Building functional and robust scaffolds for engineered biological tissue requires a nanoscale mechanistic understanding of how cells use the scaffold for their growth and development. A vast majority of the scaffolds used for cardiac tissue engineering are based on polymer materials, the matrices of nanofibers. Attempts to load the polymer fibers of the scaffold with additional sophisticated features, such as electrical conductivity and controlled release of the growth factors or other biologically active molecules, as well as trying to match the mechanical features of the scaffold to those of the extracellular matrix, cannot be efficient without a detailed knowledge of how the cells are attached and strategically positioned with respect to the scaffold nanofibers at micro and nanolevel. Studying single cell - single fiber interactions with the aid of confocal laser scanning microscopy (CLSM), scanning probe nanotomography (SPNT), and transmission electron microscopy (TEM), we found that cardiac cells actively interact with substrate nanofibers, but in different ways. While cardiomyocytes often create a remarkable "sheath" structure, enveloping fiber and, thus, substantially increasing contact zone, fibroblasts interact with nanofibers in the locations of focal adhesion clusters mainly without wrapping the fiber.

We found that cardiomyocytes grown on electrospun polymer nanofibers often create a striking "sheath" structure, enveloping fiber with the formation of a very narrow (∼22 nm) membrane gap leading from the fiber to the extracellular space. This wrapping makes the entire fiber surface available for cell attachment. This finding gives a new prospective view on how scaffold nanofibers may interact with growing cells. It may play a significant role in effective design of novel nanofiber scaffolds for tissue engineering concerning mechanical and electrical properties of scaffolds as well as controlled drug release from "smart" biomaterials.

Heparan sulfate proteoglycans, syndecan-1 and glypican, are low-affinity receptors for <em>fibroblast</em> <em>growth</em> <em>factor</em> 2 (FGF2). Since FGF2 stimulates skeletal muscle cell proliferation but inhibits differentiation, differences in syndecan-1 and glypican expression might affect muscle development and <em>growth</em> by changing the intensity of FGF2 signaling. In the present study, the pectoralis major muscle from 14 to 24-day-old-embryos, and from 1 to 16-week-old birds from a turkey line (F) selected for increased 16-week bodyweight and its genetic control line (RBC2), were used to address how syndecan-1 and glypican are expressed during skeletal muscle formation. The expression of syndecan-1 and glypican was measured by semiquantitative reverse transcription polymerase chain reaction. For males, the F-line embryos expressed more syndecan-1 (days 14 and 16) and glypican (days 14 and 18) than the RBC2 line. Similar line differences for males were observed during posthatch development. The male embryos from both lines expressed more syndecan-1 at days 18 through <em>22</em> and more glypican at days 20 and <em>22</em> than the corresponding females. The temporal and spatial distribution of syndecan-1 and glypican was detected by in situ hybridization. Syndecan-1 was identified in all muscle fibers at all embryonic stages studied, whereas the glypican was detected from embryonic day 18. The data from the current study provided new information about the expression of syndecan-1 and glypican as it relates to skeletal muscle <em>growth</em> properties.

OBJECTIVE

Recent reports of animal models have shown that growth factors have stimulating effect on brain perfusion via the development of blood vessels. However, studies on the effect of growth factors on brain perfusion in humans are lacking. The aim of our study was to prospectively investigate in humans the relation between growth factors and brain perfusion.

METHODS

We analyzed circulating levels of vascular endothelial growth factor (VEGF), granulocyte-macrophage colony-stimulating growth factor (GM-CSF), tumor necrosis factor alpha (TNFalpha) and basic fibroblast growth factor (bFGF) in 121 consecutive patients (99 men and 22 women, age 58+/-10 years) who were enrolled in a prospective cohort study of patients with symptomatic atherosclerotic disease. In all patients regional cerebral blood flow (rCBF; in mL/min/100g) measurements were performed with arterial spin labeling magnetic resonance imaging. Cerebrovascular risk factors were assessed by means of a questionnaire and physical, ultrasonographic and laboratory examination.

RESULTS

Increasing levels of TNFalpha were significantly associated with a higher rCBF (beta=7.0; 95% confidence interval 0.7; 13.9), independent of the presence of cerebrovascular risk factors. No significant association was found for VEGF, GM-CSF and bFGF.

CONCLUSIONS

Increasing levels of TNFalpha are associated with increased rCBF, independent of the presence of cerebrovascular risk factors.

OBJECTIVE

Urinary excretion of several pro-angiogenic and antiangiogenic substances has been correlated with malignant tumor growth. The aim of this study was to assay angiogenic activity in urine from patients with cancer of the prostate and benign prostatic hyperplasia (BPH).

