Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) inhibits radiation-induced apoptosis, and radioprotects haematopoietic, cartilage <em>growth</em> plate, pulmonary and gastrointestinal tissues. Conversely, chronic overexpression of bFGF may promote fibrosis. We measured the endogenous circulating bFGF in blood of patients undergoing conditioning TBI. Twenty-six patients with haematopoietic malignancies were conditioned with cyclophosphamide/TBI for allogeneic BMT. Daily blood samples were collected each morning prior to, during, and for several days after TBI. bFGF levels in plasma of normal volunteers are 0.8-26 pg/ml. bFGF was below detectability in <em>22</em>%, 30% and 45% of patients pre-TBI, during TBI or post-TBI respectively. Mean circulating plasma levels of bFGF decreased from a median of 52 pg/ml pre-TBI to 26 pg/ml during TBI, and to 5 pg/ml post-TBI. Among the 26 patients, 13 had more than one non-detectable plasma bFGF level, an additional five had at least one non-detectable level, and only eight patients had detectable levels in all daily samples. Naturally high levels of bFGF were observed in some patients undergoing fractionated TBI. In contrast, as many as 79% of patients had low bFGF levels in one or more samples. The impact of endogenous bFGF on the tolerance of normal tissues to irradiation is unknown, and warrants further study.

BACKGROUND

We recently reported that ramipril more than doubled maximum walking times in patients with peripheral artery disease with intermittent claudication.

OBJECTIVE

Our aim was to conduct exploratory analyses of the effects of ramipril therapy on circulating biomarkers of angiogenesis/arteriogenesis, thrombosis, inflammation, and leukocyte adhesion in patients with intermittent claudication.

RESULTS

One hundred sixty-five patients with intermittent claudication (mean, 65.3 [SD, 6.7] years) were administered ramipril 10 mg per day (n=82) or matching placebo (n=83) for 24 weeks in a randomized, double-blind study. Plasma biomarkers of angiogenesis/arteriogenesis (vascular endothelial <em>growth</em> <em>factor</em>-A, <em>fibroblast</em> <em>growth</em> <em>factor</em>-2), thrombosis (D-dimer, von Willebrand <em>factor</em>, thrombin-antithrombin III), inflammation (high-sensitivity C-reactive protein, osteopontin), and leukocyte adhesion (soluble vascular cell adhesion molecule-1, soluble intracellular adhesion molecule-1) were measured at baseline and 24 weeks. Relative to placebo, ramipril was associated with increases in vascular endothelial <em>growth</em> <em>factor</em>-A by 38% (95% confidence interval [CI], 34%-42%) and <em>fibroblast</em> <em>growth</em> <em>factor</em>-2 by 64% (95% CI, 44-85%; P<0.001 for both), and reductions in D-dimer by 24% (95% CI, -30% to -18%), von Willebrand <em>factor</em> by <em>22</em>% (95% CI, -35% to -9%), thrombin-antithrombin III by 16% (95% CI, -19% to -13%), high-sensitivity C-reactive protein by 13% (95% CI, -14% to -9%), osteopontin by 12% (95% CI, -14% to -10%), soluble vascular cell adhesion molecule-1 by 14% (95% CI, -18% to -10%), and soluble intracellular adhesion molecule-1 by 15% (95% CI, -17% to -13%; all P<0.001). With the exception of von Willebrand <em>factor</em>, all the above changes correlated significantly with the change in maximum walking time (P=0.02-0.001) in the group treated with ramipril.

CONCLUSIONS

Ramipril is associated with an increase in the biomarkers of angiogenesis/arteriogenesis and reduction in the markers of thrombosis, inflammation, and leukocyte adhesion. This study informs strategies to improve mobility in patients with intermittent claudication.

UNASSIGNED

http://clinicaltrials.gov. Unique identifier: NCT00681<em>22</em>6.

OBJECTIVE

To characterize the presence of mitogen(s) in the peritoneal fluid (PF).

METHODS

Aliquots of PF aspirated at laparoscopy were assessed for mitogenic activity by the rate of [3H]-thymidine incorporation into the deoxyribonucleic acid of various cell lines.

METHODS

Peritoneal fluids were obtained from patients having a laparoscopy for the investigation of infertility or pelvic pain or for tubal ligation.

METHODS

Seven women with laparoscopic evidence of endometriosis (stage I and II) and six without evidence of endometriosis.

METHODS

None.

METHODS

After 24 hours of culture in absence of serum, NIH/3T3 mouse embryo <em>fibroblasts</em>, (KLE) human endometrial adenocarcinoma cells, and primary cultures of rabbit endometrial cells were incubated in the presence of appropriate amounts of PF or bovine serum albumin (control) for <em>22</em> hours before adding the [3H]-thymidine for 2 hours.

