Angiogenesis is crucial for tumour <em>growth</em> and formation of metastasis. Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) is known to have potent angiogenic activity and has been identified in a wide variety of malignancies including head and neck squamous cell carcinomas (HNSCC). Clarification of its localization in HNSCC is important for the understanding of angiogenesis. Cryosiat sections of 27 HNSCC were immunostained for bFGF using a standard streptavidin-biotin complex procedure. Western blot analysis revealed three immunoreactive bFGF isoforms of 18, <em>22</em> and 24 kDa. Immunohistochemical and -cytochemical localization of bFGF was studied at light and electron microscopic levels. bFGF was mainly localized within the focal tumoral areas rather than in the tumour stroma. On a subcellular level, the ultrastructural investigation showed electron-dense bFGF localization in the cytosol of the carcinoma cells, but bFGF labelling within the nuclei of HNSCC cancer cells was one prominent finding of this study. In conclusion, the presence of bFGF isoforms in most of the cancer cells supports the theory of a direct paracrine mechanism of bFGF from cancer cells on tumour angiogenesis. The nuclear localization of bFGF in the HNSCC cells supports its activity as a transcriptional <em>factor</em>.

Alveolar macrophages and their products are thought to be important mediators of the inflammatory lesions and consequent interstitial fibrosis caused by inhalation of inorganic particles. Identification of a homolog of platelet-derived <em>growth</em> <em>factor</em> (PDGF) produced by rat alveolar macrophages that were stimulated with carbonyl iron particles and asbestos fibers motivated our studies on the biologic activity of this potent cytokine. Macrophage-derived PDGF (MD-PDGF) competes for specific membrane receptors on rat lung <em>fibroblasts</em>, initiating DNA synthesis and cell replication. The present report demonstrates that purified human PDGF and the MD-PDGF are chemotactic for early passage rat lung <em>fibroblasts</em>, but not for lung macrophages. Rat lung <em>fibroblasts</em> exhibit a typical bell-shaped, dose-related curve and respond optimally between 2 and 4 ng/ml PDGF. We found that alveolar macrophage-conditioned medium (AMCM), fractionated by gel filtration in 1 M acetic acid, induced a clear chemotactic response in the same fractions (20 to <em>22</em> ml) where PDGF was identified by enzyme immunoassay. In contrast, AMCM fractionated by gel filtration in phosphate-buffered saline did not induce any chemotactic activity unless the fractions were treated further with 1 M acetic acid. In this case, chemotactic activity was observed in those fractions with molecular weights of 150 and greater than 200 kD. All chemotactic activity observed with fractionated AMCM was blocked greater than 90% by an anti-PDGF antibody. These observations demonstrate that MD-PDGF is chemotactic for rat lung <em>fibroblasts</em> if it first is released from its binding protein, alpha-macroglobulin (alpha-M), which is secreted into the medium along with PDGF.(ABSTRACT TRUNCATED AT 250 WORDS)

The aim of the present study was to identify the biomarkers involved in the development of hepatocellular adenoma (HCA) through integrated analysis of gene expression and methylation microarray. The microarray dataset GSE7473, containing HNF1α-mutated HCA and their corresponding non-tumor livers, 5 HNF1α-mutated HCA and 4 non-related non-tumor livers, was downloaded from the Gene Expression Omnibus (GEO) database. The DNA methylation profile GSE43091, consisting of 50 HCA and 4 normal liver tissues, was also downloaded from the GEO database. Differentially expressed genes (DEGs) were identified by the limma package of R. A t-test was conducted on the differentially methylated sites. Functional enrichment analysis of DEGs was performed through the Database for Annotation, Visualization and Integrated Analysis. The genes corresponding to the differentially methylated sites were obtained by the annotation files of methylation chip platform. A total of 182 DEGs and 3,902 differentially methylated sites were identified in HCA. In addition, 238 enriched GO terms, including organic acid metabolic process and carboxylic acid metabolic process, and 14 KEGG pathways, including chemical carcinogenesis, were identified. Furthermore, 12 DEGs were identified to contain differentially methylated sites, among which, 8 overlapped genes, including pregnancy zone protein and solute carrier family <em>22</em> member 1 (SLC<em>22</em>A1), exhibited inverse associations between gene expression levels and DNA methylation levels. The DNA methylation levels may be potential targets of HCA. The present study revealed that the 8 overlapped genes, including annexin A2, chitinase 3-like 1, <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 4, mal, T-cell differentiation protein like, palladin, cytoskeletal associated protein, plasmalemma vesicle associated protein and SLC<em>22</em>A1, may be potential therapeutic targets of HCA.

