The <em>fibroblast</em> <em>growth</em> <em>factors</em> (FGFs) are involved in hematopoiesis and tumorigenesis. However, little is known about the contribution of the FGFs identified within the past 10 years to leukemogenesis. To elucidate whether these FGFs (FGF-8, -9, -10, -11, -12, -13, -14, -16, -17, -18, -<em>19</em>, -20, and -21) are expressed in leukemic cells, we performed RT-PCR analyses using 28 cell lines. The members of a fetal-oncogenic subfamily, FGF-8/-17/-18, were often expressed (53.5%, 25.0%, and 32.1%) with the co-expression of their receptors. Realtime quantitative-PCR analysis showed that FGF-8/-17 were aberrantly expressed in patients with acute leukemia. Moreover, cell proliferation assays revealed the proliferation activity of FGF-17 on leukemic cells expressing its receptors. These results demonstrated that certain recently identified FGFs play an important role in the <em>growth</em> of leukemic cells, possibly with an autocrine mode of action, and that these FGFs will become novel biomarkers for hematopoietic tumors.

Proteoglycans, once thought to primarily serve as structural components of extracellular matrix, are now being focused on for their role in tissue and cell regulation, particularly angiogenesis. Many <em>growth</em> <em>factors</em>, notably the <em>fibroblast</em> <em>growth</em> family (FGF) which now numbers <em>19</em> members, bind to heparin and heparan sulfate proteoglycans and this binding has been shown to have a significant impact on the availability and activity of these <em>growth</em> <em>factors</em>. Proteoglycans can serve as both temporal and spatial regulators and effective inhibitor design may depend on disruption of these interactions. We have developed a simple assay for evaluating small inhibitors of proteoglycan-ligand binding. The assay is based on cell-free incubation of the reactants and filtration across a cationic membrane. Conditions were established that allow one to semiquantitatively determine binding constants for both direct proteoglycan as well as soluble inhibitor affinity. The assay has been demonstrated using a model heparan sulfate proteoglycan preparation (perlecan from cultured bovine endothelial cells) and FGF-2. Protamine sulfate, sucrose octasulfate, and heparin were analyzed as model inhibitor molecules. This type of assay may have wide application as a fast and easy screening tool for small potential agonists and antagonists of proteoglycan-protein interactions.

Secreted from intestine, human <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (hFGF<em>19</em>) is an endocrine metabolic regulator that controls bile acid synthesis in the liver. Earlier studies have suggested that hFGF<em>19</em> at 10-100 nM levels signals through FGF receptor 4 (FGFR4) in the presence of a co-receptor, betaKlotho, but its activity and receptor specificity at physiological concentrations (picomolar levels) remain unclear. Here we report that hFGF<em>19</em> at picomolar levels require sulfated glycosaminoglycans (sGAGs), such as heparan sulfate, heparin, and chondroitin sulfates, for its signaling via human FGFR4 in the presence of human betaKlotho. Importantly, sGAGs isolated from liver are highly active in enhancing the picomolar hFGF<em>19</em> signaling. At nanomolar levels, in contrast, hFGF<em>19</em> activates all types of human FGFRs, i.e. FGFR1c, FGFR2c, FGFR3c, and FGFR4 in the co-presence of betaKlotho and heparin and activates FGFR4 even in the absence of betaKlotho. These results show that sGAGs play crucial roles in specific and sensitive hFGF<em>19</em> signaling via FGF receptors and suggest that hepatic sGAGs are involved in the highly potent and specific signaling of picomolar hFGF<em>19</em> through FGFR4 and betaKlotho. The results further suggest that hFGF<em>19</em> at pathological concentrations may evoke aberrant signaling through various FGF receptors.

Chick ciliary ganglion neurons have a cholinergic membrane component that binds alpha-bungarotoxin with high affinity but has no known function. The component is different from the nicotinic ACh receptor on the neurons that mediates cholinergic transmission through the ganglion. Ciliary neuronotrophic <em>factor</em> (CNTF) has been shown to enhance the survival of ciliary ganglion neurons in cell culture and has been postulated to act as a target-derived trophic <em>factor</em> for the neurons in vivo. We show here that a <em>factor</em> indistinguishable from CNTF specifically down-regulates alpha-bungarotoxin binding sites on the neurons while increasing cell <em>growth</em> and the number of ACh receptors on the cells. Similar effects, though reduced in magnitude, are seen with chick sympathetic neurons. CNTF has no effect on the number of ACh receptors found on chick myotubes in culture. The down-regulation of alpha-bungarotoxin binding sites on neurons caused by CNTF occurs with a half-time of about <em>19</em> hr and is largely reversed within a 4 d period following CNTF removal. It is distinct from the down-regulation caused by cholinergic agonists. Nerve <em>growth</em> <em>factor</em> and <em>fibroblast</em> <em>growth</em> <em>factor</em> have no apparent effect on the number of alpha-bungarotoxin binding sites on the neurons, though <em>fibroblast</em> <em>growth</em> <em>factor</em> does stimulate neuronal <em>growth</em>. The results indicate that the effects of CNTF on the alpha-bungarotoxin binding component are both novel for a <em>growth</em> <em>factor</em> and specific, and they suggest a relationship between the component and the regulation of <em>growth</em> by the target tissue.