RESULTS

Aliquots of PF containing greater than 100 micrograms/mL total protein concentration stimulated the proliferation of all cell types. This mitogenic effect was dose-dependent and was greatest on endometrial-like cells. The mitogenic activity was fully recovered after adsorption on dextran-coated charcoal and was sensitive to tryptic digestion and to heat. It could not be retained on cartridge of C18 silica and was calculated to have a molecular weight greater than 30,000 by gel permeation chromatography.

CONCLUSIONS

Peritoneal fluid from women with or without endometriosis contain growth factor(s) that are proteinaceous in nature and capable of stimulating the proliferation of endometrial-like cells and fibroblasts. These mitogens could play a role in the pathogenesis of endometriosis.

Hyaluronic acid (HA), type III procollagen, fibronectin, and <em>fibroblast</em> <em>growth</em> <em>factors</em> (FGF) were measured in 43 bronchoalveolar lavage fluid (BALF) specimens obtained from 38 patients with farmer's lung (FL) and in BALF of 9 nonexposed normal control subjects. Bronchoalveolar lavage was done in 21 farmers with acute FL (acute) and in <em>22</em> with a history of previous FL (Ex) who were still in daily contact with dairy barns. All farmers from the acute and Ex groups had a lymphocytic alveolitis, respectively, 62.7 (3.5) percent (mean [SEM]) and 48.1 (4.3) percent. Hyaluronic acid, type III procollagen, fibronectin, and FGF were all highly increased in acute disease. These substances were also increased in the BALF of subjects of the Ex group who had no clinical symptoms or signs of acute disease at the time of lavage, but were actively farming. The increase in type III procollagen, however, was less in this group than in the subjects with acute disease. These observations suggest that the fibrosing activities and potentialities of the allergic alveolitis of FL are fully expressed at the time of clinical presentation and also in the subclinical phase of the disease in susceptible farmers who remain exposed after an initial acute phase of the disease.

Scaffolds, <em>growth</em> <em>factors</em>, and cells are three essential components in regenerative medicine. Nonwoven filters, which capture cells, provide a scaffold that localizes and concentrates cells near injured tissues. Further, the cells captured on the filters are expected to serve as a local supply of <em>growth</em> <em>factors</em>. In this study, we investigated the <em>growth</em> <em>factors</em> produced by cells captured on nonwoven filters. Nonwoven filters made of polyethylene terephthalate (PET), biodegradable polylactic acid (PLA), or chitin (1.2-<em>22</em> μm fiber diameter) were cut out as 13 mm disks and placed into cell-capturing devices. Human mesenchymal stem cells derived from adipose tissues (h-ASCs) and peripheral blood cells (h-PBCs) were captured on the filter and cultured to evaluate <em>growth</em> <em>factor</em> production. The cell-capture rates strongly depended on the fiber diameter and the number of filter disks. Nonwoven filter disks were composed of PET or PLA fibers with fiber diameters of 1.2-1.8 μm captured over 70% of leukocytes or 90% of h-ASCs added. The production of vascular endothelial <em>growth</em> <em>factor</em> (VEGF), transforming <em>growth</em> <em>factor</em> β1, and platelet-derived <em>growth</em> <em>factor</em> AB were significantly enhanced by the h-PBCs captured on PET or PLA filters. h-ASCs on PLA filters showed significantly enhanced production of VEGF. These enhancements varied with the combination of the nonwoven filter and cells. Because of the enhanced <em>growth</em> <em>factor</em> production, the proliferation of human <em>fibroblasts</em> increased in conditioned medium from h-PBCs on PET filters. This device consisting of nonwoven filters and cells should be investigated further for possible use in the regeneration of impaired tissues.

OBJECTIVE

To investigate the relationship of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) and proliferation of endogenous neural stem cells (NSCs) after human cerebral infarction.

METHODS

Paraffin-embedded brain tissues of 22 human fatal cases of CI from the brain tissues around subventricular zone and subgranular layer zone were stained with HE and immunohistochemistry stain. The endogenous neural stem cells were marked by nestin. The expression changes of EGF, bFGF and nestin in the perihematomal tissues were analysed with the SPSS 13.0 system.

RESULTS

(1) Compared with the controls, the number of nestin-positive cells increased at 24 - 72 h (14 +/- 6)/HP in the ipsilateral SVZ and began to rise at 4.5 - 10 h (11 +/- 5)/HP in the ipsilateral SGZ, reached maximum at 120 - 144 h ((38 +/- 7)/HP in the SVZ, (54 +/- 17)/HP in the SGZ, and decreased markedly at 216 - 336 h, but it was still elevated compared with the controls (P < 0.05). (2) The number of bFGF-positive cells increased at 4.5 - 10 h (8.1 +/- 2.9)/HP in the SVZ, (19.0 +/- 8.2)/HP in the SGZ, reached maximum at 24 - 70 h (15.6 +/- 3.5)/HP in the SVZ, (32.0 +/- 5.7)/HP in the SGZ and decreased at 72 - 96 h, but it was still elevated compared with the controls (P < 0.05). (3) The number of EGF-positive cells increased at 4.5 - 10 h (4.3 +/- 1.6)/HP in the SVZ, (7.0 +/- 3.7)/HP in the SGZ, reached maximum at 120 - 144 h (27.0 +/- 1.4)/HP in the SVZ, (51.5 +/- 4.9)/HP in the SGZ and decreased at 216 - 336 h, but it was still elevated compared with the controls (P < 0.05).

