Zoharia Evron - Academia.edu (original) (raw)

Papers by Zoharia Evron

European Journal of Cancer Supplements, 2010

Background: Most data on the therapeutic potential and expression of TRAIL in colorectal cancer h... more Background: Most data on the therapeutic potential and expression of TRAIL in colorectal cancer has come from in vitro studies using tumour cell lines. To gain a clearer understanding about the susceptibility of patient tumours to TRAIL, we derived primary human cancer epithelial cells [1]. Increased apoptosis was observed in both primary PAP60 and MIH55 after treatment with SuperKiller TRAIL. Treating patient tumour xenograft/SCID mouse models with Killer TRAIL in vivo for 5 consecutive days suppressed tumour growth, although less efficiently compared to in vitro experiments. RAS oncogenes sensitise cells to TRAIL induced apoptosis [2]. We have presented evidence that this effect is usually mediated by TRAIL receptor DR4 and DR5 overexpression and/or redistribution in cell models [4]. Materials and Methods: Primary colorectal tumour cells, colorectal cell lines bearing RAS and BRAF mutant oncogenes, mouse xenographs and colorectal clinical samples were either treated with recombinant TRAIL and/or analysed for the presence of RAS and BRAF oncogenic mutations and DR4, DR5 expression. Results: We present evidence that BRAF oncogenes sensitise cells to TRAIL induced apoptosis via TRAIL receptor DR5 [3]. We have also shown that DR5 is the most frequently upregulated DR in clinical samples of colon cancer. Furthermore, the presence of K-RAS and BRAF mutations in the tumour may directly or indirectly enhance DR expression [5]. Discussion: Mutations on K-RAS and BRAF oncogenes have been shown in many studies to be associated with resistance to EGFR targeted therapeutics and combinations. TRAIL-based therapeutics, other as mono-or combination therapy could provide a promising alternative for K-RAS/BRAF bearing colorectal tumours.

Journal of Orthopaedics and Traumatology, 2007

One method to exogenously enhance the repair of articular cartilage is the local application of g... more One method to exogenously enhance the repair of articular cartilage is the local application of growth factors. This method is based on the chondrogenic effects of some agents and their potential ability to enhance cell migration. Human fetal chondrocytes were isolated and cultured. Their proliferation under the influence of different agents was microscopically evaluated. Fetal calf serum at 5% and

Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of... more Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of nondiabetic (n = 5) and diabetic (n = 5) age-matched subjects. The discs used were discarded material from operations. To clarify the reason for the higher risk of disc prolapse in diabetic patients. The pathogenesis of diabetes results from a combination of neurologic dysfunctions and a yet undefined metabolic failure, which leads to an abnormal proteoglycan profile. The following methods were used to determine the proteoglycan profile: the measurement of 35S-sulfate uptake per gram wet tissue into sulfated glycosaminoglycan using fresh tissue explants; extraction of proteoglycans by 4 M guanidinium chloride containing protease inhibitors, with further purification by ultracentrifugation on cesium chloride buoyant density gradient under dissociative conditions; total uronic acid and protein contents in the various gradient fractions; assessing the length of sugar side chains of isolated 35Sulfate-glycosaminoglycan molecules by separation of the glycosaminoglycan molecules on a Sepharose 6B-CL column; and paper chromatography of the final digest products of glycosaminoglycan molecules obtained by chondroitinase ABC, a glycosaminoglycan-degrading enzyme. The findings show that discs from normal nondiabetic subjects have 15 times the rate of 35Sulfate incorporation into glycosaminoglycan molecules than do discs of diabetic patients. The proteoglycans of diabetic patients are banded at a lower buoyant density, indicating a lowered glycosylation rate and a lower number of sugar side chains per core protein. In discs of diabetic patients, there is a slight increase in the chain length of chondroitin sulfate. Further analysis of the glycosaminoglycan chains showed a decreased amount of keratan sulfate, compared with that in nondiabetic subjects. However, the total uronic acid content of the disc tissues and the ratio of uronic acid to protein of each fraction were unchanged in diabetic patients versus that in control subjects. Discs in patients with diabetes have proteoglycans with lower buoyant density and substantially undersulfated glycosaminoglycan, which with the specific neurologic damage in these patients, might lead to increased susceptibility to disc prolapse.

