Anna Babinska | SUNY: Downstate Medical Center (original) (raw)
Papers by Anna Babinska
Cancer Cell International, Aug 11, 2023
Background The F11R/JAM-A cell adhesion protein was examined as the therapeutic target in triple ... more Background The F11R/JAM-A cell adhesion protein was examined as the therapeutic target in triple negative breast cancer (TNBC) with the use of the peptide antagonist to F11R/JAM-A, that previously inhibited the early stages of breast cancer metastasis in vitro. Methods The online in silico analysis was performed by TNMPlot, UALCAN, and KM plotter. The in vitro experiments were performed to verify the effect of peptide 4D (P4D) on human endothelial cell lines EA.hy926 and HMEC-1 as well as on human TNBC cell line MDA-MB-231. The cell morphology upon P4D treatment was verified by light microscopy, while the cell functions were assessed by colony forming assay, MTT cell viability assay, BrdU cell proliferation assay, and Transepithelial/Endothelial Electrical Resistance measurements. The in vivo experiments on 4T1 murine breast cancer model were followed by histopathological analysis and a series of quantitative analyses of murine tissues. Results By in silico analysis we have found the elevated gene expression in breast cancer with particular emphasis on TNBC. The elevated F11R expression in TNBC was related with poorer survival prognosis. Peptide 4D has altered the morphology and increased the permeability of endothelial monolayers. The colony formation, viability, and proliferation of MDA-MB-231 cells were decreased. P4D inhibited the metastasis in 4T1 breast cancer murine model in a statistically significant manner that was demonstrated by the resampling bootstrap technique. Conclusions The P4D peptide antagonist to F11R/JAM-A is able to hinder the metastasis in TNBC. This assumption needs to be confirmed by additional 4T1 mouse model study performed on larger group size, before making the decision on human clinical trials.
Breast Cancer Research and Treatment, Oct 24, 2019
Purpose To examine the involvement of the F11R/JAM-A protein in breast cancer metastasis, we util... more Purpose To examine the involvement of the F11R/JAM-A protein in breast cancer metastasis, we utilized the F11R/JAM-A antagonistic peptide 4D (P4D) in experiments of transendothelial migration (TEM) of breast cancer cells. Methods Experiments were conducted in the mouse 4T1 breast cancer model utilizing the human mammary epithelial cell and endothelial cell lines. The levels of soluble F11R/JAM-A (sJAM-A) in the murine plasmas were measured by ELISA. Levels of F11R/JAM-A mRNA and protein in cell lines were assessed by qRT-PCR and Western blot, respectively. Cell surface expression of F11R/JAM-A was demonstrated by flow cytometry. Functional tests included the TEM of breast cancer cells and adhesion of breast cancer cells to the endothelium. The endothelial permeability was studied by fluorescent tracer assay and by the Real-Time Cell Analysis (RTCA). Results The tumor inducers Tβ4 and TGF-β1 reduced the levels of sJAM-A in murine plasma, and reduced the F11R/JAM-A protein levels in the human microvascular endothelial cell line HMEC-1. The adhesion and TEM measured between breast cancer cells and inflamed or Tβ4-treated endothelium were inhibited by P4D. The presence of P4D did not destabilize the pre-existing tight junctions in the endothelial monolayer. The barrier-protecting effect of P4D was stronger than that of forskolin, when a booster dose of P4D was applied to the inflamed endothelium. Conclusions F11R/JAM-A protein can be considered as a novel target in the treatment of breast cancer metastasis. In vivo and clinical studies are needed to further investigate the effectiveness of F11R/JAM-A-derived peptide as a possible antimetastatic drug.
