<b>Background & Aims:</b> Liver cancer stem cells (LCSCs) mediate therapeutic resistance and correlate with poor outcomes in patients with hepatocellular carcinoma (HCC). <em>Fibroblast</em> <em>growth</em> <em>factor</em> (FGF)-<em>19</em> is a crucial oncogenic driver gene in HCC and correlates with poor prognosis. However, whether FGF<em>19</em> signaling regulates the self-renewal of LCSCs is unknown. <b>Methods:</b> LCSCs were enriched by serum-free suspension. Self-renewal of LCSCs were characterized by sphere formation assay, clonogenicity assay, sorafenib resistance assay and tumorigenic potential assays. Ca²⁺ image was employed to determine the intracellular concentration of Ca²⁺. Gain- and loss-of function studies were applied to explore the role of FGF<em>19</em> signaling in the self-renewal of LCSCs. <b>Results:</b> FGF<em>19</em> was up-regulated in LCSCs, and positively correlated with certain self-renewal related genes in HCC. Silencing FGF<em>19</em> suppressed self-renewal of LCSCs, whereas overexpressing FGF<em>19</em> facilitated CSCs-like properties <i>via</i> activation of FGF receptor (FGFR)-4 in none-LCSCs. Mechanistically, FGF<em>19</em>/FGFR4 signaling stimulated store-operated Ca²⁺ entry (SOCE) through both the PLCγ and ERK1/2 pathways. Subsequently, SOCE-calcineurin signaling promoted the activation and translocation of nuclear <em>factors</em> of activated T cells (NFAT)-c2, which transcriptionally activated the expression of stemness-related genes (<i>e.g., NANOG</i>, <i>OCT4</i> and <i>SOX2</i>), as well as <i>FGF<em>19</em></i>. Furthermore, blockade of FGF<em>19</em>/FGFR4-NFATc2 signaling observably suppressed the self-renewal of LCSCs. <b>Conclusions:</b> FGF<em>19</em>/FGFR4 axis promotes the self-renewal of LCSCs <i>via</i> activating SOCE/NFATc2 pathway; in turn, NFATc2 transcriptionally activates FGF<em>19</em> expression. Targeting this signaling circuit represents a potential strategy for improving the therapeutic efficacy of HCC.

<strong class="sub-title"> Keywords: </strong> FGF<em>19</em>; NFATc2, LCSCs; SOCE; self-renewal.

Sorafenib (Sor) is clinical standard therapy for advanced hepatocellular carcinoma (HCC). However, detailed molecular mechanism behind Sor-exerted pharmacological effect remains unknown. In this study, sera samples, staged hepatic cancer tissues from Sor-treated patients with advanced HCC were harvested for a group of biochemical tests and immunoassays. Compared to non-treated control, blood contents of alanine transaminase (ALT), aspartate transaminase (AST), alphafetoprotein (AFP), <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) were decreased in Sor-treated HCC patients, while the level of interleukin 10 (IL-10) were increased. As well, reduced triglyceride (TG), total cholesterol (T-CHOL), interferon gamma (IFN-γ), and tumor necrosis <em>factor</em> alpha (TNF-α) levels in sera were checked in Sor-treated HCC patients. In comparison with non-treated cancer sections, Sor-treated HCC cells showed decreased positive cells of proliferative marker for proliferating cell nuclear antigen (PCNA) and metastasized biomarker for cytokeratin <em>19</em> (CK<em>19</em>). In addition, elevated immunofluorescence-labeled cells of endoplasmic reticulum (ER)-stress markers of activating transcription <em>factor</em> 6 (ATF6), eukaryotic initiation <em>factor</em> 2α kinase (eIF2α), glucose-regulated protein (GRP-78), X-box binding protein 1 (XBP1) were observed in Sor-treated HCC livers. Further, validated data from Western blot assay exhibited that hepatocellular expressions of ATF6, eIF2α, GRP78, XBP1 in Sor-treated HCC liver cells were up-regulated. Briefly, our present clinicopathologic findings indicate that Sor-induced ER stress may be responsible for therapeutic mechanism against advanced HCC. In addition, induction of intracellular ER stress functions as a promising strategy for treating advanced HCC.

Ursodeoxycholic acid (UDCA) treatment can reduce itch and lower endogenous serum bile acids in intrahepatic cholestasis of pregnancy (ICP). We sought to determine how it could influence the gut environment in ICP to alter enterohepatic signalling. The gut microbiota and bile acid content were determined in faeces from 35 pregnant women (14 with uncomplicated pregnancies and 21 with ICP, 17 receiving UDCA). Faecal bile salt hydrolase activity was measured using a precipitation assay. Serum <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>) and 7α-hydroxy-4-cholesten-3-one (C4) concentrations were measured following a standardised diet for 21 hours. Women with a high ratio of Bacteroidetes to Firmicutes were more likely to be treated with UDCA (Fisher's exact test p = 0.0178) than those with a lower ratio. Bile salt hydrolase activity was reduced in women with low Bacteroidetes:Firmicutes. Women taking UDCA had higher faecal lithocholic acid (p < 0.0001), with more unconjugated bile acids than women with untreated ICP or uncomplicated pregnancy. UDCA-treatment increased serum FGF<em>19</em>, and reduced C4 (reflecting lower bile acid synthesis). During ICP, UDCA treatment can be associated with enrichment of the gut microbiota with Bacteroidetes. These demonstrate high bile salt hydrolase activity, which deconjugates bile acids enabling secondary modification to FXR agonists, enhancing enterohepatic feedback via FGF<em>19</em>.