Retinoic acid has important actions on cell differentiation and osteoblastic function, and some of these actions may be mediated by changes in the insulin-like <em>growth</em> <em>factor</em> (IGF) axis. Skeletal cells synthesize IGF I and II and the six known IGF binding proteins (IGFBP). IGFBP-6 binds IGF II with high affinity and prevents IGF II-mediated effects. In <em>fibroblasts</em>, IGFBP-6 levels are regulated by retinoic acid, and we postulated that retinoic acid may regulate IGF II in bone by altering IGFBP-6 synthesis. We examined the effect of retinoic acid on IGFBP-6 expression in cultures of osteoblast-enriched cells from <em>22</em>-day fetal rat calvariae (Ob cells). Retinoic acid caused a time- and dose-dependent increase in IGFBP-6 mRNA levels, as determined by Northern blot analysis. The effect was maximal after 48 h of treatment and observed with retinoic acid at concentrations of 10 nM to 1 microM. Retinoic acid increased IGFBP-6 polypeptide levels in the culture medium, as determined by Western immunoblot analysis. Cycloheximide at 3.6 microM slightly decreased IGFBP-6 transcripts but did not prevent the stimulatory effect of retinoic acid. The decay of IGFBP-6 mRNA in transcriptionally arrested Ob cells was similar in control and retinoic acid-treated cells, and retinoic acid increased the rates of IGFBP-6 transcription, as determined by nuclear run on assays. In conclusion, retinoic acid enhances IGFBP-6 expression in Ob cells by transcriptional mechanisms. Since IGFBP-6 prevents the effects of IGF II, increased synthesis of IGFBP-6 could mediate selected actions of retinoic acid in bone.

Insulin-like <em>growth</em> <em>factor</em> 1 (IGF-1) is a potent mitogen and differentiation <em>factor</em> with particular relevance to orthopedic tissue engineering. A biologically based Ca2+-alginate microcapsule vehicle, utilizing genetically modified primary normal human <em>fibroblasts</em> (NHFs), was developed and characterized for localized synthesis and delivery of human IGF-1 (hIGF-1). Normal human <em>fibroblasts</em> were transfected to overexpress the hIGF-1 gene, leading to cells that expressed 4 ng of hIGF-1 per 10(6) cells per 24 hours. Encapsulation within alginate led to a six-fold enhancement in the generation and release of hIGF-1 to <em>22</em> ng of hIGF-1 per 10(6) cells per 24 hours. Release was constitutive, predictable, and exhibited highly repeatable first-order kinetics with no initial burst. Released <em>growth</em> <em>factor</em> was biologically active and exhibited a proliferative effect comparable to commercially available recombinant hIGF-1. The magnitude of hIGF-1 release met the requirements of orthopedic tissue generation, and this approach is considered an attractive alternative to other proposed methods of <em>growth</em> <em>factor</em> delivery.

BACKGROUND

Oncogenic osteomalacia is an acquired form of hypophosphatemic osteomalacia where the tumour resection may lead to cure of the disease. Tumours originating from the musculoskeletal region form an important subgroup of oncogenic osteomalacia.

METHODS

This was a retrospective study conducted at a tertiary care centre in south India where we analyzed the hospital records of all the patients with musculoskeletal oncogenic osteomalacia from January 2010-April 2016.

RESULTS

A total number of 73 patients were diagnosed to have adult onset hypophosphatemic osteomalacia out of which 13 patients (M: F = 6:7; mean age: 45.38 ± 18.23 years) with musculoskeletal oncogenic osteomalacia were included in the study. Common presenting symptoms were bony pains, proximal myopathy and fractures. Mean duration of symptoms from the initial hospital visit was 58.46 ± 64.48 months. The initial mean <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) 23 levels being 828.86 ± 113.<em>22</em> RU/ml (Normal range: <em>22</em>-91). Imaging modalities used for localization of the tumour: DOTATATE PET/CT (8 patients), FDG PET/CT (3 patients), 1 patient (Both DOTATATE PET/CT and FDG PET/CT) and whole body Tc 99 m Red blood cell (RBC) blood pool scintigraphy (2 patients). 9 patients underwent surgery and all achieved remission. 4 patients denied surgical consent.

CONCLUSIONS

Musculoskeletal oncogenic osteomalacia is a major subgroup of oncogenic osteomalacia which need more extensive whole body imaging for the localization of the tumour. Surgical excision often leads to remission of the disease.

