LGF («LIVER GROWTH FACTOR») como factor de proliferación, migración y diferenciación de las células madre neurales y su posible utilidad en enfermedad de Parkinson / LGF («LIVER GROWTH FACTOR») as a factor involved in the proliferation, migration and differentiation of neural stem cells: Potential use in Parkinson’s disease

El factor de crecimiento de hígado, LGF, es un factor de proliferación y regeneraciónen distintos tipos celulares, y estudios preliminares fundamentan suposible uso en terapias neuroregenerativas. El objetivo del trabajo ha sido comprobarsi el LGF estimula la proliferación celular y la neurogénesis en el modeloexperimental de enfermedad de Parkinson (EP) de lesión con 6-OHDA. La administraciónintraventricular de LGF incrementó la incorporación de bromodeoxiuridina(BrdU) y la expresión de los marcadores de células madre neurales(CMN) nestina y proteina ácida de la glía, observándose células doblemente marcadaspara BrdU y nestina en la zona subventricular y el parénquima estriatal.Sin embargo, no se encontraron células BrdU-positivas/ b tubulina III-positivas,sugiriendo que el LGF no es un factor neurogénico para las CMN de la zona subventricular.La infusión intraestriatal de LGF no afectó a la proliferación celular,pero aumentó significativamente el número de terminales tirosina hidroxilasapositivosen el estriado denervado y mejoró la conducta rotacional provocada porapomorfina. Las conclusiones son: 1) la infusión intraventricular de LGF estimulala proliferación y migración de las CMN; y 2) la infusión intraestriatal deLGF estimula la regeneración de los terminales dopaminérgicos dañados, lo quepodría explicar la discreta mejoría conductual observada en estos animales

Liver growth factor (LGF) promotes the proliferation and regeneration ofdifferent cell types, and preliminary studies indicate that LGF might be usefulfor neuroregenerative therapies. The aim of this work was to study the cell proliferativeand neurogenic effects of LGF in a 6-OHDA Parkinson’s disease (PD)experimental model. Intraventricular infusion of LGF increased bromodeoxyuridine(BrdU) incorporation in the subventricular zone and striatum. In addition,the striatum ipsilateral to LGF infusion showed a higher expression ofneural stem cell (NSC) markers nestin and glial fibrillary acidic protein, andthe presence of cells double-labeled for BrdU and nestin. However, no BrdU-positive/B tubulin III-positive cells were observed, suggesting that LGF is not aneurogenic factor. Intrastriatal LGF infusion significantly increased the numberof tyrosine hydroxylase-positive fibers in the striatum of dopamine-denervatedrats. Moreover, apomorphine-induced behavioral rotations improved inthese animals. We conclude that: 1) Intraventricular LGF infusion stimulatesneural stem cell proliferation and migration; and 2) Intraestriatal LGF infusionstimulates the regeneration of dopaminergic terminals in the striatum,which might explain the partial behavioral recovery observed in these animals

In vitro and in vivo characterization of neural stem cells.

Neural stem cells are defined as clonogenic cells with self-renewal capacity and the ability to generate all neural lineages (multipotentiality). Cells with these characteristics have been isolated from the embryonic and adult central nervous system. Under specific conditions, these cells can differentiate into neurons, glia, and non-neural cell types, or proliferate in long-term cultures as cell clusters termed "neurospheres". These cultures represent a useful model for neurodevelopmental studies and a potential cell source for cell replacement therapy. Because no specific markers are available to unequivocally identify neural stem cells, their functional characteristics (self-renewal and multipotentiality) provide the main features for their identification. Here, we review the experimental and ultrastructural studies aimed at identifying the morphological characteristics and the antigenic markers of neural stem cells for their in vitro and in vivo identification.

Developmental expression of fibroblast growth factor (FGF) receptors in neural stem cell progeny. Modulation of neuronal and glial lineages by basic FGF treatment.

