific Membrane Antigen—PSMA) is an important marker in the diagnosis of prostate cancer, however, relatively little is known about its biochemical and structure-function characteristics. We have expressed mutant forms of PSMA and have started to address the roles of three putative domains of PSMA in its cellular localization and peptidase activity. Three mutants,
    Estrogen is a key regulator of vascular responses and angioadaptation in multiple organs and tissues, including brain. However, the consequences of a loss of ovarian steroid hormone secretion on the status of microvascular networks in... more
    Estrogen is a key regulator of vascular responses and angioadaptation in multiple organs and tissues, including brain. However, the consequences of a loss of ovarian steroid hormone secretion on the status of microvascular networks in brain and meninges are largely unknown. Here, using the perfused dura mater model coupled with high-resolution digital epifluorescence and laser scanning confocal microscopy and computer-assisted morphometric analysis, we demonstrate that cessation of ovarian hormone production causes dramatic vascular remodeling in meningeal microvascular networks characterized by a threefold decrease in microvessel density and capillary rarefaction and an almost fourfold increase in vascular permeability. These changes were accompanied by a significant decrease in angiopoietin-1 (Ang-1) expression and Ang-1/Tie-2 ratio (1.4-fold, P < 0.01, and 1.5-fold, P < 0.05, respectively) in ovariectomized animals compared with intact females, but no changes were detected ...
    Poster from:<br>Y. M. Kassim, V. B. S. Prasath, O. Glinskii, V. Glinsky, V. Huxley, K. Palaniappan. <i>Microvasculature segmentation of arterioles using deep CNN</i>. Machine Learning in Science and Engineering, Carnegie... more
    Poster from:<br>Y. M. Kassim, V. B. S. Prasath, O. Glinskii, V. Glinsky, V. Huxley, K. Palaniappan. <i>Microvasculature segmentation of arterioles using deep CNN</i>. Machine Learning in Science and Engineering, Carnegie Mellon University, Pittsburgh, PA, USA, Jun 6 - 8, 2018. <br><br>This poster is related to the following paper: Y. M. Kassim, V. B. S. Prasath, O. V. Glinskii, V. V. Glinsky, V. H. Huxley, K. Palaniappan. <strong>Microvasculature segmentation of arterioles using deep CNN</strong>. IEEE International Conference on Image Processing (ICIP), Beijing, China. Proc. IEEE, pp. 580-584, 2017.<br>. <br>
    Bone is a common site of metastasis for breast cancer and the mechanisms of metastasis are not fully elucidated. The purpose of our study was to characterize temporal and molecular dynamics of adhesive interactions between human breast... more
    Bone is a common site of metastasis for breast cancer and the mechanisms of metastasis are not fully elucidated. The purpose of our study was to characterize temporal and molecular dynamics of adhesive interactions between human breast cancer cells (HBCC) and human bone marrow endothelium (HBME) with piconewton resolution using atomic force microscopy (AFM). In adhesion experiments, a single breast cancer cell, MDA-MB-231 (MB231) or MDA-MB-435 (MB435) was attached to the AFM cantilever and brought into contact with a confluent HBME monolayer for different time periods (0.5 to 300 sec). The forces required to rupture individual molecular interactions and completely separate interacting cells were analyzed as measures of cell-cell adhesion. Adhesive interactions between HBME and either MB231 or MB435 cells increased progressively as cell-cell contact time was prolonged from 0.5 to 300 sec due to the time-dependent increase in the number and frequency of individual adhesive events, as ...
    Commonly used drawing tools for interactive image segmentation and labeling include active contours or boundaries, scribbles, rectangles and other shapes. Thin vessel shapes in images of vascular networks are difficult to segment using... more
    Commonly used drawing tools for interactive image segmentation and labeling include active contours or boundaries, scribbles, rectangles and other shapes. Thin vessel shapes in images of vascular networks are difficult to segment using automatic or interactive methods. This paper introduces the novel use of a sparse set of user-defined seed points (supervised labels) for precisely, quickly and robustly segmenting complex biomedical images. A multiquadric spline-based binary classifier is proposed as a unique approach for interactive segmentation using as features color values and the location of seed points. Epifluorescence imagery of the dura mater microvasculature are difficult to segment for quantitative applications due to challenging tissue preparation, imaging conditions, and thin, faint structures. Experimental results based on twenty epifluorescence images is used to illustrate the benefits of using a set of seed points to obtain fast and accurate interactive segmentation compared to four interactive and automatic segmentation approaches.

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