We examined various parameters of platelet adhesion to the vascular walls in mice in both in the ... more We examined various parameters of platelet adhesion to the vascular walls in mice in both in the arterioles and venules in the presence of a peptide antagonist of the F11 Receptor (peptide 4D), utilizing the intravital microscopy system. To induce an inflammatory phenotype of the vascular endothelium, mice were injected intraperitoneally with both murine recombinant TNF-alpha and murine recombinant IFN-gamma prior to the initiation of the measurement of platelet adhesion. The F11R peptide 4D was injected to the peptide-treated group of animals in 3 consecutive applications. The control, vehicle- treated animal group received injections of equal volumes of saline at each time point. At the time of measurements conducted to assess platelet adhesion to the vasculature, mice were anesthetized and injected with the platelet-specific fluorescent anti-GPIbβ antibody. Video imaging was carried out at the magnification of 200x for 40 s with an exposure time of 200 ms in at least three venules and at least three arterioles in each mouse. The analysis of platelets tethering to the endothelium cells in the recorded movies was performed with the use of TrackMate plugin that is implemented in FIJI software. Three parameters, each characterizing platelet interactions with the vascular wall, were calculated: 1. the time of interaction (referred to as the 'adhesion time'), 2. the reciprocal distance (i.e. the distance in a given view field that was overcome by a rolling platelet), which may be regarded as a measure of an overall platelet 'immobilization' on endothelium integrated through time, and 3. the integrated measure of these two parameters, the so called reciprocal 'velocity' of a platelet sliding on the vessel wall (referred to as 'adhesion time*distance-1'). These results demonstrate that the platelets were sliding with a higher velocity in the arterioles of the peptide-treated group and suggest that peptide 4D significantly inhibits platelets' ability to interact with endothelial layer of arterioles (Representative vide [...]
Atherosclerosis, 2019
Background and aims: The F11 Receptor (F11R), AKA Junctional Adhesion Molecule-A (JAM-A) (F11R/JA... more Background and aims: The F11 Receptor (F11R), AKA Junctional Adhesion Molecule-A (JAM-A) (F11R/JAM-A), is an adhesion protein constitutively expressed on the membrane surface of circulating platelets and the luminal surface of inflamed endothelial cells (EC). Platelet adhesion to an inflamed endothelium is one of the early steps of atherosclerotic plaque formation. Our previous studies, conducted with cultured EC in vitro, have demonstrated the expression of F11R/JAM-A on the luminal surface of inflamed EC, platelet adhesion to inflamed EC through F11R/JAM-A interactions, and inhibition of this interaction by the presence of F11R/JAM-A antagonistic peptide (F11Rpeptide 4D). In the present study, we examined in vivo the overall health-benefits and cardiovascular effects of long-term treatment of animals prone to atherosclerosis, ApoE-/mice, with F11R-peptide 4D.
MOJ Proteomics & Bioinformatics, 2018
One top research assay used to validate the efficacy of pharmacological agents used in antiplatel... more One top research assay used to validate the efficacy of pharmacological agents used in antiplatelets aggregation therapy is mass spectrometry coupled with high throughput profiling of changes in the proteomic expression in the activated platelets exposed to inhibitors or agonists of platelets signaling. Herein we present the development of a new, bottom-up proteomics platform that enabled monitoring of changes in protein expression profiles of healthy human platelets, ex-vivo activated with thrombin, using as reference the proteome of resting platelets. The assay employed the platelets cryogenic lysis and protein solubilization under denaturant conditions, enabling the extraction of key membranebound proteins, in addition to the cytosolic and organelles-derived proteins secreted in microparticles, and found in the platelets releasates. Nano LC-ESI/MS/MS sequencing of tryptic/Glu-C/Lys-C peptides from the "in solution" one step digestion of the whole platelets proteomes (2x10 8 platelets/patient sample) was performed on a Q-Exactive quadrupole orbitrap mass spectrometer coupled with label free quantification (LFQ) analysis. The changes in proteomic landscapes were qualitatively and quantitatively analyzed using a combination of systems biology and bioinformatics approaches empowered by the ingenuity pathway analysis (IPA), and the screening of identified proteins and genes entities against curated databases containing platelets proteomes, including the "Adhesome", "Exocarta", "Reactome" and "Platelet Web". The label-free global proteomics analysis retrieved a total of 924 proteins (FDR <1.3% for proteins and <0.8% for peptides) out of which 330 were shared between resting and thrombin-activated platelets. About 50% of the total proteome of thrombin-activated platelets was represented by up-regulated and previously validated protein biomarkers, involved in the pathways mediating the protease and thrombin activated receptor (PARs), integrin signaling, the actin and rhoA linked to cytoskeleton signaling, and activated kinases pathways regulating the cellular motility, aggregation, procoagulation and degranulation. In conclusion, the systems biology approaches validated our newly developed LFQ proteomic assay as a reliable tool for monitoring the changes in protein expression profiles in thrombin-activated platelets, and further advocate for employment of these approaches in assessing the efficacy of antiplatelets drugs (inhibitors of platelets aggregation) during the prevention and treatment of cardiovascular and cerebrovascular diseases.