Neural stem cells (NSCs) are self-renewable, multipotential cells capable of differentiating into the three major neural cell types, but the mechanisms which regulate their development are not fully understood. Both basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) promote the proliferation of NSCs. However, studies on the role of FGFs in the differentiation of EGF-expanded NSCs are still incomplete. We have studied the expression of distinct FGF receptors (FGFRs) in the progeny of EGF-expanded NSCs isolated from E15 rat striatum. In situ hybridization analysis and immunocytochemistry showed a developmentally related expression pattern and a cell lineage-specific distribution of these receptors. FGFR1 and FGFR2 were identified in many early precursors and in the oligodendrocyte lineage. The latter receptor was also present in a subpopulation of astrocytes. FGFR3 was detected in a restricted population of early precursors, in oligodendroglial progenitors, and in neurons and protoplasmic astrocytes of late-term cultures. Basic FGF treatment of the progeny of NSCs increased the proliferative rate of precursors and the number of oligodendrocytes generated, whereas the number of differentiating neurons was significantly reduced. Together these data provide evidence that FGFs modulate the development of EGF-expanded NSCs, and that this is at least partly determined by a cell lineage-specific expression of multiple FGFRs.

Abolished angiogenicity and tumorigenicity of rat glioma by 1-naphthalenemonosulfonate.

Suramins and suradistas, an important group of potential anti-cancer agents, inhibit fibroblast growth factor (FGF) mitogenic activity. It has been shown that naphthalenesulfonates, with a common chemical function to the family of suramins and suradistas, mimic their inhibitory activity, abolishing FGF-induced angiogenesis in vivo, and inducing apoptosis of C6 glioma cells in culture. In the present report, we show that intratumoral administration of 1-naphthalenemonosulfonate induces a considerable regression of gliomas in rats, significantly enhances apoptosis, and attenuates tumor angiogenesis. These findings may lead to new approaches for the treatment of glioblastoma, a most common primary malignant brain tumor of very poor prognosis, as well as of other angiogenesis-dependent malignancies.

Sacroiliac joint dysfunction: a long-term follow-up study.

Sixty-nine patients with sacroiliac joint dysfunction were prospectively evaluated and treated with a structured physical therapy program. Follow-up clinical outcome was obtained from a patient questionnaire administered by an independent reviewer a minimum of 2 years after treatment. Average patient age was 40 years, and 80% were women. Ninety-five percent rated their result as good or excellent, while 5% believed their outcome was fair or poor. A structured physical therapy program can produce good long-term results in most patients; however, 5% continue to be symptomatic. This small subset may be candidates for more invasive evaluation.

Inhibition of fibroblast growth factor/fibroblast growth factor receptor activity in glioma cells impedes tumor growth by both angiogenesis-dependent and -independent mechanisms.

We undertook a series of systematic studies to address the role of fibroblast growth factor/fibroblast growth factor receptor (FGF/FGFR) activity in tumor growth and angiogenesis. We expressed dominant-negative FGFR2 (FGFR2-DN) or FGFR1 (FGFR1-DN) in glioma C6 cells by using constitutive or tetracycline-regulated expression systems. Anchorage-dependent or independent growth was inhibited in FGFR-DN-expressing cells. Tumor development after xenografting FGFR-DN-expressing cells in immunodeficient mice or after transplantation in rat brain was strongly inhibited. Quantification of microvessels demonstrated a significant decrease in vessel density in tumors derived from FGFR-DN-expressing cells. Furthermore, in a rabbit corneal assay, the angiogenic response after implantation of FGFR-DN-expressing cells was decreased. In tumors expressing FGFR-DN, vascular endothelial growth factor expression was strongly inhibited as compared with control tumor. These results indicate that inhibition of FGF activity may constitute a dominant therapeutic strategy in the treatment of FGF-producing cerebral malignancies and may disrupt both angiogenesis-dependent and -independent signals required for glioma growth and invasion.

Acidic fibroblast growth factor inhibits junctional communication of Schwann cells in culture.