The majority of cells are cultured with Dulbecco's modified Eagle's medium (DMEM) or RPMI supplemented with fetal bovine serum (FBS), which contains numerous <em>factors</em>, including cytokines, nutrients and unknown <em>growth</em> <em>factors</em>. These <em>factors</em> may affect cell <em>growth</em>, apoptosis and differentiation. The serum-free medium, STK2, has been previously reported as suitable for the cell culture of human mesenchymal stem cells. However, how STK1 or STK2 affect the cell proliferation of normal and cancer cells remains unknown. The present study examined the <em>growth</em> of the human gingival <em>fibroblast</em> (HGF-1) cell-line and the HSC-3, CA9-<em>22</em> and MSTO cancer cell-lines, cultured with STK1 and STK2. STK1 increased the cell proliferation of HGF-1 compared to DMEM by assessment with the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)- 2H-tetrazolium (MTS) assay, whereas STK1 and STK2 markedly inhibited the cell proliferation of HSC-3 and MSTO. The cell proliferation rate of CA9-<em>22</em> cultured with STK1 or STK2 for 96 h was ~2-fold higher than the rate for 24 h culture. The shape of the HSC-3 cells was also found to have changed to round when cultured with STK2. These results indicate that STK1 increased the cell proliferation of HGF-1 compared to DMEM, whereas the proliferation of HSC-3 and MSTO was inhibited by STK1 and STK2. Thus, STK1 and STK2 had different affects on the cell <em>growth</em> of HGF-1, CA9-<em>22</em>, HSC-3 and MSTO.

The 8p11 myeloproliferative syndrome is an aggressive neoplasm associated with chromosomal abnormalities involving rearrangement of the <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 (FGFR1) gene. We report herein a rare case of a t(1;8)(q25;p11.2) with a TPR-FGFR1 rearrangement, in which the patient presented with myeloproliferative neoplasm-like symptoms and T-lymphoblastic lymphoma. Sequence analysis revealed a fusion transcript with exon <em>22</em> of the TPR gene joined to exon 13 of the FGFR1 gene, which is a novel breakpoint for the TPR gene in the TPR-FGFR1 rearrangement.

Biomechanical remodeling of stroma by cancer-associated <em>fibroblasts</em> (CAF) in early stages of cancer is critical for cancer progression, and mechanical cues such as extracellular matrix stiffness control cell differentiation and malignant progression. However, the mechanism by which CAF activation occurs in low stiffness stroma in early stages of cancer is unclear. Here, we investigated the molecular mechanism underlying CAF regulation by SPIN90 and microtubule acetylation under conditions of mechanically soft matrices corresponding to normal stromal rigidity. SPIN90 was downregulated in breast cancer stroma but not tumor, and this low stromal expression correlated with decreased survival in breast cancer patients. Spin90 deficiency facilitated recruitment of mDia2 and APC complex to microtubules, resulting in increased microtubule acetylation. This increased acetylation promoted nuclear localization of YAP, which upregulated expression of myofibroblast marker genes on soft matrices. Spin90 depletion enhanced tumor progression, and blockade of microtubule acetylation in CAF significantly inhibited tumor <em>growth</em> in mice. Together, our data demonstrate that loss of SPIN90-mediated microtubule acetylation is a key step in CAF activation in low stiffness stroma. Moreover, correlation among these <em>factors</em> in human breast cancer tissue supports the clinical relevance of SPIN90 and microtubule acetylation in tumor development. Cancer Res; 77(17); 4710-<em>22</em>. ©2017 AACR.

Adult neurogenesis and the neurogenic niche in the dentate gyrus are subjects of much research interest. Enhancing our knowledge of this niche process and the role played by this unique microenvironment would further our understanding of plasticity and its relevance for cognition in health and disease. The complex three-dimensional (3D) nature of the niche microenvironment is poorly recapitulated in current cell culture experimental procedures. Neural precursor cells (NPCs) are cultured either on two-dimensional (2D) surfaces, where cells quickly reach confluency and passaging is required, or as 3D neurospheres, with the limitation of poor diffusion of nutrients and thus partial differentiation of cells over time. Herein, we culture NPCs on microscale scaffolds termed microcarriers, composed of poly(ethylene glycol) and heparin, designed to more closely represent the 3D environment of the neurogenic niche. The interconnected macroporous structure of the microcarriers allows NPCs to attach to their pore walls with subsequent continuous proliferation (analyzed up to 28 days) without formation of a necrotic core. Removal of basic <em>fibroblast</em> <em>growth</em> <em>factor</em> and epidermal <em>growth</em> <em>factor</em> from the culture medium results in differentiation of the NPCs. Unlike 2D culture, a high percentage of neurons was achieved on the microcarriers (<em>22</em>% MAP2 positive cells) indicating that these 3D microscale scaffolds give a more conducive environment for neuronal differentiation. Microcarrier culture of NPCs allows long-term cell expansion and better differentiation, which provides superior culture conditions for studying/modelling the neurogenic niche.

