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Abstract Objective To determine the efficacy of fibroblast growth factor‐2 (FGF‐2) in treating chronic nonhealing tympanic membrane (TM) perforations. Method Double‐blinded, randomized placebo controlled phase 2 clinical trial for patients with chronic TM perforations of more than 3 months duration with a cross‐over arm. Patients received either FGF‐2 or placebo (sterile water) saturated gelatin sponge in the perforation after rimming the perforation under topical anesthesia. The perforation was then covered with Tisseel fibrin glue. The primary endpoint was complete closure of the TM perforation. Secondary end points included change in hearing and partial TM closure rates. The TM was examined every 3 weeks with otoendoscopy for closure. The treatment was repeated if there was incomplete closure every 3 weeks up to a total of three treatments per arm. Results Seventy four patients were recruited for the study. Fifty seven met eligibility criteria and fifty four completed the study. Ten of 14 perforations closed completely in the placebo group (71.4%) and 23 of 40 perforations closed completely in the FGF‐2 treatment group (57.5%), P value = .36. Pure tone averages and word recognition scores were not statistically significantly different between study groups post‐treatment. After initial complete closure, re‐perforation occurred in seven FGF‐2 treated patients and two placebo patients making the effective final closure rate 40% for FGF and 57% for placebo, respectively. Conclusion No statistically significant difference in tympanic membrane perforation closure rate was found between the FGF‐2 and placebo groups. There were no differences in hearing outcomes between the groups. Level of evidence 1b.

The purpose of this study was to investigate the effect of recombinant human basic fibroblast growth factor (rhbFGF) on nerve regeneration immediately after nerve injury. Partial defects were created in the inferior alveolar nerve of rabbits and were treated with (Experimental group) or without (Control group) rhbFGF for 1–3 weeks. Injured tissues were then characterized immunohistochemically using primary antibodies to S-100; a marker for Schwann cells, and to neurofilament (NF); a marker for axons. The number of S-100-immunopositive cells in the experimental group was significantly increased compared with the control group at weeks 2 and 3 (P < 0.05). The number of NF-immunopositive cells in the experimental group was also significantly increased compared with the control group at week 3 (P < 0.05). These results suggest that application of rhbFGF immediately after partial damage of an inferior alveolar nerve might be effective to promote nerve regeneration. [ABSTRACT FROM AUTHOR]

Objective: To determine the efficacy of fibroblast growth factor‐2 (FGF‐2) in treating chronic nonhealing tympanic membrane (TM) perforations. Method: Double‐blinded, randomized placebo controlled phase 2 clinical trial for patients with chronic TM perforations of more than 3 months duration with a cross‐over arm. Patients received either FGF‐2 or placebo (sterile water) saturated gelatin sponge in the perforation after rimming the perforation under topical anesthesia. The perforation was then covered with Tisseel fibrin glue. The primary endpoint was complete closure of the TM perforation. Secondary end points included change in hearing and partial TM closure rates. The TM was examined every 3 weeks with otoendoscopy for closure. The treatment was repeated if there was incomplete closure every 3 weeks up to a total of three treatments per arm. Results: Seventy four patients were recruited for the study. Fifty seven met eligibility criteria and fifty four completed the study. Ten of 14 perforations closed completely in the placebo group (71.4%) and 23 of 40 perforations closed completely in the FGF‐2 treatment group (57.5%), P value =.36. Pure tone averages and word recognition scores were not statistically significantly different between study groups post‐treatment. After initial complete closure, re‐perforation occurred in seven FGF‐2 treated patients and two placebo patients making the effective final closure rate 40% for FGF and 57% for placebo, respectively. Conclusion: No statistically significant difference in tympanic membrane perforation closure rate was found between the FGF‐2 and placebo groups. There were no differences in hearing outcomes between the groups. Level of evidence: 1b. [ABSTRACT FROM AUTHOR]

Sclerosing stromal tumours (SSTs) are rare benign tumours and are generally observed in individuals in their teens to 20 s. Many cases are suspected as malignant tumours pre-operatively, owing to their high vascularity on diagnostic imaging. Herein, we aimed to evaluate the expressions of various angiogenic factors in SSTs. The expressions of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) were examined by immunohistochemical staining. Upon immunohistochemistry, overexpressions of VEGF, bFGF, and HGF in the ovarian stroma were observed in SSTs. In the normal ovary, the expressions were strong around the vessels and weak in the stroma of the ovary, while a Sertoli-Leydig cell tumour showed weak staining with VEGF, locally strong staining with bFGF and negative staining with HGF. This is the first report about angiogenic factors such as bFGF, and HGF in SST. Impact statement What is already known on this subject? Sclerosing stromal tumour (SST) is a rare benign tumour with high vascularity. The high vascularity of SSTs are reported to be related to the overexpression of vascular endothelial growth factor (VEGF). But there is no study on the expressions of other angiogenic factors in SST. What do the results this study add? In the immunohistochemical analysis, VEGF, bFGF and HGF were found to be widely stained in SSTs. This results suggest the possibility that the high vascularity seen on diagnostic imaging, and the many small vessels observed microscopically may be related to the expressions of VEGF, bFGF and HGF in SSTs. What are the implications of these findings for clinical practice and/or further research? Our results suggest the possibility that the combined expression pattern of VEGF, bFGF, and HGF may be used as marker of SSTs. [ABSTRACT FROM AUTHOR]