CONCLUSIONS

Perhaps the increased expression of EGF and bFGF after CI was a reaction of endogenous reparation and it correlated with the proliferation and endogenous of neural stem cells in human.

OBJECTIVE

To investigate the role of transforming growth factor beta1 (TGFbeta1) in the development of Helicobacter pylori (H.pylori)-associated non-metaplastic atrophic gastritis.

METHODS

The expressions of TGFbeta1, CD68 and smooth muscle actin(SMA) were detected immunohistochemically in 10 patients with mild non-atrophic gastritis, 30 patients with mild non-metaplastic atrophic gastritis, and 32 patients with severe non-metaplastic atrophic gastritis having H.pylori infecion. Meanwhile, three cases of mild non-atrophic gastritis and 4 cases of severe non-metaplastic atrophic gastritis were observed with electron microscope.

RESULTS

The count of TGFbeta1 positive cells per high-power field (HPF) in severe non-metaplastic atrophic gastritis group (53 +/- 22) was significantly higher than that in mild non-atrophic gastritis group(22+/-9/HPF) and mild non-atrophic gastritis group(0-3/HPF, P<0.01). The count of CD68 positive cells in severe non-metaplastic atrophic gastritis group (23+/-7/HPF) was significantly higher than that in mild non-atrophic gastritis group (13+/-6/HPF) and mild non-atrophic gastritis group(0-3/HPF, P<0.01). Correlation analysis showed that the expressions of TGFbeta1 and CD68 had a moderate correlation in each group (r=0.634, P<0.01; r=0.699, P<0.01). Compared with mild non-atrophic gastritis, SMA-positive myofibroblasts and smooth muscle cells in the lamina propria increased in mild and severe non-metaplastic atrophic gastritis. Ultrastructurally, the proliferation of fibroblasts in gastric lamina propria was observed in mild non-atrophic gastritis, while the proliferation of fibroblasts and presence of myofibroblasts could be observed in mild non-metaplastic atrophic gastritis, and there was a parallel phenomenon between myofibroblasts and fibroblasts, as well as smooth muscle cells.

CONCLUSIONS

Our findings indicate that TGFbeta1 expression increases with severity of H.pylori- associated non-metaplastic atrophic gastritis, suggesting that TGFbeta1 might play an important role in the development of non-metaplastic atrophic gastritis.

The LNCaP human prostate cancer cell line is androgenand stromal-dependent for in vivo <em>growth</em>. We co-inoculated LNCaP cells with human fetal <em>fibroblasts</em>, isolated from prostate, bone (male), and lung (male and female) derived from 18- to <em>22</em>-week-old human fetal tissue, into non-castrate male nude mice. Co-inoculation of LNCaP with fetal prostatic <em>fibroblasts</em> resulted in high tumor take rates (27 of 30, or 90%) 6 to 8 weeks after subcutaneous co-inoculation. Serum prostate specific antigen (PSA) values correlated strongly with wet tumor weight (r=0.86). The fetal <em>fibroblast</em> enhancement of tumor take rates in vivo was neither gender- nor organ-specific. Fetal <em>fibroblast</em>-conditioned medium (CM) did not have a significant proliferative effect on LNCaP cell <em>growth</em> in vitro. Areas of angiogenesis were demonstrable in all tumors, with blood vessels arising at the interface between stromal and tumor cells. The fetal <em>fibroblasts</em>, but not the LNCaP cells, expressed significant amounts of the mRNA and protein for vascular endothelial cell <em>growth</em> <em>factor</em> (VEGF). Treatment of tumor-bearing animals with neutralizing antibodies to VEGF resulted in significant tumor <em>growth</em> suppression. These findings indicate that VEGF is an important mediator of stromal-induced enhancement of human prostate cancer cell <em>growth</em> in vivo.