PLOS ONE, Sep 28, 2015
Introduction Based on a single rat study by Lillemoe et al, the consensus has been formed to impl... more Introduction Based on a single rat study by Lillemoe et al, the consensus has been formed to implicate sorbitol rather than sodium polystyrene sulfonate (SPS) as the culprit for colon necrosis in humans treated with SPS and sorbitol. We tested the hypothesis that colon necrosis by sorbitol in the experiment was due to the high osmolality and volume of sorbitol rather than its chemical nature. Methods 26 rats underwent 5/6 nephrectomy. They were divided into 6 groups and given enema solutions under anesthesia (normal saline, 33% sorbitol, 33% mannitol, SPS in 33% sorbitol, SPS in normal saline, and SPS in distilled water). They were sacrificed after 48 hours of enema administration or earlier if they were very sick. The gross appearance of the colon was visually inspected, and then sliced colon tissues were examined under light microscopy. Results 1 rat from the sorbitol and 1 from the mannitol group had foci of ischemic colonic changes. The rats receiving SPS enema, in sorbitol, normal saline, distilled water, had crystal deposition with colonic necrosis and mucosal erosion. All the rats not given SPS survived until sacrificed at 48 h whereas 11 of 13 rats that received SPS in sorbitol, normal saline or distilled water died or were clearly dying and sacrificed sooner. There was no difference between sorbitol and mannitol when given without SPS. Conclusions In a surgical uremic rat model, SPS enema given alone or with sorbitol or mannitol seemed to cause colon necrosis and high mortality rate, whereas 33% sorbitol without SPS did not.
F11-receptor (F11R/JAM) mediates platelet adhesion to endothelial cells: Role in inflammatory thr... more F11-receptor (F11R/JAM) mediates platelet adhesion to endothelial cells: Role in inflammatory thrombosis
gene are required for an initial step of atherogenesis induced by inflammatory cytokines
Frontiers in Bioscience, 2018
Blood, 2000
This study demonstrates that the human platelet F11 receptor (F11R) functions as an adhesion mole... more This study demonstrates that the human platelet F11 receptor (F11R) functions as an adhesion molecule, and this finding is confirmed by the structure of the protein as revealed by molecular cloning. The F11R is a 32-/35-kd protein duplex that serves as the binding site through which a stimulatory monoclonal antibody causes platelet aggregation and granule secretion. A physiological role for the F11R protein was demonstrated by its phosphorylation after the stimulation of platelets by thrombin and collagen. A pathophysiological role for the F11R was revealed by demonstrating the presence of F11R-antibodies in patients with thrombocytopenia. Adhesion of platelets through the F11R resulted in events characteristic of the action of cell adhesion molecules (CAMs). To determine the structure of this protein, we cloned the F11R cDNA from human platelets. The predicted amino acid sequence demonstrated that it is an integral membrane protein and an immunoglobulin superfamily member containin...