Despite the regenerative capability of bone, treatment of large defects often requires bone grafts. The challenge for bone grafting is to establish rapid and sufficient vascularization. Three-dimensional (3D) multicellular spheroids consisting of the relevant cell types can be used as "mini tissues" to study the complexity of angiogenesis. We investigated two-dimensional (2D) expansion, differentiation and characterization of primary osteoblasts as steps toward the establishment of 3D multicellular spheroids. Supplementation of cell culture medium with vitamin D(3) induces the osteocalcin expression of osteoblasts. An increased osteocalcin concentration of 10.8 ± 0.58 ng/ml could be measured after <em>19</em> days in supplemented medium. Vitamin D(3) has no influence on the expression of alkaline phosphatase or the deposition of calcium. Expression of these additional osteogenic markers requires addition of a cocktail of osteogenic <em>factors</em> that, conversely, have no influence on the expression of osteocalcin. Supplementation of the cell culture medium with both vitamin D(3) and a cocktail of osteogenic <em>factors</em> is recommended to produce an osteoblast phenotype that secretes osteocalcin, expresses alkaline phosphatase and deposits calcium. In such a supplemented medium, a mean osteocalcin concentration of 11.63 ± 4.85 ng/ml was secreted by the osteoblasts. Distinguishing osteoblasts and <em>fibroblasts</em> remains a challenge. Neither differentiated nor undifferentiated osteoblasts can be distinguished from <em>fibroblasts</em> by the expression of CD90, ED-A-fibronectin or α-smooth muscle actin; however, these cell types exhibit clear differences in their <em>growth</em> characteristics. Osteoblasts can be arranged as 3D spheroids by coating the bottom of the cell culture device with agarose. The cellular composition of 3D multicellular spheroids can be evaluated quantitatively using vital fluorescence labeling techniques. Spheroids are a promising tool for studying angiogenic and osteogenic phenomena in vivo and in vitro.

OBJECTIVE

The dual pathway model of urothelial carcinogenesis does not fully explain grade and stage progression in patients with initial low-grade, non-muscle invasive urothelial carcinomas. Fibroblast growth factor receptor 3 (FGFR3) mutations are a hallmark of the low-grade pathway, with subsequent progression to muscle invasion occurring when FGFR3 mutant tumours exhibit a homozygous CDKN2A deletion. We hypothesized that grade heterogeneity represents the morphological manifestation of molecular changes associated with disease progression.

RESULTS

We identified retrospectively 29 non-muscle invasive papillary urothelial carcinomas with grade heterogeneity (<20% high grade). Nineteen had sufficient material for immunohistochemistry, CDKN2A fluorescence in-situ hybridization and FGFR3 mutation analysis. Eight pure low-grade urothelial carcinomas (PLGUC) were also analysed. FGFR3 mutation was seen in 10 of 19 cases. A homozygous CDKN2A deletion was identified in the low-grade areas of eight of nine (88%) technically suitable FGFR3 mutant cases (including five pTa cancers), in five of nine FGFR3 wild-type carcinomas and in none of the PLGUC. Increased MIB-1 expression was seen in low-grade areas of 12 of 19, in high-grade areas of 17 of 19 cases with grade heterogeneity and in none of the PLGUC. p53 staining was increased in one of 19 low-grade and seven of 19 high-grade areas.

CONCLUSIONS

Our findings show that grade heterogeneity in urothelial carcinoma is characterized by increased MIB-1 labelling, and particularly in the FGFR3 mutant pathway, with homozygous deletions of CDKN2A in low- and high-grade areas. This would suggest that CDKN2A deletion occurs prior to grade progression and supports the current convention to assign the highest grade to urothelial carcinomas with grade heterogeneity.

BACKGROUND

The cause of early-accelerated atherosclerosis development observed in Chronic Kidney Disease (CKD) is not fully understood. The determination of the relationship between the levels of fibroblast growth factor 23 (FGF-23) and the development of endothelial dysfunction, left ventricular hypertrophy, and myocardial infarction lends support to the possibility that FGF-23 plays a role in the development of atherosclerosis in CKD. Only a few studies, however, have been conducted that analyze the relationship between FGF-23 levels in the progression of CKD and the development of atherosclerosis, and these studies have generally been limited to those patients receiving dialysis therapy due to end stage renal disease (ESRD).