Tetrahedron, 2010

Eight novel thiazole and oxazole containing cyclic peptides, microcyclamides GL616, GL582, GL614A... more Eight novel thiazole and oxazole containing cyclic peptides, microcyclamides GL616, GL582, GL614A, GL614B, GL614C, GL546A, GL546B, and GL628, as well as the known microcyclamide A, were isolated from the hydrophylic extract of a Microcystis sp. water-bloom collected in Gilboa reservoir, Valley of Armageddon, Israel. The planar structure of the compounds was determined by homonuclear and inverseheteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers of the amino acids was studied using Marfey's method for HPLC following ozonolysis, hydrolysis and derivatization with Marfey's reagent. This is the first example where acidic and modified amino acids are incorporated in this group of ribosomally biosynthesized metabolites.

International Orthopaedics, 2006

Background: The purpose of this study is to illustrate the routes of migration of precartilaginou... more Background: The purpose of this study is to illustrate the routes of migration of precartilaginous cells from the perichondrial ring of LaCroix, as a potential reservoir for growth-plate germ cells. Methods: Chondrocytes derived from the ring of LaCroix of young chicks' proximal tibia were cultured in vitro and transfected with adenovirus vector containing the gene encoding for Escherichia coli (beta)-galactosidase (lacZ) gene, which allows assessment of the migratory routes of these cells. The lacZ-transfected cells were injected back into the perichondrial ring of LaCroix of young chicks' proximal tibias. Four weeks later the migration root was assessed microscopically. Results: Injection of cells derived from the ring of LaCroix of neonate chicks, transfected in culture with adenoviruses containing LacZ reporter gene, allows the assessment of migratory potential of these cells. Stained cells were found at the outer layer of the epiphysis, particularly in areas adjacent to the perichondrial ring. Further longitudinal histopathological studies along the bone axis demonstrated a condensed layer of the stained cells arranged horizontally along parts of the physis.

International Journal of Experimental Pathology, 2001

Primary synovial chondromatosis (PSC) is a rare disorder of the synovium typi®ed by cartilaginous... more Primary synovial chondromatosis (PSC) is a rare disorder of the synovium typi®ed by cartilaginous nodule formation within the synovial membrane. Fibroblast growth factor receptor 3 (FGFR3) is a recently described speci®c marker of mesenchymal precartilaginous stem cells. Expression patterns of FGFR3 and its speci®c ligand, fibroblast growth factor 9 (FGF 9), were evaluated both in situ and in cell cultures. Histologically, cells at the periphery of the cartilage nodules express FGFR3 and PCNA (proliferating cell nuclear antigen). Elevated levels of FGF 9, its speci®c ligand, have been found in synovial¯uids of patients with synovial chondromatosis. Synoviocytes but not chondrocytes from affected patients express FGF9 in culture. This pattern is absent in normal synovium and cartilage. Downregulation of FGF9 may provide a possible nonoperative therapy for PSC.

International Journal of Experimental Pathology, 2008

The different clinical entities of osteochondromas, hereditary multiple exostoses (HME) and non-f... more The different clinical entities of osteochondromas, hereditary multiple exostoses (HME) and non-familial solitary exostosis, are known to express localized exostoses in their joint metaphyseal cartilage. In the current study biopsies of osteochondromas patients were screened with respect to a number of cellular and molecular parameters. Specifically, cartilaginous biopsy samples of nine HME patients, 10 solitary exostosis patients and 10 articular cartilages of control subjects were collected and cell cultures were established. Results obtained showed that one of the two HME samples that underwent DNA sequencing analysis (HME-1) had a novel mutation for an early stop codon, which led to an aberrant protein, migrating at a lower molecular weight position. The EXT-1 mRNA and protein levels in chondrocyte cultures derived from all nine HME patients were elevated, compared with solitary exostosis patients or control subjects. Furthermore, cell cultures of HME patients had significantly decreased pericellular heparan sulphate (HS) in comparison with cultures of solitary exostosis patients or control subjects. Immunohistochemical staining of tissue sections and Western blotting of cell cultures derived from HME patients revealed higher levels of heparanase compared with solitary exostosis patients and of control subjects. Further investigations are needed to determine whether the low pericellular HS levels in HME patients stem from decreased biosynthesis of HS, increased degradation or a combination of both. In conclusion, it appears that due to a mutated glycosyltransferase, the low content of pericellular HS in HME patients leads to the anatomical deformations with exostoses formation. Hence, elevation of HS content in the pericellular regions should be a potential molecular target for correction.