BACKGROUND

Endogenous pancreatic islets are supported by a dense sinusoidal capillary system which is disrupted following isolation and culture in vitro. A rapid and accurate revascularization is therefore crucial for the survival and functioning of the transplanted islet. Although a blood flow is established in islet grafts within 1-2 weeks, these islets show poor development of intra-islet capillaries. To improve the revascularization process and the arrangement of the new blood vessels, islet production of the factors governing these processes needs to be further characterized.

OBJECTIVE

To study the expression of factors which regulate angiogenesis in cultured rat islets.

METHODS

Rat islets were isolated and cultured for one week. After 6 hours of exposure to normoxic (21% O2) or hypoxic (1% O2) conditions, mRNA expression was evaluated by the GEArray Angiogenesis 1 and 2 systems. The expression of the vascular endothelial growth factor (VEGF), the tyrosine kinase with immunoglobulin and epidermal growth factor homology domains 1 (Tie1) and acidic fibroblast growth factor (aFGF), was further evaluated by semi-quantitative RT-PCR.

RESULTS

We found the expression of 19 genes that code for factors either promoting or preventing angiogenesis. Only VEGF and Tie1 were upregulated in response to hypoxia.

CONCLUSIONS

Hypoxia-induced islet vascularization may involve VEGF and Tie-induced signaling events. The results also show that cultured islets express genes which prevent angiogenesis concurrently with genes coding for factors stimulating angiogenesis. The balance between these factors is probably of vital importance for the revascularization process in transplanted islets. Thus, pharmacologic or genetic attenuation of islet-derived angiostatic factors may prove beneficial in promoting islet revascularization in future transplantation trials.

As a key humoral regulator of phosphate homeostasis and its involvement in the pathogenesis of human disease, human <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (hFGF23) has become a particularly attractive therapeutic target. To prepare soluble and bioactive recombinant human FGF23 to meet the increasing demand in its pharmacological application, small ubiquitin-related modifier (SUMO)-FGF23 fusion gene and FGF23 non-fusion gene were amplified by standard PCR methods and cloned into vector pET-22b and pET-3c, then transformed into Escherichia coli Rosetta (DE3) and BL21 (DE3). The best combination of plasmid and host strain was screened, and only Rosetta (DE3)/pET-SUMO-FGF23 was screened for rhFGF23 protein expressed. The average bacterial yield and the soluble expression level of recombinant hFGF23 of three batches attained 687 ± 18 g and 30 ± 1.5%, respectively, after treatment with 0.4 mM isopropyl-thio-β-galactopyranoside for <em>19</em> h at 16 °C in a 30-L fermentor, after which it was purified by DEAE Sepharose FF and nickel nitrilotriacetic acid affinity chromatography. Once cleaved by the SUMO protease, the recombinant human FGF23 was released from the fusion protein. The purity of rFGF23 was shown by high performance liquid chromatography to be greater than 90% and the yield was 60 ± 1.5 mg/L. In vitro data showed that the purified rFGF23 can induce the phosphorylation of mitogen-activated protein kinases in the glioma U251 cell. The results of in vivo animal experiments also showed that rFGF23 could decrease the concentration in the plasma of normal rats fed with a fixed formula diet.

The <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 3 (Fgfr3) protein is a tyrosine kinase receptor involved in the signal transduction of various <em>fibroblast</em> <em>growth</em> <em>factors</em>. Recent studies suggest its important role in normal development. In humans, mutation in Fgfr3 is responsible for <em>growth</em> disorders such as achondroplasia, hypoachondroplasia, and thanatophoric dysplasia. Here, we report the complete genomic organization of the mouse Fgfr3 gene. The murine gene spans approximately 15 kb and consists of <em>19</em> exons and 18 introns. One major and one minor transcription initiation site were identified. Position +1 is located 614 nucleotides upstream from the ATG initiation codon. The translation initiation and termination sites are located in exons 2 and <em>19</em>, respectively. Five Sp1 sites, two AP2 sites, one Zeste site, and one Krox 24 site were observed in the 5'-flanking region. The Fgfr3 promoter appears to be contained within a CpG island and, as is common in genes having multiple Sp1-binding sites, lacks a TATA box.