Elastofibroma is a rare tumour that occurs in the subscapular space, and it typically presents in middle-aged and older individuals. The aetiology of elastofibroma remains unknown. Recent, sporadic reports have shown, immunohistologically, that fibroblasts in elastofibroma may produce abnormal elastic and collagen fibres through the action of transforming growth factor-beta (TGF-β), a factor that promotes fibroblast proliferation. However, that finding lacked quantitative measurements and controls. Therefore, in this study, we performed quantitative, immunohistochemical analyses of TGF-β1 and basic fibroblast growth factor (bFGF) in three elastofibromas, and we compared them to ten dermatofibromas and keloids, and five normal skin. In elastofibroma specimens, 16-59 % fibroblasts were positive for TGF-β1 in the cytoplasm, compared to 96 % in dermatofibroma, 93 % in keloid and 2 % in normal dermis specimens. Also, in elastofibroma specimens, 26-67 % of fibroblasts were positive for bFGF in the cytoplasm, compared to 97 % in dermatofibroma, 97 % in keloid, and 22 % in normal dermis specimens. Intriguingly, the tumour size and growth rate were proportional to the percentage of cells positive for bFGF. Finally, greater levels of bFGF expressions in fibroblasts were associated with larger sized elastofibromas. These results suggested that elastofibroma development depended on high expression of TGF-β1 and bFGF. [ABSTRACT FROM AUTHOR]

Conclusion: Our results demonstrate for the first time a potentially enhanced basal secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin (IL)-17A-stimulated secretion of IL-6 from nasal polyp fibroblasts, enhanced basal secretion of IL-6 from eosinophilic nasal polyp fibroblasts, and a remarkable up-regulation of IL-9 and granulocyte colony-stimulating factor (G-CSF) from nasal fibroblasts by IL-17A stimulation. Objectives: The fibroblast, one of the main cell types making up nasal polyps, is thought to be a target cell of various cytokines. Methods: Subcultured fibroblasts were established from human polyp biopsy tissues. Simultaneous quantification of 27 kinds of cytokines and chemokines in culture supernatants in unstimulated and IL-17A-stimulated conditions was performed with a human multiplex cytokine assay system. Results: The IL-17A receptor was expressed at similar levels in all three groups. In the eosinophilic group, basal secretion levels of IL-6 were significantly higher than those in the control and non-eosinophilic groups. Basal secretion of MCP-1 in both the non-eosinophilic and eosinophilic groups was also higher than that of the control group. Both IL-9 and G-CSF secretion were remarkably enhanced by IL-17A stimulation in all three groups. The receptor-mediated response by IL-17A significantly up-regulated IL-6 release alone in the non-eosinophilic and eosinophilic groups as compared with the control group. [ABSTRACT FROM AUTHOR]

Lesions of the nigrostriatal pathway are known to induce a compensatory up-regulation of various neurotrophic factors. In this study we examined protein content of basic fibroblast growth factor (FGF-2) in tissue samples taken from the ventral midbrain and striatum at two different time points following a neurotoxic lesion of the nigrostriatal pathway in two different rat strains, the outbred Sprague–Dawley (SD) and inbred F344 × Brown Norway F1 hybrid (F344BNF1). Despite both rat strains having comparable lesions of the nigrostriatal pathway, we observed a difference in the temporal up-regulation of FGF-2 in ventral midbrain samples taken from the side ipsilateral to the lesion. Basic FGF was significantly up-regulated in ventral midbrain in SD rats 1 week post-lesion while we did not observe an up-regulation of FGF-2 in the lesioned ventral midbrain of F344BNF1 at this same time point. However, both strains showed a significant up-regulation of FGF-2 in the lesioned ventral midbrain 3 weeks post-lesion. Sprague–Dawley rats also appeared to be more sensitive to the lesion in terms of up-regulating FGF-2 expression. The differences reported here suggest currently unknown genetic differences between these two strains may be important factors for regulating the compensatory release of neurotrophic factors, such as FGF-2, in response to a neurotoxic lesion of the nigrostriatal pathway. [ABSTRACT FROM AUTHOR]