METHODS

In the present study, carotid artery intima-media thicknesses (IMT) were measured ultrasonically as a marker of atherosclerosis in 91 patients with CKD stage 3 - 4 (61 female and 30 male, age between 19 - 65 years, glomerular filtration rate [GFR] 15 - 60 mL/min 1.73 m2, CKD was not related to diabetes mellitus, and without cardiovascular-cerebral disease) in contrast to 36 healthy volunteers (26 female and 10 male, age between 19 - 65 years, GFR>> 90 mL/min 1.73 m2, and without any diagnoses of acute or chronic disease), and a possible role of FGF-23 on atherosclerosis was analyzed.

RESULTS

Patients were similar to controls with respect to age, gender, smoking status, body mass index, and plasma glucose and lipid profile. On the other hand, IMT measurements (p < 0.00001) and FGF-23 levels (p = 0.00012) were significantly higher in patients than controls. IMT was measured above the subclinical atherosclerosis limit of 0.750 mm in 54% of the patients. Multivariate regression analysis showed that patients' age, high sensitive c-reactive protein (hsCRP), and FGF-23 levels were independent predictors of IMT (p < 0.00001, r = 0.559). Independent of other variables, every 1 μmol/L increase in FGF-23 levels resulted in 0.444 mm increase of IMT measurements in patients with CKD.

CONCLUSIONS

Our findings suggest that monitoring serum FGF-23 may be useful as a non-invasive indicator of subclinical atherosclerosis in patients with chronic kidney disease.

The regenerative capacity of peripheral nerves is limited after nerve injury. A number of <em>growth</em> <em>factors</em> modulate many cellular behaviors, such as proliferation and migration, and may contribute to nerve repair and regeneration. Our previous study observed the dynamic changes of genes in L4-6 dorsal root ganglion after rat sciatic nerve crush using transcriptome sequencing. Our current study focused on upstream <em>growth</em> <em>factors</em> and found that a total of <em>19</em> upstream <em>growth</em> <em>factors</em> were dysregulated in dorsal root ganglions at 3, 9 hours, 1, 4, or 7 days after nerve crush, compared with the 0 hour control. Thirty-six rat models of sciatic nerve crush injury were prepared as described previously. Then, they were divided into six groups to measure the expression changes of representative genes at 0, 3, 9 hours, 1, 4 or 7 days post crush. Our current study measured the expression levels of representative upstream <em>growth</em> <em>factors</em>, including nerve <em>growth</em> <em>factor</em>, brain-derived neurotrophic <em>factor</em>, <em>fibroblast</em> <em>growth</em> <em>factor</em> 2 and amphiregulin genes, and explored critical signaling pathways and biological process through bioinformatic analysis. Our data revealed that many of these dysregulated upstream <em>growth</em> <em>factors</em>, including nerve <em>growth</em> <em>factor</em>, brain-derived neurotrophic <em>factor</em>, <em>fibroblast</em> <em>growth</em> <em>factor</em> 2 and amphiregulin, participated in tissue remodeling and axon <em>growth</em>-related biological processes Therefore, the experiment described the expression pattern of upstream <em>growth</em> <em>factors</em> in the dorsal root ganglia after peripheral nerve injury. Bioinformatic analysis revealed <em>growth</em> <em>factors</em> that may promote repair and regeneration of damaged peripheral nerves. All animal surgery procedures were performed in accordance with Institutional Animal Care Guidelines of Nantong University and ethically approved by the Administration Committee of Experimental Animals, China (approval No. 20170302-017) on March 2, 2017.

BACKGROUND

Fibroblast growth factor-2 (FGF-2) protein level has been shown to be elevated in aged mice muscle compared to adult mice. It activates the satellite cell quiescence, which leads to satellite cell depletion and may accelerate aging process. The purpose of this study was to see the effect of resistance exercise on skeletal muscle FGF-2 protein level in aged mice.

METHODS

This study included eight young adult control C57BL/6 male mice (age 12 weeks, YCON group) and 14 aged C57BL/6 male mice (age 19 months), randomly divided into two groups (old control, OCON, n = 7; old resistance exercise, ORT, n = 7). Resistance ladder climbing exercise was conducted 3 d/wk for 12 weeks. Soleus and tibialis anterior muscles were collected for body composition, relative grip strength, and muscle wet weight and for enzyme-linked immunosorbent assay protein analysis.

RESULTS

Relative soleus muscle wet weight and hindlimb lean mass showed a significant increase in ORT group compared to OCON group (p = 0.013 and p = 0.015, respectively). In relative grip strength, both OCON and ORT showed a significant decrease compared to YCON (p < 0.001 and p = 0.011, respectively). However, ORT showed a significant increase compared to OCON (p = 0.02). OCON showed a significant increase in skeletal muscle FGF-2 protein level compared to YCON in soleus (p = 0.035), and ORT showed a significant decrease compared to OCON in soleus muscle (p = 0.045). FGF-2 protein level was significantly decreased in tibialis anterior muscle in the ORT group compared to OCON (p = 0.022). Correlation analysis showed a negative correlation between FGF-2 protein level and soleus and tibialis anterior muscle weight (r2 = 0.514, p = 0.0035; r2 = 0.312, p = 0.025, respectively).