Platelet-derived <em>growth</em> <em>factor</em> (PDGF), an important bone cell mitogen, exists as a homo- or heterodimer product of the PDGF-A and -B genes. Normal unstimulated cells of the osteoblast lineage express the PDGF-A gene, but it is not known whether they express the PDGF-B gene. We examined the expression of PDGF-B messenger RNA (mRNA) levels in cultures of osteoblast-enriched cells from <em>22</em>-day-old fetal rat calvariae (Ob cells) and determined whether they were modified by transforming <em>growth</em> <em>factor</em>-beta 1 (TGF beta 1), basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), insulin-like <em>growth</em> <em>factor</em> I (IGF-I), and PDGF-BB. Ob cells expressed PDGF-B transcripts of 3.5 kilo-bases, as determined by Northern blot analysis. Treatment of Ob cells with TGF beta 1 at 0.01-1.2 nM caused a dose-dependent increase in steady state PDGF-B mRNA, an effect that was initially observed after 2 h and was maximal after 6h. Cycloheximide induced PDGF-B transcripts and decreased the effect of TGF beta 1. TGF beta 1 did not modify the half-life of PDGF-B mRNA in transcriptionally arrested Ob cells and increased the rate of PDGF-B gene transcription in nuclear run-on assays. In contrast, treatment with PDGF-BB at 3.3 nM, bFGF at 6 nM, or IGF-I at 100 nM for 2-24 h did not modify PDGF-B mRNA levels in Ob cells. In conclusion, normal Ob cells express the PDGF-B gene, and TGF beta 1 induces its transcription, whereas bFGF, IGF-I, and PDGF-BB do not enhance the levels of PDGF-B mRNA. PDGF-BB may act not only as a systemic but also as a local regulator of bone cell function.

In our studies of the <em>growth</em>-promoting effect of a cytokine, interleukin-1 (IL-1), on cultured porcine granulosa cells, we found that the potency of IL-1 action correlated with the serum concentration in the culture medium and that IL-1 acted synergistically with insulin to increase the number of cells in the presence of low serum concentrations (0.1-1%). With granulosa cells maintained in a quiescent state under serum-free conditions, we therefore examined the effects of combined treatment with IL-1 and peptide <em>growth</em> <em>factors</em>, including insulin, on [3H]thymidine incorporation by these cells. IL-1 by itself enhanced [3H]thymidine incorporation in a concentration-dependent manner. Moreover, IL-1 acted synergistically with insulin, epidermal <em>growth</em> <em>factor</em> (EGF), or <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) to enhance [3H]thymidine incorporation. Combinations of maximally effective concentrations of insulin (1 micrograms/ml), EGF (1 ng/ml), or FGF (50 ng/ml) with the maximally effective concentration of IL-1 (10 ng/ml) increased the levels of [3H]thymidine incorporation to 10-, <em>22</em>-, and 20-fold, respectively, over the control values. Whereas IL-2 (0.1-100 ng/ml) did not affect [3H]thymidine incorporation, tumor necrosis <em>factor</em> alpha (TNF alpha) stimulated [3H]thymidine incorporation by itself and reproduced the actions of IL-1 to act synergistically with insulin, EGF, or FGF. When IL-1 and TNF alpha were added together in relatively low concentrations (1 ng/ml each), the combination had synergistic effects in enhancing [3H]thymidine incorporation. The present study demonstrates that cytokines and peptide <em>growth</em> <em>factors</em> act synergistically to markedly enhance porcine granulosa cell <em>growth</em> in vitro.

Bovine pituitary <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) stimulates the incorporation of [3H]thymidine into DNA in serum-depleted cultures of some but not other human melanoma cells. The melanotic malignant melanoma cell line MIRW exhibited a 40% increase in [3H]thymidine incorporation into DNA and a 48% increase in cell number in response to 3.73 x 10(-9) M FGF. This same concentration of FGF produced a <em>22</em>% increase in [3H]thymidine incorporation in the melanotic melanoma cell line Hs0294. However, FGF had no effect on the amelanotic melanoma cell line Hs0675, early-passage cultures of a human amelanotic melanoma (W-1), or early-passage cultures of a congenital nevus (N-1).

The <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) family consists of <em>22</em> ligands in mice and humans. FGF signaling is vital for embryogenesis and, when dysregulated, can cause disease. Loss-of-function genetic analysis in the mouse has been crucial for understanding FGF function. Such analysis has revealed that multiple Fgfs sometimes function redundantly. Exploring such redundancy between Fgf3 and Fgf4 is currently impossible because both genes are located on chromosome 7, about 18.5 kb apart, making the frequency of interallelic cross-over between existing mutant alleles too infrequent to be practicable. Therefore, we retargeted Fgf3 and Fgf4 in cis, generating an Fgf3 null allele and a conditional Fgf4 allele, subject to Cre inactivation. To increase the frequency of cis targeting, we used an F1 embryonic stem cell line that contained 129/SvJae (129) and C57BL/6J (B6) chromosomes and targeting constructs isogenic to the 129 chromosome. We confirmed cis targeting by assaying for B6/129 allele-specific single-nucleotide polymorphisms. We demonstrated the utility of the Fgf3(Δ)-Fgf4(flox)-cis mouse line by showing that the caudal axis extension defects found in the Fgf3 mutants worsen when Fgf4 is also inactivated. This Fgf3(Δ)-Fgf4(flox)-cis line will be useful to study redundancy of these genes in a variety of tissues and stages in development.

OBJECTIVE

To investigate whether autologous transplantation of adult stem cells could improve post-infarcted heart function.

