The aim of this study was to compare the serum levels of the anti-angiogenic factor endostatin (S-endostatin) as a potential marker of vasculogenesis after autologous cell therapy (ACT) versus percutaneous transluminal angioplasty (PTA) in diabetic patients with critical limb ischemia (CLI). A total of 25 diabetic patients with CLI treated in our foot clinic during the period 2008-2014 with ACT generating potential vasculogenesis were consecutively included in the study; 14 diabetic patients with CLI who underwent PTA during the same period were included in a control group in which no vasculogenesis had occurred. S-endostatin was measured before revascularization and at 1, 3, and 6 months after the procedure. The effect of ACT and PTA on tissue ischemia was confirmed by transcutaneous oxygen pressure (TcPO2) measurement at the same intervals. While S-endostatin levels increased significantly at 1 and 3 months after ACT (both P < 0.001), no significant change of S-endostatin after PTA was observed. Elevation of S-endostatin levels significantly correlated with an increase in TcPO2 at 1 month after ACT ( r = 0.557; P < 0.001). Our study showed that endostatin might be a potential marker of vasculogenesis because of its significant increase after ACT in diabetic patients with CLI in contrast to those undergoing PTA. This increase may be a sign of a protective feedback mechanism of this anti-angiogenic factor.

• Keywords
• angiogenic factors, autologous cell therapy, critical limb ischemia, endostatin,
• MeSH
• Angioplasty * MeSH
• Antigens, CD34 analysis   MeSH
• Transplantation, Autologous MeSH
• Cell- and Tissue-Based Therapy MeSH
• Diabetes Mellitus, Type 2 blood   complications   MeSH
• Diabetic Foot blood   therapy   MeSH
• Endostatins blood   MeSH
• Neovascularization, Physiologic MeSH
• Ischemia blood   therapy   MeSH
• Stem Cells cytology   MeSH
• Extremities blood supply   MeSH
• Middle Aged MeSH
• Humans MeSH
• Peripheral Vascular Diseases therapy   MeSH
• Aged MeSH
• Stem Cell Transplantation * MeSH
• Treatment Outcome MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, CD34 MeSH
• Endostatins MeSH

INTRODUCTION: Multiple myeloma (MM) is still incurable due to resistance against various therapies. Thus, the identification of biomarkers predicting progression is urgently needed. Here, we evaluated four biomarkers in bone marrow and peripheral blood of MM patients for their prognostic significance. MATERIALS & METHODS: Bone marrow- and peripheral blood plasma levels of FLT3-L, soluble TIE2, endostatin, and osteoactivin were determined in patients with monoclonal gammopathy of undetermined significance (MGUS, n = 14/n = 4), patients with newly diagnosed MM (NDMM, n = 42/n = 31) and patients with relapsed/refractory MM (RRMM, n = 27/n = 16) by sandwich ELISA. RESULTS: Median FLT3-L expression increased from MGUS (58.77 pg/ml in bone marrow; 80.40 pg/ml in peripheral blood) to NDMM (63.15 pg/ml in bone marrow; 85.05 pg/ml in peripheral blood) and was maximal in RRMM (122 pg/ml in bone marrow; 160.47 pg/ml in peripheral blood; NDMM vs. RRMM p<0.001). A cut-off value of FLT3-L >92 pg/ml in bone marrow and >121 pg/ml in peripheral blood was associated with relapse or refractoriness in MM patients. FLT3-L was found to be a high predictive marker for discrimination between NDMM and RRMM as well in bone marrow as in peripheral blood (AUC 0.75 in bone marrow; vs 0.84 in peripheral blood). CONCLUSION: High levels of FLT3-L in bone marrow and peripheral blood of MM patients identify patients with progressive disease and are associated with relapse or refractoriness in MM patients. FLT3-L could be useful as a marker to identify RRMM patients and should be evaluated as target for future therapies.