Thrombosis and Haemostasis, 2018
It remains unclear whether plasma phospholipid transfer protein (PLTP) is involved in hyper-coagu... more It remains unclear whether plasma phospholipid transfer protein (PLTP) is involved in hyper-coagulation or hypo-coagulation. This study investigated the direct effect of PLTP on platelet aggregation and the underlying mechanism. Washed platelets from humans or mice and mouse platelet-rich plasma and human recombinant PLTP were isolated. PLTP is present in human platelets. We assessed adenosine diphosphate (ADP)-, collagen- and thrombin-induced platelet aggregation, phosphatidylserine externalization and photothrombosis-induced cerebral infarction in mice. PLTP over-expression increased platelet aggregation, while PLTP deficiency had the opposing reaction. Human recombinant PLTP increased both mouse and human platelet aggregation in a dose-dependent manner. Phosphatidylserine externalization provides a water/lipid surface for the interaction of coagulation factors, which accelerates thrombosis. Compared with wild-type controls, platelets from PLTP transgenic mice had significantly mo...
Biopolymers, 2014
Peptides with enhanced resistance to proteolysis, based on the amino acid sequence of the F11 rec... more Peptides with enhanced resistance to proteolysis, based on the amino acid sequence of the F11 receptor molecule (F11R, aka JAM-A/Junctional adhesion molecule-A), were designed, prepared, and examined as potential candidates for the development of anti-atherosclerotic and anti-thrombotic therapeutic drugs. A sequence at the N-terminal of F11R together with another sequence located in the first Ig-loop of this protein, were identified to form a steric active-site operating in the F11R-dependent adhesion between cells that express F11R molecules on their external surface. In silico modeling of the complex between two polypeptide chains with the sequences positioned in the active-site was used to generate peptide-candidates designed to inhibit homophilic interactions between surface-located F11R molecules. The two lead F11R peptides were modified with D-Arg and D-Lys at selective sites, for attaining higher stability to proteolysis in vivo. Using molecular docking experiments we tested ...
Hoppe-Seyler´s Zeitschrift für physiologische Chemie, 1983
The complex formation between fi-competitive inhibition parameters such as C/ max bronectin and d... more The complex formation between fi-competitive inhibition parameters such as C/ max bronectin and denatured collagen is associated Generally, an alteration of the fibronectin antiwith a significant decrease of antigenic expression genie structure is similar to that observed after of the fibronectin epitopes. This conclusion is thermic denaturation of the molecule, evidenced by critical changes in the value of Durch Komplexbindung an Kollagen verursachte Antigenveränderungen von menschlichem Fibronektin Zusammenfassung: Durch Komplexbindung von für die kompetitive Hemmung hin. Die so ver-Fibronektin mit denaturiertem Kollagen ergibt ursachte Änderung der Antigenstruktur des Fisich eine deutliche Verminderung der Antigen-bronektins hat den gleichen Charakter, wie bei expression der Fibronektindeterminanten. Darauf der Thermodenaturierung dieses Proteins, deuten wesentliche Änderungen der Parameter
European Journal of Biochemistry, 1995
Antisense phosphodiester and phosphorothioate oligodeoxynucleotides (23-residue or 24-residue oli... more Antisense phosphodiester and phosphorothioate oligodeoxynucleotides (23-residue or 24-residue oligodeoxynucleotides) were constructed for sequences of type-1 plasminogen-activator-inhibitor mRNA to assess their capability to modulate type-1 plasminogen-activator-inhibitor-mediated fibrinolysis. Antisense oligodeoxynucleotides were targeted at the mRNA sequence coding a signal peptide, at a part of the reactive center Ile342-Pro349, and at an internally translated segment Asn265-Leu272. The effect of antisense oligonucleotides on the concentration of type-1 plasminogen activator inhibitor in conditioned media and human endothelial cells was determined by the activity test with fibrin as a substrate, and by immunoprecipitation after metabolic labelling of cells with [35S]methionine. Three phosphorothioate oligodeoxynucleotides were specifically inhibitory while phosphodiester oligodeoxynucleotides with the same sequence did not show any activity. Phosphorothioate oligodeoxynucleotides 2, 4 and 6 inhibited the synthesis of type-1 plasminogen activator inhibitor in endothelial cells in a time-dependent and concentration-dependent manner. These data suggest that antisense oligodeoxynucleotides may be useful in the design of antithrombolytic therapeutics.