METHODS

Bone marrow mononuclear cells (MNCs) were isolated from adult rabbits' tibias after coronary ligation. These cells were exposed to 5-azacytidine 10 micromol/L for 24 h on the third day of culture. After being labeled with bromodeoxyuridine (BrdU), the cells were auto-transplanted into bordering zone of the infarcted area at 2 weeks after injury. The animals were killed at 3 days, 2 weeks, 1 month, and 2 months after transplantation, respectively. The left ventricular functions, capillary density, and cardiac nerve density were measured and the differentiation of the engrafted cells was determined by immunostaining.

RESULTS

BrdU-labeled MNCs were well aligned with the host cardiomyocytes. Parts of them were incorporated into capillary and arteriolar vessel walls. In addition to inducing angiogenic ligands (basic <em>fibroblast</em> <em>growth</em> <em>factor</em>, vascular endothelial <em>growth</em> <em>factor</em>) and inflammation cytokines (interleukin 1-beta) during the early period of MNCs implantation, MNCs induced 2.0-fold increase in capillary density as well. Moreover, GAP43-positive and TH-positive nerve density were markedly higher in the MNCs-treated groups than that in the non-treated hearts. Left ventricular ejection fraction, LV+dp/dt(max), and LV-dp/dt(max) were 47%, 67%, and 55% in MNCs-treated heart respectively, which was higher than that of the control heart, whereas left ventricular end-diastolic volume, left ventricular end-diastolic diameter, and left ventricular end-diastolic pressure were 45%, <em>22</em>%, and 50% respectively in MNCs-treated heart, which was lower than that of the control heart at 2 months after cell transplantation.

CONCLUSIONS

Autologous transplantation of MNCs induced angiogenesis and nerve sprouting and improved left ventricular diastolic function.

Brain ribonuclease (BRB) is a member of the ribonuclease A superfamily that is constitutively expressed in a range of tissues and is the functional homolog of human ribonuclease 1. This study was designed to characterize BRB gene expression in granulosa cells (GCs) during development of bovine dominant ovarian follicles and to determine the hormonal regulation of BRB in GCs. Estrous cycles of Holstein cows (n = 18) were synchronized, and cows were ovariectomized on either day 3 to 4 or day 5 to 6 after ovulation during dominant follicle <em>growth</em> and selection. Ovaries were collected, follicular fluid (FFL) was aspirated, and GCs were collected for RNA isolation and quantitative polymerase chain reaction. Follicles were categorized as small (1-5 mm; pooled per ovary), medium (5-8 mm; individually collected), or large (8.1-17 mm; individually collected) based on surface diameter. Estradiol (E2) and progesterone (P4) levels were measured by radioimmunoassay (RIA) in FFL. Abundance of BRB messenger RNA (mRNA) in GCs was 8.6- to 11.8-fold greater (P < 0.05) in small (n = 31), medium (n = 66), and large (n = 33) subordinate E2-inactive (FFL E2 < P4) follicles than in large (n = 16) dominant E2-active (FFL E2>> P4) follicles. In the largest 4 follicles, GCs BRB mRNA abundance was negatively correlated (P < 0.01) with FFL E2 (r = -0.65) and E2:P4 ratio (r = -0.46). In experiment 2, GCs from large (8-<em>22</em> mm diameter) and small (1-5 mm diameter) follicles were treated with insulin-like <em>growth</em> <em>factor</em> 1 (IGF1; 0 or 30 ng/mL) and/or tumor necrosis <em>factor</em> alpha (0 or 30 ng/mL); IGF1 increased (P < 0.05) BRB mRNA abundance, and tumor necrosis <em>factor</em> alpha decreased (P < 0.001) the IGF1-induced BRB mRNA abundance in large-follicle GCs. In experiment 3 to 6, E2, follicle-stimulating hormone, <em>fibroblast</em> <em>growth</em> <em>factor</em> 9, cortisol, wingless 3A, or sonic hedgehog did not affect (P>> 0.10) abundance of BRB mRNA in GCs; thyroxine and luteinizing hormone increased (P < 0.05), whereas prostaglandin E2 (PGE2) decreased (P < 0.05) BRB mRNA abundance in small-follicle GCs. Treatment of small-follicle GCs with recombinant human RNase1 increased (P < 0.05) GCs numbers and E2 production. In conclusion, BRB is a hormonally and developmentally regulated gene in bovine GCs and may regulate E2 production during follicular <em>growth</em> in cattle.

