Vascular endothelial growth factor-A165 (VEGF-A165) and fibroblast growth factor-2 (FGF-2) are currently used for the functionalization of biomaterials designed for tissue engineering. We have developed a new simple method for heterologous expression and purification of VEGF-A165 and FGF-2 in the yeast expression system of Pichia pastoris. The biological activity of the growth factors was assessed in cultures of human and porcine adipose tissue-derived stem cells (ADSCs) and human umbilical vein endothelial cells (HUVECs). When added into the culture medium, VEGF-A165 stimulated proliferation only in HUVECs, while FGF-2 stimulated the proliferation of both cell types. A similar effect was achieved when the growth factors were pre-adsorbed to polystyrene wells. The effect of our recombinant growth factors was slightly lower than that of commercially available factors, which was attributed to the presence of some impurities. The stimulatory effect of the VEGF-A165 on cell adhesion was rather weak, especially in ADSCs. FGF-2 was a potent stimulator of the adhesion of ADSCs but had no to negative effect on the adhesion of HUVECs. In sum, FGF-2 and VEGF-A165 have diverse effects on the behavior of different cell types, which maybe utilized in tissue engineering.

• MeSH
• buněčná adheze účinky léků   MeSH
• endoteliální buňky pupečníkové žíly (lidské) cytologie   metabolismus   MeSH
• fibroblastový růstový faktor 2 chemie   genetika   farmakologie   MeSH
• kmenové buňky cytologie   metabolismus   MeSH
• lidé MeSH
• prasata MeSH
• proliferace buněk účinky léků   MeSH
• rekombinantní proteiny chemie   farmakologie   MeSH
• vaskulární endoteliální růstový faktor A chemie   genetika   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Cardiac atrophy is the most common complication of prolonged application of the left ventricle (LV) assist device (LVAD) in patients with advanced heart failure (HF). Our aim was to evaluate the course of unloading-induced cardiac atrophy in rats with failing hearts, and to examine if increased isovolumic loading obtained by intraventricular implantation of an especially designed spring expander would attenuate this process. Heterotopic abdominal heart transplantation (HTx) was used as a rat model of heart unloading. HF was induced by volume overload achieved by creation of the aorto-caval fistula (ACF). The degree of cardiac atrophy was assessed as the weight ratio of the heterotopically transplanted heart (HW) to the control heart. Isovolumic loading was increased by intraventricular implantation of a stainless steel three-branch spring expander. The course of cardiac atrophy was evaluated on days 7, 14, 21, and 28 after HTx Seven days unloading by HTx in failing hearts sufficed to substantially decrease the HW (-59 ± 3%), the decrease progressed when measured on days 14, 21, and 28 after HTx Implantation of the spring expander significantly reduced the decreases in whole HW at all the time points (-39 ± 3 compared with -59 ± 3, -52 ± 2 compared with -69 ± 3, -51 ± 2 compared with -71 ± 2, and -44 ± 2 compared with -71 ± 3%, respectively; P<0.05 in each case). We conclude that the enhanced isovolumic heart loading obtained by implantation of the spring expander attenuates the development of unloading-induced cardiac atrophy in the failing rat heart.

• MeSH
• aorta chirurgie   MeSH
• atriální natriuretický faktor genetika   metabolismus   MeSH
• atrofie metabolismus   patofyziologie   prevence a kontrola   chirurgie   MeSH
• biologické markery metabolismus   MeSH
• design vybavení MeSH
• experimentální implantáty MeSH
• exprese genu MeSH
• fibroblastový růstový faktor 2 genetika   metabolismus   MeSH
• heterotopická transplantace MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• píštěle MeSH
• potkani inbrední LEW MeSH
• přenašeč glukosy typ 1 genetika   metabolismus   MeSH
• sarkoplazmatická Ca2+-ATPáza genetika   metabolismus   MeSH
• srdce patofyziologie   MeSH
• srdeční komory patofyziologie   chirurgie   MeSH
• srdeční selhání metabolismus   patofyziologie   chirurgie   terapie   MeSH
• tkáňové expandéry * MeSH
• transplantace srdce * MeSH
• vena cava superior chirurgie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Fibroblast growth factors (FGFs) serve numerous regulatory functions in complex organisms, and their corresponding therapeutic potential is of growing interest to academics and industrial researchers alike. However, applications of these proteins are limited due to their low stability. Here we tackle this problem using a generalizable computer-assisted protein engineering strategy to create a unique modified FGF2 with nine mutations displaying unprecedented stability and uncompromised biological function. The data from the characterization of stabilized FGF2 showed a remarkable prediction potential of in silico methods and provided insight into the unfolding mechanism of the protein. The molecule holds a considerable promise for stem cell research and medical or pharmaceutical applications.