Cancer Cell International, Aug 11, 2023
Background The F11R/JAM-A cell adhesion protein was examined as the therapeutic target in triple ... more Background The F11R/JAM-A cell adhesion protein was examined as the therapeutic target in triple negative breast cancer (TNBC) with the use of the peptide antagonist to F11R/JAM-A, that previously inhibited the early stages of breast cancer metastasis in vitro. Methods The online in silico analysis was performed by TNMPlot, UALCAN, and KM plotter. The in vitro experiments were performed to verify the effect of peptide 4D (P4D) on human endothelial cell lines EA.hy926 and HMEC-1 as well as on human TNBC cell line MDA-MB-231. The cell morphology upon P4D treatment was verified by light microscopy, while the cell functions were assessed by colony forming assay, MTT cell viability assay, BrdU cell proliferation assay, and Transepithelial/Endothelial Electrical Resistance measurements. The in vivo experiments on 4T1 murine breast cancer model were followed by histopathological analysis and a series of quantitative analyses of murine tissues. Results By in silico analysis we have found the elevated gene expression in breast cancer with particular emphasis on TNBC. The elevated F11R expression in TNBC was related with poorer survival prognosis. Peptide 4D has altered the morphology and increased the permeability of endothelial monolayers. The colony formation, viability, and proliferation of MDA-MB-231 cells were decreased. P4D inhibited the metastasis in 4T1 breast cancer murine model in a statistically significant manner that was demonstrated by the resampling bootstrap technique. Conclusions The P4D peptide antagonist to F11R/JAM-A is able to hinder the metastasis in TNBC. This assumption needs to be confirmed by additional 4T1 mouse model study performed on larger group size, before making the decision on human clinical trials.
Breast Cancer Research and Treatment, Oct 24, 2019
Purpose To examine the involvement of the F11R/JAM-A protein in breast cancer metastasis, we util... more Purpose To examine the involvement of the F11R/JAM-A protein in breast cancer metastasis, we utilized the F11R/JAM-A antagonistic peptide 4D (P4D) in experiments of transendothelial migration (TEM) of breast cancer cells. Methods Experiments were conducted in the mouse 4T1 breast cancer model utilizing the human mammary epithelial cell and endothelial cell lines. The levels of soluble F11R/JAM-A (sJAM-A) in the murine plasmas were measured by ELISA. Levels of F11R/JAM-A mRNA and protein in cell lines were assessed by qRT-PCR and Western blot, respectively. Cell surface expression of F11R/JAM-A was demonstrated by flow cytometry. Functional tests included the TEM of breast cancer cells and adhesion of breast cancer cells to the endothelium. The endothelial permeability was studied by fluorescent tracer assay and by the Real-Time Cell Analysis (RTCA). Results The tumor inducers Tβ4 and TGF-β1 reduced the levels of sJAM-A in murine plasma, and reduced the F11R/JAM-A protein levels in the human microvascular endothelial cell line HMEC-1. The adhesion and TEM measured between breast cancer cells and inflamed or Tβ4-treated endothelium were inhibited by P4D. The presence of P4D did not destabilize the pre-existing tight junctions in the endothelial monolayer. The barrier-protecting effect of P4D was stronger than that of forskolin, when a booster dose of P4D was applied to the inflamed endothelium. Conclusions F11R/JAM-A protein can be considered as a novel target in the treatment of breast cancer metastasis. In vivo and clinical studies are needed to further investigate the effectiveness of F11R/JAM-A-derived peptide as a possible antimetastatic drug.