The effect of acidic fibroblast growth factor (aFGF) was investigated on junctional communication of rat Schwann cells (SC) in culture. As measured by dye transfer, the incidence of coupling between SC was very low during the phase of proliferation and increased slowly and progressively with time under culture conditions that induced the myelinating phenotype. Treatment with aFGF alone or in combination with heparin decreased markedly coupling between SC in both culture stages. The coupling inhibition was rapid, the earliest effects being apparent 5-15 min after addition of growth factor, and was transient with a slower recovery of coupling at 1-3 h. The uncoupling effect of aFGF could be prevented by an inhibitor of protein-tyrosine kinase. Addition of heparin to cultures decreased the most effective aFGF concentration by 100-fold, from 100 ng ml-1 to 1 ng ml-1. The dose-response curves exhibited a characteristic window-shape. The results suggest that FGF might be involved in the regulation of the junctional communication between rat SC via tyrosine kinases.

Fibroblast growth factor-1 inhibits medial smooth muscle cells apoptosis after balloon injury.

We performed balloon injury in the rat common carotid artery and identified apoptosis of vascular smooth muscle cells (VSMCs) by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, 2 h after injury. Balloon injury induces apoptosis and loss of bcl-X protein in the innermost layers of the media of the common carotid artery. Treatment with acidic fibroblast growth factor (aFGF, FGF-1) attenuated by 56% the balloon angioplasty-induced apoptosis. In addition, FGF-1 treatment also induces expression of the bcl-X anti-apoptotic protein in the same site of the media showing VSMC apoptosis. These data suggest that the anti-apoptotic effect of FGF in injured vascular wall was mediated by a bcl-X pathway and identified FGF as an important factor in vascular remodeling.

Cardioprotection from ischemia by fibroblast growth factor: role of inducible nitric oxide synthase.

Growing evidence from both animal experiments and clinical observations indicates that fibroblast growth factor (FGF) plays a protective role in myocardial reperfusion injury. The molecular and cellular mechanisms that lead to this postischemic myocardial protection, however, remain largely unexplored. We studied the cardioprotective effects of human recombinant acidic fibroblast growth factor (aFGF, FGF-1) in a rat model of myocardial reperfusion injury, induced by 20 minutes of left coronary artery occlusion followed by 24 hours of reperfusion. Intravenous FGF-1 administration at the onset of heart reperfusion attenuated both the functional impairment and the histological changes of ischemia/reperfusion injury. FGF-1 increases more than twice the left ventricular contractile function (p <0.005) compared to vehicle-treated rats. As shown by histology, myocardial tissue is better preserved with FGF-1 treatment. The infarct size, normalized for the area at risk, was significantly smaller in the FGF-1 group (p <0.01) than in the vehicle group. Furthermore, FGF-1 administration resulted in expression of inducible nitric oxide synthase (iNOS) in the area at risk. Since increased expression of iNOS could potentiate cardioprotection against myocardial reperfusion injury, our findings support a new non-mitogenic role for FGF and add a clinical interest for this protein in increasing myocardial ischemic tolerance.

Apoptosis of glioma cells induced by the fibroblast growth factor inhibitor 1,3,6-naphthalenetrisulfonate.

Fibroblast growth factors (FGFs) are powerful angiogenic polypeptides that are involved in the autocrine growth stimulation of gliomas. We report here that addition to glioma cell cultures of 1,3,6-naphthalenetrisulfonate (NTS), an inhibitor of the mitogenic activity of FGFs, significantly enhanced apoptosis, as assessed by terminal deoxynucleotidyl transferase (TdT) assay. The pro-apoptotic effect of NTS was time-dependent. These findings suggest that FGF may play a pivotal role in the survival of glioma cells, and support a clinical interest of NTS as a leading compound for the development of new antitumorals.

Fibroblast growth factor protects the kidney against ischemia-reperfusion injury.

Ischemia-reperfusion injury, a common source of renal dysfunction in adults, is associated with tubular epithelial cell damage. Since fibroblast growth factors (FGF) attenuated tissue injury after transient myocardial ischemia, we hypothesized that acidic fibroblast growth factor (aFGF; FGF-1) would attenuate renal ischemia-reperfusion injury. We studied the effects of FGF-1 in a rat model of acute renal failure induced by bilateral renal ischemia (60 min) and 1, 2 or 7 days reperfusion. After FGF-1 administration at the onset of renal reperfusion, there was less functional impairment of the kidneys. The histological changes were not as severe as in controls. Increases in serum creatinine and blood urea nitrogen 24 h after reperfusion were attenuated by 35% (p< 0.01) and by 53% (p< 0.001), respectively, in FGF-1-treated animals compared to vehicle-treated rats. The ischemia/reperfusion-induced increase in tissue myeloperoxidase, a marker of neutrophil infiltration, was mitigated (67% reduction, p< 0.05) with FGF-1 treatment. As shown by histology, neutrophil infiltration and tubular cell necrosis in medulla were less pronounced (p< 0.0001 and p< 0.05, respectively) in animals receiving FGF-1. Furthermore, ischemia-induced apoptosis, prevalent in tubular cells of the cortex, was also attenuated by FGF-1-treatment (83% reduction, p< 0.0001). Pretreatment of animals with Nw-nitro-L-arginine (L-NNA), an inhibitor of nitric oxide synthase, abolished the attenuating effects of FGF-1 on neutrophil infiltration, suggesting that nitric oxide might participate in the anti-inflammatory effects of FGF-1 in this experimental design. Our data support a role for FGF-1 in attenuation of renal damage or failure after ischemia-reperfusion injury of the kidney, in part at least by inhibition of neutrophil infiltration.

Inhibition of intra-tumoral angiogenesis and glioma growth by the fibroblast growth factor inhibitor 1,3,6-naphthalenetrisulfonate.

1,3,6-naphthalenetrisulfonate (NTS) can inhibit the proliferation in vitro of cells of various origin including glioma. We have studied the effects of NTS on intra-tumoral angiogenesis and tumor growth in the rabbit cornea after implantation of C6 rat glioma cells. It was found that neovascularization and glioma growth were abolished by topical administration of NTS. This effect could be mediated by both induction of programmed cell death and inhibition of growth, in endothelium and in tumor cells, most likely as a consequence of the disruption of the autocrine and paracrine effects of FGF released from endothelial and tumor cells. The results suggest that NTS is a promising candidate to lead the development of new angiogenesis inhibitors for the treatment of cancer and other diseases whose progression is dependent upon the development of new blood vessels.

Acidic fibroblast growth factor rescues gerbil hippocampal neurons from ischemic apoptotic death.

Neurotrophic factors have been shown to support the survival of injured neurons and promote their recovery. Here, we investigated whether acidic fibroblast growth factor (aFGF) could modify programmed cell death caused by transient forebrain ischemia in the gerbil. The data show that systemic administration of 2.6 microg aFGF after 5 min ischemia followed by 7 days of brain reperfusion significantly (p < 0.05) reduced the occurrence of apoptotic cell death in CA1 neurons. These data suggest that aFGF would contribute to brain protection after acute stroke.

Physical activity and fitness in patients with headache disorders.

Recent studies indicate that aerobic activities might reduce severity and/or frequency of migraine attacks. The present study was intended to investigate whether physical activities and fitness (aerobic endurance, flexibility, and muscle strength endurance) as well as body composition are different in patients with headache disorders and healthy control subjects. The study included 56 patients (aged 17-64 years) with headache disorders (migraine, tension-type, cluster, analgetics abuse, and other types of headache) and 145 age-matched volunteers without history of recurrent or chronic headache. A standardized questionnaire revealed similar self-esteem of physical activities in both groups. Objective physical fitness testing in a representative sample of 22 patients and 36 control subjects showed significantly reduced aerobic endurance in female and male patients as well as reduced flexibility in female patients as compared to control subjects, whereas muscle strength endurance was not significantly different between both groups. Female patients presented with a significantly higher total body fat as compared to control subjects. In conclusion, headache patients turned out to be less physically fit than control subjects. There was a discrepancy between self-esteem and objective test results regarding physical activity and fitness in patients with headache disorders.

Fibroblast growth factor-1 prevents myocardial apoptosis triggered by ischemia reperfusion injury.

BACKGROUND: Apoptosis is a constant feature of reperfusion injury in ischemic cardiac myocytes, leading to late cell death. Since fibroblast growth factors (FGFs) inhibit apoptosis in differentiated cells, we hypothesized that FGF-1 (acidic FGF), in its native form, and a non-mitogenic isoform would attenuate myocardial ischemia-reperfusion- induced apoptosis. METHODS AND RESULTS: The effect of native and non-mitogenic fibroblast growth factor-1 mutein (FGF-1 and m-FGF-1) on apoptosis assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method was tested in a rat model of 20 min regional myocardial ischemia and 24h reperfusion. Myocardial ischemia followed by reperfusion resulted in a high myocardial apoptosis rate in the area at risk. When given as a systemic bolus inmediately after myocardial ischemia, both FGF-1 and m-FGF-1 significantly reduced apoptosis (by 60 and 61.2, respectively; p<0.0001). CONCLUSIONS: The programed myocyte cell death triggered by ischemia-reperfusion injury is attenuated by FGF-1 in its native or non mitogenic isoforms, suggesting that this effect does not depend on the mitogenic properties of this protein. FGF-1 would contribute to the functional preservation of the myocardium after acute myocardial infarction.

Loss of basic fibroblast growth factor in the subcommissural organ of old spontaneously hypertensive rats.

The immunocytochemical localization of basic fibroblast growth factor (bFGF) was studied in subcommissural organ (SCO) of aged-matched normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SH) rats at 10, 14 and 18 months of age using a polyclonal antibody against bFGF. The bFGF-like immunoreactivity (bFGF-ir) was observed in SCO ependymal cells of young and old normotensive rats. However, a progressive loss of bFGF-immunopositive ependymal SCO cells was observed with age in SH rats (27.24, 57.5 and 96.9% in 10, 14 and 18 months old respectively) compared with aged-matched WKY normotensive rats. Considering the potential role of the SCO in sleep regulation and sodium homeostasis, which are altered in essential hypertension, these data show a new neuroendocrine anomaly to be added to the many others previously observed in this rat strain, when they develop hypertension as they get old.

Correction of hypertension by normalization of endothelial levels of fibroblast growth factor and nitric oxide synthase in spontaneously hypertensive rats.

Acidic and basic fibroblast growth factors (FGFs) share a wide range of diverse biological activities. To date, low levels of FGF have not been correlated with a pathophysiologic state. We report that blood vessels of spontaneously hypertensive rats are shown to be associated with a marked decrement in endothelial basic FGF content. This decrement correlates both with hypertension and with a decrease in the endothelial content of nitric oxide synthase. Restoration of FGF to physiological levels in the vascular wall, either by systemic administration or by in vivo gene transfer, significantly augmented the number of endothelial cells with positive immunostaining for nitric oxide synthase, corrected hypertension, and ameliorated endothelial-dependent responses to vasoconstrictors. These results suggest an important role for FGFs in blood pressure homeostasis and open new avenues for the understanding of the etiology and treatment of hypertension.

Immunohistochemical localization of basic fibroblast growth factor in choroid plexus of the rat.

In the present study, we report that an intense bFGF-immunoreactivity has been detected in the choroid plexus of the brain ventricles of adult rats. These results suggest that epithelial choroid plexus cells may be the source of the cerebrospinal fluid bFGF.

The tooth-removable partial denture interface.

During the metal finishing phase of removable partial denture construction, metal is removed from all aspects of the casting. The amount removed from areas designed to contact prepared surfaces of abutments is critical to the resulting fit of the partial. Castings were evaluated before and during finishing procedures to measure metal loss at the tooth-removable partial denture interface. Finishing and fitting techniques for controlling the loss of metal are presented.

Ischemic reperfusion injury in rabbit spinal cord: protective effect of superoxide dismutase on neurological recovery and spinal infarction.

The potential role of superoxide dismutase (SOD), a specific superoxide anion radical scavenger, in treating spinal cord ischemia was investigated in rabbits subjected to aortic occlusion for 20 min. SOD treatment, targeted to the early reperfusion period, reduced both motor dysfunction and incidence of spinal infarcts at 7 days after ischemia. Present results suggest that oxygen-derived free radicals play a role in the pathogenesis of infarcts developing in the spinal cord after ischemia and reperfusion injuries.