CONCLUSIONS

Twelve weeks of resistance exercise reduced soleus and tibialis anterior muscle FGF-2 protein level in aged mice with improvement in appendicular lean mass and muscle strength. Thus, resistance exercise could be an efficient intervention for preventing age-related increase of FGF-2 in skeletal muscle.

The rising prevalence of non-alcoholic fatty liver disease (NAFLD) parallels the global increase in the number of people diagnosed with obesity and metabolic syndrome. The gut-liver axis (GLA) plays an important role in the pathogenesis of NAFLD/non-alcoholic steatohepatitis (NASH). In this review, we discuss the clinical significance and underlying mechanisms of action of gut-derived secretory <em>factors</em> in NAFLD/NASH, focusing on recent human studies. Several studies have identified potential causal associations between gut-derived secretory <em>factors</em> and NAFLD/NASH, as well as the underlying mechanisms. The effects of gut-derived hormone-associated drugs, such as glucagon-like peptide-1 analog and recombinant variant of <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em>, and other new treatment strategies for NAFLD/NASH have also been reported. A <em>growing</em> body of evidence highlights the role of GLA in the pathogenesis of NAFLD/NASH. Larger and longitudinal studies as well as translational research are expected to provide additional insights into the role of gut-derived secretory <em>factors</em> in the pathogenesis of NAFLD/NASH, possibly providing novel markers and therapeutic targets in patients with NAFLD/NASH.

Purpose: High circulating levels of the hormone prolactin (PRL) protect against experimental diabetic retinopathy (DR) due to the retinal accumulation of vasoinhibin, a PRL fragment that inhibits blood vessel permeability and growth. A phase 2 clinical trial is investigating a new therapy for DR based on elevating serum PRL levels with levosulpiride, a prokinetic dopamine D2 receptor blocker. Here, we tested whether levosulpiride-induced hyperprolactinemia elevates PRL and vasoinhibin in the vitreous of volunteer patients with proliferative DR (PDR) undergoing elective pars plana vitrectomy.

Methods: Patients were randomized to receive placebo (lactose pill, orally TID; n = 19) or levosulpiride (25 mg orally TID; n = 18) for the 7 days before vitrectomy. Vitreous samples from untreated non-diabetic (n = 10) and PDR (n = 17) patients were also studied.

Results: Levosulpiride elevated the systemic (101 ± 13 [SEM] vs. 9.2 ± 1.3 ng/mL, P < 0.0001) and vitreous (3.2 ± 0.4 vs. 1.5 ± 0.2 ng/mL, P < 0.0001) levels of PRL, and both levels were directly correlated (r = 0.58, P < 0.0002). The vitreous from non-diabetic patients or from PDR patients treated with levosulpiride, but not from placebo-treated PDR patients, inhibited the basic fibroblast growth factor (bFGF)- and vascular endothelial growth factor (VEGF)-induced proliferation of endothelial cells in culture. Vasoinhibin-neutralizing antibodies reduced the vitreous antiangiogenic effect. Matrix metalloproteases (MMPs) in the vitreous cleaved PRL to vasoinhibin, and their activity was higher in non-diabetic than in PDR patients.

Conclusions: Levosulpiride increases the levels of PRL in the vitreous of PDR patients and promotes its MMP-mediated conversion to vasoinhibin, which can inhibit angiogenesis in DR.

Translational relevance: These findings support the potential therapeutic benefit of levosulpiride against vision loss in diabetes.

Keywords: 16K prolactin; antiangiogenic factor; dopamine D2 receptor blocker; hyperprolactinemia; proliferative diabetic retinopathy.

The novel Coronavirus Disease 20<em>19</em> (COVID-<em>19</em>) crisis is now present in more than 200 countries. It started in December 20<em>19</em> and has, so far, led to more than 149, 470,968 cases, 3,152,121 deaths, and 127,133,013 survivors recovered by 28 April 2021. COVID-<em>19</em> has a high morbidity, and mortality of 2%, on average, whereas most people are treated after a period of time. Some people who recover from COVID-<em>19</em> are left with 20 to 30% decreased lung function. In this context, exercise focused on skeletal muscle with minimal lung involvement could potentially play an important role. Regular exercise protects against diseases associated with chronic low-grade systemic inflammation. This long-term effect of exercise may be ascribed to the anti-inflammatory response elicited by an acute bout of exercise, which is partly mediated by muscle-derived myokines. The isometric training system seems to have this feature, because this system is involved with the skeletal muscle as the target tissue. However, no studies have examined the effect of exercise on the treatment and recovery of COVID-<em>19</em>, and, more importantly, "muscle-lung cross-talk" as a mechanism for COVID-<em>19</em> treatment. It is suggested that this theoretical construct be examined by researchers.

<strong class="sub-title"> Keywords: </strong> COVID-<em>19</em>; anti-inflammatory treatment; chronic diseases; cytokines; exercise; fibroblast growth factor 21; immune response; myokines; oxidative stress.