The aim of the study was to assess the specificity of albumin-messenger RNA (mRNA)-positive cells in peripheral blood as an indicator for circulating malignant hepatocytes. Albumin-mRNA-positive cells in the peripheral blood mononuclear cell (PMNC) fraction were detected by reverse-transcription polymerase chain reaction (RT-PCR). Albumin-mRNA-positive cells in PMNC were found in 12 of <em>19</em> (63%) patients with hepatocellular carcinoma, but also in 22 of 25 (88%) patients with chronic hepatitis without evidence for hepatoma, and in 18 of <em>19</em> (94%) of patients with acute viral hepatitis. In addition, 8 of 28 (28%) of healthy control individuals had also albumin-mRNA-positive cells in peripheral blood. PMNC known to be spontaneously negative for albumin-mRNA could be induced in vitro to transcribe albumin-mRNA after activation with a variety of substances such as interleukin-1 (IL-1) plus concanavalin A (Con A), IL-2, bacterial lipopolysaccharide, platelet derived <em>growth</em> <em>factor</em>, alpha-<em>fibroblast</em> <em>growth</em> <em>factor</em>, or hepatocyte <em>growth</em> <em>factor</em>. These results show that the majority of patients with acute and chronic liver disease without evidence for hepatocellular carcinoma has albumin-mRNA-positive cells in their PMNC fraction indicating the nonspecificity of that parameter for the presence of circulating malignant hepatocytes. In addition, in vitro experiments suggest that PMNC are capable of transcribing mRNA for albumin at a low level after activation. In vivo-activated PMNC are likely to be the source of positivity in healthy controls, patients with nonmalignant acute and chronic liver disease, and patients with hepatocellular carcinoma.

A new human functional tumor cell line, designated as T3M-3, has been established from a xenotransplanted choriocarcinoma grown in nude mice. One of the biggest problems of the in vitro culture of these tumor cells using the xenotransplanted tumors had been the dense contamination of <em>fibroblasts</em> of host nude mouse origin. In the present study, these <em>fibroblasts</em> were completely removed by incubating the cells with antiserum raised against nude mouse spleen cells. The cell line established from the remaining tumor cells has been successfully propagated in vitro for as long as 4 years. These cells show the morphology of epithelioid cells containing a prominent nucleus with one or two large nucleoli. The cells grow in a monolayered sheet with the population-doubling time of <em>19</em> hr. The cells show perfect tumor takes when they are reinoculated into nude mice. Chromosomal analysis revealed that the cell is a human aneuploid one with a hypotriploid mode. These cultured cells maintained well the function of secreting large amounts of human chorionic gonadotropin, progesterone, and estrogen. The secretion of human chorionic gonadotropin and progesterone by these cells is enhanced by stimulation with tumor promoters, such as 12-O-tetradecanoylphorbol-13-acetate and teleocidin B, or with epidermal <em>growth</em> <em>factor</em> in a dose-and time-dependent manner. Interestingly, however, the tumor promoters did not exert a marked effect on the cellular binding of epidermal <em>growth</em> <em>factor</em>, indicating that the receptors for these reagents in T3M-3 cells are not shared by epidermal <em>growth</em> <em>factor</em>.

Acidic <em>fibroblast</em> <em>growth</em> <em>factor</em> (aFGF) is a potent <em>growth</em> <em>factor</em> for vascular smooth muscle cells and may mediate vasculopathy in cardiac allografts subjected to chronic immunological injury. Therefore, we examined cardiac expression of aFGF, the number of rejection episodes, and other potential risk <em>factors</em> in 32 heart transplant patients who underwent intravascular ultrasound (IVUS) for detection of cardiac allograft vasculopathy (CAV). As defined by IVUS, CAV was present in 21 patients and absent in 11 patients (follow-up time: 52 +/- 21 vs. 51 +/- 12 months, respectively, P = NS). The level of aFGF in myocardial biopsies obtained at the time of IVUS was measured by semiquantitative reverse transcriptase polymerase chain reaction and expressed as the aFGF:GAPDH ratio. Higher level of aFGF were associated with CAV (mean aFGF:GAPDH ratio was 1.45 +/- 0.99 in patients with vs. 0.18 +/- 0.12 in patients without CAV [P < 0.001]). A strong association was found between high levels of cardiac aFGF and CAV, as 18 of <em>19</em> patients (95%) with high levels of aFGF (aFGF:GAPDH>> 1) but only 3 of 13 patients with low levels of aFGF had CAV (P < 0.001). The relative risk of high level of aFGF for CAV was 4.1. Untreated low grade rejection (ISHLT I), but not treated high grade rejection (ISHLT>> 2), was also associated with CAV (average number of untreated low grade rejection episodes was 3.5 +/- 1.8 in patients with vs. 2.1 +/- 1.0 in patients without CAV [P = 0.04]). Among other risk <em>factors</em> examined (age, sex, serum cholesterol, blood pressure, CMV infection, dose of immunosuppressants, and ischemic time), only triglycerides were higher in patients with than those without CAV (P = 0.003). We conclude that increased cardiac production of aFGF is significantly associated with CAV, which suggests that aFGF may serve as an important mediator in CAV. Untreated low grade rejection also poses an increased risk for CAV.