<em>Fibroblast</em> <em>growth</em> <em>factor</em>-23 (FGF-23) secreted by osteocytes is known as a circulating <em>factor</em> that is essential for phosphate homeostasis. Recent studies have implicated FGF-23 in the nociceptive signalling of peripheral sensory neurons. However, the relevant mechanisms underlying this effect are not known. In this study, we determine the role of FGF-23 in regulating T-type Ca2+ channels (T-type channels) in small-diameter dorsal root ganglion (DRG) neurons in mice. Our results show that FGF-23 increases T-type channel currents in a concentration-dependent manner. This FGF-23-induced response was dependent on FGF type 1 receptor (FGFR1) and was accompanied by a depolarizing shift in the steady-state inactivation curve. Pretreatment of neurons with the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 prevented the FGF-23-mediated T-type channel response. Analysis of phospho-Akt (p-Akt) revealed that FGF-23 significantly activated Akt, but Akt inhibition did not affect the FGF-23-induced T-type channel current increase. The cell-permeable protein kinase A (PKA) inhibitor KT-5720 pretreatment and intracellular application of PKI 6-<em>22</em> both abolished the stimulatory effects of FGF-23 on T-type channels, but inhibition of PKC had no effect. In summary, these findings indicate that FGF-23 stimulates T-type channel activity via activation of FGFR1, which is coupled to the PI3K-dependent PKA signalling cascade in small DRG neurons.

Rat stromal bone-marrow cells cultured in the presence of dexamethasone, ascorbic acid, beta-glycerophosphate, and <em>fibroblast</em> <em>growth</em> <em>factor</em>-2 (FGF-2) express the osteogenic phenotype (Pitaru et al., J. Bone Miner. Res. 8:919-929, 1993). The purpose of this study was to establish a long-term homogeneous culture expressing the osteogenic phenotype. The cultures were routinely passaged every 5 days in the absence or presence of either or both dexamethasone and FGF-2, and the cumulative doubling number and the expression of the osteogenic phenotype were determined. Cultures treated with dexamethasone (10(-7) M) ceased proliferation and only upon addition of FGF-2 (3 ng/ml) was a spontaneous immortalization achieved, as expressed by sustained proliferation for about 1 year, with a doubling time of <em>22</em> h and more than 300 doublings in 72 passages. Both FGF-2 and dexamethasone are required and act synergistically to maintain cell propagation, alkaline phosphatase expression, and osteocalcin secretion; however, protein content was FGF-2 dependent and the mineralization was dexamethasone dependent. Repetitive single-cell cloning tested the homogeneity and stability of the cells expressing the osteogenic phenotype in these long-term cultures. It was shown that 25% to 50% of subclones derived from clones with an osteogenic phenotype do not further express the osteogenic phenotype. In conclusion, we have established a spontaneously immortalized dexamethasone- and FGF-2-dependent rat stromal bone-marrow-derived long-term culture expressing the osteogenic phenotype. The cultures tend to lose the osteogenic phenotype, and dexamethasone supports the long-term preservation of the osteogenic phenotype.

The cooperative cell kinetic actions of ET-1 with TGF-alpha or EGF in normal rat kidney <em>fibroblasts</em> (NRK-49F) and KNRK cells (Kirsten MSV transformed) were analyzed by [3H]-thymidine incorporation assay and flow cytometry. A marked synergistic effect of TGF-alpha and ET-1 (or EGF and ET-1) on DNA synthesis and G1 to S transition was observed in NRK cells; 15-20% S for TGF-alpha and 12% S for ET-1 alone but 45-50% S in combination. There was no detectable effect on cell cycle kinetics by TGF-alpha (1 ng/ml) or EGF (1 ng/ml) plus ET-1 (1 ng/ml) in KNRK cells treated for <em>22</em> hours. Insulin, insulin-like <em>growth</em> <em>factor</em> I (IGF-I), <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF), platelet derived <em>growth</em> <em>factor</em> (PDGF), and transforming <em>growth</em> <em>factor</em> beta (TGF-beta) were also tested and found to have no significant synergistic effects on ET-1 actions. Our findings suggest that the combination of TGF-alpha (EGF) and ET-1 is an important part of an intricate network which coordinates progression of G1 to S phase in normal cells.