METHODS

Urine specimens from 22 healthy male volunteers (control), 33 patients with BPH and 29 with organ confined prostate cancer were analyzed for angiogenic activity in a bovine capillary endothelial cell proliferation assay. In parallel the concentration of basic fibroblast growth factor and vascular endothelial growth factor was determined by enzyme immunoassay in the corresponding urine specimens.

RESULTS

Urine samples from patients with BPH and prostate cancer increased bovine capillary endothelial cell proliferation by 13.1% and 15.1%, respectively, whereas urine from the control group showed a significantly lower angiogenic activity, increasing endothelial cell proliferation by only 0.7% (p = 0.001). Urinary basic fibroblast growth factor and vascular endothelial growth factor were highest in patients with BPH and lowest in the group with prostate cancer (p = 0.0001).

CONCLUSIONS

Urine from patients with BPH and prostate cancer stimulates endothelial cell proliferative activity. The degree of endothelial cell stimulation does not correlate with the concentration of basic fibroblast growth factor or vascular endothelial growth factor. Whether the observed pro-angiogenic activity is due to an increased production or release of (an) other angiogenic factor(s) and/or loss of (an) angiogenesis inhibitor(s), deserves further investigation.

To gain insights into the role of iodine deficiency in favoring thyroid tumorigenesis (particularly of the follicular histotype), <em>22</em> Sicilian patients with thyroid tumors were selected for having lived permanently in either one of two areas of different iodine availability. Eleven patients (age 46.1 +/- 14.6 years, mean +/- SD; 10 females and 1 male) were from the iodine-deficient (ID) areas of the provinces of Messina and Catania (mean urinary excretion of iodine = 48.1 micrograms/24 hours). Thyroid tumors were follicular or Hürthle cell adenomas (no. = 3), follicular carcinomas (FC, no. = 4), papillary carcinomas (PC, no. = 2) and anaplastic carcinomas (no. = 2). Eleven patients (age 47.1 +/- 15.2 years; 10 females and 1 male) were from the metropolitan area of Messina, an area of relative iodine-sufficiency (IS) (urinary excretion of iodine = 95.2 micrograms/24 hours). These 11 patients had serum levels of TSH that were significantly lower than the corresponding values of the 11 patients from the ID area (0.76 +/- 0.33 vs 1.80 +/- 1.<em>22</em> mU/l, p = 0.01) The tumors of the 11 patients from the IS area were: follicular or Hürthle cell adenomas (no. = 6), Hürthle cell carcinoma (no. = 1), FC (no. = 2), PC (no. = 2). Molecular biology studies revealed that both the normal as well as the tumor tissue of all <em>22</em> patients did not harbor any of the three classical activating mutations (codons 12, 13 and 61) in any of the three ras oncogenes. Similar negative results were obtained as far as loss of heterozygosity of the retinoblastoma (Rb) anti-oncogene is concerned. Immunohistochemistry studies were performed to investigate expression of c-met and basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) proto-oncogenes. Only one Hürthle cell carcinoma and the two PC from the IS group, and one FC and the two PC from the ID group stained for the c-met oncogene. Expression of c-met was greater (3+) in the four PC (concerning 70-80% of the tumor cells) than in the other two cancers (1+; < 5% of the tumor cells). In the IS group, positivity for bFGF was detected in 3/6 adenomas, 1/2 FC, the Hürthle cell carcinoma and the two PC. In the ID group, positivity for bFGF was observed in 2/3 adenomas, 2/4 FC, the two PC and the two anaplastic carcinomas. The 8 positive cases from the ID group had a greater level of bFGF expression than the 7 positive cases from the IS group (intensity of staining = 2.0+ vs 1.57+). Interestingly, the greatest expression of bFGF was seen in the cases with peri-tumoral lymphocytic infiltration from either group. In the ID group correlations between (i.) pre-intervention serum TSH and intensity of tumoral staining for bFGF, (ii.) serum TSH and per cent of tumoral cells reactive with anti-bFGF and (iii.) between intensity of staining for bFGF and per cent of tumoral cells bFGF +ve were higher than in the IS group. We conclude that activating mutations of ras, loss of DNA from the Rb locus and over-expression of both c-met and bFGF are of no pathogenetic relevance in driving thyroid tumorigenesis of iodine-deficient areas.