We examined various parameters of platelet adhesion to the vascular walls in mice in both in the ... more We examined various parameters of platelet adhesion to the vascular walls in mice in both in the arterioles and venules in the presence of a peptide antagonist of the F11 Receptor (peptide 4D), utilizing the intravital microscopy system. To induce an inflammatory phenotype of the vascular endothelium, mice were injected intraperitoneally with both murine recombinant TNF-alpha and murine recombinant IFN-gamma prior to the initiation of the measurement of platelet adhesion. The F11R peptide 4D was injected to the peptide-treated group of animals in 3 consecutive applications. The control, vehicle- treated animal group received injections of equal volumes of saline at each time point. At the time of measurements conducted to assess platelet adhesion to the vasculature, mice were anesthetized and injected with the platelet-specific fluorescent anti-GPIbβ antibody. Video imaging was carried out at the magnification of 200x for 40 s with an exposure time of 200 ms in at least three venules and at least three arterioles in each mouse. The analysis of platelets tethering to the endothelium cells in the recorded movies was performed with the use of TrackMate plugin that is implemented in FIJI software. Three parameters, each characterizing platelet interactions with the vascular wall, were calculated: 1. the time of interaction (referred to as the 'adhesion time'), 2. the reciprocal distance (i.e. the distance in a given view field that was overcome by a rolling platelet), which may be regarded as a measure of an overall platelet 'immobilization' on endothelium integrated through time, and 3. the integrated measure of these two parameters, the so called reciprocal 'velocity' of a platelet sliding on the vessel wall (referred to as 'adhesion time*distance-1'). These results demonstrate that the platelets were sliding with a higher velocity in the arterioles of the peptide-treated group and suggest that peptide 4D significantly inhibits platelets' ability to interact with endothelial layer of arterioles (Representative vide [...]
Atherosclerosis, 2019
Background and aims: The F11 Receptor (F11R), AKA Junctional Adhesion Molecule-A (JAM-A) (F11R/JA... more Background and aims: The F11 Receptor (F11R), AKA Junctional Adhesion Molecule-A (JAM-A) (F11R/JAM-A), is an adhesion protein constitutively expressed on the membrane surface of circulating platelets and the luminal surface of inflamed endothelial cells (EC). Platelet adhesion to an inflamed endothelium is one of the early steps of atherosclerotic plaque formation. Our previous studies, conducted with cultured EC in vitro, have demonstrated the expression of F11R/JAM-A on the luminal surface of inflamed EC, platelet adhesion to inflamed EC through F11R/JAM-A interactions, and inhibition of this interaction by the presence of F11R/JAM-A antagonistic peptide (F11Rpeptide 4D). In the present study, we examined in vivo the overall health-benefits and cardiovascular effects of long-term treatment of animals prone to atherosclerosis, ApoE-/mice, with F11R-peptide 4D.
MOJ Proteomics & Bioinformatics, 2018
One top research assay used to validate the efficacy of pharmacological agents used in antiplatel... more One top research assay used to validate the efficacy of pharmacological agents used in antiplatelets aggregation therapy is mass spectrometry coupled with high throughput profiling of changes in the proteomic expression in the activated platelets exposed to inhibitors or agonists of platelets signaling. Herein we present the development of a new, bottom-up proteomics platform that enabled monitoring of changes in protein expression profiles of healthy human platelets, ex-vivo activated with thrombin, using as reference the proteome of resting platelets. The assay employed the platelets cryogenic lysis and protein solubilization under denaturant conditions, enabling the extraction of key membranebound proteins, in addition to the cytosolic and organelles-derived proteins secreted in microparticles, and found in the platelets releasates. Nano LC-ESI/MS/MS sequencing of tryptic/Glu-C/Lys-C peptides from the "in solution" one step digestion of the whole platelets proteomes (2x10 8 platelets/patient sample) was performed on a Q-Exactive quadrupole orbitrap mass spectrometer coupled with label free quantification (LFQ) analysis. The changes in proteomic landscapes were qualitatively and quantitatively analyzed using a combination of systems biology and bioinformatics approaches empowered by the ingenuity pathway analysis (IPA), and the screening of identified proteins and genes entities against curated databases containing platelets proteomes, including the "Adhesome", "Exocarta", "Reactome" and "Platelet Web". The label-free global proteomics analysis retrieved a total of 924 proteins (FDR <1.3% for proteins and <0.8% for peptides) out of which 330 were shared between resting and thrombin-activated platelets. About 50% of the total proteome of thrombin-activated platelets was represented by up-regulated and previously validated protein biomarkers, involved in the pathways mediating the protease and thrombin activated receptor (PARs), integrin signaling, the actin and rhoA linked to cytoskeleton signaling, and activated kinases pathways regulating the cellular motility, aggregation, procoagulation and degranulation. In conclusion, the systems biology approaches validated our newly developed LFQ proteomic assay as a reliable tool for monitoring the changes in protein expression profiles in thrombin-activated platelets, and further advocate for employment of these approaches in assessing the efficacy of antiplatelets drugs (inhibitors of platelets aggregation) during the prevention and treatment of cardiovascular and cerebrovascular diseases.