Abstract: Recently, we proposed that rabbit embryonic stem (ES) cells can be stable mammalian ES cells and can be a small animal model for human ES cell research. However, the signaling pathways controlling rabbit ES cell pluripotency remain largely unknown. Here we report that bFGF can maintain the undifferentiated status of rabbit ES cells and found that Activin/Nodal signaling through Smad2/3 activation is necessary to maintain the pluripotent status of rabbit ES cells. We further show that in spite of STAT3 in rabbit ES cells, LIF is dispensable for maintenance of undifferentiated status in rabbit ES cells. Although phosphorylation of Janus Kinase signal transducer and activator (JAK/STAT) disappeared after JAK-inhibitor treatment, OCT4 is constantly produced. When rabbit ES cells were cultured for more than 40 passages in the absence of LIF, expression of stem cell markers and teratoma formation were observed. Additionally, treatment with Rho-associated kinase (ROCK) inhibitor, Y27632, to rabbit ES cells significantly enhanced cell growth. These findings suggest that molecular mechanisms underlying rabbit ES cell self-renewal and pluripotency are similar to primate ES cells. Rabbit ES cells may provide a translational research model for the study of human diseases in vitro and applications to transplantation therapy. [Copyright &y& Elsevier]

Objectives: The aim of the present study was to investigate whether porous carbonate apatite (CA) containing basic FGF (bFGF) enhances bone augmentation as compared with porous CA alone. Material and methods: Thirty-six disc-shaped porous CA implants were inserted into femur defects in 36 Wister rats. Porous CA containing 0, 5, or 50 ng of bFGF was placed in the defect. Bone augmentation was evaluated radiographically using micro-CT and histologically by conventional staining at 2 and 12 weeks. Results: At 2 weeks postoperatively, new bone was evident in the defect sites with porous CA containing either 5 or 50 ng of bFGF. The area of regenerated bone, however, was significantly higher with porous CA containing 5 ng of bFGF than with porous CA containing 50 ng. At 12 weeks postoperatively, no differences in new bone formation were seen between porous CA containing 5 and 50 ng bFGF. Porous CA containing bFGF, however, markedly increased the amount of bone newly formed as compared with porous CA without bFGF. Regardless of the implantation period, TRAP-positive cells were visible around the material, indicating that osteoclast-like cells were resorbing porous CA. In accordance with the evidence, bioresorption of the material increased over time: 6.3–7.8% at 2 weeks to 9.1–11.5% at 12 weeks. Conclusions: Porous CA is useful as a growth factor carrier and scaffold for bone augmentation. [ABSTRACT FROM AUTHOR]

Objective: To evaluate whether basic fibroblast growth factor (FGF) benefits the in vitro development of human primordial follicles. Design: Human ovarian tissue was placed in organ culture for 4 weeks with basic FGF and either fetal calf serum or a serum-free combination. Control groups were cultured with a neutralizing antibody against basic FGF. Setting: Major tertiary care and referral academic centers. Patient(s): Fourteen women/girls undergoing various gynecological operations and two fetuses from women undergoing pregnancy terminations. Intervention(s): None. Main Outcome Measure(s): Follicular counts, immunohistochemistry for proliferating cell nuclear antigen and bromodeoxyuridine incorporation and measurement of 17β E2 production. Result(s): Only in the serum-free culture system was the number of developing follicles in samples cultured with basic FGF significantly higher than in uncultured specimens. The E2 production increased significantly in the second week, and there was a significant reduction in E2 secretion with the addition of the neutralizing antibody against basic FGF. The percentage of granulosa cells (GCs) that stained for proliferating cell nuclear antigen or bromodeoxyuridine was significantly higher in developing follicles than in primordial follicles, regardless of treatment. Conclusion(s): Basic FGF apparently plays a role in the E2 production of early follicles. High doses of basic FGF enhanced follicular development in serum-free media. [Copyright &y& Elsevier]

Fibroblast growth factor 2 (FGF-2) is expressed in isoforms of different molecular masses from one mRNA species by alternative start of translation. The higher molecular mass isoforms (FGF-221 and 23) contain an arginine-rich N-terminus organized in RG-motifs followed by the 18 kDa FGF-2 (FGF-218) core which is common to all isoforms. Both isoforms localize differentially to the nucleus. Here, we analyzed the nuclear localization of FGF-221. Surprisingly, the lack of one RG-motif in FGF-221 resulted in the nucleolar distribution characteristic of FGF-218. We have previously shown that 23 kDa FGF-2 (FGF-223) interacts specifically with the survival of motoneuron (SMN) protein, an assembly protein for small nuclear ribonucleoprotein particles. For this assembly, Sm-proteins methylated by protein arginine methyltransferase 5 (PRMT5) are required. In our study, we aimed to analyze whether FGF-223 is also a substrate for symmetrical methylation by PRMT5. We could confirm that both proteins exist in a common complex. Moreover, PRMT5 methylates FGF-223 in vitro, whereas mutated inactive PRMT5 does not. FGF-223 is therefore a new substrate of PRMT5. With regard to function, inhibition of methyltransferase activity in HEK293T cells leads to cytoplasmic enrichment of FGF-2, indicating the importance of arginine methylation for shuttling of FGF-223 to the nucleus. [ABSTRACT FROM AUTHOR]