<em>Fibroblast</em> <em>growth</em> <em>factor</em>-2 is a member of a large family of proteins that bind heparin and heparan sulfate and modulate the function of a wide range of cell types. FGF-2 occurs in several isoforms resulting from alternative initiations of traslation: an 18 kDa cytoplasmic isoform and four larger molecular weight nuclear isoforms (<em>22</em>, <em>22</em>.5, 24 and 34 kDa). It acts mainly through a paracrine/autocrine mechanism involving high affinity transmembrane receptors and heparan sulfate proteoglycan low affinity receptors. It is expressed mostly in tissues of mesoderm and neuroectoderm origin, and plays an important role in mesoderm induction, stimulates the <em>growth</em> and development of the new blood vessels (angiogenesis), normal wound healing and tissue development. FGF-2 positively regulates hematopoiesis by acting on various cellular targets: stromal cells, early and committed hematopoietic progenitors and possibly some mature blood cells. FGF-2 is a potent hematopoietic <em>growth</em> <em>factor</em> that is likely to play an important role in physiological and pathological hematopoiesis.

Rat prostate extracts contain an abundant 20-<em>22</em> kilodalton heparin-binding protein with near identical chromatographic properties, but only 0.2-1% of the mitogenic activity, of bovine brain heparin-binding <em>growth</em> <em>factor</em>-1 (acidic <em>fibroblast</em> <em>growth</em> <em>factor</em>). Amino terminal amino acid sequence (met-met-thr-asp-lys-asn-leu-lys-lys-lys-ile-glu-gly-asn-trp-arg-thr-val -tyr- leu-ala-ala-ser-?-val-glu-lys-ile-asn-glu-gly-ser-pro) and immunochemical analysis revealed that the protein is identical to the androgen-dependent protein "probasin".

The common culture system of rhesus monkey embryonic stem (rES) cells depends largely on feeder cells and serum, which limits the research and application of rES cells. This study reports a feeder layer-free and serum-free system for culture of rES cells. rES cells could be cultured through at least <em>22</em> passages on laminin in medium supplemented with serum replacement (SR), basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) and transforming <em>growth</em> <em>factor</em> beta1 (TGFbeta1), and maintained stable proliferation rates and normal karyotypes, while displaying all the embryonic stem cell characteristics including morphology, alkaline phosphatase (AKP), Oct-4, cell surface markers SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81, and formed cystic embryoid bodies in vitro. In addition, the studies showed that TGFbeta1, bFGF and laminin are necessary for maintaining the undifferentiated <em>growth</em> of rES cells in long-term culture. Moreover, withdrawal of TGFbeta1 increased the differentiation rate by decreasing the expression of integrins. Therefore, this system would provide a well-defined culture system for rES cells, and would facilitate research into self-renewal and differentiation mechanisms of rES cells.

Axons damaged in the adult mammalian central nervous system are able to regenerate when their inhibitory glial environment is replaced with a more permissive substrate. Here, we have used long oblique "bridge" grafts of <em>fibroblast</em> <em>growth</em> <em>factor</em>-4-transfected RN-<em>22</em> schwannoma cells to allow mechanically lesioned nigrostriatal axons to regenerate back to their original target in the adult rat brain. Regenerated axons were able to leave the bridge graft to form terminal arborizations and increase the density of tyrosine hydroxylase-immunoreactive fibres within the striatum. Bridge grafting also resulted in an increase in the number of neurons within the substantia nigra pars compacta taking up the fluorescent retrograde tracer Fluoro-Gold from the striatum. Animals which had received RN-<em>22</em> bridge grafts showed lower rates of amphetamine-induced rotation 10 weeks after a mechanical lesion of the nigrostriatal tract compared to lesioned controls, the magnitude of the behavioural effect being related to the number of regenerated axons, and this comparative reduction was reversed by mechanical section of the bridge graft. It is concluded that our bridge grafting strategy allowed the partial anatomical and functional regeneration of the mechanically lesioned nigrostriatal tract, an unmyelinated central axon bundle, and that bridge grafting therefore represents a realistic approach to the repair of central nervous system lesions involving axon tract damage.

Caudal appendage is a rare but reported finding seen in association with craniosynostosis. We report a newborn with caudal appendage secondary to sacrococcygeal eversion, a cloverleaf skull, choanal atresia, and a heterozygous mutation of Y375C in the juxtamembrane domain (exon 11) of <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 2 (FGFR2). Further support of this association are <em>22</em> other cases of craniosynostosis with caudal appendage or sacrococcygeal eversion in the literature. Of these, 19 had detectable mutations in FGFR2; 5, the same mutation; and 5, a similar substitution of cysteine for serine. We hypothesize that the association of craniosynostosis and caudal appendage is due to abnormal expression of FGFR2 in the tail bud of a developing embryo based on animal models. Our case and those reported in the literature suggest that in patients with caudal appendage and craniosynostosis, FGRF2 analysis should include regions outside the commonly tested exons 8 and 10, particularly the juxtamembrane domain.