The human fetal adrenal cortex is one of the largest fetal organs and synthesizes precursors for placental estrogen production as part of the feto-placental unit. The <em>factors</em> controlling the rapid <em>growth</em> of the human fetal adrenal cortex during the second and third trimesters are not known. Placental regulation of the <em>growth</em> of human fetal adrenocortical cell cultures from second trimester fetuses was studied. A placental-derived mitogenic <em>factor</em> (PDMF) was detected in tissue homogenates of 14 to <em>22</em> week human placentas and stimulated adrenocortical cell number and [3H]thymidine incorporation into DNA 5-8 fold. PDMF has been partially purified by ammonium sulfate precipitation and anion exchange chromatography. PDMF is a heat sensitive protein with disulfide bonds required for activity. The <em>growth</em> stimulation by PDMF was significantly greater than that for basic or acidic <em>fibroblast</em> <em>growth</em> <em>factor</em> by 25-50% and epidermal <em>growth</em> <em>factor</em> by 3-4 fold. The placental hormones, progesterone, estriol, estradiol, placental lactogen and chorionic gonadotropin, either alone or in combination did not stimulate fetal adrenocortical cell <em>growth</em>, except for a 41% cell number increase by progesterone. Platelet-derived <em>growth</em> <em>factor</em> and insulin-like <em>growth</em> <em>factors</em> I and II were not mitogenic for these cells. These results show that the placenta contains a potent <em>growth</em> <em>factor</em> for human fetal adrenocortical cell cultures. This implies a direct role for the placenta in control of this fetal organ's <em>growth</em>, which would make the human feto-placental unit a bi-directional relationship.

The peripheral myelin protein <em>22</em> (PMP<em>22</em>) is a tetraspan membrane protein which is localised in the compact myelin of the peripheral nerves. In <em>fibroblasts</em>, where it was originally identified as <em>growth</em> arrest related <em>factor</em> 3 (Gas3), PMP<em>22</em> has been shown to modulate cell proliferation; in the peripheral nervous system its roles are still debated. The duplication of PMP<em>22</em> is the most common cause of the demyelinating form of the autosomal dominant Charcot-Marie-Tooth neuropathy (CMT1A); rarer missense mutations of PMP<em>22</em> also cause CMT1A or severe dehypomyelinating neuropathies of infancy grouped under the heading of Dejerine-Sottas syndrome (DSS). Here, a sporadic patient affected with DSS is described; nerve biopsy disclosed a picture of hypomyelination/amyelination with basal laminae onion bulbs and no florid demyelination and it was consistent with congenital hypomyelination neuropathy (CHN); molecular analysis disclosed a novel point mutation of PMP<em>22</em> that causes a non-conservative arginine for cysteine substitution at codon 109, in the third transmembrane domain. CHN is the rarest and severest form of DSS and it is thought to reflect dysmyelination rather than demyelination. The reported case suggests that missense point mutations may alter a putative role of PMP<em>22</em> in modulating Schwann cell <em>growth</em> and differentiation.

OBJECTIVE

Translocations involving the fibroblast growth factor receptor 1 (FGFR1) gene are associated with the 8p11 myeloproliferative syndrome (EMS), a rare neoplasm that following a usually short chronic phase progresses into acute myeloid or lymphoid leukemia. The treatment commonly involves chemotherapy and, if possible, allogeneic stem cell transplantation which is the only therapeutic option for long-term survival. Given the aggressive course of EMS, we here evaluated tyrosine kinase inhibitors as treatment options to delay disease progression.

METHODS

We described a new case of EMS and used chromosome analyses, PCR, and sequencing to investigate the underlying genetic aberrations. The sensitivity to several tyrosine kinase inhibitors was tested in vitro on the EMS cell line KG1 and on primary cells from the newly diagnosed EMS patient.

RESULTS

A translocation involving chromosomes 8 and 22 was detected, and a BCR/FGFR1 fusion gene was confirmed and characterized by sequencing. KG1 cells and primary EMS cells displayed distinct sensitivity to dovitinib, ponatinib, and dasatinib as compared to normal bone marrow control cells.

CONCLUSIONS

These results suggest that treatment with tyrosine kinase inhibitors may be beneficial for patients with EMS during the search for a suitable stem cell donor and for those not eligible for transplantation.

p53 is the main intrinsic <em>factor</em> inducing apoptosis by recognizing the external stimuli and activating the p53 responsive genes to an irreversible series of events. P53 activates the transcription of specific proapoptotic genes called p53 target genes. A <em>growing</em> number of p53 responsive genes have been identified and numerous studies have demonstrated that p53 proapoptotic <em>factors</em> such as Noxa, Puma and Perp play cell type specific roles in p53's mediated response to certain stimuli. Perp (p53 apoptosis effector related to PMP-<em>22</em>) is a direct proapoptotic target gene encoding a tetraspan protein. Perp is highly expressed in cells undergoing apoptosis compared to cells under G1 arrest and its overexpression is sufficient to cause cell death in <em>fibroblasts</em>. Noxa is another member of the preapoptotic p53 genes family. When expressed Noxa acts in a BH3 motif-dependent localization to mitochondria, causing structural changes, activation of caspase 9 and release of cytochrome c from mitochondria to cytosol. Puma (p53 mutant of apoptosis) is another critical mediator of p53-dependent apoptosis. P53 binds to Puma-promoter gene sites, leading to puma production. The mtCLIC, a member of intracellular chloride channels, is a cytoplasmic and mitochondrial protein positively regulated by p53. Caspase 10 is induced in p53-dependent manner leading to cellular apoptosis. Other newly announced <em>factors</em> are also involved in p53-regulated apoptosis such as brain-specific angiogenesis inhibitor-1 (BSAI1), MSOD and GPX genes. A global discussion on this topic is attempted in the present review article.