Abstract: Introduction: We have designed an engineered graft fabricated from a biodegradable scaffold using chondrocytes and applied this construct to augment repair of tracheal stenosis. This study investigated the feasibility of using such tissue-engineered airways with autologous chondrocytes in a rabbit model. Material and Methods: Chondrocytes were isolated and expanded from the auricular cartilage of New Zealand white rabbits, then seeded onto composite 3-layer scaffolds consisting of a collagen sheet, a polyglycolic acid mesh, and a copolymer (l-lactide/ɛ-caprolactone) coarse mesh. The engineered grafts were implanted into a 0.5 × 0.8-cm defect created in the midventral portion of the cervical trachea. Gelatin sponges that slowly released basic fibroblast growth factor (b-FGF) were then placed on the constructs, which were retrieved 1 or 3 months after implantation. Results: The biodegradable scaffold seeded with chondrocytes could maintain airway structure up to 3 months after implantation. Tracheal epithelial regeneration occurred in the internal lumen of this composite scaffold. Three months after implantation, staining of the sections showed cartilage accumulation in the engineered tracheal wall. Conclusion: This composite biodegradable scaffold may be useful for developing engineered trachea. A gelatin sponge slowly releasing b-FGF might enhance chondrogenesis. [Copyright &y& Elsevier]

In general, tumors induce angiogenic factors specific to them, which leads to angiogenesis with tumor progression. However, angiogenesis in uterine endometrial cancers is complicated because the hormone dependency in their growth also modifies the angiogenic potential. Therefore, angiogenic potential in uterine endometrial cancers must be thoroughly analyzed. The upstream of the vascular endothelial growth factor ( VEGF) gene conserves estrogen-responsive elements. Progesterone primed with estrogen induces thymidine phosphorylase (TP) in the uterine endometrium. Sex steroid-dependent VEGF and TP are highly expressed in early-stage and well-differentiated uterine endometrial cancers, and basic fibroblast growth factor (bFGF) is highly expressed in advanced and poorly differentiated uterine endometrial cancers. A transcriptional factor for angiogenesis, E26 transformation specific (ETS-1), is linked to VEGF in well-differentiated uterine endometrial cancers, and to bFGF in poorly differentiated uterine endometrial cancers. Therefore, even if dedifferentiation and angiogenic switching occur due to tumor progression and long-term hormone therapy, the inhibition of ETS-1, along with inhibition of the main angiogenic factors, may be an effective strategy to suppress uterine endometrial cancers as a novel antiangiogenic therapy. [ABSTRACT FROM AUTHOR]

Abstract: Two members of the fibroblast growth factor (FGF) family, basic FGF (bFGF) and FGF-18, have been implicated in the regulation of articular and intervertebral disc (IVD) cartilage homeostasis. Studies on bFGF from a variety of species have yielded contradictory results with regards to its precise role in cartilage matrix synthesis and degradation. In contrast, FGF-18 is a well-known anabolic growth factor involved in chondrogenesis and articular cartilage repair. In this review, we examined the biological actions of bFGF and FGF-18 in articular and IVD cartilage, the specific cell surface receptors bound by each factor, and the unique signaling cascades and molecular pathways utilized to exert their biological effects. Evidence suggests that bFGF selectively activates FGF receptor 1 (FGFR1) to exert degradative effects in both human articular chondrocytes and IVD tissue via upregulation of matrix-degrading enzyme activity, inhibition of matrix production, and increased cell proliferation resulting in clustering of cells seen in arthritic states. FGF-18, on the other hand, most likely exerts anabolic effects in human articular chondrocytes by activating FGFR3, increasing matrix formation and cell differentiation while inhibiting cell proliferation, leading to dispersed cells surrounded by abundant matrix. The results from in vitro and in vivo studies suggest the potential usefulness of bFGF and FGFR1 antagonists, as well as FGF-18 and FGFR3 agonists, as potential therapies to prevent cartilage degeneration and/or promote cartilage regeneration and repair in the future. [Copyright &y& Elsevier]