• MeSH
• bodová mutace MeSH
• design s pomocí počítače * MeSH
• embryonální kmenové buňky cytologie   metabolismus   MeSH
• fibroblastový růstový faktor 2 chemie   genetika   metabolismus   MeSH
• lidé MeSH
• počítačová simulace MeSH
• proteinové inženýrství * MeSH
• řízená evoluce molekul MeSH
• sbalování proteinů MeSH
• sekvence aminokyselin MeSH
• stabilita proteinů * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Ventral root avulsion (VRA) triggers a strong glial reaction which contributes to neuronal loss, as well as to synaptic detachment. To overcome the degenerative effects of VRA, treatments with neurotrophic factors and stem cells have been proposed. Thus, we investigated neuroprotection elicited by human embryonic stem cells (hESC), modified to overexpress a human fibroblast growth factor 2 (FGF-2), on motoneurons subjected to VRA. Lewis rats were submitted to VRA (L4-L6) and hESC/FGF-2 were applied to the injury site using a fibrin scaffold. The spinal cords were processed to evaluate neuronal survival, synaptic stability, and glial reactivity two weeks post lesion. Then, qRT-PCR was used to assess gene expression of β2-microglobulin (β2m), TNFα, IL1β, IL6 and IL10 in the spinal cord in vivo and FGF2 mRNA levels in hESC in vitro. The results indicate that hESC overexpressing FGF2 significantly rescued avulsed motoneurons, preserving synaptic covering and reducing astroglial reactivity. The cells were also shown to express BDNF and GDNF at the site of injury. Additionally, engraftment of hESC led to a significant reduction in mRNA levels of TNFα at the spinal cord ventral horn, indicating their immunomodulatory properties. Overall, the present data suggest that hESC overexpressing FGF2 are neuroprotective and can shift gene expression towards an anti-inflammatory environment.

• MeSH
• doxycyklin terapeutické užití   MeSH
• fibrinová tkáňová adheziva toxicita   MeSH
• fibroblastový růstový faktor 2 genetika   metabolismus   MeSH
• genetické vektory fyziologie   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• lidské embryonální kmenové buňky metabolismus   transplantace   MeSH
• míšní kořeny patologie   MeSH
• modely nemocí na zvířatech MeSH
• motorické neurony metabolismus   patologie   MeSH
• neuroglie účinky léků   metabolismus   MeSH
• pohyb buněk MeSH
• potkani inbrední LEW MeSH
• proteiny nervové tkáně metabolismus   MeSH
• radikulopatie chemicky indukované   chirurgie   MeSH
• regulace genové exprese účinky léků   genetika   MeSH
• tkáňová adheziva toxicita   MeSH
• viabilita buněk účinky léků   genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Elevated levels of circulating angiogenic cytokines and increased expression of genes encoding angiogenic factors have been reported in recent years in patients with chronic lymphocytic leukemia (CLL) but data regarding prognostic and predictive significance are still limited. Therefore, in the present study based upon our prior pilot results, we measured mRNA expressions of angiopoietin-2 (Ang-2), fibroblast growth factor-2 (FGF-2) and endoglin (CD105) by reverse transcription quantitative PCR in purified CD19+ cells from 70 untreated CLL patients (median age, 63 years; males, 64%; Rai III/IV stages, 29 %; unmutated IgVH genes, 60 %) and evaluated their possible association with established prognostic factors and clinical course of the disease. Higher expression of Ang-2 was significantly associated with unmutated IgVH genes (n = 55, p= 0.003). Higher CD105 expression was significantly associated with unmutated IgVH genes (n = 55, p < 0.001), high CD38 expression (n = 66, p = 0.022), high ZAP-70 expression (n = 66, p = 0.010), Rai stage I-IV (n = 70, p < 0.001), progressive clinical course of CLL (n = 70, p = 0.001) and shorter time to treatment (n = 70; p < 0.001). Expression of FGF-2 was not significantly associated with any of the prognostic markers. These results indicate that elevated expression of Ang-2 and in particular CD105 by CLL cells is associated with unfavorable prognostic features and clinical outcome; thus, both cytokines appear to play an important role in biology and progression of CLL and warrant further investigation.