<AbstractText>Dysembryoplastic neuroepithelial tumors (DNTs) are slow-<em>growing</em> glioneuronal tumors, and their genetic backgrounds are getting unveiled. Recently, <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 internal tandem duplication (FGFR1-ITD) of the tyrosine kinase domain (TKD) has been demonstrated by whole-genome sequencing.</AbstractText><AbstractText>Here, we analyzed <em>22</em> DNTs using multiplex ligation-dependent probe amplification (MLPA) with formalin-fixed paraffin-embedded specimens and found a copy number gain in TKD of FGFR1 (13 cases, 59%), which suggested the presence of FGFR1-ITD. Another 5 DNTs harbored FGFR1 hot spot mutations including a double mutant case, and FGFR1 alterations were detected in 18 DNTs (82%). The BRAF V600E mutation, another important mutation in DNTs, was not observed.</AbstractText><AbstractText>With recent findings of less frequent or absent FGFR1-ITD in pilocytic astrocytomas or rosette-forming glioneuronal tumors, the analysis of FGFR1 aberrations, especially FGFR1-ITD, was suggested to be helpful to discriminate DNTs from their histological mimics.</AbstractText>

The <em>growth</em> <em>factors</em> basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) and insulin-like <em>growth</em> <em>factor</em> 1 (IGF-I) have been implicated in the pathophysiology of atherosclerosis and restenosis. The Tibetan herbal preparation PADMA-28 (a mixture of <em>22</em> plants which is used as an anti-atherosclerosis agent) was tested for its ability to inhibit the mitogenic activity of bFGF and IGF-I, <em>growth</em> <em>factors</em> involved in restenosis, atherosclerosis and tumour progression. DNA synthesis and proliferation of vascular smooth muscle cells, in response to serum bFGF, thrombin, or combinations thereof, were abrogated in the presence of microgram amounts of both the aqueous and organic, partially purified, extracts of PADMA-28. These fractions also inhibited IGF-I-mediated proliferation, migration and invasion of tumour cells responsive to IGF-I. The inhibition by PADMA 28 was reversible upon removal of the PADMA extracts, indicating that the effects were not related to cell toxicity. These and other properties (i.e., anti-oxidant activity) of PADMA-28 may be responsible for its beneficial effect as an anti-atherosclerotic agent, suggesting that this herbal preparation may have potential applications in the prevention of intimal hyperplasia and arterial stenosis secondary to coronary angioplasty and bypass surgery, as well as in the prevention and treatment of other vascular diseases and tumour <em>growth</em> and metastasis.

Bovine adrenal cortical cells <em>growth</em> on extracellular matrix-coated dishes in the presence of F-12 medium supplemented with high density lipoprotein (30 micrograms protein/ml), insulin (50 ng/ml), transferrin (1 microgram/ml), and <em>fibroblast</em> <em>growth</em> <em>factor</em> (100 ng/ml) expressed high affinity low density lipoprotein (LDL) receptors (Kd = 2.0 X 10(-8) M) present at a density of 3 X 10(4) receptors/cell. The density of LDL receptors per cell could be increased 3-fold (9 X 10(4) receptors/cell) by incubating the cells for 24 h with cholera toxin (CT; 10 ng/ml), but not with insulin (100 ng/ml). This correlated with increased LDL internalization (10-fold) and degradation (5-fold). 11-Deoxycortisol (11 DOC) release was increased by 2.7-fold. The addition of insulin (100 ng/ml) together with CT (10 ng/ml) markedly potentiated the effects of CT on LDL binding, internalization, and degradation as well as on steroid release. Preincubation (24 h) of the cells with insulin (100 ng/ml) together with CT resulted in a 12.7-fold increase in high affinity LDL receptor density (3.6 X 10(5) receptors/cell), while LDL internalization and degradation were increased by 45- and <em>22</em>-fold, respectively. This correlated with a 9.6-fold increase in the 11 DOC released into the medium. The effects of insulin on the induction of high affinity LDL receptors as well as increased internalization and degradation of [125I]LDL were both time and concentration dependent in cultures exposed to CT. Secretion of 11 DOC paralleled in a time-dependent manner the amount of internalized [125I]LDL. Exposure of the cells to chloroquine (100 microM) resulted in a 95% inhibition of [125I]LDL degradation correlating with an 80% decrease in 11 DOC released in cells preexposed to CT and insulin. The results of these studies suggest that in bovine adrenal cortex cells grown in chemically defined medium, insulin modulates steroidogenic cAMP-induced response by increasing both the LDL receptor pathway activity and 11 DOC release.