Mesenchymal stromal cells (MSCs) derived from human bone marrow are expected to be utilized for the purpose of tissue engineering, because of their extensive self-renewal or proliferation capability. The capability decreases after several passages, however. Basic fibroblast growth factor (bFGF) is commonly used for culture of various cells including bone marrow-derived MSCs. With the aim of conferring higher capability on human bone marrow MSCs, we introduced the bFGF gene into the passaged cells by retroviral system. The bFGF-expressing MSCs, even at 7 to 8 passages after the infection, showed consistent proliferation capability. The capability was not detected in control cells even in culture media containing the bFGF protein. Thus, we could not mimic the effect of forced expression of bFGF by exogenously adding the bFGF protein in culture media. Although we expressed the shortest isoform of bFGF, which was considered to be mostly cytosolic, we found the protein mostly in the nucleus. Our observations demonstrate not only an effective way to maintain proliferation potentials of MSCs, but also a possibility that there may be mechanistic and functional differences in the signal transduction events between endogenously expressed and exogenously added bFGF protein in MSCs. [ABSTRACT FROM PUBLISHER]

Conclusion. Up-regulated expression of acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) in nasal polyps may be related to epithelial proliferation and neoangiogenesis associated with nasal polyp growth. Background and objectives. aFGF and bFGF are polypeptides that effect mitogenesis, neoangiogenesis and tissue repair. The purpose of this study was to investigate the expression of aFGF and bFGF at both the mRNA and protein levels in nasal polyps. Materials and methods. Tissue samples of nasal polyp and inferior turbinate mucosae were obtained. Reverse transcriptase (RT)-PCR, in situ hybridization, and Western blotting for aFGF and bFGF were performed in nasal polyp and nasal inferior turbinate. Results. RT-PCR showed 50% aFGF and 29% bFGF expression positivity in 14 nasal polyps; however, 13 turbinates revealed no PCR products. aFGF and bFGF expression was localized to some inflammatory cells of nasal polyps by in situ hybridization. Western blotting identified aFGF and bFGF molecular masses of 18 kDa and 24 kDa, respectively, in nasal polyps, and these levels were higher than those observed in nasal turbinates. aFGF and bFGF proteins were also localized in some inflammatory cells of nasal polyps by immunostaining. [ABSTRACT FROM AUTHOR]

Tumor growth is currently viewed as a phenomenon associated with neovascularization and sustained production of angiogenic factors, but whether a transient angiogenic switch may trigger tumor growth remains unclear. Here, we report that leukemia cells (MOLT-3) were poorly angiogenic and remained dormant when injected s.c. into immunodeficient mice. However, progressive growth of lymphoid tumors was invariably recorded when irradiated angiogenic cells from Kaposi's sarcoma (KS-IMM) were locally coinjected with MOLT-3 cells or administered later. The persistence of KS-IMM cells in vivo was tracked by flow cytometry and real-time PCR analysis, and it was limited to a few days, during which angiogenesis was induced and preceded tumor growth. The engraftment of other types of poorly tumorigenic cancer cells was also greatly improved by irradiated KS-IMM cells. Moreover, short-term treatment with angiogenic factors, including basic FGF or VEGF, either given as recombinant factors or delivered by retroviral vectors, also accelerated tumor growth. These findings may emphasize that tumor angiogenesis is a process requiring a higher amount of angiogenic factors for its induction than maintenance. [ABSTRACT FROM AUTHOR]

A novel HIV-1 transgenic rat model of childhood HIV-1–associated nephropathy. Background. A characteristic finding of human immunodeficiency virus (HIV)-associated nephropathy (HIVAN) is the presence of heavy proteinuria, focal or global glomerulosclerosis, and microcystic tubular dilatation leading to renal enlargement, and rapid progression to end-stage renal disease (ESRD). Methods. We have recently developed the first HIV-1 transgenic rat model that carry a noninfectious HIV-1 DNA construct lacking 3.1 kb of sequence overlapping the gag and pol sequences, and develop many of the clinical lesions seen in HIV-infected patients, including HIVAN. To gain further insight into the pathogenesis of childhood HIVAN, we followed the clinical and renal pathologic outcome of 165 HIV-1 transgenic (HIV-Tg) rats and their respective control littermates for a period of 18 months. Results. HIV-1 Tg rats progressively developed proteinuria and renal histologic lesions similar to those seen in children with HIVAN, leading to chronic renal failure. By in situ hybridization, HIV-1 genes were detected in glomerular and tubular epithelial cells and infiltrating mononuclear cells, which also expressed the HIV-1 envelop protein gp120. The development of HIVAN was associated with the accumulation of basic fibroblast growth factor (bFGF) in the kidney. Conclusion. These data support the notion that HIV-1 plays a direct role in the pathogenesis of HIVAN, by affecting the function and growth of renal epithelial cells, inducing the recruitment of mononuclear cells, and accumulating bFGF in the kidney, even in the absence of viral replication. These rats may provide an excellent model system to study the pathogenesis of childhood HIVAN. [ABSTRACT FROM AUTHOR]