Chronic myelogenous leukemia (CML) is characterized by a translocation involving the c-abl protein-tyrosine kinase gene. A chimeric mRNA is formed containing sequences from a chromosome <em>22</em> gene (bcr) at its 5' end and all but the variable exon 1 of c-abl sequence. The product of this mRNA, p210bcr-abl, has constitutively high protein-tyrosine kinase activity. We examined K562 cells and other lines established from CML patients for the presence of phosphotyrosine (P-Tyr)-containing proteins which might be p210bcr-abl substrates. Two-dimensional gel separation of 32P-labeled proteins followed by phosphoamino acid analysis of 25 phosphoproteins, which comprised the major alkali-stable phosphoproteins, indicated that three related proteins of 41 kDa are the most prominent P-Tyr-containing proteins detected by this method. The 41-kDa phosphoproteins are found in two other CML lines that we examined but not in lines of similar lineage isolated from patients with distinct leukemic disease. A protein that comigrates with the major form of pp41 (pp41A) and contains P-Tyr is also found in murine <em>fibroblasts</em> and B-lymphoid cells transformed by Abelson murine leukemia virus, which encodes the v-abl protein, and in platelet-derived <em>growth</em> <em>factor</em>-treated <em>fibroblasts</em>, in which it has been described previously. We analyzed three pairs of Epstein-Barr virus-immortalized B-cell lines from individual CML patients and found that only the lines in which active p210bcr-abl was present contained detectable pp41. We also performed immunoblotting with anti-P-Tyr antibodies on the same CML cell lines and detected at least four other putative substrates of p210bcr-abl, which were undetected with use of the two-dimensional gel technique.

<AbstractText>Klotho, a single-pass transmembrane protein associated with premature aging, acts as a tumor suppressor gene by inhibiting insulin/insulin-like <em>growth</em> <em>factor</em>-1 and <em>fibroblast</em> <em>growth</em> <em>factor</em> pathways. Downregulated Klotho expression is reported in melanoma, mesothelioma, bladder, breast, gastric, cervix, lung, and kidney cancers and is associated with a poor prognosis. Klotho expression and Klotho promoter hypermethylation are predictive <em>factors</em> for patient prognosis.</AbstractText><AbstractText>To investigate the potential role of Klotho in glioblastoma-multiforme (GBM), <em>22</em> GBM samples were collected from the Sheba Tumor Bank and examined.</AbstractText><AbstractText>We found that increased Klotho messenger ribonucleic acid (RNA) expression predicted longer survival (P = 0.03) of GBM patients. Methylation analysis was performed on bisulfite-treated deoxyribonucleic acid from the GBM patient samples using ionization time-of-flight mass spectrometry according to the Sequenom EpiTYPER protocols. Klotho promoter hypermethylation was detected in 65% of the GBM samples and correlated significantly with improved survival (P < 0.04). We found 3 major Klotho promotor hypermethylation sites located 585-579 bp, 540-533 bp, and 537-534 bp upstream of the transcription start site. Methylated deoxyribonucleic acid immunoprecipitation studies confirmed these results. Notably, the messenger RNA expression in these GBM samples revealed an unexpected linear correlation with methylation of these 3 hypermethylation sites identified in the Klotho promotor. Thus Klotho expression and methylation could predict prognosis in patients with GBM.</AbstractText><AbstractText>Epigenetic regulation in GBM appears to be complicated. Specific CpG islands affect genes or micro RNAs that interact to control Klotho expression. The diverse effects of these islands may be due to unique <em>factors</em> of GBM.</AbstractText>

Regulation of cell migration by cell <em>growth</em> <em>factors</em> is critical in tissue regeneration such as angiogenesis, wound healing, and bone formation. In this work, basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) with a density varying between 0 and 295 ng/cm2 was conjugated on heparinized glass slides. The amount of conjugated bFGF was determined by immunofluorescent staining. The mobility of vascular smooth muscle cells (VSMCs) was largely dominated by the bFGF density, whereas that of mesenchymal stem cells (MSCs) and endothelial cells (ECs) was slightly influenced. The migration rate of VSMCs increased initially and then decreased along with the increase of bFGF density. The fastest rate (<em>22</em> μm/h) was found on the bFGF surface with a density of 83 ng/cm2. The intrinsic mechanisms of the diverse migration behaviors of the VSMCs, MSCs, and ECs were revealed by studying the expression of bFGF receptors and migration-related proteins. The results show that the cell mobility is regulated by complex and synergetic intracellular signals in a cell type-dependent manner.