Alternative RNA processing of the human <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor-1 transcript results in receptor forms that vary in their affinity for <em>fibroblast</em> <em>growth</em> <em>factor</em>. An alternative RNA processing event involving recognition of the alpha-exon is deregulated during neoplastic transformation of glial cells. We have previously established a splicing reporter/transfection cell culture model system to identify sequences involved in recognition of this exon. In this study, the system was used to identify two sequence elements that differentially function to regulate splicing of this exon. Exclusion of the alpha-exon in glioblastoma cells specifically required the downstream intron sequence comprising the 5'-splice site. Replacement or mutation of this sequence increasing complementarity to U1 RNA resulted in enhanced exon recognition in SNB-<em>19</em> glioblastoma cells. Sequences within the exon were found to be required for alpha-exon inclusion. Deletion and gain-of-function experiments identified a 69-nucleotide exon sequence that was specifically required for alpha-exon inclusion. These studies indicate that multiple sequences are required for the regulated recognition of the alpha-exon.

We attempted to determine whether the plasma concentration of basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) was elevated in brain tumor patients. The plasma concentration of bFGF was measured in 55 brain tumor patients and 17 normal subjects by enzyme immunoassay. Upper limit of plasma bFGF in the normal subjects was 1.6 pg/ml. Elevated plasma bFGF levels were observed in 28 patients including 13 of 17 glioma patients, eight of <em>19</em> benign tumor patients, and seven of <em>19</em> malignant tumor patients. Twelve of 14 malignant glioma patients had elevated plasma bFGF levels. There was a good correlation (r = 0.42, p < 0.05) between the elevated plasma bFGF level and the tumor volume. Further studies are needed to determine whether the plasma bFGF level is a clinically useful diagnostic and prognostic marker for patients with brain tumors.

<AbstractText>The ileum-derived <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>) plays key roles in hepatic glucose homeostasis in animals in an insulin-independent manner. Here, we analyzed the association of FGF<em>19</em> with glucose effectiveness (GE, the insulin-independent glucose regulation), as well as hepatic glucose production (HGP) in Chinese subjects.</AbstractText><p><div><b>METHODS</b></div>GE was measured by frequently sampled intravenous glucose tolerance test (FSIVGTT) in normal glucose tolerance (NGT), isolated-impaired glucose tolerance (I-IGT), and isolated-impaired fasting glucose (I-IFG) subjects. The oral glucose tolerance test-derived surrogate of GE (oGE) was determined in NGT, I-IFG, combined glucose intolerance (CGI), and type 2 diabetes (T2DM) subjects. HGP was assessed by labeled ([3-³H]-glucose) hyperinsulinemic-euglycemic clamp in NGT subjects. Insulin secretion and sensitivity were calculated by the hyperglycemic and hyperinsulinemic-euglycemic clamps in a subgroup of NGT, I-IGT, and I-IFG subjects. Serum FGF<em>19</em> levels were determined by ELISA.</p><AbstractText>FGF<em>19</em> positively correlated with GE (r = 0.29, P = 0.004) as determined by FSIVGTT. The result was further confirmed by oGE (r = 0.261, P < 0.001). FGF<em>19</em> was negatively associated with FPG (r = - 0.228, P = 0.025), but the association no longer existed after adjusting for GE (r = - 0.177, P = 0.086). FGF<em>19</em> was negatively associated with basal HGP (r = - 0.697, P = 0.006). However, the correlation between FGF<em>19</em> and insulin secretion and sensitivity were not found.</AbstractText><AbstractText>FGF<em>19</em> levels are associated positively with GE and negatively with HGP. The increase of FPG in human is at least partially due to the decrease of FGF<em>19</em> in an insulin-independent manner.</AbstractText>

BACKGROUND

Bile acid (BA) synthesis is regulated by BA signalling in the liver and by <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>), synthesized and released from the intestine. In morbid obesity, faecal excretion and hepatic synthesis of BAs and cholesterol are strongly induced and caloric restriction reduces their faecal excretion considerably. We hypothesized that the high intestinal food mass in morbidly obese subjects promotes faecal excretion of BAs and cholesterol, thereby creating a shortage of both BAs and cholesterol in the liver.

METHODS

Ten morbidly obese women (BMI 42 ± 2.6 kg m-2 ) were monitored on days 0, 3, 7, 14 and 28 after beginning a low-calorie diet (800-1100 kcal day-1 ). Serum was collected and liver size and fat content determined. Synthesis of BAs and cholesterol was evaluated from serum markers, and the serum levels of lipoproteins, BAs, proprotein convertase subtilisin/kexin type 9 (PCSK9), insulin, glucose and FGF<em>19</em> were monitored. Fifty-four nonobese women (BMI <25 kg m-2 ) served as controls.

RESULTS

At baseline, synthesis of both BAs and cholesterol and serum levels of BAs and PCSK9 were elevated in the obese group compared to controls. Already after 3 days on a low-calorie diet, BA and cholesterol synthesis and serum BA and PCSK9 levels normalized, whereas LDL cholesterol increased. FGF<em>19</em> and triglyceride levels were unchanged, and liver volume was reduced by 10%.