• MeSH
• Diagnosis, Differential MeSH
• Adult MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Endostatins metabolism   MeSH
• Bone Marrow metabolism   MeSH
• Middle Aged MeSH
• Humans MeSH
• Membrane Glycoproteins metabolism   MeSH
• Membrane Proteins metabolism   MeSH
• Multiple Myeloma metabolism   MeSH
• Biomarkers, Tumor metabolism   MeSH
• Prognosis MeSH
• Receptor, TIE-2 metabolism   MeSH
• Recurrence MeSH
• ROC Curve MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Endostatins MeSH
• flt3 ligand protein MeSH Browser
• GPNMB protein, human MeSH Browser
• Membrane Glycoproteins MeSH
• Membrane Proteins MeSH
• Biomarkers, Tumor MeSH
• Receptor, TIE-2 MeSH

The release of proangiogenic cytokines into the circulation after stem cell (SC) therapy and compensatory increase of angiogenesis inhibitors may reflect local vasculogenesis but also can increase the risk of side effects. The aim of our study was to evaluate serum levels of angiogenic cytokines with regard to the assessment of local and systemic vasculogenesis in diabetic patients with no-option critical limb ischemia (NO-CLI). Twenty-five diabetic patients with NO-CLI treated with SCs isolated from bone marrow or stimulated peripheral blood were included in the study. Serum levels of proangiogenic cytokines (VEGF, bFGF, Ang-1, PDGF-AA, and PDGF-BB) and an antiangiogenic cytokine (endostatin) were assessed 6 months after cell treatment, compared to baseline values, and correlated with the number of injected CD34(+) cells. The clinical effect of SC therapy (assessed by changes in TcPO2) and potential systemic vasculogenesis (assessed by eye fundus examination) were evaluated after 6 months. Serum levels of angiogenic inhibitor endostatin increased significantly after 1 and 3 months (p = 0.0003), but no significant increase in serum levels of proangiogenic cytokines was observed. A significant correlation between number of injected CD34(+) cells and serum levels of endostatin was observed (r = 0.41, p < 0.05); however, proangiogenic cytokines did not correlate with CD34(+) cells. No correlation between increase in TcPO2 after treatment and serum levels of any of the angiogenic cytokines were seen, and no signs of systemic vasculogenesis in the retina were observed after 6 months. Despite the significant increase in the levels of the angiogenic inhibitor endostatin following SC treatment, there was no risk of systemic vasculogenesis after SC therapy as documented by serum levels of proangiogenic cytokines or changes in the retina.

• MeSH
• Antigens, CD34 metabolism   MeSH
• Transplantation, Autologous MeSH
• Cytokines blood   MeSH
• Diabetes Mellitus blood   therapy   MeSH
• Endostatins blood   MeSH
• Neovascularization, Physiologic * MeSH
• Ischemia blood   complications   therapy   MeSH
• Oxygen metabolism   MeSH
• Middle Aged MeSH
• Humans MeSH
• Partial Pressure MeSH
• Aged MeSH
• Stem Cell Transplantation * MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Antigens, CD34 MeSH
• Cytokines MeSH
• Endostatins MeSH
• Oxygen MeSH

We investigated whether a genetic modification of BCR-ABL-transformed mouse cells that resulted in endostatin (ES) production altered their oncogenic potential. Mouse B210 cells, which express p210bcr-abl fusion protein and induce leukemia-like disease and extremely rarely solid tumors after intravenous (i.v.) administration, were used. The cells were transfected with a plasmid carrying genes for mouse ES and resistance to blasticidine. Transduced cells were isolated in media supplemented with blasticidine. Production of ES was determined by Western blotting. For further tests, two clones were selected, and their pathogenicity after i.v. inoculation was tested. Compared with the parental B210 cells, the capability of both gene-modified cell clones to induce lethal leukemia was reduced. However, mice that did not succumb to leukemia subsequently developed solid tumors. They were composed of poorly differentiated cells with irregular nuclei and roughly granular chromatin and were well vascularized. FISH revealed the presence of the BCR-ABL fusion gene both in tumors and spleens. Immunohistological investigation of the tumors demonstrated the production of ES in vivo and the cell lines derived from the tumors produced detectable amounts of ES, this demonstrating that the formation of solid tumors was not associated with the loss or silencing of the ES gene.