Thrombospondin-1 (TSP-1) is a multifunctional matrix protein implicated in cancer cell adhesion, migration, and invasion, inhibition of angiogenesis, and activation of latent transforming <em>growth</em> <em>factor</em>-beta (TGF-beta). The effect of cell density was investigated on the production of TSP-1, basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), and vascular endothelial <em>growth</em> <em>factor</em> (VEGF) by two glioblastoma cell lines. The effect of TGF-beta was also examined. The amount of intracellular TSP-1 protein decreased significantly as the cell density increased in cultures of both T98G and A172 cells. The amount of intracellular TSP-1 was highest in sparse tumor cell cultures and lowest in densely confluent tumor cell cultures. The maximum reduction of TSP-1 protein production was 56.8% and 44.6% in T98G and A172 cells, respectively. The cell density did not affect the production of bFGF or VEGF. TGF-beta2 treatment did not affect the production of TSP-1, bFGF, or VEGF proteins. Treatment with excess TGF-beta2 resulted in a slight but significant decrease (<em>22</em>%; P < 0.02) of TGF-beta2 production by A172 cells, but not by T98G cells. The present results indicate that the production of TSP-1 protein is regulated by cell density of glioblastoma cells, while that of angiogenic <em>factors</em> is not affected by tumor cell density. This suggests that high tumor cell density may tilt the angiogenic balance in favor of angiogenesis.

<em>Fibroblast</em> <em>growth</em> <em>factor</em>-23 (FGF-23) accumulates in blood of patients with chronic kidney disease (CKD) and is associated both with cardiovascular complications and disease progression. However, our knowledge of the sites and mechanisms that regulate plasma FGF-23 is still incomplete. We measured plasma intact FGF-23 across the kidney, splanchnic organs, and lung in 11 patients [estimated glomerular filtration rate (eGFR) 60 ± 6 ml/min] during elective diagnostic cardiac catheterizations. In these patients FGF-23 was removed by the kidney, with a fractional extraction (FE) of ∼<em>22</em>%. The FE of FGF-23 across the kidney was similar to that of creatinine (∼17%, P = NS). In addition, the FGF-23 FE by the kidney was significantly directly related to eGFR (r = 0.709 P = 0.018) and to kidney creatinine FE (r = 0.736 P = 0.013) but only as a trend to plasma phosphate levels (r = 0.55, P = 0.18). There was no difference in FGF-23 levels in blood perfusing splanchnic organs and cardiopulmonary bed. However, the arterial-venous difference of FGF-23 across the lung was directly related to FGF-23 pulmonary artery levels, suggesting that the lung, and possibly the heart, participate in the homeostasis of plasma FGF-23 when its systemic levels are increased. Our data show that the human kidney is the only site for FGF-23 removal from blood and suggest that FGF-23 is predominantly removed by glomerular filtration. The kidney ability to remove FGF-23 from the circulation likely accounts for the early increase in blood of FGF-23 in patients with CKD.

BACKGROUND

The average population age is increasing and the incidence of age-related vascular complications is rising in parallel. Impaired wound healing and disordered ischemia-mediated angiogenesis are key contributors to age-impaired vascular complications that can lead to amputation. High-density lipoproteins (HDL) have vasculo-protective properties and augment ischemia-driven angiogenesis in young animals. We aimed to determine the effect of reconstituted HDL (rHDL) on aged mice in a murine wound healing model and the hindlimb ischemia (HLI) model.

METHODS

Murine wound healing model-24-month-old aged mice received topical application of rHDL (50 μg/wound/day) or PBS (vehicle control) for 10 days following wounding. Murine HLI model-Femoral artery ligation was performed on 24-month-old mice. Mice received rHDL (40 mg/kg) or PBS, intravenously, on alternate days, 1 week pre-surgery and up to 21 days post ligation. For both models, blood flow perfusion was determined using laser Doppler perfusion imaging. Mice were sacrificed at 10 (wound healing) or 21 (HLI) days post-surgery and tissues were collected for histological and gene analyses.