PLOS ONE, Sep 28, 2015
Introduction Based on a single rat study by Lillemoe et al, the consensus has been formed to impl... more Introduction Based on a single rat study by Lillemoe et al, the consensus has been formed to implicate sorbitol rather than sodium polystyrene sulfonate (SPS) as the culprit for colon necrosis in humans treated with SPS and sorbitol. We tested the hypothesis that colon necrosis by sorbitol in the experiment was due to the high osmolality and volume of sorbitol rather than its chemical nature. Methods 26 rats underwent 5/6 nephrectomy. They were divided into 6 groups and given enema solutions under anesthesia (normal saline, 33% sorbitol, 33% mannitol, SPS in 33% sorbitol, SPS in normal saline, and SPS in distilled water). They were sacrificed after 48 hours of enema administration or earlier if they were very sick. The gross appearance of the colon was visually inspected, and then sliced colon tissues were examined under light microscopy. Results 1 rat from the sorbitol and 1 from the mannitol group had foci of ischemic colonic changes. The rats receiving SPS enema, in sorbitol, normal saline, distilled water, had crystal deposition with colonic necrosis and mucosal erosion. All the rats not given SPS survived until sacrificed at 48 h whereas 11 of 13 rats that received SPS in sorbitol, normal saline or distilled water died or were clearly dying and sacrificed sooner. There was no difference between sorbitol and mannitol when given without SPS. Conclusions In a surgical uremic rat model, SPS enema given alone or with sorbitol or mannitol seemed to cause colon necrosis and high mortality rate, whereas 33% sorbitol without SPS did not.
F11-receptor (F11R/JAM) mediates platelet adhesion to endothelial cells: Role in inflammatory thr... more F11-receptor (F11R/JAM) mediates platelet adhesion to endothelial cells: Role in inflammatory thrombosis
gene are required for an initial step of atherogenesis induced by inflammatory cytokines
Frontiers in Bioscience, 2018
Blood, 2000
This study demonstrates that the human platelet F11 receptor (F11R) functions as an adhesion mole... more This study demonstrates that the human platelet F11 receptor (F11R) functions as an adhesion molecule, and this finding is confirmed by the structure of the protein as revealed by molecular cloning. The F11R is a 32-/35-kd protein duplex that serves as the binding site through which a stimulatory monoclonal antibody causes platelet aggregation and granule secretion. A physiological role for the F11R protein was demonstrated by its phosphorylation after the stimulation of platelets by thrombin and collagen. A pathophysiological role for the F11R was revealed by demonstrating the presence of F11R-antibodies in patients with thrombocytopenia. Adhesion of platelets through the F11R resulted in events characteristic of the action of cell adhesion molecules (CAMs). To determine the structure of this protein, we cloned the F11R cDNA from human platelets. The predicted amino acid sequence demonstrated that it is an integral membrane protein and an immunoglobulin superfamily member containin...