Uncommitted mesenchymal stem cells (MSC), upon commitment and differentiation give rise to several mature mesenchymal lineages. Although the involvement of specific growth factors, including FGF2, in the development of committed MSC is known, the effect of FGF2 on uncommitted progenitors remains unclear. We have analyzed on a comparative basis, the subcellular distribution and mitogenic effect of FGF2 in committed and uncommitted MSC prepared from human bone marrow. Indirect immunofluorescence studies showed strong nuclear FGF2 staining in both progenitors; however, cytoplasmic staining was only detected in committed cells. Western blot analysis revealed the presence of 22.5 and 21–22 kDa forms of FGF2 in the nucleus of both progenitors; however, their relative content was higher in uncommitted than in committed cells. Exogenous FGF2 stimulated proliferation and sustained quiescence in committed and uncommitted cells, respectively. These results show that both type of progenitors, apart from morphological and proliferative differences, display specific patterns of response to FGF2. [ABSTRACT FROM AUTHOR]

Neural precursor cell (NPC) dysfunction has been consistently implicated in autism. Induced pluripotent stem cell (iPSC)-derived NPCs from two autism groups (three idiopathic [I-ASD] and two 16p11.2 deletion [16pDel]) were used to investigate if proliferation is commonly disrupted. All five individuals display defects, with all three macrocephalic individuals (two 16pDel, one I-ASD) exhibiting hyperproliferation and the other two I-ASD subjects displaying hypoproliferation. NPCs were challenged with bFGF, and all hyperproliferative NPCs displayed blunted responses, while responses were increased in hypoproliferative cells. mRNA expression studies suggest that different pathways can result in similar proliferation phenotypes. Since 16pDel deletes MAPK3, P-ERK was measured. P-ERK is decreased in hyperproliferative but increased in hypoproliferative NPCs. While these P-ERK changes are not responsible for the phenotypes, P-ERK and bFGF response are inversely correlated with the defects. Finally, we analyzed iPSCs and discovered that 16pDel displays hyperproliferation, while idiopathic iPSCs were normal. These data suggest that NPC proliferation defects are common in ASD., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Basic fibroblast growth factor (bFGF, FGF–2), a classical transforming factor, mitogen, and survival factor in multiple cell types, and has a paradoxic role in mammary epithelial cell transformation and proliferation. We have also demonstrated that recombinant FGF–2 uncharacteristically promotes cell death in MCF–7 human breast cancer cells. In this study, we investigated the effects of FGF–2 overexpression on survival in the same MCF–7 cells. In eight breast cancer cell lines and two nontransformed mammary epithelial cell lines, we demonstrated that high levels of Bcl–2 are only expressed in cells with undetectable levels of FGF–2 on western blot. In retrovirally transduced MCF–7 cells expressing both cytoplasm– and nucleus–localizing FGF–2 species and ones expressing only cytoplasm–localizing FGF–2 species, Bcl–2 levels were strongly decreased at both the mRNA and protein levels. Immunoprecipitation of Bax demonstrated a decreased association of Bax with Bcl–2 in these cells. Levels of Bax did not correlate with expression of FGF–2 in the 10 cell lines or in MCF–7 cells. The clonogenic potential of MCF–7 cells in tissue culture was decreased by the expression of FGF–2 and was additively suppressed by the chemotherapeutic agents etoposide and 5–fluorouracil in a dose and time dependent manner. MCF–7 cells overexpressing FGF–2 had a greater rate of programmed cell death at baseline and in response to etoposide and 5–fluorouracil in a TUNEL assay by immunofluorescent microphotography and by flow cytometric quantitation. The pro–apoptotic effect of FGF–2 overexpression on the chemosensitivity of these cells was confirmed by quantitative morphologic determination. These data demonstrate that the expression of FGF–2 downregulates Bcl–2 and promotes programmed cell death in MCF–7 human breast cancer cells. [ABSTRACT FROM AUTHOR]