BACKGROUND

Fibroblast growth factor (FGF) 23 is one of the most recently discovered FGFs. This phosphaturic hormone produced in bones is a risk factor for cardiovascular diseases and thus mortality. Klotho is an essential coreceptor for FGF23 and at the same time it is known as a "longevity" hormone. There are no data considering FGF23 and Klotho roles in heart transplant (HT) recipients. The aim of this study was to assess Klotho and FGF23 serum concentration in heart transplant recipients depending on immunosuppressive therapy regimen and comorbidities.

METHODS

Eighty-four stable heart transplant recipients were enrolled in the study; 22 healthy volunteers served as control subjects. FGF23 and Klotho protein concentration, markers of renal function, such as cystatin C and neutrophil gelatinase-associated lipocalin (NGAL), and heart failure markers, such as copeptine and N-termiinal pro-B-type natriuretic peptide (NT-proBNP), were evaluated.

RESULTS

FGF23 concentration was significantly higher in the HT group whereas Klotho protein was significantly lower. FGF23 correlated with creatinine level (r = 0.72; P < .001), estimated glomerular filtration rate (eGFR; r = -0.32; P < .01), cystatin C (r = 0.36; P < .01), NGAL (r = 0.51; P < .001), hemoglobin (r = -0.39; P < .001), NT-proBNP (r = 0.51; P < .001), high-density lipoprotein (HDL; r = 0.27; P < .05), intraventricular septum thickness (r = 0.42; P < .01) and right ventricular systolic pressure (r = 0.34; P < .05). Klotho protein correlated only with age (r = -0.21; P < .05), creatinine (r = -0.21; P < .05), and eGFR (r = -0.31; P < .01). FGF23 concentration was significantly higher in patients with eGFR <60 mL/min whereas Klotho protein was significantly lower. FGF23 predictors were renal function (creatinine concentration; β = 0.45; P = .0001), HDL (β = 0.33; P = .003), intraventricular septum thickness (β = 0.38; P = .0003), and right ventricular systolic pressure (β = 0.34; P = .003), explaining 70% of FGF23 variability.

CONCLUSIONS

FGF23/Klotho system disorders in HT recipients are related to cardiovascular system function and kidney failure and could cause increased risk of cardiovascular disease.

In a microassay for anchorage-independent <em>growth</em> in soft agar, NR6 cells form colonies in a dose-dependent manner in the presence of <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF). Using this assay system, the ability of thin sequential slices of embryonic chick limb bud to promote colony formation was investigated. A functional gradient of colony-promoting ability along the proximo-distal axis of the developing chick limb bud (stages <em>22</em>-26) was observed. The highest number of colonies was observed in the presence of the most distal slices, and colony number decreased progressively at proximal levels. This gradient was specifically eliminated by the addition of anti-FGF antibody to the assay, indicating that it was caused by a functional gradient of an FGF-like molecule. Limbs of stages 21-26 were assayed: before this time limb buds are too small to slice in the proximo-distal axis in the required manner. The FGF-like gradient was observed at stages <em>22</em> to 26.

BACKGROUND

Data about angiogenic factors in diabetic foot syndrome (DFS) are insufficient. Therefore, in the present study we focus on circulating endothelial progenitor cells (EPCs) and two major angiogenic factors: vascular endothelial growth factor (VEGF-A) and fibroblast growth factor (FGF-2) in patients with DFS.

METHODS

We included 75 subjects: 45 patients with type 2 diabetes and 30 controls. The study group was divided into 2 subgroups: 23 patients with diabetic foot and 22 patients without diabetic complications. The concentration of VEGF-A, soluble VEGF receptor 2 (sVEGF-R2) and FGF-2 were measured in plasma samples. The number of circulating EPCs was determined in peripheral venous blood. The number of endothelial progenitor cells was measured with FACSCalibur flow cytometer using monoclonal antibodies directed against antigens specific for EPCs.

RESULTS

In our study we observed significant higher levels of VEGF-A and FGF-2 and lower sVEGF-R2 concentration in patients with T2DM compared to healthy subjects. The conducted analysis showed decreased levels of VEGF-A and elevated levels of FGF-2 in patients with DM complicated DFS compared to diabetic patients without DFS. Increased circulating EPCs number was reported in patients with DFS, and the difference was almost statistically significant.

CONCLUSIONS

The high concentration of VEGF-A and FGF-2, and a positive correlation between them indicate their participation in the process of angiogenesis in T2DM. Decreased sVEGF-R2 may result from inactivation of VEGF-A during complexes formation.