The bioassay of platelet-derived <em>growth</em> <em>factor</em> (PDGF) in platelets was established using the rat <em>fibroblast</em> cell line 3Y1. 3Y1 cells (5 X 10(3)/well) were cultured in 24-well microculture plates with RPMI 1640 medium using 1% of PDGF-free serum and 5% of platelet extracts for 4 days. Cell proliferation was measured by the increase of DNA content. This assay made it possible to measure the biological PDGF activity. PDGF activity of platelet concentrates kept the same level as that of fresh platelet samples during preservation for up to 120 h at <em>22</em> degrees C; cryopreserved platelets also retained PDGF activity well.

OBJECTIVE

Pharmacologic modulation of the contents of the pericardial space has been shown to influence the response of coronary arteries to balloon injury. Endoluminal (EL) local delivery of various drugs into coronaries has been found to be limited by short residence time, as well as by highly variable deposited agent concentration. We hypothesized that compounds placed into the pericardial space (P) would penetrate into coronary tissue with greater consistency than seen after EL delivery and provide for prolonged coronary exposure to agents.

RESULTS

125I-labeled basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), platelet-derived <em>growth</em> <em>factor</em> (PDGF), albumin, or 131I-labeled diazeniumdiolated albumin (NONO-albumin) were delivered as model/therapeutic proteins into the porcine pericardial space (n = 15 pigs) or into coronaries using an EL delivery catheter (n = 48 arteries). In subjects receiving 125I-labeled proteins, the delivery target or mid-regions of the left anterior descending (LAD) and left circumflex (LCx) arteries were harvested at 1 h or 24 h for gamma-counting and autoradiography, and fractional intramural delivery (FID) or retention measured as percent agent in 100 mg artery/agent in infusate for both time points. In the animals receiving 131I-labeled NONO-albumin, serial gamma imaging was employed to evaluate the rate of redistribution in individual animals following either pericardial or endoluminal delivery. At 1 h, FID values ranged from 0.00064 to 0.0052% for P delivery (median 0.00<em>22</em>%), and from 0.00021 to 6.7 for EL delivery (median 0.27%). At 24 h, FID values ranged from 0.00011 to 0.003 for P delivery (median 0.0013), and from 0.0002 to 1.4 for EL delivery. The estimated T1/2 for bFGF redistribution from the vascular tissue was <em>22</em> h (P) and 7 h (EL), respectively, while the directly determined T1/2 values for NONO-albumin redistribution from the delivery region were <em>22</em>.2 h (P) and 2.5 h (EL).

CONCLUSIONS

These data show that pericardial fluid contents can access coronary arteries with intramural concentrations which typically vary by 10-15-fold, while EL delivery results in a remarkably wide intramural concentration range with up to 33,000-fold variability. The apparent redistribution rate is more rapid following EL delivery, possibly due to sustained diffusive tissue loading from the pericardial space. Pericardial delivery appears to offer substantial advantages over EL administration with respect to residence time and reproducibility.

<em>Fibroblast</em> <em>growth</em> <em>factor</em>-2 (FGF-2) is a potent mitogen and angiogenic <em>factor</em> normally absent from the adult circulation. We have previously shown that it appears in normal maternal serum and that circulating FGF-2 levels are elevated in pregnancies complicated by diabetes. This study was performed to determine whether serum FGF-2 is more abundant in pregnant diabetic women with retinopathy than in those without. Serum was collected monthly between 14-30 weeks gestation and every 2 weeks from then until delivery (35-38 weeks) from 36 women with type 1 diabetes. FGF-2 was extracted by heparin-Sepharose affinity chromatography and quantified by specific RIA. Patients were divided according to the White classification of diabetes. In 17 women without retinopathy (White groups B, C, and D0), immunoreactive FGF-2 was detectable at 14 weeks (mean +/- SEM, 154 +/- 39 pmol/L), was maximal after 26 weeks (306 +/- 38 pmol/L), after which values steadily declined to term (212 +/- 48 pmol/L). In 19 women with simplex or proliferative retinopathy (White groups D+ and R), circulating levels of FGF-2 were significantly greater between <em>22</em>-32 weeks gestation (<em>22</em> weeks, 480 +/- 102 vs. 239 +/- 38 pmol/L; P < 0.05). Serum FGF-2 was significantly correlated with hemoglobin A1c levels at <em>22</em>, 30, and 34 weeks gestation. The mean birth weight of the infants did not significantly differ between groups. Macroalbuminuria was absent in all patients, and creatinine clearance and blood pressure did not significantly differ between the two groups. The results suggest that serum FGF-2 is substantially elevated in pregnant diabetic women with retinopathy in second and early third trimesters. It is unlikely that in these patients this was primarily due to altered FGF-2 clearance, but may relate to excessive production by the utero-placental compartment. The high circulating levels of FGF-2 may be causally related to the development of diabetic retinopathy.