Experimental and Molecular Pathology, 2003

Hereditary osteochondromas are often caused by mutation in the EXT1 gene. The lesions are typifie... more Hereditary osteochondromas are often caused by mutation in the EXT1 gene. The lesions are typified by formation of a "pseudo" growth plate like lesion growing at 60 degrees to the normal growth direction of the bone. Such lesions can be mimicked surgically by reverting the position--the polarity of the zone of LaCroix. The current study attempts to compare the pathology between EXT1 gene expression in humans and surgically created osteochondroma pathology in a rat model. Tissues of human bunion, human embryonal tissue, and human adult cartilage as well as normal rat epiphyses served as controls. Rats were operated on and a 60 degree span of the ring of LaCroix was inverted as described by Delgado (Delgado, E., Rodriguez, J. I., Serada, A., Tellez, M., and Pariagoa, R.. Clin. Orthop. 201, 251-258 (1985)). The surgically created osteochondromas were assessed by histology, histochemistry, and immunohistochemistry. The findings show that the surgically created lesions contain only a small amount of FGF receptor 3 (FGFR3) expressed on mesenchymal stem cells located in the perichondrium, as compared to the cell population carrying FGFR3 in the contralateral limb. Indian hedgehog and Bcl2 are downregulated, while BMP-2 is overexpressed in the operated limb, compared to the LaCroix ring of the contralaetral limb. The shortage, as well as the disturbed migration routes of the residual mesenchymal stem cells in surgically created osteochondromas leads eventually to resorption of the pathological elements. In search of additional markers characterizing such pathological structures composed of mesenchymal stem cells and cartilaginous and bony cells, EXT1 gene was found to be expressed in the surgically created osteochondromas, like in normal growth plates. Nitric oxide synthase was also expressed like in adult cartilage, though tumor necrosis factor alpha typifying Bunion formation was absent. In summary, surgically created osteochondromas lack the massive and continuous population of mesenchymal stem cells with Bcl2 expression. However, the small residual mesenchymal cell population gives rise to short-lived EXT1-expressing cells that disappear eventually due to spontaneous resorption.

European Journal of Heart Failure, 2003

To evaluate whether satellite cells injected into infarct areas in rabbits remain viable during 6... more To evaluate whether satellite cells injected into infarct areas in rabbits remain viable during 6 weeks follow-up and can improve cardiac function as assessed by echocardiography. Myocardial infarction was induced in 16 New Zealand white rabbits, by ligation of the marginalis sinistra artery. Tissue from gluteus muscle biopsies was dissected into small pieces and cultured. Within 2-3 weeks the cells were expanded by 2-3 orders of magnitude and were fluorescent labeled. Single cell pellets for resuspension at >10(6)/1 ml were directly injected into the infarct areas in 8 rabbits. In 8 additional rabbits, 1 ml saline was injected (control). Regional left ventricular function was assessed weekly by 2-D echocardiography until animals were sacrificed. Analysis was performed blind and independently by two experienced echocardiographers, based on the American Society of Echocardiography scheme. Six treated and five control rabbits completed the study. One week after the artery occlusion, left ventricular function scoring did not differ between groups, mean 8.7+/-1.6 vs 8.3+/-1.9 (P=0.74). At 6 weeks post-injection, echocardiographic score was significantly better in the treated group, mean 2.6+/-0.9 vs 6.9+/-2.1 (P=0.002). The treated group showed significant gradual segmental improvement between the first week up to week 6. After sacrifice, macro and microscopic transmural areas showed typical changes of myocardial infarction. Histochemical staining identified viable grafted cells in high density 6 weeks post-transplantation in all grafted hearts. Autologous satellite cells (skeletal myofiber), can be successfully grafted into rabbit hearts following myocardial infarction and may induce improved regional left ventricular function.