Subcutaneous implantation of bone morphogenetic protein (BMP) combined with a fibrous glass membrane (FGM) induces cartilage formation in the entire inner area of the membrane within 2 wk. It has been hypothesized that a tight FGM network (1 μm exclusion size) provides immature cells with spaces for penetrating into the membrane, but not for vascular formation, at least until 2 wk. To test this hypothesis, basic fibroblast growth factor (bFGF), known to be a potent stimulant of capillary formation, was applied to the implant. BMP was combined with FGM in the presence or absence of bFGF, and then implanted subcutaneously into the backs of rats. The bFGF-supplemented implant caused 1.3 times higher alkaline phosphatase activity and 3 times higher calcium contents at 2 wk, whereas type II collagen contents decreased, thus indicating that bFGF enhances bone formation in BMP/FGM implants. These results suggest that bFGF induces faster and stronger invasion of capillaries into the FGM and destroys its tight network, resulting in acceleration of the ossification process. [ABSTRACT FROM AUTHOR]

Increased expression of basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) A chain, and tissue plasminogen activator (tPA) by smooth muscle cells (SMC) has been postulated to mediate the progression of intimal hyperplasia. We tested whether heparin would suppress the expression of these genes in stimulated human saphenous vein SMC. Quiescent cultured human saphenous vein SMC were stimulated for 4 h with heat-inactivated fetal bovine serum (10% by vol) in the presence or absence of heparin (1 to 250 µg/ml). Heparin (50 µg/ml) attenuated the induction by serum of bFGF mRNA, tPA mRNA, and tPA secretion. Nonanticoagulant heparin also attenuated serum induction of bFGF and tPA mRNA levels. To further study the role of second messenger signaling, a more specific mode of SMC stimulation was used with thrombin (3 U/ml) in the presence or absence of dibutyryl cyclic AMP (Bu-cAMP; 0.5 m M). In contrast to heparin, which had no effect on PDGF expression, Bu-cAMP decreased the induction by thrombin of PDGF-A chain mRNA levels. In thrombin-stimulated SMC, Bu-cAMP significantly decreased secretion of PDGF-AA protein. Thrombin, however, caused an increase in bFGF mRNA levels which was potentiated by Bu-cAMP with associated potentiation by Bu-cAMP of intracellular bFGF protein levels. The induction of tPA mRNA and tPA secretion by thrombin was sharply blocked by Bu-cAMP. These results suggest that heparin reduces intimal hyperplasia at least partly via partial inhibition of SMC gene expression. [ABSTRACT FROM AUTHOR]

Objective: The collateral circulation plays a critical role in the prognosis of ischaemic heart disease, with a clear correlation between neovascularisation, infarction, and tissue recovery. The aim of the study was to address the question of whether endogenous acidic and basic fibroblast growth factors (FGF) participate in ischaemia induced collateral enlargement and development in the myocardium, and also which cells may represent the source of these growth factors. Methods: Eight pigs received an ameroid constrictor around the left circumflex coronary artery which, by slow coronary occlusion, induces ischaemia and collateral growth in the left ventricle. The degree of stenosis and development of collaterals were determined angiographically. About 14 d after constrictor implantation pigs were killed and hearts were excised and prepared for western blot analysis and immunohistochemistry. Monoclonal antibodies were raised against acidic and basic FGF, characterised for their specificity, and used for the localisation of growth factors in sections of ischaemic and normal pig heart. Results: The eight pigs included in this study showed a gradual 70-100% left circumflex coronary stenosis about 14 d after implantation of the constrictor. Out of four pigs with a complete occlusion, three developed visible collaterals. Antibody staining to acidic FGF was detected only in hearts from pigs with complete occlusion of the coronary artery. The growth factor was localised in cardiomyocytes close to small necrotic tissue patches. In control tissue no acidic FGF staining was evident. Basic FGF could not be detected in ischaemic or in normal perfused pig hearts. Conclusions: These data show that (1) complete occlusion of the left circumflex coronary artery in pig hearts induces collateral growth; (2) cardiomyocytes are able to produce acidic FGF in response to ischaemia, thus providing a mitogen for endothelial cells; (3) endogenous basic FGF, which was not detected in normal or ischaemic pig hearts, appears not to play an important role in ischaemia induced collateral growth at the chosen time point of investigation.Cardiovascular Research 1993;27:1220-1228 [ABSTRACT FROM PUBLISHER]