RESULTS

Daily topical application of rHDL increased the rate of wound closure by Day 7 post-wounding (25 %, p < 0.05). Wound blood perfusion, a marker of angiogenesis, was elevated in rHDL treated wounds (Days 4-10 by <em>22</em>-25 %, p < 0.05). In addition, rHDL increased wound capillary density by 52.6 %. In the HLI model, rHDL infusions augmented blood flow recovery in ischemic limbs (Day 18 by 50 % and Day 21 by 88 %, p < 0.05) and prevented tissue necrosis and toe loss. Assessment of capillary density in ischemic hindlimb sections found a 90 % increase in rHDL infused animals. In vitro studies in <em>fibroblasts</em> isolated from aged mice found that incubation with rHDL was able to significantly increase the key pro-angiogenic mediator vascular endothelial <em>growth</em> <em>factor</em> (VEGF) protein (25 %, p < 0.05).

CONCLUSIONS

rHDL can promote wound healing and wound angiogenesis, and blood flow recovery in response to ischemia in aged mice. Mechanistically, this is likely to be via an increase in VEGF. This highlights a potential role for HDL in the therapeutic modulation of age-impaired vascular complications.

Background: Methylcitric acid (MCA) analysis has been mainly utilized for the diagnosis of propionate disorders or as a second tier test in newborn screening, but its utility for patients monitoring still need to be established. We explored the potential contribution of MCA in the long-term management of organic acidurias.

<strong class="sub-title"> Patients and methods: </strong> We prospectively evaluated plasma MCA and its relationship with disease biomarkers, clinical status and disease burden in <em>22</em> patients, 13 with propionic acidemia (PA) and 9 with methylmalonic acidemia (MMA) on standard treatment and/or after transplantation. Samples were collected at scheduled routine controls or during episodes of metabolic decompensation (MD), 10 patients were evaluated after transplantation (6 liver, 2 combined liver and kidney, 2 kidney).

Results: MCA levels were higher in PA compared to MMA and its levels were not influenced by the clinical status (MD vs well state). The disease burden showed a direct correlation with MCA and FGF21, for both diseases. In MMA, MCA was higher in elder patients and, along with FGF21 and plasma methylmalonic acid, negatively correlated with GFR. In both diseases, MCA correlated with ammonia, glycine, lysine, C3, and the C3/C2, C3/C16 ratios. All transplanted patients showed a significant reduction of MCA in comparison to baseline values, with some differences dependent on the type of transplantation.

Conclusions: Our study provided new insights in understanding the disease pathophysiology, showing similarities between MCA and FGF21 in predicting disease burden, long-term complications and in evaluating the impact of organ transplantation. This article is protected by copyright. All rights reserved.

Keywords: Methylcitric acid; fibroblast growth factor 21; follow-up; methylmalonic acidemia; propionic acidemia; transplantation.

OBJECTIVE

To explore the effect of basic fibroblast growth factor (bFGF) on focal cerebral ischemic injury and antioxidant enzyme activities.

METHODS

Rats underwent 24-h middle cerebral artery occlusion by intraluminal suture. Infarction volume was shown with staining and quantitated by image analysis system. Neurologic deficit scores were determined with a 0-5 grade scale. Antioxidant enzyme activities of forebrains were detected.

RESULTS

bFGF (45 micrograms.kg-1.h-1 i.v. for 3 h, started 5 min after the onset of ischemia) showed potent neuroprotective effects. Infarction volumes were decreased from 272 mm3 +/- 22 mm3 (saline-treated) to 201 mm3 +/- 30 mm3 (bFGF-treated). Neurologic deficit scores were decreased from 3.6 +/- 1.5 (saline-treated) to 2.3 +/- 1.6 (bFGF-treated). Focal cerebral ischemia induced an increase in the activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), but a decrease in the level of glutathione s-transferase (GSH-ST). Treatment of bFGF further increased the CAT activity but had no effect on the activities of SOD, GSH-PX, and GSH-ST.

CONCLUSIONS

bFGF has a neuroprotective effect against focal cerebral ischemic injury. The elevation of CAT activity by bFGF may be involved in this effect.