ACS Medicinal Chemistry Letters, 2011

The cytotoxic activity of aloe-emodin (AE), a natural anthranoid that readily permeates anthracyc... more The cytotoxic activity of aloe-emodin (AE), a natural anthranoid that readily permeates anthracycline-resistant tumor cells, was improved by the attachment of an amino-sugar unit to its anthraquinone core. The new class of AE glycosides (AEGs) showed a significant improvement in cytotoxicity-up to more than 2 orders of magnitude greater than those of AE and the clinically used anthracycline doxorubicin (DOX)-against several cancer cell lines with different levels of DOX resistance. Incubation with the synthetic AEGs induced cell death in less than one cell cycle, indicating that these compounds do not directly target the cell division mechanism. Confocal microscopy provided evidence that unlike DOX, AEGs accumulated in anthracycline-resistant tumor cells in which resistance is conferred by P-glycoprotein efflux pumps. The results of this study demonstrate that AEGs may serve as a promising scaffold for the development of cytotoxic agents capable of overcoming anthracycline resistance in tumor cells.

Connective Tissue Research, 2012

The familial disease of hereditary multiple exostoses is characterized by abnormal skeletal defor... more The familial disease of hereditary multiple exostoses is characterized by abnormal skeletal deformities requiring extensive surgical procedures. In hereditary multiple exostoses patients there is a shortage in the pericellular glycosaminoglycan (GAG) of heparan sulfate (HS), related to defective activity of HS glycosyltransferases, mainly in the pericellular regions of chondrocytes. This study searched for a novel approach employing xylosides with different aglycone groups priming a variety of GAG chains, in attempting to alter the GAG compositional profile. Cell cultures of patients with osteochondroma responded to p-nitrophenyl β-d-xyloside by a significant increase in total GAG synthesis, expressed mainly in the extracellular domains, limited to chondroitin sulfate). The different β-d-xylosides, in addition to increasing the synthesis of extracellular GAGs, led to a significant depletion of the intracellular GAG domains. In mouse chondrocyte cultures, β-d-xylosides with different aglycones created a unique distribution of the GAG pools. Of special interest was the finding that the naphthalene methanol β-d-xyloside showed the highest absolute levels of HS-GAGs in both extracellular and intra-pericellular moieties compared with other β-d-xylosides and with controls without xyloside. In summary, β-d-xylosides can be utilized in chondrocyte cultures to modify the distribution of GAGs between the extracellular and intracellular compartments. In addition, xylosides may alter the profile of specific GAG chains in each moiety.

European Journal of Cancer Supplements, 2010

Background: Most data on the therapeutic potential and expression of TRAIL in colorectal cancer h... more Background: Most data on the therapeutic potential and expression of TRAIL in colorectal cancer has come from in vitro studies using tumour cell lines. To gain a clearer understanding about the susceptibility of patient tumours to TRAIL, we derived primary human cancer epithelial cells [1]. Increased apoptosis was observed in both primary PAP60 and MIH55 after treatment with SuperKiller TRAIL. Treating patient tumour xenograft/SCID mouse models with Killer TRAIL in vivo for 5 consecutive days suppressed tumour growth, although less efficiently compared to in vitro experiments. RAS oncogenes sensitise cells to TRAIL induced apoptosis [2]. We have presented evidence that this effect is usually mediated by TRAIL receptor DR4 and DR5 overexpression and/or redistribution in cell models [4]. Materials and Methods: Primary colorectal tumour cells, colorectal cell lines bearing RAS and BRAF mutant oncogenes, mouse xenographs and colorectal clinical samples were either treated with recombinant TRAIL and/or analysed for the presence of RAS and BRAF oncogenic mutations and DR4, DR5 expression. Results: We present evidence that BRAF oncogenes sensitise cells to TRAIL induced apoptosis via TRAIL receptor DR5 [3]. We have also shown that DR5 is the most frequently upregulated DR in clinical samples of colon cancer. Furthermore, the presence of K-RAS and BRAF mutations in the tumour may directly or indirectly enhance DR expression [5]. Discussion: Mutations on K-RAS and BRAF oncogenes have been shown in many studies to be associated with resistance to EGFR targeted therapeutics and combinations. TRAIL-based therapeutics, other as mono-or combination therapy could provide a promising alternative for K-RAS/BRAF bearing colorectal tumours.