CONCLUSIONS

The results suggest that hepatic BAs and cholesterol are deficient in morbid obesity. Caloric restriction rapidly counteracts these deficiencies, normalizing BA and cholesterol synthesis and circulating PCSK9 levels, indicating that overproduction of cholesterol in enlarged peripheral tissues cannot explain this phenotype. We propose that excessive food intake promotes faecal loss of BAs and cholesterol contributing to their hepatic deficiencies.

OBJECTIVE

The objective of this study was to determine the concentration of fibroblast growth factor (FGF) and soluble intracellular adhesions molecule (sICAM-1) in serum and follicular fluid (FF) of polycystic ovary (PCO), endometriosis and tubal factor infertility and male factor infertility patients, and to investigate the relationship between these parameters and the outcome of intracytoplasmic sperm injection (ICSI).

METHODS

The concentration of FGF and sICAM-1 in serum and FF were determined in patients undergoing controlled ovarian hyperstimulation (COH) for ICSI therapy for various etiology of infertility and the results of cytokines concentration and ICSI outcome were compared between the groups. Twenty patients with PCO (G.I), 17 with endometriosis (G.II), 19 with tubal damage (G.III) and 19 with male factor infertility (G.IV) were enrolled in this study. Quantitative determination of levels of FGF and sICAM-1 was performed using enzyme-linked immunosorbent assays (ELISAs).

RESULTS

The FGF level in serum of PCO patients (G.I) were 4.8 +/- 2.3 and in FF were 104.0 +/- 39.0 pg/mL. The corresponding values in the endometriosis patients group (G.II) were 5.9 +/- 3.1 and 125.4 +/- 74.9 pg/mL. The concentration of FGF in tubal factor infertility group (G.III) in serum was significantly higher (P = 0.009) than those observed in the PCO group (G.I) 7.4 +/- 4.5 pg/mL, whereas the concentration in FF was at the same level like the other groups investigated, 128.7 +/- 75.9 pg/mL. Besides, the sICAM-1 (pg/ml) concentration in FF showed a significant difference between the groups investigated (G.I, 175.3 +/- 52.8; G.II 194.4 +/- 32.2; G.III 233.1 +/- 54.3; and G.IV 215.1 +/- 54.4 ng/mL; P = 0.003). The sICAM-1 levels in serum were not significantly different between the groups (217.0 +/- 42.9; 216.3 +/- 73.6; 254.8 +/- 79.6; 237.56 +/- 78.4 ng/ml; P = 0.267). The fertilization rate was significantly higher in G.III (66.0 +/- 23.89%) in comparison to G.II (38.8 +/- 33.9%; P = 0.014) or G.IV (38.7 +/- 22.7%; P = 0.012). The pregnancy rates were similar in all groups (30, 35.3 and 35.0, 38.6%, respectively).

CONCLUSIONS

Both, FGF and sICAM-1 are present in serum and FF of patients undergoing controlled ovarian hyperstimulation for ICSI therapy. The FGF concentration in serum differs significantly between the groups investigated, whereas, no significant difference could be observed in the FF concentration of FGF. On the other hand, the sICAM in serum showed no significant difference between the groups, whereas, sICAM in FF demonstrated a significant difference between the patient groups investigated. On the whole, the ICSI outcome was not related to serum or FF concentrations of FGF or sICAM-1. Therefore, the mean concentration of FGF and sICAM-1 in serum and in FF could not be used to predict the fertilization rate in an ICSI program.

<AbstractText><em>Fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>) is a postprandial hormone which plays diverse roles in the regulation of bile acid, glucose, and lipid metabolism. Administration of FGF<em>19</em> to obese/diabetic mice lowers body weight, improves insulin sensitivity, and enhances glycemic control. The primary target organ of FGF<em>19</em> is the liver, where it regulates bile acid homeostasis in response to nutrient absorption. In contrast, the broader pharmacologic actions of FGF<em>19</em> are proposed to be driven, in part, by the recruitment of the thermogenic protein uncoupling protein 1 (UCP1) in white and brown adipose tissue. However, the precise contribution of UCP1-dependent thermogenesis to the therapeutic actions of FGF<em>19</em> has not been critically evaluated.</AbstractText><AbstractText>Using WT and germline UCP1 knockout mice, the primary objective of the current investigation was to determine the in vivo pharmacology of FGF<em>19</em>, focusing on its thermogenic and anti-obesity activity.</AbstractText><AbstractText>We report that FGF<em>19</em> induced mRNA expression of UCP1 in adipose tissue and show that this effect is required for FGF<em>19</em> to increase caloric expenditure. However, we demonstrate that neither UCP1 induction nor an elevation in caloric expenditure are necessary for FGF<em>19</em> to induce weight loss in obese mice. In contrast, the anti-obesity action of FGF<em>19</em> appeared to be associated with its known physiological role. In mice treated with FGF<em>19</em>, there was a significant reduction in the mRNA expression of genes associated with hepatic bile acid synthesis enzymes, lowered levels of hepatic bile acid species, and a significant increase in fecal energy content, all indicative of reduced lipid absorption in animals treated with FGF<em>19</em>.</AbstractText><AbstractText>Taken together, we report that the anti-obesity effect of FGF<em>19</em> occurs in the absence of UCP1. Our data suggest that the primary way in which exogenous FGF<em>19</em> lowers body weight in mice may be through the inhibition of bile acid synthesis and subsequently a reduction of dietary lipid absorption.</AbstractText>