Objective: Acidic and basic fibroblast growth factors (FGF) are potent mitogens for vascular endothelial and smooth muscle cells and also promote angiogenesis. Given that atheroma is associated with the proliferation of a range of cell types, including smooth muscle cells, it has been proposed that these growth factors may play a part in the genesis and/or maintenance of atheromatous lesions. The aim of our study was to examine the distribution of acidic and basic FGF and their known receptor, fibroblast growth factor receptor 1 (FGFR-1), in normal and atherosclerotic arteries. Methods: Formalin fixed wax embedded archival material from a wide range of vessels was examined using a three stage avidin-biotin immunoperoxidase technique and specific polyclonal antibodies to acidic and basic FGF and FGFR-1. Results: In normal arteries basic FGF immunoreactivity was found in medial smooth muscle cells and adventitial blood vessels, the luminal endothelium being non-reactive. Acidic FGF expression was observed predominantly in adventitial fibroblasts, while FGFR-1 expression was confined to the adventitial microvasculature. In early, simple, and advanced lesions acidic and basic FGF were expressed in macrophages and smooth muscle cells, the principal cell types involved in atherosclerotic lesion formation. Increased expression of basic FGF and FGFR-1 was particularly associated with neovascularisation of the atheromatous lesions. Conclusions: Acidic and basic FGF are expressed in early, simple and advanced atherosclerotic plaques. This suggests that in vivo these growth factors may have a role in the genesis of this disease. Basic FGF and FGFR-1 may potentially be involved in plaque neovascularisation. Such FGF driven angiogenic events may be central to the life threatening complications of atheroma and provide new options for therapeutic intervention.Cardiovascular Research 1993;27:1214-1219 [ABSTRACT FROM PUBLISHER]

Primary Rhodamine fibrosarcoma (RdF) cells from rats were shown to grow in serum-free medium supplemented with basic fibroblast growth factor (bFGF), albumin, and transferrin, all of which were purified from RdF tissue. Their growth rate with these supplements was similar to that of cells in medium supplemented with calf serum. bFGF purified from RdF tissue (Rd-bFGF), which was previously designated as DNA synthesis factor, stimulated the growth of primary RdF cells maximally at 30 ng/ml in the presence of the other two proteins. Albumin and transferrin were separated from partially purified tumor growth stimulating activity which was previously shown to stimulate growth of primary RdF cells in serum-free medium. The albumin (RdA) and transferrin (RdT) found in the extract of RdF tissue were not due simply to contamination of the tissue with blood, but to their accumulations in the tissue. The growth stimulatory activities of RdA and RdT on primary RdF cells in serum-free medium were maximal at 30 and 10 μg/ml, respectively. These results suggest that Rd-bFGF, RdA, and RdT, all of which accumulate in the tumor tissue, are essential for growth of RdF cells in the tissue. [ABSTRACT FROM AUTHOR]

Basic fibroblast growth factor (FGF) is thought to be involved in carcinogenesis and, to clarify its clinical significance, the study of its blood level in cancer patients is important. Plasma levels of basic FGF are reported to be elevated in some cancers. However, little is known of basic FGF levels in plasma in hepatocellular carcinoma (HCC). In this study, we measured basic FGF plasma levels in patients with chronic liver disease and compared the levels in chronic hepatitis (CH), liver cirrhosis (LC), and HCC. We also examined whether these levels were related to serum levels of asparate aminotransferase, alanine aminotransferase, γ-glutamyl transpeptidase, alkaline phosphatase, leucine aminopeptidase, total bilirubin, total protein, and albumin, and to the indocyanine green test (i.e., liver function tests) and to type III procollagen, 7S domain of IV type collagen, and hyaluronic acid (i.e., markers of liver fibrosis). Levels of basic FGF. determined by a quantitative “sandwich” enzyme immunoassay, were significantly elevated with the progression of liver disease; being 3.67 ± 2.37 (mean ± SD), 7.78 ± 6.61, and 12.37 ± 7.67pg/ml in the CH, LC, and HCC groups, respectively. FGF levels were elevated to a greater extent in the HCC patients than in the CH ( P<0.0001) and LC patients ( P=0.0117). Levels were higher in LC than in CH ( P=0.0204). None of the liver function test findings or levels of markers of liver fibrosis were correlated with levels of basic FGF. These results suggest that circulating basic FGF could serve as a new indicator of the progression of chronic liver disease. The extremely elevated plasma of level basic FGF in the HCC group suggests that basic FGF may be related to the development of HCC. [ABSTRACT FROM AUTHOR]

Neural stem progenitor cells (NSPCs) from E13.5 mouse embryos can be maintained in culture under proliferating conditions. Upon growth-factor removal, they may differentiate toward either neuronal or glial phenotypes or both. Exosomes are small extracellular vesicles that are part of the cell secretome; they may contain and deliver both proteins and genetic material and thus play a role in cell–cell communication, guide axonal growth, modulate synaptic activity and regulate peripheral nerve regeneration. In this work, we were interested in determining whether NSPCs and their progeny can produce and secrete extracellular vesicles (EVs) and if their content can affect cell differentiation. Our results indicate that cultured NSPCs produce and secrete EVs both under proliferating conditions and after differentiation. Treatment of proliferating NSPCs with EVs derived from differentiated NSPCs triggers cell differentiation in a dose-dependent manner, as demonstrated by glial- and neuronal-marker expression. [ABSTRACT FROM AUTHOR]