Journal of Orthopaedics and Traumatology, 2007

One method to exogenously enhance the repair of articular cartilage is the local application of g... more One method to exogenously enhance the repair of articular cartilage is the local application of growth factors. This method is based on the chondrogenic effects of some agents and their potential ability to enhance cell migration. Human fetal chondrocytes were isolated and cultured. Their proliferation under the influence of different agents was microscopically evaluated. Fetal calf serum at 5% and

Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of... more Characterization of the analytic profile of proteoglycans in the intervertebral discs at L4-L5 of nondiabetic (n = 5) and diabetic (n = 5) age-matched subjects. The discs used were discarded material from operations. To clarify the reason for the higher risk of disc prolapse in diabetic patients. The pathogenesis of diabetes results from a combination of neurologic dysfunctions and a yet undefined metabolic failure, which leads to an abnormal proteoglycan profile. The following methods were used to determine the proteoglycan profile: the measurement of 35S-sulfate uptake per gram wet tissue into sulfated glycosaminoglycan using fresh tissue explants; extraction of proteoglycans by 4 M guanidinium chloride containing protease inhibitors, with further purification by ultracentrifugation on cesium chloride buoyant density gradient under dissociative conditions; total uronic acid and protein contents in the various gradient fractions; assessing the length of sugar side chains of isolated 35Sulfate-glycosaminoglycan molecules by separation of the glycosaminoglycan molecules on a Sepharose 6B-CL column; and paper chromatography of the final digest products of glycosaminoglycan molecules obtained by chondroitinase ABC, a glycosaminoglycan-degrading enzyme. The findings show that discs from normal nondiabetic subjects have 15 times the rate of 35Sulfate incorporation into glycosaminoglycan molecules than do discs of diabetic patients. The proteoglycans of diabetic patients are banded at a lower buoyant density, indicating a lowered glycosylation rate and a lower number of sugar side chains per core protein. In discs of diabetic patients, there is a slight increase in the chain length of chondroitin sulfate. Further analysis of the glycosaminoglycan chains showed a decreased amount of keratan sulfate, compared with that in nondiabetic subjects. However, the total uronic acid content of the disc tissues and the ratio of uronic acid to protein of each fraction were unchanged in diabetic patients versus that in control subjects. Discs in patients with diabetes have proteoglycans with lower buoyant density and substantially undersulfated glycosaminoglycan, which with the specific neurologic damage in these patients, might lead to increased susceptibility to disc prolapse.

Tetrahedron, 2010

Eight novel thiazole and oxazole containing cyclic peptides, microcyclamides GL616, GL582, GL614A... more Eight novel thiazole and oxazole containing cyclic peptides, microcyclamides GL616, GL582, GL614A, GL614B, GL614C, GL546A, GL546B, and GL628, as well as the known microcyclamide A, were isolated from the hydrophylic extract of a Microcystis sp. water-bloom collected in Gilboa reservoir, Valley of Armageddon, Israel. The planar structure of the compounds was determined by homonuclear and inverseheteronuclear 2D NMR techniques as well as high-resolution mass spectrometry. The absolute configuration of the asymmetric centers of the amino acids was studied using Marfey's method for HPLC following ozonolysis, hydrolysis and derivatization with Marfey's reagent. This is the first example where acidic and modified amino acids are incorporated in this group of ribosomally biosynthesized metabolites.

International Orthopaedics, 2006

Background: The purpose of this study is to illustrate the routes of migration of precartilaginou... more Background: The purpose of this study is to illustrate the routes of migration of precartilaginous cells from the perichondrial ring of LaCroix, as a potential reservoir for growth-plate germ cells. Methods: Chondrocytes derived from the ring of LaCroix of young chicks' proximal tibia were cultured in vitro and transfected with adenovirus vector containing the gene encoding for Escherichia coli (beta)-galactosidase (lacZ) gene, which allows assessment of the migratory routes of these cells. The lacZ-transfected cells were injected back into the perichondrial ring of LaCroix of young chicks' proximal tibias. Four weeks later the migration root was assessed microscopically. Results: Injection of cells derived from the ring of LaCroix of neonate chicks, transfected in culture with adenoviruses containing LacZ reporter gene, allows the assessment of migratory potential of these cells. Stained cells were found at the outer layer of the epiphysis, particularly in areas adjacent to the perichondrial ring. Further longitudinal histopathological studies along the bone axis demonstrated a condensed layer of the stained cells arranged horizontally along parts of the physis.