Cell therapy raises hope to reduce the harmful effects of acute myocardial ischemia. Stem and progenitor cells (SPCs) may be a valuable source of trophic <em>factors</em>. In this study, we assessed the plasma levels of selected trophic <em>factors</em> in patients undergoing application of autologous bone marrow (BM)-derived, lineage-negative (Lin^-) stem/progenitor cells into the coronary artery in the acute phase of myocardial infarction. The study group consisted of 15 patients with acute myocardial infarction (AMI) who underwent percutaneous revascularization and, afterwards, Lin^- stem/progenitor cell administration into the infarct-related artery. The control group consisted of <em>19</em> patients. BM Lin^- cells were isolated using immunomagnetic methods. Peripheral blood was collected on day 0, 2, 4, and 7 and after the first and third month to assess the concentration of selected trophic <em>factors</em> using multiplex fluorescent bead-based immunoassays. We found in the Lin^- group that several angiogenic trophic <em>factors</em> (vascular endothelial <em>growth</em> <em>factor</em>, Angiopoietin-1, basic <em>fibroblast</em> <em>growth</em> <em>factor</em>, platelet-derived <em>growth</em> <em>factor</em>-aa) plasma level significantly increased to the 4th day after myocardial infarction. In parallel, we noticed a tendency where the plasma levels of the brain-derived neurotrophic <em>factor</em> were increased in the Lin^- group. The obtained results suggest that the administered SPCs may be a valuable source of angiogenic trophic <em>factors</em> for damaged myocardium, although this observation requires further in-depth studies.

OBJECTIVE

Chronic liver disease induces an acquired deficiency of S-adenosyl-L-methionine (SAMe) leading to impairment of detoxifying processes in the liver. Ursodeoxycholic acid (UDCA) represents the standard treatment in primary biliary cholangitis (PBC). As both compounds exert their hepatoprotective effects by different mechanisms, it is conceivable that when used together their effect might be additive. The aim of this study was to analyse the effect of SAMe supplementation on liver biochemistry and health-related quality of life (HRQoL) in patients with PBC, treated with UDCA.

METHODS

In this prospective pilot, proof of the principle, non-randomized and open label study we enrolled 24 patients with PBC treated with UDCA for at least 6 months. They had received both UDCA in a standard dose of 13-15 mg/kg b.w. and SAMe in the dose of 1200 mg daily over a period of 6 months. A group of 24 patients with PBC treated with UDCA served as control for liver biochemistry (Study registered on the platform ClinicalTrials.gov under ID: NCT02557360).

RESULTS

We observed a significant decrease of ALP, GGT and total cholesterol in non-cirrhotic patients treated with SAMe. There was also a significant improvement of fatigue and pruritus in PBC-40 questionnaire and amelioration of anxiety in STAI 2 questionnaire in the SAMe group. Treatment with SAMe neither increased sulfation capacity of the liver nor had an effect on <em>fibroblast</em> <em>growth</em> <em>factor</em>-<em>19</em> serum levels.

CONCLUSIONS

Our pilot study demonstrates a positive effect of adding SAMe to UDCA in non-cirrhotic patients with PBC.

Sevelamer, a non-absorbable amine-based resin used for treatment of hyperphosphataemia, has been demonstrated to have a marked bile acid-binding potential alongside beneficial effects on lipid and glucose metabolism. The aim of this study was to investigate the glucose-lowering effect and mechanism(s) of sevelamer in patients with type 2 diabetes.

In this double-blinded randomized controlled trial, we randomized 30 patients with type 2 diabetes to sevelamer (n = 20) or placebo (n = 10). Participants were subjected to standardized 4-hour liquid meal tests at baseline and after 7 days of treatment. The main outcome measure was plasma glucagon-like peptide-1 excursions as measured by area under the curve. In addition, blood was sampled for measurements of glucose, lipids, glucose-dependent insulinotropic polypeptide, C-peptide, glucagon, <em>fibroblast</em> <em>growth</em> <em>factor</em>-<em>19</em>, cholecystokinin and bile acids. Assessments of gastric emptying, resting energy expenditure and gut microbiota composition were performed.

Sevelamer elicited a significant placebo-corrected reduction in plasma glucose with concomitant reduced <em>fibroblast</em> <em>growth</em> <em>factor</em>-<em>19</em> concentrations, increased de novo synthesis of bile acids, a shift towards a more hydrophilic bile acid pool and increased lipogenesis. No glucagon-like peptide-1-mediated effects on insulin, glucagon or gastric emptying were evident, which points to a limited contribution of this incretin hormone to the glucose-lowering effect of sevelamer. Furthermore, no sevelamer-mediated effects on gut microbiota composition or resting energy expenditure were observed.

Sevelamer reduced plasma glucose concentrations in patients with type 2 diabetes by mechanisms that seemed to involve decreased intestinal and hepatic bile acid-mediated farnesoid X receptor activation.

Disruption of protein:protein interactions (PPIs) that regulate the function of voltage-gated Na⁺ (Nav) channels leads to neural circuitry aberrations that have been implicated in numerous channelopathies. One example of this pathophysiology is mediated by dysfunction of the PPI between Nav1.6 and its regulatory protein <em>fibroblast</em> <em>growth</em> <em>factor</em> 14 (FGF14). Thus, peptides derived from FGF14 might exert modulatory actions on the FGF14:Nav1.6 complex that are functionally relevant. The tetrapeptide Glu-Tyr-Tyr-Val (EYYV) mimics surface residues of FGF14 at the β8-β9 loop, a structural region previously implicated in its binding to Nav1.6. Here, peptidomimetics derived from EYYV (<b>6</b>) were designed, synthesized, and pharmacologically evaluated to develop probes with improved potency. Addition of hydrophobic protective groups to <b>6</b> and truncation to a tripeptide (<b>12</b>) produced a potent inhibitor of FGF14:Nav1.6 complex assembly. Conversely, addition of hydrophobic protective groups to <b>6</b> followed by addition of an <i>N</i>-terminal benzoyl substituent (<b><em>19</em></b>) produced a potentiator of FGF14:Nav1.6 complex assembly. Subsequent functional evaluation using whole-cell patch-clamp electrophysiology confirmed their inverse activities, with <b>12</b> and <b><em>19</em></b> reducing and increasing Nav1.6-mediated transient current densities, respectively. Overall, we have identified a negative and positive allosteric modulator of Nav1.6, both of which could serve as scaffolds for the development of target-selective neurotherapeutics.

Keywords: fibroblast growth factor 14 (FGF14); molecular docking; neurotherapeutics; peptidomimetics; protein:protein interactions (PPIs); voltage-gated Na+ (Nav) channels.

Regional changes to the intestinal microenvironment brought about by Roux-en-Y gastric bypass (RYGB) surgery may contribute to some of its potent systemic metabolic benefits through favorably regulating various local cellular processes. Here, we show that the intestinal contents of RYGB-operated compared with sham-operated rats region-dependently confer superior glycemic control to recipient germ-free mice in association with suppression of endotoxemia. Correspondingly, they had direct barrier-stabilizing effects on an intestinal epithelial cell line which, bile-exposed intestinal contents, were partly farnesoid X receptor (FXR)-dependent. Further, circulating <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> levels, a readout of intestinal FXR activation, negatively correlated with endotoxemia severity in longitudinal cohort of RYGB patients. These findings suggest that various host- and/or microbiota-derived luminal <em>factors</em> region-specifically and synergistically stabilize the intestinal epithelial barrier following RYGB through FXR signaling, which could potentially be leveraged to better treat endotoxemia-induced insulin resistance in obesity in a non-invasive and more targeted manner.

Keywords: Cell Biology; Human Metabolism; Molecular Biology.

UNASSIGNED

<em>Fibroblast</em> <em>growth</em> <em>factor</em> (FGF)<em>19</em> has been shown to improve glycemic homeostasis and lipid metabolism in animal models. In humans, decreased FGF<em>19</em> level has been described in diabetes. The present study aimed to investigate the expression of FGF<em>19</em> in gestational diabetes mellitus (GDM) patients.

METHODS

Samples for measurement were obtained from 20 women with GDM and 25 healthy controls. The messenger ribonucleic acid (mRNA) and protein expression levels of FGF<em>19</em>, FGF21 and co-receptor β-klotho (KLB) in the placenta, rectus muscle and subcutaneous fat tissues were quantified by real-time quantitative polymerase chain reaction, western blot and immunohistochemistry, respectively.

RESULTS

Women with GDM had significantly lower mRNA and protein expressions of FGF<em>19</em> than control women in the placenta (mRNA 0.33 ± 0.05 vs 0.72 ± 0.09; protein 0.34 ± 0.13 vs 0.85 ± 0.20) and rectus muscle (mRNA 0.83 ± 0.11 vs 1.28 ± 0.<em>19</em>; protein 0.78 ± 0.24 vs 1.23 ± 0.39). However, there were no significant differences between GDM women and controls with respect to the expression levels of FGF21 and β-klotho in the placenta and rectus muscle. There were almost no detectable FGF<em>19</em> and FGF21 expressions in subcutaneous fat tissue. Furthermore, β-klotho expression levels were not different between the GDM and control group in subcutaneous fat.

CONCLUSIONS

FGF<em>19</em> expressions are decreased in the placenta and rectus muscle of women with GDM. This might contribute to the pathophysiology or development of GDM.