<AbstractText><em>Fibroblast</em> <em>growth</em> <em>factor</em> (FGF)2 is an important stimulatory modulator of satellite cells in skeletal muscle. Satellite cells play a cardinal role in muscle <em>growth</em> and repair.</AbstractText><AbstractText>We evaluated whether skeletal muscle expression of FGF2 and muscle <em>growth</em> and differentiation <em>factors</em> are reduced in patients with hypogonadotropic hypogonadism (HH) and whether testosterone replacement therapy results in their restoration.</AbstractText><AbstractText>This is a secondary analysis of a previously completed trial of testosterone replacement in men with type 2 diabetes and HH.</AbstractText><AbstractText>Clinical Research Center at a university.</AbstractText><AbstractText>Twenty-two men with HH and 20 eugonadal men were compared at baseline.</AbstractText><AbstractText>Twelve men with HH were treated with intramuscular injections of 250 mg testosterone every 2 weeks for <em>22</em> weeks, and 10 men received placebo injections. Quadriceps muscle biopsies and blood samples were obtained before and after testosterone therapy.</AbstractText><AbstractText>The expression of FGF2 and FGF receptor (FGFR)2 in skeletal muscle of men with HH was significantly lower than that in eugonadal men by 57% and 39%, respectively (P < 0.05). After <em>22</em> weeks of testosterone, the expression of FGF2 increased, whereas that of myogenic regulatory <em>factor</em> (MRF)4 and myostatin decreased significantly. There was no change in expression of FGFR2, myogenin, or myogenic differentiation protein in the skeletal muscle. Plasma FGF2 and IGF-1 concentrations increased after testosterone therapy.</AbstractText><AbstractText>These data show that testosterone is a major modulator of FGF2, MRF4, and myostatin expression in skeletal muscle. These effects may contribute to the increase in muscle mass after testosterone therapy.</AbstractText>

Previous studies have indicated that medium conditioned by 21-day-old fetal rat calvariae contains bioactive proteins termed bone-derived <em>growth</em> <em>factors</em> (BDGF) I and II. In the present studies we have purified the nontransforming BDGF II by dialysis, molecular sieving, three reverse phase HPLC steps, and preparative polyacrylamide gel electrophoresis. The second HPLC step (HPLC-2) yielded a recovery of <em>22</em>% of the biological activity and achieved a 1500-fold purification, resulting in 20 micrograms protein/liter calvarial conditioned medium; the third HPLC step was of limited value in the purification of BDGF. Analytical PAGE revealed that the majority of the protein in HPLC-2-purified BDGF migrated with a relative molecular mass (Mr) of 11,000 and two additional proteins were seen at a Mr of <em>22</em>,000-23,000. On preparative PAGE, the material migrating with a Mr of 11,000 stimulated parameters of bone and <em>fibroblast</em> <em>growth</em> in vitro, whereas the material with a Mr of <em>22</em>,000-23,000 had less biological activity. Isoelectric focusing revealed that BDGF had an isoelectric point (pI) of 5. BDGF enhanced the incorporation of [3H]thymidine into DNA in <em>fibroblast</em> and calvarial cultures and of [3H]proline into collagen and noncollagen protein in calvariae. In conclusion, fetal rat calvariae secrete a BDGF with an estimated Mr of 11,000 and a pI of 5; this material stimulates bone and <em>fibroblast</em> <em>growth</em> in vitro.

OBJECTIVE

Emerging evidence shows that <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> (FGF<em>22</em>) plays a critical role in the etiology of depression. However, the molecular mechanisms of FGF<em>22</em> are not fully comprehended. Here, the effect of FGF<em>22</em> in depression and its relationship with interleukin-1β (IL-1β) were investigated in clinical, animal, and cell experiments.

METHODS

Serum from depressive patients was collected, and the levels of FGF<em>22</em> and IL-1β were analyzed by ELISA. The chronic unpredictable mild stress (CUMS) model was established, and primary hippocampal neuronal cells were cultured to examine changes in FGF<em>22</em> and IL-1β levels in rat hippocampus.

RESULTS

The results revealed a negative correlation between serum FGF<em>22</em> levels and serum IL-1β levels. The expression of IL-1β in the CUMS rat hippocampus decreased, and the apoptosis of hippocampal cells improved after the injection of lentiviral vector-mediated FGF<em>22</em> (LV-FGF<em>22</em>). Further tests in primary hippocampal neuronal cells also showed a reduction in IL-1β and the cell apoptosis rate after treatment with FGF<em>22</em>.

CONCLUSIONS

In conclusion, the results revealed that FGF<em>22</em> plays a role in alleviating depression, which may be mediated by reduced expression of IL-1β.