International Journal of Experimental Pathology, 2001

Primary synovial chondromatosis (PSC) is a rare disorder of the synovium typi®ed by cartilaginous... more Primary synovial chondromatosis (PSC) is a rare disorder of the synovium typi®ed by cartilaginous nodule formation within the synovial membrane. Fibroblast growth factor receptor 3 (FGFR3) is a recently described speci®c marker of mesenchymal precartilaginous stem cells. Expression patterns of FGFR3 and its speci®c ligand, fibroblast growth factor 9 (FGF 9), were evaluated both in situ and in cell cultures. Histologically, cells at the periphery of the cartilage nodules express FGFR3 and PCNA (proliferating cell nuclear antigen). Elevated levels of FGF 9, its speci®c ligand, have been found in synovial¯uids of patients with synovial chondromatosis. Synoviocytes but not chondrocytes from affected patients express FGF9 in culture. This pattern is absent in normal synovium and cartilage. Downregulation of FGF9 may provide a possible nonoperative therapy for PSC.

International Journal of Experimental Pathology, 2008

The different clinical entities of osteochondromas, hereditary multiple exostoses (HME) and non-f... more The different clinical entities of osteochondromas, hereditary multiple exostoses (HME) and non-familial solitary exostosis, are known to express localized exostoses in their joint metaphyseal cartilage. In the current study biopsies of osteochondromas patients were screened with respect to a number of cellular and molecular parameters. Specifically, cartilaginous biopsy samples of nine HME patients, 10 solitary exostosis patients and 10 articular cartilages of control subjects were collected and cell cultures were established. Results obtained showed that one of the two HME samples that underwent DNA sequencing analysis (HME-1) had a novel mutation for an early stop codon, which led to an aberrant protein, migrating at a lower molecular weight position. The EXT-1 mRNA and protein levels in chondrocyte cultures derived from all nine HME patients were elevated, compared with solitary exostosis patients or control subjects. Furthermore, cell cultures of HME patients had significantly decreased pericellular heparan sulphate (HS) in comparison with cultures of solitary exostosis patients or control subjects. Immunohistochemical staining of tissue sections and Western blotting of cell cultures derived from HME patients revealed higher levels of heparanase compared with solitary exostosis patients and of control subjects. Further investigations are needed to determine whether the low pericellular HS levels in HME patients stem from decreased biosynthesis of HS, increased degradation or a combination of both. In conclusion, it appears that due to a mutated glycosyltransferase, the low content of pericellular HS in HME patients leads to the anatomical deformations with exostoses formation. Hence, elevation of HS content in the pericellular regions should be a potential molecular target for correction.

Experimental and Molecular Pathology, 2003

Hereditary osteochondromas are often caused by mutation in the EXT1 gene. The lesions are typifie... more Hereditary osteochondromas are often caused by mutation in the EXT1 gene. The lesions are typified by formation of a "pseudo" growth plate like lesion growing at 60 degrees to the normal growth direction of the bone. Such lesions can be mimicked surgically by reverting the position--the polarity of the zone of LaCroix. The current study attempts to compare the pathology between EXT1 gene expression in humans and surgically created osteochondroma pathology in a rat model. Tissues of human bunion, human embryonal tissue, and human adult cartilage as well as normal rat epiphyses served as controls. Rats were operated on and a 60 degree span of the ring of LaCroix was inverted as described by Delgado (Delgado, E., Rodriguez, J. I., Serada, A., Tellez, M., and Pariagoa, R.. Clin. Orthop. 201, 251-258 (1985)). The surgically created osteochondromas were assessed by histology, histochemistry, and immunohistochemistry. The findings show that the surgically created lesions contain only a small amount of FGF receptor 3 (FGFR3) expressed on mesenchymal stem cells located in the perichondrium, as compared to the cell population carrying FGFR3 in the contralateral limb. Indian hedgehog and Bcl2 are downregulated, while BMP-2 is overexpressed in the operated limb, compared to the LaCroix ring of the contralaetral limb. The shortage, as well as the disturbed migration routes of the residual mesenchymal stem cells in surgically created osteochondromas leads eventually to resorption of the pathological elements. In search of additional markers characterizing such pathological structures composed of mesenchymal stem cells and cartilaginous and bony cells, EXT1 gene was found to be expressed in the surgically created osteochondromas, like in normal growth plates. Nitric oxide synthase was also expressed like in adult cartilage, though tumor necrosis factor alpha typifying Bunion formation was absent. In summary, surgically created osteochondromas lack the massive and continuous population of mesenchymal stem cells with Bcl2 expression. However, the small residual mesenchymal cell population gives rise to short-lived EXT1-expressing cells that disappear eventually due to spontaneous resorption.