The root of Actinidia valvata dunn has been widely used in the treatment of hepatocellular carcinoma (HCC), proved to be beneficial for a longer and better life in China. In present work, total saponin from root of Actinidia valvata Dunn (TSAVD) was extracted, and its effects on hepatoma H<em>22</em>-based mouse in vivo were observed. Primarily transplanted hypodermal hepatoma H<em>22</em>-based mice were used to observe TSAVD effect on tumor <em>growth</em>. The microvessel density (MVD), vascular endothelial <em>growth</em> <em>factor</em> (VEGF), basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) are characterized <em>factors</em> of angiogenesis, which were compared between TSAVD-treated and control groups. Antimetastasis effect on experimental pulmonary metastasis hepatoma mice was also observed in the study. The results demonstrated that TSAVD can effectively inhibit HCC <em>growth</em> and metastasis in vivo, inhibit the formation of microvessel, downregulate expressions of VEGF and bFGF, and retrain angiogenesis of hepatoma <em>22</em> which could be one of the reasons.

Feeding a modified fish diet has been suggested to improve insulin sensitivity in bottlenose dolphins; however, insulin sensitivity was not directly measured. Since demonstrating an improvement in insulin sensitivity is technically difficult in dolphins, we postulated that directional changes in the hormone axis: <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21)/Adiponectin/Ceramide (Cer), could provide further support to this hypothesis. We measured 2-h post-prandial serum FGF21, total adiponectin, percent unmodified adiponectin, ceramide, and sphingosine levels from dolphins fed a diet rich in heptadecanoic acid (C17:0) over 24 weeks. Serum FGF21 was quantified by ELISA with an observed range of 129-1599 pg/ml, but did not significantly change over the 24-week study period. Total adiponectin levels (mean ± SD) significantly increased from 776 ± 400 pmol/ml at week 0 to 1196 ± 467 pmol/ml at week 24. The percent unmodified adiponectin levels (mean ± SD) decreased from 23.8 ± 6.0% at week 0 to 15.2 ± 5.2% at week 24. Interestingly, although FGF21 levels did not change, there was a good correlation between FGF21 and total adiponectin (ρ = 0.788, P < 0.001). We quantified the abundances of serum ceramides and sphingosines (SPH) because adiponectin has a defined role in sphingolipid metabolism through adiponectin receptor-mediated activation of ceramidases. The most abundant ceramide in dolphin sera was Cer 24:1 comprising 49% of the ceramides measured. Significant reductions were observed in the unsaturated Cer 18:1, Cer 20:1, and Cer 24:1, whereas significant increases were observed in saturated Cer <em>22</em>:0, Cer 24:0, and Cer 26:0. However, total serum ceramides did not change. Significant elevations were detected for total sphingosine, dihydrosphingosine, sphingosine-1-phosphate, and dihydrosphingosine-1-phosphate. Proteomic analysis of the serum proteins revealed few changes in serum proteins over the study period. In conclusion, shifting the dolphin diet to fishes rich in odd chain saturated fatty acids, such as C17:0, resulted in increased serum levels of the insulin sensitizing hormone adiponectin and serum SPH consistent with an insulin-sensitizing phenotype. It is still unclear whether FGF21 plays a role in the regulation of adiponectin in dolphins, similar to that shown in laboratory animal models.

Specific <em>growth</em> <em>factors</em> induce formation and differentiation of excitatory and inhibitory synapses, and are essential for brain development and function. <em>Fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> (FGF<em>22</em>) is important for specifying excitatory synapses during development, including in the hippocampus. Mice with a genetic deletion of FGF<em>22</em> (FGF<em>22</em>KO) during development subsequently have fewer hippocampal excitatory synapses in adulthood. As a result, FGF<em>22</em>KO mice are resistant to epileptic seizure induction. In addition to playing a key role in learning, the hippocampus is known to mediate mood and anxiety. Here, we explored whether loss of FGF<em>22</em> alters affective, anxiety or social cognitive behaviors in mice. We found that relative to control mice, FGF<em>22</em>KO mice display longer duration of floating and decreased latency to float in the forced swim test, increased immobility in the tail suspension test, and decreased preference for sucrose in the sucrose preference test, which are all suggestive of a depressive-like phenotype. No differences were observed between control and FGF<em>22</em>KO mice in other behavioral assays, including motor, anxiety, or social cognitive tests. These results suggest a novel role for FGF<em>22</em> specifically in affective behaviors.