During the peri-implantation period, ruminant conceptuses go through rapid elongation, followed by their attachment to the uterine endometrial epithelial cells, during which interferon-tau (IFNT), a trophectodermal cytokine required for the process of maternal recognition of pregnancy, is expressed in a temporal and spatial manner. On day <em>22</em> (day 0 = day of estrus), 2 to 3 d after the initiation of bovine conceptus attachment to the uterine epithelium, when IFNT production begins to subside, the expression of molecules related to epithelial-mesenchymal transition, zinc finger E-box binding homeobox 1, snail family transcriptional repressor 2, N-cadherin, and vimentin was found in the trophectoderm. Through the use of in vitro coculture system with bovine trophoblast CT-1 and endometrial epithelial cells, a series of experiments have been conducted to elucidate mechanisms associated with the regulation of IFNT gene transcription and conceptus implantation, including epithelial-mesenchymal transition processes. Expression of IFNT, both up- and downregulation, during the peri-implantation period is tightly controlled. Cytokines and cell adhesion molecules such as epidermal <em>growth</em> <em>factor</em>, basic <em>fibroblast</em> <em>growth</em> <em>factor</em>, transforming <em>growth</em> <em>factor</em> beta, activin A, L-selectin-podocalyxin, and vascular cell adhesion molecule 1-integrin α4 expressed in utero all contribute to the initiation of epithelial-mesenchymal transition in the trophectoderm. These results indicate that conceptus implantation to the uterine endometrium proceeds while elongated conceptuses and endometria express cell adhesion molecules and their receptors, and the trophectoderm experiences epithelial-mesenchymal transition. Data accumulated suggest that while the conceptus and the endometrial epithelium adhere, trophectodermal cells must gain more flexibility for binucleate and possibly trinucleate cell formation during the peri-implantation period, and that understanding and constructing the conditions throughout implantation processes is key to improving ruminants' fertility.

Accumulating evidence has indicated that long non‑coding RNAs (lncRNAs) have crucial roles in wound healing and that vascular lesions in diabetic wounds are frequently difficult to heal. However, the role of angiogenesis pathway‑associated lncRNAs in wound healing in diabetic patients has remained to be fully elucidated. In the present study, human skin <em>fibroblasts</em> were cultured under high‑glucose conditions in vitro to mimic a diabetic environment and the angiogenesis pathway‑associated lncRNA expression profile in the high‑ and normal‑glucose groups was examined. The microarray data indicated that 14 lncRNAs and <em>22</em> mRNAs were differentially expressed. Several candidate lncRNAs and mRNAs were then analyzed by reverse transcription‑quantitative PCR and the results were consistent with the microarray data. Furthermore, the University of California Santa Cruz Genome Browser was used to identify mRNAs linked to angiogenesis pathways near the transcriptional region of lncRNAs. The results suggested that lncRNAs RP4‑791C19.1 and CTD‑2589O24.1 may act on their target genes epidermal <em>growth</em> <em>factor</em> receptor and p21 (RAC1) activated kinase 1, respectively, as enhancers and cis‑regulate their expression. Therefore, the present study confirmed that several angiogenesis pathway‑associated lncRNAs were differentially expressed under high‑glucose conditions, which may have a key role in wound healing in patients with diabetes.

Background: Hyperphosphatemia confers adverse cardiovascular outcomes, and commonly occurs in late-stage CKD. Fibroblast growth factor 7 (FGF7) is a phosphaturic peptide which decreases renal phosphate transport in vitro and in vivo. Serum FGF7 concentrations are reduced in hyperphosphatemic patients with hypophosphatasia and are elevated in some hypophosphatemic patients with tumor-induced osteomalacia. No data, however, are available on whether circulating FGF7 concentrations increase to compensate for phosphate retention in CKD patients.

Methods: This was a cross-sectional study performed among 85 adult patients with varying estimated glomerular filtration rates (eGFR). We measured serum intact FGF7 (iFGF7) concentration using an iFGF7 immunoassay and determined its associated factors. Relationships between eGFR and mineral metabolism biomarkers [phosphate, iFGF7, iFGF23, parathyroid hormone (PTH), and 1,25-dihydroxyvitamin D (1,25(OH)₂D)] were explored.