European Journal of Heart Failure, 2003

To evaluate whether satellite cells injected into infarct areas in rabbits remain viable during 6... more To evaluate whether satellite cells injected into infarct areas in rabbits remain viable during 6 weeks follow-up and can improve cardiac function as assessed by echocardiography. Myocardial infarction was induced in 16 New Zealand white rabbits, by ligation of the marginalis sinistra artery. Tissue from gluteus muscle biopsies was dissected into small pieces and cultured. Within 2-3 weeks the cells were expanded by 2-3 orders of magnitude and were fluorescent labeled. Single cell pellets for resuspension at >10(6)/1 ml were directly injected into the infarct areas in 8 rabbits. In 8 additional rabbits, 1 ml saline was injected (control). Regional left ventricular function was assessed weekly by 2-D echocardiography until animals were sacrificed. Analysis was performed blind and independently by two experienced echocardiographers, based on the American Society of Echocardiography scheme. Six treated and five control rabbits completed the study. One week after the artery occlusion, left ventricular function scoring did not differ between groups, mean 8.7+/-1.6 vs 8.3+/-1.9 (P=0.74). At 6 weeks post-injection, echocardiographic score was significantly better in the treated group, mean 2.6+/-0.9 vs 6.9+/-2.1 (P=0.002). The treated group showed significant gradual segmental improvement between the first week up to week 6. After sacrifice, macro and microscopic transmural areas showed typical changes of myocardial infarction. Histochemical staining identified viable grafted cells in high density 6 weeks post-transplantation in all grafted hearts. Autologous satellite cells (skeletal myofiber), can be successfully grafted into rabbit hearts following myocardial infarction and may induce improved regional left ventricular function.

ACS Medicinal Chemistry Letters, 2011

The cytotoxic activity of aloe-emodin (AE), a natural anthranoid that readily permeates anthracyc... more The cytotoxic activity of aloe-emodin (AE), a natural anthranoid that readily permeates anthracycline-resistant tumor cells, was improved by the attachment of an amino-sugar unit to its anthraquinone core. The new class of AE glycosides (AEGs) showed a significant improvement in cytotoxicity-up to more than 2 orders of magnitude greater than those of AE and the clinically used anthracycline doxorubicin (DOX)-against several cancer cell lines with different levels of DOX resistance. Incubation with the synthetic AEGs induced cell death in less than one cell cycle, indicating that these compounds do not directly target the cell division mechanism. Confocal microscopy provided evidence that unlike DOX, AEGs accumulated in anthracycline-resistant tumor cells in which resistance is conferred by P-glycoprotein efflux pumps. The results of this study demonstrate that AEGs may serve as a promising scaffold for the development of cytotoxic agents capable of overcoming anthracycline resistance in tumor cells.

Connective Tissue Research, 2012

The familial disease of hereditary multiple exostoses is characterized by abnormal skeletal defor... more The familial disease of hereditary multiple exostoses is characterized by abnormal skeletal deformities requiring extensive surgical procedures. In hereditary multiple exostoses patients there is a shortage in the pericellular glycosaminoglycan (GAG) of heparan sulfate (HS), related to defective activity of HS glycosyltransferases, mainly in the pericellular regions of chondrocytes. This study searched for a novel approach employing xylosides with different aglycone groups priming a variety of GAG chains, in attempting to alter the GAG compositional profile. Cell cultures of patients with osteochondroma responded to p-nitrophenyl β-d-xyloside by a significant increase in total GAG synthesis, expressed mainly in the extracellular domains, limited to chondroitin sulfate). The different β-d-xylosides, in addition to increasing the synthesis of extracellular GAGs, led to a significant depletion of the intracellular GAG domains. In mouse chondrocyte cultures, β-d-xylosides with different aglycones created a unique distribution of the GAG pools. Of special interest was the finding that the naphthalene methanol β-d-xyloside showed the highest absolute levels of HS-GAGs in both extracellular and intra-pericellular moieties compared with other β-d-xylosides and with controls without xyloside. In summary, β-d-xylosides can be utilized in chondrocyte cultures to modify the distribution of GAGs between the extracellular and intracellular compartments. In addition, xylosides may alter the profile of specific GAG chains in each moiety.