• MeSH
• angiopoetin-2 * genetika   MeSH
• CD antigeny * genetika   MeSH
• chronická lymfatická leukemie * metabolismus   MeSH
• dospělí MeSH
• endoglin MeSH
• fibroblastový růstový faktor 2 * genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• messenger RNA * analýza   MeSH
• mutace MeSH
• receptory buněčného povrchu * genetika   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• těžké řetězce imunoglobulinů genetika   MeSH
• variabilní oblast imunoglobulinu genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH

Transplantation of mesenchymal stem cells (MSC) improves functional recovery in experimental models of spinal cord injury (SCI); however, the mechanisms underlying this effect are not completely understood. We investigated the effect of intrathecal implantation of human MSC on functional recovery, astrogliosis and levels of inflammatory cytokines in rats using balloon-induced spinal cord compression lesions. Transplanted cells did not survive at the lesion site of the spinal cord; however, functional recovery was enhanced in the MSC-treated group as was confirmed by the Basso, Beattie, and Bresnahan (BBB) and the flat beam test. Morphometric analysis showed a significantly higher amount of remaining white matter in the cranial part of the lesioned spinal cords. Immunohistochemical analysis of the lesions indicated the rearrangement of the glial scar in MSC-treated animals. Real-time PCR analysis revealed an increased expression of Irf5, Mrc1, Fgf2, Gap43 and Gfap. Transplantation of MSCs into a lesioned spinal cord reduced TNFα, IL-4, IL-1β, IL-2, IL-6 and IL-12 and increased the levels of MIP-1α and RANTES when compared to saline-treated controls. Intrathecal implantation of MSCs reduces the inflammatory reaction and apoptosis, improves functional recovery and modulates glial scar formation after SCI, regardless of cell survival. Therefore, repeated applications may prolong the beneficial effects induced by MSC application.

• MeSH
• chemokin CCL5 genetika   metabolismus   MeSH
• fibroblastový růstový faktor 2 genetika   metabolismus   MeSH
• gliový fibrilární kyselý protein genetika   metabolismus   MeSH
• interferonové regulační faktory genetika   metabolismus   MeSH
• interleukiny genetika   metabolismus   MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• lokomoce MeSH
• mezenchymální kmenové buňky metabolismus   MeSH
• poranění míchy metabolismus   terapie   MeSH
• potkani Wistar MeSH
• protein GAP-43 genetika   metabolismus   MeSH
• receptory imunologické genetika   metabolismus   MeSH
• TNF-alfa genetika   metabolismus   MeSH
• transplantace mezenchymálních kmenových buněk * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Fibroblast growth factor 2 (FGF2) is one of the most studied growth factors to date. Most attention has been dedicated to the smallest, 18 kDa FGF2 variant that is released by cells and acts through activation of cell-surface FGF-receptor tyrosine kinases. There are, however, several higher molecular weight (HMW) variants of FGF2 that rarely leave their producing cells, are retained in the nucleus and act independently of FGF-receptors (FGFR). Despite significant evidence documenting the expression and intracellular trafficking of HMW FGF2, many important questions remain about the physiological roles and mechanisms of action of HMW FGF2. In this review, we summarize the current knowledge about the biology of HMW FGF2, its role in disease and areas for future investigation.

• MeSH
• fenotyp MeSH
• fibroblastový růstový faktor 2 genetika   metabolismus   MeSH
• lidé MeSH
• malý jaderný ribonukleoprotein U2 metabolismus   MeSH
• molekulová hmotnost MeSH
• protein - isoformy genetika   chemie   metabolismus   MeSH
• proteinový komplex SMN metabolismus   MeSH
• receptory fibroblastových růstových faktorů genetika   chemie   metabolismus   MeSH
• regulace genové exprese MeSH
• ribozomální proteiny metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH
• Research Support, N.I.H., Extramural MeSH

The basic fibroblast growth factor (bFGF) is considered to be one of the candidate genes in the processes of tumour growth and angiogenesis. The aim of the present investigation was to find possible association of new polymorphisms in bFGF with proliferative diabetic retinopathy (PDR) and determine the plasma level in PDR. Allele, genotype and haplotype frequencies were determined in the association study comprising three groups of Caucasian subjects (n=488) (diabetics with/ PDR/ and without retinopathy/ non-PDR/ and non-diabetics/ non-DM/) in order to identify genetic marker for PDR. The plasma level of the bFGF protein was analysed by ELISA method. Significantly higher frequencies of 754C allele of the new 754C/G polymorphisms was found between PDR and non-DM group (p=0.05, OR=1.38). The comparison of plasma level of the bFGF showed statistically significant difference among studied groups (p=0.001). The bFGF plasma level in PDR group was significantly higher than in the groups of non-PDR and non-DM (p=0.017, p=0.001, respectively) and was significantly higher for CC and GC genotypes of 754C/G polymorphism in PDR group (p=0.006). Increased plasma level of the bFGF confirmed the importance of this candidate gene in the formation of PDR. However, the regulatory mechanisms of the bFGF level need further examinations.