Results: For eGFRs of ≥ 60 (n = 31), 45-59 (n = 16), 30-44 (n = 11), 15-29 (n = 15), and < 15 mL/min/1.73 m² (n = 12), median (IQ25-75) iFGF7 concentrations were 46.1 (39.2-56.9), 43.1 (39.0-51.5), 47.3 (38.3-66.5), 47.7 (37.7-55.8), and 49.6 (42.5-65.6) pg/mL, respectively (P = 0.62). Significant increases in serum iFGF23, PTH, and phosphate were observed at eGFRs of < 33 (95 % CI, 26.40-40.05), < 29 (95 % CI, 22.51-35.36), and < 22 mL/min/1.73 m² (95 % CI, 19.25-25.51), respectively, while significant decreases in serum 1,25(OH)₂D were observed at an eGFR of < 52 mL/min/1.73 m² (95 % CI, 42.57-61.43). No significant correlation was found between serum iFGF7 and phosphate, iFGF23, PTH or 1,25(OH)₂D. In multivariable analyses, body mass index (per 5 kg/m² increase) was independently associated with the highest quartile of serum iFGF7 concentration (OR, 1.20; 95 % CI, 1.12-1.55).

Conclusions: Compensatory decreases in circulating 1,25(OH)₂D and increases in circulating iFGF23 and PTH, but not iFGF7, facilitate normalization of serum phosphate concentration in early stages of CKD. Whether other circulating phosphaturic peptides change in response to phosphate retention in CKD patients deserves further study.

Keywords: Chronic kidney disease; Fibroblast growth factor; Parathyroid hormone; Phosphate; Vitamin D.

2D Autocorrelation, comparative molecular field analysis (CoMFA), and comparative molecular similarity indices analysis (CoMSIA) were undertaken for a series of substituted pyrido[2,3-d]pyrimidine derivatives to correlate platelet-derived <em>growth</em> <em>factor</em> receptor (PDGFR), <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor (FGFR), and c-Src tyrosine kinases' inhibition with 2D and 3D structural properties of <em>22</em> known compounds. QSAR models with considerable internal as well as external predictive ability were obtained. The relevant 2D autocorrelation descriptors for modeling each protein tyrosine kinase (PTK) inhibitory activity were selected by genetic algorithm (GA) and multiple linear regression (MLR) approach. The 2D autocorrelation space brings different descriptors for each PTK inhibition and suggests the atomic properties relevant for the inhibitors to interact with each PTK active site. CoMFA and CoMSIA were developed with a focus on interpretative ability using coefficient contour maps. CoMSIA produced significantly better results for all correlations. The results indicate a strong correlation between the inhibitory activity of the modeled compounds and the hydrophobic and H-bond donor fields around them.

What is the central question of this study? <em>Fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) plays important therapeutic roles in metabolic diseases but it is associated with bone loss, through insulin-like <em>growth</em> <em>factor</em> binding protein 1 (IGFBP1) in animals. However, the FGF21-IGFBP1 axis on age-related bone loss has not been explored in humans. What is the main finding and its importance? Using 'genetically-linked' parent and child family pairs, we show that FGF21, but not IGFBP1 concentration, is higher in older than younger adults. Our results suggest that the age-associated decline in bone mineral density is associated with FGF21 and increased bone turnover but unlikely through IGFBP1 in healthy humans. ABSTRACT: Introduction Bone fragility increases with age. The <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21)-insulin-like <em>growth</em> <em>factor</em> binding protein 1 (IGFBP1) axis regulates bone loss in animals. However, the role of FGF21 in mediating age-associated bone fragility in humans remains unknown. Purpose This study explored the FGF21-regulatory axis in bone turnover and age-related decline in bone mineral density (BMD). Methods Twenty 'genetically-linked' family (parent and child) pairs were recruited. Younger adults were <em>22</em>-39 years old and older adults were 60-71 years old. BMD and serum concentrations of FGF21, IGFBP1, receptor activator of nuclear <em>factor</em> kappa-B ligand (RANKL), tartrate-resistant acid phosphatase 5b (TRAP5b) and bone-specific alkaline phosphatase (BAP) were measured. Results Older adults had 10-18% lower BMD at the hip and spine (p < 0.008) and 2-fold higher FGF21 concentration (p < 0.001). IGFBP1 concentration was similar in younger and older adults (p = 0.961). RANKL concentration was 44% lower (p = 0.006) while TRAP5b and BAP concentrations were 36% and 31% higher (p = 0.01 and p = 0.004) in older adults than in younger adults. Adjusting for sex did not affect these results. FGF21 concentration was negatively correlated with BMD at the spine (r = -0.460,p = 0.003), but not with IGFBP1 concentration (r = -0.144,p = 0.374). IGFBP1 concentration was not correlated with BMD at the hip or spine (all p > 0.05). Conclusions In humans, FGF21 may be involved in the age-associated decline in BMD, especially at the spine, through increased bone turnover. IGFBP1 is unlikely the downstream effector of FGF21 in driving age-associated decline in BMD and in RANKL-associated osteoclast differentiation. This article is protected by copyright. All rights reserved.