Thrombosis and Haemostasis, 2018
It remains unclear whether plasma phospholipid transfer protein (PLTP) is involved in hyper-coagu... more It remains unclear whether plasma phospholipid transfer protein (PLTP) is involved in hyper-coagulation or hypo-coagulation. This study investigated the direct effect of PLTP on platelet aggregation and the underlying mechanism. Washed platelets from humans or mice and mouse platelet-rich plasma and human recombinant PLTP were isolated. PLTP is present in human platelets. We assessed adenosine diphosphate (ADP)-, collagen- and thrombin-induced platelet aggregation, phosphatidylserine externalization and photothrombosis-induced cerebral infarction in mice. PLTP over-expression increased platelet aggregation, while PLTP deficiency had the opposing reaction. Human recombinant PLTP increased both mouse and human platelet aggregation in a dose-dependent manner. Phosphatidylserine externalization provides a water/lipid surface for the interaction of coagulation factors, which accelerates thrombosis. Compared with wild-type controls, platelets from PLTP transgenic mice had significantly mo...
Biopolymers, 2014
Peptides with enhanced resistance to proteolysis, based on the amino acid sequence of the F11 rec... more Peptides with enhanced resistance to proteolysis, based on the amino acid sequence of the F11 receptor molecule (F11R, aka JAM-A/Junctional adhesion molecule-A), were designed, prepared, and examined as potential candidates for the development of anti-atherosclerotic and anti-thrombotic therapeutic drugs. A sequence at the N-terminal of F11R together with another sequence located in the first Ig-loop of this protein, were identified to form a steric active-site operating in the F11R-dependent adhesion between cells that express F11R molecules on their external surface. In silico modeling of the complex between two polypeptide chains with the sequences positioned in the active-site was used to generate peptide-candidates designed to inhibit homophilic interactions between surface-located F11R molecules. The two lead F11R peptides were modified with D-Arg and D-Lys at selective sites, for attaining higher stability to proteolysis in vivo. Using molecular docking experiments we tested ...
Hoppe-Seyler´s Zeitschrift für physiologische Chemie, 1983
The complex formation between fi-competitive inhibition parameters such as C/ max bronectin and d... more The complex formation between fi-competitive inhibition parameters such as C/ max bronectin and denatured collagen is associated Generally, an alteration of the fibronectin antiwith a significant decrease of antigenic expression genie structure is similar to that observed after of the fibronectin epitopes. This conclusion is thermic denaturation of the molecule, evidenced by critical changes in the value of Durch Komplexbindung an Kollagen verursachte Antigenveränderungen von menschlichem Fibronektin Zusammenfassung: Durch Komplexbindung von für die kompetitive Hemmung hin. Die so ver-Fibronektin mit denaturiertem Kollagen ergibt ursachte Änderung der Antigenstruktur des Fisich eine deutliche Verminderung der Antigen-bronektins hat den gleichen Charakter, wie bei expression der Fibronektindeterminanten. Darauf der Thermodenaturierung dieses Proteins, deuten wesentliche Änderungen der Parameter
European Journal of Biochemistry, 1995
Antisense phosphodiester and phosphorothioate oligodeoxynucleotides (23-residue or 24-residue oli... more Antisense phosphodiester and phosphorothioate oligodeoxynucleotides (23-residue or 24-residue oligodeoxynucleotides) were constructed for sequences of type-1 plasminogen-activator-inhibitor mRNA to assess their capability to modulate type-1 plasminogen-activator-inhibitor-mediated fibrinolysis. Antisense oligodeoxynucleotides were targeted at the mRNA sequence coding a signal peptide, at a part of the reactive center Ile342-Pro349, and at an internally translated segment Asn265-Leu272. The effect of antisense oligonucleotides on the concentration of type-1 plasminogen activator inhibitor in conditioned media and human endothelial cells was determined by the activity test with fibrin as a substrate, and by immunoprecipitation after metabolic labelling of cells with [35S]methionine. Three phosphorothioate oligodeoxynucleotides were specifically inhibitory while phosphodiester oligodeoxynucleotides with the same sequence did not show any activity. Phosphorothioate oligodeoxynucleotides 2, 4 and 6 inhibited the synthesis of type-1 plasminogen activator inhibitor in endothelial cells in a time-dependent and concentration-dependent manner. These data suggest that antisense oligodeoxynucleotides may be useful in the design of antithrombolytic therapeutics.