The inflammatory response of the lung to noxious <em>factors</em> contributes to the pathogenesis of chronic lung injury. Inflammatory mediators regulate the insulin-like <em>growth</em> <em>factor</em> (IGF) system, a key modulator of lung <em>fibroblast</em> proliferation. The activity of IGFs is regulated by IGF-binding proteins (IGFBPs) secreted by lung cells. To investigate the regulation of lung <em>fibroblast</em> IGFBPs by cytokines, we exposed <em>19</em>-d fetal rat lung <em>fibroblasts</em> to various pro- and anti-inflammatory mediators. IGFBP abundance in conditioned medium (CM) was measured by ligand blot and RNA transcript abundance by RNase protection assays. Fetal rat lung <em>fibroblasts</em> exposed to interleukin (IL)-1beta or tumor necrosis <em>factor</em> (TNF)-alpha for 48 h demonstrated increased abundance of CM IGFBP-3 (5.9- and 4.7-fold increases for IL-1beta and TNF-alpha, respectively) and IGFBP-4 (5.7- and 7.4-fold increases for IL-1beta and TNF-alpha, respectively) that was accompanied by a small increase in IGFBP-4 mRNA and a larger increase in IGFBP-3 mRNA abundance. IGFBP-4 specific proteolysis was examined in CM collected from fetal rat lung <em>fibroblasts</em> after incubation with serum-free medium (SFM), IL-1beta, or TNF-alpha for 48 h. Cell-free aliquots of SFM-CM incubated at 37C for 24 h showed a 65% decrease in IGFBP-4 abundance that was inhibited by 1,10-phenanthroline. In contrast, CM from cells exposed to IL-1beta or TNF-alpha incubated at 37 degrees C for 24 h did not show a significant decrease in IGFBP-4 abundance unless IGF-I was present during the cell-free incubation. Addition of IGFBP-3 to aliquots of SFM-CM reversed the IGF-I-mediated acceleration of IGFBP-4 proteolysis. Similarly, addition of IGFBP-3 to cells in culture increased the accumulation of CM IGFBP-4. These results demonstrate that cytokines regulate IGFBP production and clearance by fetal lung cells and suggest a mechanism by which cytokines regulate cell proliferation following lung injury.

OBJECTIVE

A strong correlation exists between tenascin-C induction, and acute inflammation. Generally increased tenascin-C concentrations are correlated with various inflammatory, and infectious diseases. In patients with diagnosis of Inflammatory Bowel Disease (IBD) presence of tenascin-C in colonic mucosa demonstrates tissue repair, and its mucosal concentrations are correlated with local disease activity Therefore plasma levels of tenascin-C have been demonstrated to be a helpful indicator of the activity of inflammatory bowel diseases. In this study, firstly in the literature, we aimed to display the correlation between tenascin-C expression, and formation of intestinal stricture.

METHODS

A total of 43 patients (male, n=<em>19</em>; 44.2%; and female, n=24, 55.8%) aged between <em>19</em>, and 63 years, with clinically, endoscopically, radiologically, and histopathologically confirmed definitive diagnosis of Crohn's disease who were examined, diagnosed, and treated in the Gastroenterology Clinic of Haydarpasa Numune Training and Research Hospital between January 2011, and April 2012 were investigated. Serum tenascin-C levels were measured using commercial sandwich enzyme-linked immunosorbent assay Human Tenascin-C Purified Protein kit (Chemicon, Millipore(R), USA). Study groups were categorized based on the type of the disease as inflammatory (n=17; 39.5%), obstructive (27.2%), and fistule formation (n=10; 23.3%) Crohn's disease. For statistical analysis SPSS (Statistical Package for Social Sciences) Statistics 15 program was used.

RESULTS

Median tenascin- C value in the obstructive group (6.57 ng/mL; range, 4.26-21.87 ng/mL) was statistically significantly higher than that detected in the inflammatory (1.74 ng/mL; range,1.29-3.16 ng/mL), and fistulizing (1.44 ng/mL; range, 0.74-2.47 ng/mL) groups (p=0.002).

CONCLUSIONS

Intestinal fibroblasts have an important role in the stricture formation process in CD. Transforming growth factor (TGF)-b1 cytokine is in the center of this process. A strong correlation exists between tenascin-C induction, and acute inflammation. As a known fact, serum tenascin-C levels can be used in the determination of activity of IBD. Starting from this point, serum tenascin-C levels can be useful in the categorization of the Crohn's disease without the need for invasive methods. In the future, studies with larger patient series investigating use of serum tenascin-C in the prediction of stricturing Crohn's disease should be conducted.

It was previously demonstrated that patients with bile acid (BA) diarrhea have low fasting serum levels of the ileal hormone--<em>fibroblast</em> <em>growth</em> <em>factor</em>-<em>19</em> (FGF-<em>19</em>). In this paper, the authors demonstrate a positive correlation between ileal transcripts of FGF-<em>19</em> and (75)SeHCAT retention, providing further evidence of FGF-<em>19</em> deficiency as an explanation for BA diarrhea. Variants in KLB and FGFR4 genes (that determine the functional re-uptake of BA in the portal circulation by hepatocytes) are also demonstrated to be associated with (75)SeHCAT retention, confirming a second potential mechanism for the development of BA diarrhea.