• MeSH
• diabetická retinopatie genetika   krev   MeSH
• fibroblastový růstový faktor 2 genetika   krev   MeSH
• financování organizované MeSH
• frekvence genu MeSH
• genetická variace MeSH
• genotyp MeSH
• jednonukleotidový polymorfismus MeSH
• lidé středního věku MeSH
• lidé MeSH
• patologická angiogeneze genetika   MeSH
• polymerázová řetězová reakce MeSH
• polymorfismus konformace jednovláknové DNA MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH

Various types of feeder cells have been adopted for the culture of human embryonic stem cells (hESCs) to improve their attachment and provide them with stemness-supporting factors. However, feeder cells differ in their capacity to support the growth of undifferentiated hESCs. Here, we compared the expression and secretion of four well-established regulators of hESC pluripotency and/or differentiation among five lines of human foreskin fibroblasts and primary mouse embryonic fibroblasts throughout a standard hESC culture procedure. We found that human and mouse feeder cells secreted comparable levels of TGF beta 1. However, mouse feeder cells secreted larger quantities of activin A than human feeder cells. Conversely, FGF-2, which was produced by human feeder cells, could not be detected in culture media from mouse feeder cells. The quantity of BMP-4 was at about the level of detectability in media from all feeder cell types, although BMP-4 dimers were present in all feeder cells. Production of TGF beta 1, activin A, and FGF-2 varied considerably among the human-derived feeder cell lines. Low- and high-producing human feeder cells as well as mouse feeder cells were evaluated for their ability to support the undifferentiated growth of hESCs. We found that a significantly lower proportion of hESCs maintained on human feeder cell types expressed SSEA3, an undifferentiated cell marker. Moreover, SSEA3 expression and thus the pluripotent hESC compartment could be partially rescued by addition of activin A. Cumulatively, these results suggest that the ability of a feeder layer to promote the undifferentiated growth of hESCs is attributable to its characteristic growth factor production.

• MeSH
• aktiviny biosyntéza   genetika   MeSH
• antigeny sacharidové asociované s nádorem genetika   MeSH
• biologické markery metabolismus   MeSH
• buněčná diferenciace MeSH
• DNA primery genetika   MeSH
• druhová specificita MeSH
• embryonální antigeny specifické pro určité stadium vývoje MeSH
• embryonální kmenové buňky cytologie   metabolismus   metabolismus   MeSH
• exprese genu MeSH
• fibroblastový růstový faktor 2 biosyntéza   genetika   MeSH
• financování organizované MeSH
• glykosfingolipidy genetika   MeSH
• kokultivační techniky metody   MeSH
• kostní morfogenetické proteiny analýza   genetika   MeSH
• kostní morfogenetický protein 4 MeSH
• kultivační média speciální analýza   MeSH
• lidé MeSH
• messenger RNA genetika   metabolismus   MeSH
• myši MeSH
• pluripotentní kmenové buňky cytologie   metabolismus   MeSH
• proliferace buněk MeSH
• sekvence nukleotidů MeSH
• transformující růstový faktor beta1 biosyntéza   genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

The mutation status of genes encoding the variable region of immunoglobulin heavy chains (IgV(H)) is a strong predictor of disease progression and survival in B-cell chronic lymphocytic leukemia (B-CLL). We investigated whether there is an association between the concentration of both vascular endothelial growth factor and basic fibroblast growth factor and IgV(H) mutation status in 49 untreated B-CLL patients.

• MeSH
• chronická lymfatická leukemie * genetika   krev   MeSH
• fibroblastový růstový faktor 2 * genetika   krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• somatická hypermutace imunoglobulinových genů genetika   MeSH
• těžké řetězce imunoglobulinů * genetika   MeSH
• variabilní oblast imunoglobulinu * genetika   MeSH
• vaskulární endoteliální růstový faktor A * genetika   krev   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH