Bovine brain-derived <em>growth</em> <em>factor</em> (BDGF), a 16-<em>17</em> kDa protein with biochemical properties resembling brain-derived acidic <em>fibroblast</em> <em>growth</em> <em>factor</em> (acidic FGF) and endothelial cell <em>growth</em> <em>factor</em>, was found to have potent chemotactic activity for bovine ligament <em>fibroblasts</em>, human skin <em>fibroblasts</em> and rat astroglial cells, maximal at 100-200 pg/ml. The chemotactic activity was completely blocked by protamine sulfate (5 ug/ml), an inhibitor of receptor-binding and mitogenic activity of BDGF. BDGF did not stimulate migration of human monocytes. These results indicate that the effects of BDGF 'in vivo' might extend to mesenchymal cell recruitment.

Owing to strong interactions between pancreatic islets and the surrounding capillary network, we hypothesized that high glucose concentrations might affect key angiogenesis <em>factors</em> from isolated human islets, thus contributing to beta-cell failure in diabetes. Human islets from eight distinct donors were studied following 96 h in culture in the presence of normal (5.5 mmol/l) or high (16.7 mmol/l) glucose concentrations. Similar studies were performed with HUVECs. Human angiogenesis-related genes and corresponding proteins were studied by real-time quantitative PCR (RT-qPCR) and protein arrays respectively. Angiogenesis and proliferation assays were also performed with HUVECs under the same culture conditions. RT-qPCR and proteome analysis of human islets incubated with 16.7 mM/l glucose revealed a significant decrease in pro-angiogenic <em>factors</em> including vascular endothelial <em>growth</em> <em>factor</em> (VEGF) mRNA by 20% and VEGF protein levels by 42% as well as additional proteins such as <em>fibroblast</em> <em>growth</em> <em>factor</em>-4 by 41%, MMP9 by 18%, monocyte chemoattractant protein-1 by 21%, and prolactin by 25%. In contrast, we observed a <em>17</em>% increase in thrombospondin-1 (TSP-1, listed as THBS1 in the HUGO database) and a 37% increase in angiotensinogen gene expression levels, but neither angiotensin-converting enzyme nor angiotensin II type 1 receptor gene expression was affected. The amounts of anti-angiogenic proteins such as TSP-1 and serpin B5/maspin were also increased by 70 and 98% respectively as well as endostatin by 63%. Angiogenesis assays of HUVECS in the presence of high glucose concentrations revealed a 30% decrease in tree-like tubular network formation. These data suggest that glucose reduces key <em>factors</em> of islet angiogenesis, which might exacerbate beta-cell failure.

Ischemia-induced angiogenic response is reduced in spontaneously hypertensive rats (SHR). To study whether exogenous basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) infusion is effective in expanding collateral circulation in frankly hypertensive SHR, femoral arteries of male SHR (weighing approximately 250 g) were kept intact (nonoccluded control; n = 9) or occluded for 4h(n = 12) or for 16 days with vehicle (n = 14) or bFGF [0.5 (n = <em>17</em>), 5.0 (n = 13), and 50.0 (n = 14) microg. kg-1. day-1 for 14 days] intraarterially. Maximal collateral-dependent blood flows (BF) to the hindlimbs were determined with 85Sr- and 141Ce-labeled microspheres during running at 20 and 25 m/min (15% grade). Preexercise heart rates (approximately 530 beats/min) and blood pressures (BP; approximately 200 mmHg) were similar across groups except in the high-dose bFGF group, where BP was reduced by approximately 12% (P < 0.05). Femoral artery occlusion for 4 h resulted in approximately 95% reduction of BF in calf muscles [199 +/- 18.7 (nonoccluded group) to 10 +/- 1.0 ml. min-1. 100 g-1; P < 0.001]. BF to calf muscles of the vehicle and low-dose bFGF (0.5 microg. kg-1. day-1) groups increased to 36 +/- 3.2 and 45 +/- 2.0 ml. min-1. 100 g-1, respectively (P < 0.001). bFGF infusion at 5.0 and 50.0 microg. kg-1. day-1 further increased (P < 0.001) BF to calf muscles (62 +/- 4.6 and 62 +/- 2.2 ml. min-1. 100 g-1, respectively). Our results show that bFGF can effectively increase BF in hypertensive rats. The reduced hypertension with high-dose bFGF suggests that a critical signal in arteriogenesis (nitric oxide bioavailability) may be restored. These findings suggest that the dulled endothelial nitric oxide synthase of SHR does not preempt collateral vessel remodeling.

We have previously reported that calcium channel antagonists can block both the <em>growth</em> of meningiomas in culture and the potent <em>growth</em> stimulation of meningioma cells by epidermal <em>growth</em> <em>factor</em> (EGF) and platelet-derived <em>growth</em> <em>factor</em> (PDGF). This study further defines the nature of this <em>growth</em> inhibition. Primary meningioma cultures were established, and cells were characterized. <em>Fibroblast</em> <em>growth</em> <em>factor</em> or insulin-like <em>growth</em> <em>factor</em>-I <em>growth</em> stimulation in the presence of calcium channel antagonists was examined. In addition, the effects of ethylene glycol-bis-(aminoethylether) N,N,N',N"-tetraacetic acid and Bay K 8644, a calcium channel agonist, on the <em>growth</em> <em>factors</em> were analyzed. <em>Growth</em> <em>factor</em> receptor immunohistochemistry was performed on the original tumors and the in vitro meningioma cells. Twelve of <em>17</em> (71%) meningiomas in this study were positive for the EGF receptor, and 14 of <em>17</em> (82%) were positive for the PDGF receptor. Five of six (83%) of the culture cells were positive for the EGF receptor, and four of five (80%) were positive for the PDGF receptor. Intracellular calcium changes were quantified using the intracellular calcium-chelating, fluorescent dye, Fura-2. The <em>growth</em> stimulation of <em>fibroblast</em> <em>growth</em> <em>factor</em> and insulin-like <em>growth</em> <em>factor</em>-I on meningioma cells in culture was decreased in a dose-dependent manner by calcium channel antagonists. The <em>growth</em> stimulation of <em>fibroblast</em> <em>growth</em> <em>factor</em> and insulin-like <em>growth</em> <em>factor</em>-I was not affected by a reduction of extracellular calcium, whereas the <em>growth</em> stimulation of EGF and PDGF was. Interestingly, intracellular calcium was not increased after exposure to <em>growth</em> <em>factors</em> but was increased after serum stimulation. This increase could be blocked by preincubation with verapamil. Calcium channel antagonists can inhibit proliferation of meningioma cells in culture after stimulation with a number of <em>growth</em> <em>factors</em>. These drugs might disrupt intracellular calcium homeostasis or interfere with key elements of the <em>growth</em> <em>factor</em> signal transduction pathways. These mechanisms as well as the potential clinical relevance of these findings are discussed.

Pleiotrophin (PTN) is a <em>growth</em> and neurite extension promoting polypeptide which is highly expressed in brain and in tissues derived from mesenchyme. The PTN gene is developmentally regulated and is closely related to the MK and RI-HB genes, both of which are developmentally regulated and induced by retinoic acid. We now have screened <em>17</em> cell lines and report that expression of the PTN gene in these cells is restricted to embryo <em>fibroblasts</em> and intestinal smooth muscle cells. However, NIH 3T3 cells stimulated by the platelet-derived <em>growth</em> <em>factor</em> (PDGF) express a marked increase in levels of PTN mRNA whereas retinoic acid failed to increase levels of PTN mRNA in NIH 3T3 cells or in F9 embryonal carcinoma cells within 72 hours of exposure. The results suggest that expression of the PTN gene is highly restricted and that the PTN gene is a new member of the PDGF-induced cytokine family.

Liposomes made by sonication of egg yolk phosphatidyl choline support the proliferation of low-density bovine vascular and corneal endothelial cells, and vascular smooth muscle cells maintained on basement laminacoated dishes and exposed to a defined medium supplemented with transferrin. The optimal <em>growth</em>-promoting effect of phosphatidyl choline was observed at concentrations of 25 micrograms/ml for low-density cultures of vascular smooth muscle cells, and 100 micrograms/ml for vascular and corneal endothelial cells. The <em>growth</em> rate and final cell density of vascular endothelial cells exposed to a synthetic medium supplemented with transferrin and either high-density lipoproteins or phosphatidyl choline has been compared. Although cultures exposed to phosphatidyl choline reached a final cell density similar to that of cultures exposed to high-density lipoproteins, they had a longer average doubling time (<em>17</em> h vs. 12 h) during their logarithmic <em>growth</em> phase and a shorter lifespan (<em>17</em> generations vs. 30 generations). Similar observations were made in the case of vascular smooth muscle cells or bovine corneal endothelial cells maintained in medium supplemented with transferrin, <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) or epidermal <em>growth</em> <em>factor</em> (EGF), and insulin and exposed to either high-density lipoproteins or phosphatidyl choline. Since phosphatidyl choline can, for the most part, replace high-density lipoproteins in supporting the proliferation of various cell types, it is likely that the <em>growth</em> stimulating signal conveyed by high-density lipoproteins is associated with its polar lipid fraction, which is composed mostly of phosphatidyl cholines.

BACKGROUND

Unlike other IL-<em>17</em> family members, the Th2-derived cytokine IL-25 (IL-<em>17</em>E) induces (promotes) Th2 responses. One or both of the two receptors for IL-25 (IL-<em>17</em>RA, IL-<em>17</em>RB) is expressed on inflammatory cells and tissue structural cells, suggesting that in addition to promoting Th2-type inflammation IL-25 may also act on structural cells at sites of Th2-type inflammation such as in the asthmatic bronchial mucosa to promote remodelling changes.

OBJECTIVE

Our previous studies showed elevated expression of IL-25 and IL-<em>17</em>RB immunoreactivity in asthmatic airways with co-localization of the latter to endothelial cells. We therefore hypothesized that IL-25 acts on endothelial cells through this receptor to induce production of the key angiogenic and remodelling cytokine basic fibroblast growth factor (bFGF).

METHODS

Polymerase chain reaction (PCR) immunocytochemistry/immunohistochemistry and ELISA were employed to detect expression of IL-<em>17</em>RB, IL-<em>17</em>RA and bFGF by human vascular endothelial cells (HUVEC) and immunoreactivity for IL-25 and bFGF in asthmatic bronchial biopsies. Receptor-blocking antibodies, PCR and an in vitro angiogenesis assay were used to investigate whether IL-25 acts on IL-<em>17</em>RB or IL-<em>17</em>RA to induce bFGF expression and angiogenesis. PCR was also employed to investigate the signalling pathways involved in IL-25-mediated bFGF expression.

RESULTS

HUVEC constitutively expressed IL-<em>17</em>RB, IL-<em>17</em>RA and bFGF. Production of the latter was further increased by IL-25, but attenuated after blockade of the IL-<em>17</em>RB, but not the IL-<em>17</em>RA receptor. Neutralization of endogenous VEGF and bFGF completely abrogated IL-25-induced angiogenesis which was also inhibited by blocking IL-<em>17</em>RB, but not IL-<em>17</em>RA. The PI3K-specific inhibitor LY294002 also completely attenuated IL-25-induced bFGF expression. Immunoreactivity for IL-25 and bFGF was elevated in the asthmatic bronchial mucosa and the expression of each correlated with the other.

CONCLUSIONS

Our data support the hypothesis that IL-25 contributes to elevated bFGF in asthmatic airways by acting on the endothelial cell IL-<em>17</em>RB receptor through PI3K-signalling pathways. Targeting the pathways might benefit therapy of airways remodelling.

Fetal skeletal dysplasias are a heterogeneous group of rare genetic disorders, affecting approximately 2.4-4.5 of 10,000 births. We performed a retrospective review of the perinatal autopsies conducted between the years 2002-2011 at our center. The study population consisted of fetuses diagnosed with skeletal dysplasia with subsequent termination, stillbirth and live-born who died shortly after birth. Of the 2002 autopsies performed, 112 (5.6%) were diagnosed with skeletal dysplasia. These 112 cases encompassed <em>17</em> of 40 groups of Nosology 2010. The two most common Nosology groups were osteogenesis imperfecta [OI, 27/112 (24%)] and the <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor type 3 (FGFR3) chondrodysplasias [27/112 (24%)]. The most common specific diagnoses were thanatophoric dysplasia (TD) type 1 [20 (<em>17</em>.9%)], and OI type 2 [20 (<em>17</em>.9%)]. The combined radiology, pathology, and genetic investigations and grouping the cases using Nosology 2010 resulted in a specific diagnosis in 96 of 112 cases.

The aim of this study was to investigate the prevalence of <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 (FGFR1) amplification by fluorescence in situ hybridization (FISH) in a lung cancer patient cohort and to correlate results with morphology, silver in situ hybridization (SISH), and patient outcome. FGFR1 FISH and SISH were performed in 406 and 385 lung cancer cases, respectively, and the results were compared. High-level FGFR1 amplification was defined as the ratio of FGFR1/centromere 8 ≥2, or tumor cell percentage with ≥15 signals ≥10%, or average number of signals/tumor cell nucleus ≥6. Low-level amplification was defined as tumor cell percentage with ≥5 signals ≥50%. Of 406 tumors tested, there were 191 squamous cell carcinomas, 28 carcinomas with focal squamous morphology, 24 large cell carcinomas with squamous immunoprofile, 115 adenocarcinomas, <em>17</em> neuroendocrine tumors, and 31 carcinomas without squamous morphology or immunoprofile. FGFR1 FISH was assessable in 368 tumors, with FGFR1 amplification identified in 50, including 48 tumors with either squamous morphology or immunoprofile (48 of 225, 21.3%), and two 'marker-null' tumors without squamous or glandular morphology or immunoprofile (2 of 143, 1.4%; P<0.0001). FGFR1 SISH was assessable in 347 tumors. All 46 FGFR1 FISH-amplified tumors with tumor available for testing showed amplification with SISH, while all other tumors were negative. There was no relationship between FGFR1 amplification status and disease-free (P=0.88, HR=1.04, 95% confidence interval (CI)=0.67-1.60) or overall survival (P=0.97, HR=1.01, 95% CI=0.65-1.58) in surgically radically treated patients with tumors with any squamous morphology or immunoprofile. FGFR1 amplification is a common abnormality in tumors with any squamous morphology or immunoprofile, but it is also present in 'marker-null' tumors. The results of FGFR1 SISH showed 1:1 correlation with the results of FGFR1 FISH, indicating that SISH may be an alternative method to detect FGFR1 amplification. No relationship was detected between patient outcome and FGFR1 amplification.

BACKGROUND

Atrial fibrillation (AF) is associated with increased mortality and morbidity, including risk for cerebral macro- and microinfarctions and cognitive decline, even in the presence of adequate oral anticoagulation. AF is strongly related to increased inflammatory activity whereby anti-inflammatory agents can reduce the risk of new or recurrent AF. However, it is not known whether anti-inflammatory therapy can also modify the deterioration of neurocognitive function in older patients with AF. In the present study, older patients with AF were treated with intensive lipid-lowering therapy with atorvastatin 40 mg and ezetimibe 10 mg, or placebo. We examined the relationship between neurocognitive functions and inflammatory burden.

RESULTS

Analysis of inflammatory markers revealed significant reductions in high sensitivity C-reactive protein (hs-CRP), <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF), granulocyte colony-stimulating <em>factor</em> (G-CSF), granulocyte-macrophage colony-stimulating <em>factor</em> (GM-CSF), interleukin-1 receptor antagonist (IL-1RA), interleukin (IL)-9, IL-13 and IL-<em>17</em>, and interferon-γ (IFNγ) in the treatment group compared to placebo. Reduction in plasma concentration of IL-1RA, IL-2, IL-9 and IL-12, and macrophage inflammatory protein-1β (MIP-1β) correlated significantly with improvement in the neurocognitive functions memory and speed. Loss of volume in amygdala and hippocampus, as determined by magnetic resonance imaging (MRI), was reduced in the treatment arm, statistically significant for left amygdala.

CONCLUSIONS

Anti-inflammatory therapy through intensive lipid-lowering treatment with atorvastatin 40 mg and ezetimibe 10 mg can modify the deterioration of neurocognitive function, and the loss of volume in certain cerebral areas in older patients with AF. TRIAL REGISTRATION ClinicalTrials.gov: NCT00449410.

BACKGROUND

Recent studies have shown the possible role of growth factors and the involvement of macrophages as a source of them in the pathogenesis of bronchiolitis obliterans (BO) after lung transplantation.

OBJECTIVE

The authors intended to determine whether depletion of recipient macrophages by gadolinium chloride (GdCl3) resulted in decreased obliterative airway disease (OAD) in a rat model of heterotopic tracheal transplantation.

METHODS

A tracheal segment of donor rats (Brown Norway) was transplanted into a subcutaneous pouch of fully major histocompatibility complex-incompatible recipient rats (Lewis). Recipients were injected intravenously with 80 mg/kg of GdCl3.6H2O or saline on days 0, 7, and 14 posttransplant. Allografts were harvested on days 7, 14, 17, and 21 and the degree of OAD resulting from fibroproliferative tissue was pathologically scored on a scale of 0 to 4. A portion of allografts was submitted to reverse-transcriptase polymerase chain reaction analysis to examine mRNA expression for platelet-derived growth factor (PDGF), basic fibroblast growth factor, and transforming growth factor-beta1.

RESULTS

Immunohistochemical studies confirmed reduction in the number of ED2+ macrophages in tracheal allografts by GdCl3 injection. GdCl3 treatment significantly decreased OAD of allografts, with the histologic score of 1.4+/-0.3 in the treated animals compared with 3.0+/-0.5 in the controls (mean+/-SE, P=0.02) at day 21 posttransplant, and this was accompanied by decreased PDGF-A and PDGF-B gene expression in the GdCl3 group at day 17 posttransplant.

CONCLUSIONS

Macrophage reduction by GdCl3 resulted in significantly decreased OAD development and reduced PDGF mRNA expression in allografts. This suggests a potential effectiveness of therapies targeting recipient macrophages in preventing BO after lung transplantation.

A strategy was developed for the purification of a biologically active polypeptide <em>growth</em> and migration <em>factor</em> from skimmed bovine milk. This 25-kDa dimeric molecule, termed milk <em>growth</em> <em>factor</em> (MGF), was isolated by a method consisting of a combination of strong cation-exchange chromatography, low-pressure hydrophobic-interaction chromatography, hydrophobic-interaction HPLC and size-exclusion HPLC steps, which separated the protein according to its properties of charge, hydrophobicity and size, respectively. On average, a total purification of 10(6)-10(7)-fold and a yield of approximately 115 +/- 78 ng/MGF/milk was obtained using the method described. All purification steps were performed with novel combinations of ethanol and volatile acidic salt (ammonium acetate) solutions in order to retain biological activity of the protein. These conditions, together with the easy removal of salt by lyophilization, facilitated the detection of biological activity in fractions collected at each step of the purification by means of a sensitive in vitro <em>fibroblast</em>-migration assay in which the half-maximal activity was obtained at a concentration of approximately <em>17</em> +/- 4 pg/ml (i.e. approximately 1 pM) of the pure protein. Biological activity of the dimeric protein was unaffected by heat treatment or exposure to acid (pH 2.0), but was lost upon reduction to its monomeric form. Amino acid composition and sequence analyses demonstrate that MGF is related to transforming <em>growth</em> <em>factor</em> type beta 2.

OBJECTIVE

Chondroitin sulfate (CS) is a glycosaminoglycan in the extracellular matrix of all vertebrates. A biocompatible, nonimmunogenic, pliable hydrogel preparation of CS has recently been produced and has shown benefit in wound healing in murine and porcine epidermis. The objective of the current experiment is to compare the wound healing properties of CS hydrogel versus no treatment in wounds of the maxillary sinus mucosa.

METHODS

Prospective investigation in an animal model.

METHODS

A 6 mm wound was created in bilateral maxillary sinuses of <em>17</em> New Zealand white rabbits. CS hydrogel (case) and no dressing (control) were randomly assigned to one side each as wound treatment. Wounds were examined ex vivo at 2, 4, 6, 10, and 14 day postinjury intervals. Wound diameter was measured microscopically by a blinded investigator. Representative specimens were examined histologically.

RESULTS

The CS disc was partially integrated into the wounds at the 4-day interval and completely integrated by the 6-day interval. The average wound diameters for the case versus control side were similar at 2 days (3.75 mm vs. 4.42 mm) but differed significantly at 4 days (2.86 mm. vs. 3.80 mm., P =.035). At 6 days, the wounds could not be discerned on either the case or control sides. However, histologic analysis revealed accelerated healing with the CS treatment. The treated wounds displayed respiratory epithelium as opposed to the squamous epithelium exhibited on the untreated sides.

CONCLUSIONS

Despite some limitations, the New Zealand white rabbit is an effective model for the study of sinonasal wound healing. CS hydrogel accelerates wound healing in sinonasal mucosa at a 4-day endpoint. We propose that the CS hydrogel acts as a surrogate extracellular matrix, serving as a repository for cytokines and growth factors produced by the regenerating mucosa. It may also provide a structural framework for fibroblasts and epithelial regeneration. Further study is necessary to establish this relationship.

<em>Fibroblast</em> <em>growth</em> <em>factor</em>-2 (FGF-2) is made by osteoblasts and modulates their function. There are high molecular weight (HMW) protein isoforms of FGF-2 that have nuclear localization sequences and a low molecular weight (LMW) 18 kDa FGF-2 protein that is exported from cells. Since FGF-2 is a trophic <em>factor</em> and potent mitogen for osteoblasts, the goal of this study was to utilize targeted overexpression of FGF-2 as a novel means of assessing different FGF-2 isoforms on osteoblastic cell viability and proliferation. Either LMW or HMW human Fgf2 cDNAs were cloned downstream of 3.6 kb alpha1(I)-collagen 5' regulatory elements (Col 3.6). A set of expression vectors, called Col3.6-Fgf2 isoforms-IRES-GFPsaph, capable of concurrently overexpressing either LMW or HMW FGF-2 isoforms concomitant with GFPsaph from a single bicistronic mRNA were built. Viable cell number in ROS <em>17</em>/2.8 cells stably transfected with Vector (Col3.6-IRES-GFPsaph) versus each of the Col3.6-Fgf2-IRES-GFPsaph constructs were compared. In the presence of 1 or 10% serum, DNA synthesis was increased in cells expressing any isoform of FGF-2 compared with vector. However, cells transfected with HMW isoform had augmented DNA synthesis in 1 or 10% serum compared with cells expressing either ALL or LMW FGF-2 isoforms. A neutralizing FGF-2 antibody significantly reduced the mitogenic response in cells harboring ALL or the LMW FGF-2 isoforms but did not block the mitogenic effect of cells harboring the HMW isoforms. In summary, overexpression of any isoform of FGF-2 protein increased viable cell number and OB proliferation in the presence of low or high concentrations of serum. However, the HMW/nuclear isoforms preferentially mediate augmented OB proliferation. We conclude that differential expression of FGF-2 proteins isoforms is important in modulating OB function.

Infant leukemia cells with 46XY,t(11; <em>17</em>)(q23; p13) karyotype and a hybrid pre B myeloid phenotype (HLA-DR, (Ia), B4 and My7-positive and CALLA and T11-negative) and immunoglobulin heavy chain gene rearrangement were maintained in long-term culture for over 10 months. The in-vitro survival and <em>growth</em> of the leukemia cells were strictly dependent upon the presence of their autologous marrow stromal cells. The latter could be replaced by the 14F1.1 clone of preadipocytes derived from mouse bone marrow. Neither heterologous human marrow or foreskin <em>fibroblasts</em> nor <em>fibroblast</em> or endothelial like cell lines from mouse stroma could mimic the effect of autologous stroma or 14F1.1 adipocytes. The leukemia cells maintained their original phenotype throughout the 10-month culture period with either their autologous stroma or the 14F1.1 adipocytes. They could be induced to differentiate in two distinct directions. Phorbol myristate acetate induced adherence of the leukemia cells and development of macrophage properties. In contrast, conditioned medium from a hybridoma producing B-cell <em>growth</em> <em>factor</em> caused aggregation of the leukemia cells and expression of CALLA antigen and surface IgM. This bipotency of the leukemia cells and their dependence upon marrow stroma are properties in common with stem cells.

Normal <em>growth</em> and differentiation of the lung depends upon mesenchymal-epithelial interactions during development. Recombination experiments using immature (Day <em>17</em>) and mature (Day 21) fetal rat lung <em>fibroblasts</em> (FRLF) revealed that the stimulatory effect of mature <em>fibroblasts</em> on fetal type II epithelial cells is blocked by immature <em>fibroblasts</em>. Similarly, conditioned medium from Day <em>17</em> FRLFs blocks the stimulatory effect (<em>fibroblast</em>-pneumonocyte <em>factor</em>) of Day 21 conditioned medium on type II epithelial cells. This blocking activity is nondialyzable, trypsin sensitive, and heat stable. Its activity is neutralized by an antibody to TGF beta, in both conditioned media and recombined cell studies, and its activity is mimicked by TGF beta. Developmentally, TGF beta-like activity is present in conditioned medium from 15- to 19-day FRLF, decreasing precipitously between 19 and 21 days gestation. Northern blot analysis of mRNAs from fetal rat lung <em>fibroblasts</em> on Days <em>17</em>, 19, and 21 revealed expression of TGF beta at all three stages of development.

OBJECTIVE

Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), a <em>17</em>- to 24-kDa protein known to be essential for the survival of neurons, induced fiber out<em>growth</em> of ganglion cells in cultures of rat retina and rescued photoreceptor cell loss in the retina of Royal College of Surgeon rats. The authors evaluated the efficacy of bFGF in rescuing the neuronal loss in rat retina after retinal ischemia.

METHODS

Retinal ischemia was induced in 29 eyes of <em>17</em> albino Lewis rats by increasing the intraocular pressure to 110 mm Hg for 45 minutes via an intracameral catheter. A total of 800 ng of bFGF was delivered into the anterior chamber at the time of induction of ischemia. Sixteen eyes of nine rats received bFGF, and 13 eyes of eight rats received heparin in phosphate-buffered saline as vehicle control. The animals were euthanized 7 or 14 days after reperfusion.

RESULTS

Morphologic examination of the retinas at both time points showed that necrosis of the retinal ganglion cells (RGCs) and thinning of the inner plexiform and inner nuclear layers were less severe in the bFGF-treated eyes than in the vehicle-treated eyes. On morphometric examination, 7 days after reperfusion, the mean thickness of the inner retinal layers and the RGC counts on flat preparations of retina in both the posterior and the peripheral portions of the retina were significantly higher in the bFGF-treated eyes than in the vehicle-treated eyes (P < 0.02). At 14 days, similar beneficial effects were noted in all morphometric parameters, except RGC counts in the posterior pole.

CONCLUSIONS

These results demonstrate that bFGF partially protects the RGCs and other inner retinal elements from ischemic injury.

The corpus luteum (CL) of the estrous cycle in the cow is a dynamic organ which has a life time of approximately <em>17</em>-18 days. The main function of the CL is to secrete a large amount of progesterone (P) thereby supporting the achievement of pregnancy. As the CL matures, the steroidogenic cells establish contact with many capillaries and the matured CL is composed of many vascular endothelial cells that account for up to 50% of all CL cells. The bovine CL produces several major angiogenic and vasoactive foctors such as vascular endothelial <em>growth</em> <em>factor</em> (VEGF), basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), angiopoietin-1 and -2 (ANPT-1 and -2), prostaglandin F(2alpha) (PGF(2alpha)), endothelin-1 (EDN1), angiotensin II (Ang II) and nitric oxide (NO). These <em>factors</em> regulate P secretion directly and/or indirectly within the CL. Moreover, different actions of PGF(2alpha) in the early cycle CL (non-luteolytic) and the mid cycle CL (luteolytic) may provide insight into the luteolysis cascade in the cow. The aim of the present review is to describe the current concepts of the local mechanisms for the cascade of development and regression of the bovine CL as regulated by luteal angiogenic and vasoactive <em>factors</em>.

Left ventricular hypertrophy is an independent risk factor for morbidity and mortality in patients with hypertensive heart disease. Cardiac hypertrophy, associated with increased cardiac fibrosis and myocardial relaxation impairment, shows gender-based differences with significantly higher mortality in men. The role of estrogen in the pathogenesis of this process is poorly understood. After our previous demonstration that cardiac myocytes and fibroblasts contain functional estrogen receptors, we therefore investigated: 1) the influence of different estrogen metabolites on cardiac fibroblast growth; 2) the influence of different estrogen metabolites on the expression of the immediate early gene c-Fos; 3) the influence of estrogen on the L-type calcium channel in cardiomyocytes.

METHODS

1) Neonatal rat cardiac fibroblasts were incubated with 17 beta-estradiol, estrone, 2-hydroxyestrone, and 2-methoxyestradiol (all 10(-9) M). Bromodeoxyuridine incorporation was measured after 24 h. 2) c-Fos expression was demonstrated by immunoblotting. 3) L-type (Ca2+) current with and without 17 beta-estradiol was assessed in adult guinea pig cardiomyocytes by whole cell patch clamp.

RESULTS

Cardiac fibroblast growth was stimulated by estrogen metabolites with 2-hydroxyestrone as the most potent activator; in addition, 10(-5) M 17 beta-estradiol reduced the L-type Ca2+ current by about 20% in cardiomyocytes.

CONCLUSIONS

Estrogen induces both short term effects (non-genomic) and long term effects (genomic) on the heart and may therefore account for gender- and age-based differences in hypertensive heart disease.

BACKGROUND

It is still being discussed if the assessment of basal markers or if adhesion molecules expression contributes additional prognostic information to the classic prognostic factors and hence should be included into standard morphologic reports.

METHODS

The aim of the study was to assess the prognostic significance of: (i) classification recommended by St Gallen experts (ii) tumor grade, expression of (iii) basal markers, (iv) adhesion molecules, and (v) matrix metalloproteinase 2 (MMP-2) in patients with T1-T2 N0M0 chemotherapy-naive ductal breast cancer.

RESULTS

In 79 patients with tumors characterized by estrogen receptor (ER) and progesterone receptor (PgR) positive, human epidermal growth factor receptor 2 negative (HER2) phenotype and MIB-1 labeling index (MIB-l) LI ≤ 15% (low-risk group) cumulative 17-year breast cancer-specific survival probability was 100% and was significantly higher than in 95 patients from the high-risk group (ER(-)/PgR(-)/HER2(-) or HER2(+) or MIB-1 LI>> 15%) (72.5%). We found that MMP-2 fibroblast expression indicated 2 subgroups with significantly different survival rates in women with grade 3 tumor (88.9% for MMP-2 positivity and 56.0% for negativity). Cox multivariate analysis revealed that both grade 3 combined with stromal fibroblast MMP-2(-) and a high-risk group according to St Gallen recommendations are independent negative prognostic factors that influence survival of patients with breast cancer.

CONCLUSIONS

To the best of our knowledge, we have shown for the first time that MMP-2(-) in stromal fibroblasts might indicate poor survivors in the group of patients with grade 3 tumors and that the cumulative effect of both above-mentioned parameters might be helpful in selecting the high-risk individuals from the group of patients with luminal B subtype/HER2(+)/triple negative phenotype identified according to St Gallen recommendations.

Endothelin-1 (Et-1) stimulated DNA synthesis in placental <em>fibroblasts</em> in a dose-dependent manner, as measured by [3H]thymidine incorporation (ED50, 0.2-0.3 ng/mL). Insulin-like <em>growth</em> <em>factor</em>-I (IGF-I) interacted synergistically with Et-1 to potentiate the stimulation of DNA synthesis. Additionally, Et-1 stimulated the turnover of phosphoinositides in a time- and concentration-dependent manner (ED50, 1 ng/mL), as measured by a 2- to 3-fold increase in the total accumulation of [3H]inositol phosphates. This was accompanied by a 2- to 3-fold rise in intracellular calcium, as measured by fura-2 fluorescence. IGF-I, however, had no potentiating effect on Et-1-stimulated phosphoinositide turnover or increase in cytosolic Ca2+. The ability of Et-1 to stimulate the production of IGF-II and IGFBPs by placental <em>fibroblasts</em> was then studied. Western ligand blot analysis using an [125I]IGF-II probe revealed the presence of six major binding proteins corresponding to 42, 38, 35, 32, 31, and 24 kilodaltons. Et-1 (50 ng/mL) stimulated all binding fractions concordantly. This was accompanied by a similar increase in immunoreactive IGF-II secretion, as assessed by a specific RIA. No increase in immunoreactive IGF-I was observed. The ability of the placenta to produce Et-1 was examined by Northern blot analysis. Placentae at 14 and <em>17</em> weeks gestation expressed small amounts Et-1 mRNA, whereas significantly higher levels of mRNA were expressed at term. These data suggest that the human placenta produces Et-1 in a developmentally regulated manner that may act via paracrine and/or autocrine mechanisms to regulate placental <em>growth</em>.

We examined the localization of basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (basic FGF) in the adult rat brain by immunohistochemical and Western blotting analysis using a specific antibody against a synthetic basic FGF fragment (N-terminal 12 residues). The antibody did not cross-react with acidic FGF. Basic FGF-like immunoreactivity was located exclusively in the neuronal elements and had very heterogeneous distribution. Immunoreactive cell bodies were observed in the paraventricular, supraoptic and circular nuclei of the hypothalamus. Numerous immunoreactive neuronal processes originating from these basic FGF-positive cells extended lateroventrally and then caudally to the internal layer of the median eminence. In addition, the neurohypophysis contained a significant number of basic FGF-like immunoreactive fibers. Western-blotting analysis revealed that the hypothalamus and the hypophysis contained a main band of basic FGF immunoreactive with an apparent molecular weight of <em>17</em> kDa. These results show that the hypothalamo-hypophyseal neuroendocrine pathway contains basic FGF.

Thalidomide (Thal) is a drug with anti-angiogenic properties. To explore whether the effect of Thal on angiogenesis is associated with a reduction of angiogenic cytokine levels in progressive multiple myeloma (MM), plasma levels of basic <em>fibroblast</em> <em>growth</em> <em>factor</em>, vascular endothelial <em>growth</em> <em>factor</em>, interleukin 6, tumour necrosis <em>factor</em>-alpha and hepatocyte <em>growth</em> <em>factor</em> (HGF) were measured in 51 patients at 0, 3 and 6 months of Thal therapy. After 6 months of treatment, 26 patients were considered to be responsive to Thal therapy, including <em>17</em> minimal responses, eight partial responses and one complete response. Only HGF (decreasing, P = 0.02) in the group of responsive patients showed a statistically significant change over a period of 6 months. Because HGF levels are known to correlate to MM tumour burden, we conclude that the mechanism of action of Thal in MM is not caused by a specific inhibition of angiogenic cytokine secretion.

Cytokines are actively secreted by the respiratory mucosa of preterm children and participate in the pathogenesis of wheezing. This study aimed to identify the <em>factors</em> that could potentially influence respiratory secretion of cytokines in these children. A nasopharyngeal aspirate (NPA) was collected from 77 preterm children 1 yr after birth. NPAs from 14 healthy, 1-yr-old term children were collected in parallel. 27 cytokines were measured in the NPAs using a multiplex assay. Multivariate stepwise regression analysis with Bonferroni correction evidenced that the variable [daycare attendance] was associated with higher levels of [monocyte chemoattractant protein-1 (MCP-1), IL-6, vascular endothelial <em>growth</em> <em>factor</em> (VEGF), IL-1β, IL-10, tumor necrosis <em>factor</em> (TNF)-α]; [male sex] with higher levels of (MCP-1, VEGF, and IL-1β); [smokers at home] was associated with higher levels of MCP-1 (p < 0.0013). In turn, [prophylaxis with palivizumab] was associated with lower levels of (IL-6, IL-7) (p < 0.0013). All these mediators participate in the pathogenesis of asthma and recurrent wheezing. Preterm children secreted higher levels of chemokines (interferon-gamma inducible protein-10, macrophage inflammatory protein-1α, Eotaxin, MCP-1), <em>growth</em> <em>factors</em> (platelet-derived <em>growth</em> <em>factor</em>-bb, VEGF, <em>fibroblast</em> <em>growth</em> <em>factor</em>-basic, granulocyte macrophage colony-stimulating <em>factor</em>), Th1 (IL12, interferon-γ), Th2 (IL-9, IL-13), Th<em>17</em> (IL-6, IL-<em>17</em>) cytokines, and immunomodulatory mediators (IL1RA and granulocyte colony-stimulating <em>factor</em>) than term children. In conclusion, we have identified for the first time a group of individual and environmental <em>factors</em> influencing respiratory secretion of cytokines in preterm children at the long term after birth. To know these <em>factors</em> could help to prevent the instauration of conditions linked to the appearance of chronic respiratory diseases such as wheezing or asthma.