One of the most severe effects of coronavirus disease 20<em>19</em> (COVID-<em>19</em>) is lung disorders such as acute respiratory distress syndrome. In the absence of effective treatments, it is necessary to search for new therapies and therapeutic targets. Platelets play a fundamental role in respiratory disorders resulting from viral infections, being the first line of defense against viruses and essential in maintaining lung function. The direct application of platelet lysate (PL) obtained from the platelet-rich plasma of healthy donors could help in the improvement of the patient due its anti-inflammatory, immunomodulatory, antifibrotic, and repairing effects. This work evaluates PL nebulization by analyzing its levels of <em>growth</em> <em>factors</em> and its biological activity on lung <em>fibroblast</em> cell cultures, besides describing a scientific basis for its use in this kind of pathology. The data of the work suggest that the molecular levels and biological activity of the PL are maintained after nebulization. Airway administration would allow acting directly on the lung tissue modulating inflammation and stimulating reparative processes on key structures such as the alveolocapillary barrier, improving the disease and sequels. The protocol developed in this work is a first step for the study of nebulized PL both in animal experimentation and in clinical trials.

<strong class="sub-title"> Keywords: </strong> COVID-<em>19</em>; <em>growth</em> <em>factors</em>; nebulization; platelet; platelet lysate; platelet-rich plasma.

OBJECTIVE

Markers of chronic inflammation, acute-phase reactants, and growth factors may be concomitantly involved in a number of pathologic processes in the general population and uremic patients. In addition, growth factors may influence peritoneal membrane transport characteristics. However, the association between plasma growth factors, markers of chronic inflammation, and peritoneal membrane transport remains largely unknown. The aim of this study was to evaluate the relationship between plasma levels of selected growth factors [basic fibroblast growth factor (bFGF), transforming growth factor beta1 (TGFbeta1), vascular endothelial growth factor (VEGF)] and markers of chronic inflammation [interleukin (IL)-6, C-reactive protein (CRP), and fibrinogen] in continuous ambulatory peritoneal dialysis (CAPD) patients. The potential link between the above substances and dialysis adequacy was also explored.

METHODS

Single-center, cross-sectional study.

METHODS

Peritoneal Dialysis Unit, Medical Faculty, Jagiellonian University Hospital, Kraków, Poland.

METHODS

32 stable end-stage renal disease patients (13 M, 19 F; mean age 53.6 +/- 13.7 years) on CAPD for a median period of 19.5 months. Patients free from signs and symptoms of any inflammatory disease (including peritonitis) for at least 3 months were included into the study. All patients underwent measurements of dialysis dose [Kt/V, weekly creatinine clearance (wCCr)] and peritoneal solute transport using a standard peritoneal equilibration test (PET).

METHODS

TGFbeta1, bFGF, VEGF, and IL-6 were measured with ELISA, CRP was assayed with immunonephelometry, and fibrinogen with Multifibren U reagent (Dade Behring Marburg GmbH, Marburg, Germany). Nephron 97 for Windows software was used to assess dialysis adequacy.

RESULTS

Significant positive correlations between plasma bFGF and IL-6, as well as fibrinogen concentrations (R = 0.36, p < 0.05 and R = 0.39, p < 0.05, respectively), were found. VEGF correlated significantly with IL-6 and CRP (R = 0.65, p < 0.0001 and R = 0.51, p < 0.005, respectively). An association between VEGF and bFGF was also found (R = 0.59, p < 0.0005). Serum level of TGFbeta1 revealed no relationship with any marker of acute-phase activation, remaining growth factors, or dialysis adequacy. Positive correlation between TGFbeta1 concentration and dialysate-to-plasma ratio for creatinine in PET (R = 0.35, p < 0.05) was found. In addition, patients with lower solute transport (low/low-average transporters) had lower serum levels of both bFGF and TGFbeta1 compared to patients with higher solute transport. Patients with total wCCr>> 60 L/ week/m2 were characterized by lower levels of bFGF and IL-6. Serum level of IL-6 and plasma levels of bFGF and VEGF were significantly lower among subjects with residual renal function (RRF)>> 2.0 mL/minute.

CONCLUSIONS

Our results indicate that systemic inflammation in peritoneal dialysis patients is associated with increased plasma VEGF and bFGF but not TGFbeta1. The negative correlation with RRF suggests that either the renal clearance of these cytokines and growth factors may contribute to their elimination, or cytokines and growth factors have a negative impact on RRF. We also suggest an association between serum levels of growth factors tested and peritoneal membrane function.

Since February 2020, a large number of patients infected with new coronavirus has been cured and discharged with the controlling of epidemic. Pulmonary fibrosis, which may be one of the sequela caused by COVID-<em>19</em>, not only brings dyspnea and deterioration of lung function, but also affects patients' life because of its high mortality and poor prognosis. Vascular endothelial <em>growth</em> <em>factor</em> receptor(VEGFR) and <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor(FGFR) can inhibit the proliferation, activation and migration of <em>fibroblasts</em> by regulating the signal transduction pathway involved in the process of pulmonary fibrosis. Chinese herbal formulas pose a good therapeutic effect on pulmonary fibrosis. Present study explores the intervention effect on pulmonary fibrosis of traditional Chinese medicine(TCM) by screening the potential inhibitors of VEGFR and FGFR. The docking models of VEGFR and FGFR were established to obtain the potential active ingredients which were filtered by the docking score. According to 2 prescriptions in the Protocol for the diagnosis and treatment of coronavirus disease 20<em>19</em>(7th edition)and 9 prescriptions in Traditional Chinese medicine prescriptions for treating blight, 959 and 1 047 potential ingredients were obtained as the inhibitors of VEGFR and FGFR respectively with the screening thres-hold set as eighty percent of the docking score of the initial ligands. The potential herbs were then filtered by the components with a hit rate higher than 30%, such as Scutellariae Radix, Adenophorae Radix, Pinelliae Rhizoma, Coicis Semen, etc. To discuss the rule of TCM in the treatment of pulmonary fibrosis, the networks of TCM-channel tropism and TCM-efficacy of the potential herbs was constructed. The potential herbs for treating pulmonary fibrosis mostly belong to lung(degree=14) and spleen(degree value=8), and the efficacy is focused on reinforcing deficiency(degree=9). Qiyin Prescription and Buzhong Yiqi Decoction contain the largest number of the potential herbs. The main symptom of COVID-<em>19</em> is damp-heat stagnating in the lung, which always causes impairment of body fluid and Qi. Clinical observation shows that patients in the recovery period are mostly at the status that the remaining virus toxicity is not exhausted while the vital Qi have not recovered. The results of this study are expected to provide references for clinical medication in preventing and treating pulmonary fibrosis caused by COVID-<em>19</em>.

<strong class="sub-title">Keywords:</strong> COVID-<em>19</em>; <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor; molecular docking; pulmonary fibrosis; traditional Chinese medicine formula; vascular endothelial <em>growth</em> <em>factor</em> receptor.

<AbstractText>Chronic inflammation in adipose tissue may play a substantial role in the pathogenesis of obesity-related metabolic disorders. The present study aims to evaluate the changes in adipocytokines, bile acids, <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF-<em>19</em>) and pro-inflammatory cytokines 6 months after laparoscopic sleeve gastrectomy (LSG).</AbstractText><p><div><b>METHODS</b></div>This prospective study included 75 obese patients with body mass index >35 kg/m² who underwent LSG. All patients were recruited preoperatively and followed up post-operatively at 6 months, with laboratory assessment of their cytokines including adiponectin, leptin, resistin, bile acid, interleukin (IL)-6, IL-8, tumour necrosis <em>factor</em>-α, monocyte chemotactic protein-1, high-sensitivity C-reactive protein, plasminogen activator inhibitor-1, serum amyloid-A and FGF-<em>19</em>.</p><AbstractText>There were statistically highly significant changes regarding anthropometric parameters (weight, body mass index and waist-to-hip ratio), blood glucose and lipid profile as well as liver enzymes at 6 months post-sleeve gastrectomy. The present study showed that the levels of serum adiponectin and FGF-<em>19</em> significantly increased at 6 months of follow-up after surgery (P < 0.001), while the levels of serum leptin, resistin, high-sensitivity C-reactive protein, plasminogen activator inhibitor-1 and serum amyloid-A significantly decreased at 6 months of follow-up after surgery (P < 0.001). There were no significant differences regarding serum bile acid, IL-6, IL-8, tumour necrosis <em>factor</em>-α and monocyte chemotactic protein-1.</AbstractText><AbstractText>Weight loss after LSG is associated with significant improvement of the adipokine levels towards anti-diabetic and anti-inflammatory profiles. Future studies should use a larger sample size and longer follow-up periods.</AbstractText>

The immunosuppressive protein transforming <em>growth</em> <em>factor</em> beta (TGF-beta) inhibits the activation of various immune effector cells including cytotoxic T lymphocytes and may therefore inhibit the efficacy of immunostimulatory interleukin-2 (IL-2) gene therapy. In this study, we investigated the effect of TGF-beta downregulation on IL-2 gene therapy in the intraperitoneal model of murine ovarian teratoma (MOT). MOT cells, like many human ovarian carcinomas, were found to produce TGF-beta. Production of TGF-beta by MOT cells was suppressed using a TGF-beta antisense plasmid vector (pCEP4/TGF-beta antisense). Subcutaneous immunization of C3H mice with a mixture of IL-2 gene-transduced <em>fibroblasts</em> and TGF-beta antisense-modified MOT cells induced significantly better protection against a subsequent intraperitoneal tumor challenge compared with immunization with unmodified MOT cells alone [11/16 (69%) vs 4/21 (<em>19</em>%) tumor-free animals, P < 0.01]. Immunization with either a mixture of IL-2 gene modified <em>fibroblasts</em> and unmodified MOT cells [2/12 (17%) tumor-free animals] or TGF-beta antisense-modified MOT cells alone (0/13 tumor free animals) failed to induce significant protection compared with immunization with unmodified MOT cells. These data show that combined TGF-beta antisense and IL-2 gene therapy is required to generate effective antitumor responses in the MOT model. Our findings suggest that tumor cell expression of immunosuppressive <em>factors</em> may inhibit cytokine immunogene therapy and may have potential implications for the development of future clinical immunogene therapy protocols.

Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) is a polypeptide that is mitogenic for a wide variety of cell types. We used Northern blot analysis and immunohistochemistry to determine if bFGF is expressed in the nasal polyp tissue; bFGF messenger RNA was detectable in the polyps examined by Northern blot analysis. Strong immunostaining for bFGF was found in blood vessels and along the basement membrane of the epithelial cell layers. Basal epithelial cells and some infiltrating mononuclear cells also stained for bFGF. Proliferating cell nuclear antigen colocalized with bFGF to basal epithelial cells, endothelial cells, and areas of focal epithelial metaplasia. The polyp tissue was double-labeled with a mouse monoclonal antitryptase, a specific mast cell marker, and anti-bFGF. A significant number (65% +/- <em>19</em>%) of the bFGF-positive mononuclear cells in the polyp tissues were positive for tryptase. These findings suggest that bFGF may contribute to the endothelial and epithelial proliferation in nasal polyp tissues and that mast cells are one source of this <em>growth</em> <em>factor</em>.

Idiopathic pes equinovarus is a congenital deformity of the foot and lower leg defined as a fixation of the foot in adduction, supination, and varus. Although the pathogenesis of clubfoot remains unclear, it has been suggested that <em>fibroblasts</em> and <em>growth</em> <em>factors</em> are involved. To directly analyze the protein composition of the extracellular matrix in contracted tissue of patients with clubfoot. A total of 13 infants with idiopathic clubfoot treated with the Ponseti method were included in the present study. Tissue samples were obtained from patients undergoing surgery for relapsed clubfeet. Contracted tissues were obtained from the medial aspect of the talonavicular joint. Protein was extracted after digestion and delipidation using zip-tip C18. Individual collagenous fractions were detected using a chemiluminescent assay. Amino acid analysis of tissue samples revealed a predominance of collagens, namely collagen types I, III, and VI. The high content of glycine and h-proline suggests a predominance of collagens I and III. A total of <em>19</em> extracellular matrix proteins were identified. The major result of the present study was the observation that the extracellular matrix in clubfoot is composed of an additional 16 proteins, including collagens V, VI, and XII, as well as the previously described collagen types I and III and transforming <em>growth</em> <em>factor</em> β. The characterization of the general protein composition of the extracellular matrix in various regions of clubfoot may help in understanding the pathogenesis of this anomaly and, thus, contribute to the development of more efficacious therapeutic approaches.

<em>Fibroblast</em> <em>growth</em> <em>factor</em> receptors (FGFRs) are important in malignant progression of several human epithelial tumors. However, little is known about FGFRs in canine or human soft tissue sarcomas. Thus, our aim was to investigate expression of FGFRs and their involvement in cell survival in sarcomas of both species. FGFR1-4 and FGFRL1 transcripts as well as IIIb/IIIc splice variants of FGFR1-3 were evaluated in 3 canine- and 6 human sarcoma cell lines and <em>19</em> spontaneous canine sarcomas by SYBRqPCR. FGFR1 protein expression was assessed by immunohistochemistry. <em>Growth</em> inhibitory effects of FGFR1 inhibitor PD166866 and dominant negative recombinant FGFR adenoviral expression constructs (dnFGFR) on tumor cell lines were analyzed. Profiling of multiple FGFR transcripts detected comparable co-expression in most of human and canine sarcoma cell lines and canine tumor specimens. This indicates existence of closely related regulation mechanisms for FGFR expression in sarcomas of both species. FGFR1 with splice variant IIIc was consistently expressed with highest transcript levels. In 88% of the spontaneous tumor samples a heterogeneous FGFR1 protein expression was observed. Significant <em>growth</em> inhibition and cell death was seen after infection with dnFGFR1 in canine and human sarcoma cells, but not with dnFGFR3 and 4. PD166866 showed selective cytotoxicity with IC50 values between 12.1 and 26.4 μM. FGFR1 inhibition blocked ligand-induced tyrosine phosphorylation of ERK1/2 mitogen-activated protein kinase isoforms. This study emphasizes the important role FGFR1, especially splice variant IIIc, likely plays in sarcomas. Inhibitory small molecules could be of potential use for targeted therapy in aggressive sarcomas of both species.