• MeSH
• Fusion Proteins, bcr-abl genetics   metabolism   MeSH
• Endostatins metabolism   pharmacology   MeSH
• Human Umbilical Vein Endothelial Cells drug effects   physiology   MeSH
• Kaplan-Meier Estimate MeSH
• Culture Media, Conditioned pharmacology   MeSH
• Histocompatibility Antigens Class I metabolism   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Cell Transformation, Neoplastic MeSH
• Cell Proliferation MeSH
• Cell Line, Transformed metabolism   pathology   transplantation   MeSH
• Neoplasm Transplantation MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Fusion Proteins, bcr-abl MeSH
• Endostatins MeSH
• Culture Media, Conditioned MeSH
• Histocompatibility Antigens Class I MeSH

Two mouse HPV16-transformed cell lines, viz. MK16 cells, which induce metastasizing tumors, and TC-1 cells, which induce non-metastasizing tumors were transduced with the gene for mouse endostatin. Two clones constitutively expressing endostatin were isolated from each of them. They were denoted ME3 and ME9, and TE2 and TE5, respectively. When inoculated into mice, ME3 cells were non-oncogenic. Nearly all mice inoculated with ME9 cells developed tumors, but considerably later than did the parental MK16 cells and metastasis formation was strongly reduced in these animals. On the other hand, TE2 and TE5 cells displayed oncogenic potential similar to that of the parental cells. To provide more information on these different effects of endostatin production, cell lysates of all six lines studied were tested for the content of 25 factors known to be involved in angiogenesis. The parental MK16 cells differed from the parental TC-1 cells and also from all endostatin producing sublines by a markedly higher production of interleukin 1alpha (IL-1alpha) and, to a lesser extent, by a higher production of several other factors tested. Additional experiments indicated that the suppression of the production of IL-1alpha by the parental MK16 caused by endostatin was due to an autocrine mechanism.

• MeSH
• Autocrine Communication MeSH
• Time Factors MeSH
• Endostatins genetics   metabolism   MeSH
• Endothelial Cells metabolism   MeSH
• Genes, ras * MeSH
• Interleukin-1alpha metabolism   MeSH
• Interleukin-2 genetics   metabolism   MeSH
• Culture Media, Conditioned metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Cell Transformation, Neoplastic * MeSH
• Lung Neoplasms genetics   immunology   metabolism   prevention & control   secondary   virology   MeSH
• Oncogene Proteins, Viral genetics   MeSH
• Papillomavirus E7 Proteins MeSH
• Cell Proliferation MeSH
• Repressor Proteins genetics   MeSH
• Transduction, Genetic MeSH
• Cell Line, Transformed MeSH
• Cell Transformation, Viral * MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• E6 protein, Human papillomavirus type 16 MeSH Browser
• Endostatins MeSH
• Interleukin-1alpha MeSH
• Interleukin-2 MeSH
• Culture Media, Conditioned MeSH
• oncogene protein E7, Human papillomavirus type 16 MeSH Browser
• Oncogene Proteins, Viral MeSH
• Papillomavirus E7 Proteins MeSH
• Repressor Proteins MeSH

The aim of the study was to determine whether VEGF, TPS, TK or Endostatin determination in tissue cytosol may have some additional value in distinguishing among different types of thyroid lesions. These markers were chosen as representatives of the 2 main pathways (angiogenesis and proliferation) involved in thyroid diseases. VEGF is the most potent angiogenic promoter and Endostatin plays an opposing role. Thymidine kinase (TK) is a marker of DNA synthesis and TPS, cytokeratin 18 fragments, is a marker of the rate of proliferation. We determined qualitatively all four markers in tissue extracts: cytosol from 157 tissue specimens (93 goitre, 12 Hashimoto's thyroiditis, 39 adenomas and 13 carcinomas). In 6 cases we were able to compare both normal and pathological tissue samples from a single patient. Statistically significant differences were found in the measured markers, but outliers were present in all groups. This fact does not permit their use in differential diagnosis. The highest levels of all markers were reached in adenomas, being higher than in carcinomas, probably explained by the higher overall metabolic rate in adenomas.

• MeSH
• Cytosol metabolism   MeSH
• Endostatins metabolism   MeSH
• Humans MeSH
• Thyroid Diseases metabolism   MeSH
• Thyroid Gland metabolism   MeSH
• Thymidine Kinase metabolism   MeSH
• Vascular Endothelial Growth Factor A metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Endostatins MeSH
• Thymidine Kinase MeSH
• Vascular Endothelial Growth Factor A MeSH