♦

Vascular calcification is strongly associated with cardiovascular disease and mortality. However, some factors related to vascular calcification in patients with end-stage renal disease receiving peritoneal dialysis (PD) remain unknown. This study aimed to evaluate the associations of osteoprotegerin (OPG), osteopontin (OPN), osteocalcin (OCN), fibroblast growth factor 23 (FGF-23), magnesium, and phosphate clearance with vascular calcification in PD subjects, assessed by plain radiographs. ♦

Simple vascular calcification scores (SVCS) obtained from plain X-rays of the pelvis and hands, and the Kauppila Index (KI) from lateral lumbar X-rays were assessed in 76 adults receiving PD for ≥ 6 months (43 women, median age 39 years, median time on PD 1.4 years). Levels of OPG, OPN, OCN, and FGF-23 were determined by luminometry. ♦

Serum OPG levels were higher in subjects with vascular calcification (n = 22 with SVCS>> 3; n = 19 with KI>> 7) compared with those with less calcification (p < 0.001). Spearman's correlation coefficients between OPG and SVCS and KI were r = 0.49 and r = 0.51, respectively (both p < 0.001). Subjects with vascular calcification had significantly lower renal phosphate clearance. Multiple regression analysis showed that vascular calcification assessed by SVCS was associated with age (r = 0.2, p = 0.042), diabetes mellitus (r = 2.4, p < 0.001), body mass index (BMI) (r = 0.09, p = 0.037), and OPG (r = 0.22, p = 0.001). Vascular calcification assessed by KI was associated with age (r = 0.16, p < 0.001), time on PD (r = 0.54, p = 0.001) and OPG (r = 0.08, p = 0.04). Osteocalcin, OPN, FGF-23, and magnesium were not associated with vascular calcification. ♦

Higher levels of OPG were consistently associated with vascular calcification in subjects on PD.

BACKGROUND

We aimed to compare the effect of unfractionated heparin (UFH) and enoxaparin used as anticoagulants during hemodialysis (HD) on circulating levels of heparin-binding, endothelial-derived, proangiogenic factors-vascular endothelial (VEGF(165)) and basic fibroblast (bFGF) growth factor.

METHODS

We enrolled 22 chronic HD patients, who were randomly assigned to either enoxaparin (n=11) or UFH (n=11) anticoagulation, and followed prospectively for 12 weeks before crossing over to the alternate therapy for further 12 weeks. The cytokines were measured by immunoassay at the start, at 10 and 180min of HD.

RESULTS

The baseline VEGF(165) and bFGF levels were comparable during enoxaparin and UFH treatment. VEGF(165) significantly decreased during both enoxaparin (chi(2) ANOVA=33.0, P<10(-6)) and UFH (chi(2) ANOVA=27.2, P<10(-6)) anticoagulated HD, while over-HD bFGF remained stable regardless of the type of heparin. The switch from enoxaparin to UFH treatment was connected with 34% VEGF(165) decrease after 180min of HD and had no impact on bFGF. During UFH-anticoagulated HD 75% VEGF(165) decrease after 10min was negatively associated with heparin dosage and was more profound in patients with ischemic heart disease.

CONCLUSIONS

The traditional UFH regimen, in contrast to enoxaparin treatment, is connected with dose-depended VEGF(165) decrease during HD procedure. The biological and possible clinical relevance of this observation requires further investigations.

BACKGROUND

Low levels of vitamin D are linked to numerous adverse clinical conditions in hemodialysis (HD) patients, including disturbances of mineral and bone metabolism and increased mortality. Klotho, a molecule involved in such processes as phosphate homeostasis and aging, exists in 2 forms: a transmembrane protein acting as a coreceptor for fibroblast growth factor 23 (FGF-23) and soluble form, which is formed by cleavage of the extracellular domain of this molecule.

OBJECTIVE

The aim of the study was to evaluate the effect of cholecalciferol supplementation on soluble Klotho levels in HD patients.

METHODS

This was a prospective, open-label trial examining the effects of cholecalciferol supplementation on selected laboratory markers in 22 patients on HD. Vitamin D deficiency was assessed by the measurement of 25-hydroxyvitamin D [25(OH)D] levels. Soluble Klotho, intact FGF-23, intact parathormone (iPTH), and markers of bone formation and resorption were measured at baseline and after 12 weeks of cholecalciferol supplementation.

RESULTS

The levels of 25(OH)D increased, while those of iPTH and cross-linked C-telopeptide of type 1 collagen decreased significantly. Cholecalciferol treatment reduced the median concentration of soluble Klotho (from 438.73 pg/ml; interquartile range, 257.99-865.51 pg/ml; to 370.94 pg/ml; 181.72-710.91 pg/ml; P <0.05). FGF‑23 levels were not affected by the treatment.

CONCLUSIONS

Supplementation with cholecalciferol in HD patients decreases soluble Klotho levels without affecting the FGF-23 concentration. Replenishment of vitamin D stores results in a decrease in iPTH levels and reduced bone resorption.