Smoking cessation reduces the risk of cardiovascular disease and improves clinical outcomes. We studied the effect of smoking cessation on plasma levels of α-klotho, which is an antiaging protein. We treated 28 smokers (male:female = 23:5, 46 ± 12 years) with varenicline (n = 14) or a transdermal nicotine patch (n = 14) as part of a 12-week smoking cessation program (the VN-SEESAW Study). Pulse rate, blood pressure, plasma levels of α-klotho, <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF)-<em>19</em>, FGF-21, hemoglobin (Hb), and expiratory carbon monoxide (CO) concentration were measured before and after the antismoking intervention. Smoking cessation significantly decreased pulse rate, α-klotho, Hb, and CO concentration, but not FGF-<em>19</em> or FGF-21 in all subjects. On the contrary, body mass index significantly increased after the intervention. Changes in α-klotho levels (values at week 12 - values at week 0) were negatively associated with α-klotho levels at week 0 and positively associated with changes in Hb levels. In addition, the successful smoking cessation group (n = 21) showed significant reductions in pulse rate, systolic blood pressure, α-klotho, Hb, and CO concentration. In conclusion, smoking cessation significantly decreased serum levels of the antiaging molecule α-klotho. Our results are consistent with a previous report that an increase in α-klotho might be a compensatory response to smoking stress.

Background and aims: Non-alcoholic fatty liver disease (NAFLD) is becoming increasingly prevalent and nutrition intervention remains the most important therapeutic approach for NAFLD. Our aim was to investigate whether low- (LP) or high-protein (HP) diets are more effective in reducing liver fat and reversing NAFLD and which mechanisms are involved.

<strong class="sub-title"> Methods: </strong> <em>19</em> participants with morbid obesity undergoing bariatric surgery were randomized into two hypocaloric (1500-1600 kcal/day) diet groups, a low protein (10E% protein) and a high protein (30E% protein), for three weeks prior to surgery. Intrahepatic lipid levels (IHL) and serum <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) were measured before and after the dietary intervention. Autophagy flux, histology, mitochondrial activity, and gene expression analyses were performed in liver samples collected during surgery.

Results: IHL levels decreased by 42.6% in the HP group, but were not significantly changed in the LP group despite similar weight loss. Hepatic autophagy flux and serum FGF21 increased by 66.7% and 42.2%, respectively, after 3 weeks in the LP group only. Expression levels of fat uptake and lipid biosynthesis genes were lower in the HP group compared with those in the LP group. RNA-seq analysis revealed lower activity of inflammatory pathways upon HP diet. Hepatic mitochondrial activity and expression of β-oxidation genes did not increase in the HP group.

Conclusions: HP diet more effectively reduces hepatic fat than LP diet despite of lower autophagy and FGF21. Our data suggest that liver fat reduction upon HP diets result primarily from suppression of fat uptake and lipid biosynthesis.

Keywords: ER-stress; FGF21; autophagy; dietary protein; liver fat; mitochondria; nutrition; obesity.

Bile acids are synthesized from cholesterol only in hepatocytes. Bile acids circulating in the enterohepatic system act as physiological detergent molecules to help solubilize biliary cholesterol and emulsify dietary lipids and fat-soluble vitamins in small intestine. Bile acids are signaling molecules that activate nuclear receptor farnesoid X receptor (FXR) and cell surface G protein-coupled receptor TGR5. FXR critically regulates bile acid homeostasis by mediating bile acid feedback inhibition of hepatic bile acid synthesis. In addition, bile acid-activated cellular signaling pathways regulate metabolic homeostasis, immunity, and cell proliferation in various metabolically active organs. In the small and large intestine, gut bacterial enzymes modify primary bile acids to generate secondary bile acids to help shape the bile acid pool composition and subsequent biological effects. In turn, bile acids exhibit anti-microbial properties and modulate gut microbiota to influence host metabolism and immunity. Currently, bile acid-based therapies including systemic and intestine-restricted FXR agonists, TGR5 agonists, <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> analogue, intestine FXR antagonists, and intestine apical sodium-bile acid transporter (ASBT) inhibitors have been developed as promising treatments for non-alcoholic steatohepatitis (NASH). These pharmacological agents improved metabolic and inflammatory disorders via distinct mechanisms of action that are subjects of extensive research interest. More recently, human and experimental alcoholic liver disease (ALD) has been associated with disrupted bile acid homeostasis. In additional, new findings showed that targeting bile acid metabolism and signaling may be promising therapeutic approaches for treating ALD.

Fibroblast growth factor family (FGFs) is a kind of cytokine that plays an important role in growth, development, metabolism and disease. During bone development, multiple FGFs and fibroblast growth factor receptors (FGFRs) play important roles. Previous reports have elucidated the great importance of FGF1, 2, 4, 6, 7, 8, 9, 10, and 18 in bone development, and FGF21 and 23 in bone homeostasis and bone regulation. FGF19 was initially found in the human foetal brain, and its gene location is related to osteoporosis pseudoglioma syndrome. Presently, gene chip detection has repeatedly found that FGF19 shows spatiotemporal specificity of gene expression in bone development and bone-related diseases, as well as differences in the protein level, indicating that FGF19 affects the skeletal system. Considering the current insufficient understanding of FGF19 and its potential function in the skeletal system, this review aims to introduce the background of FGF19 in bone, summarise the research progress of FGF19 in the skeletal system, and discuss the role and therapeutic potential of FGF19 in bone development and bone-related diseases.

<strong class="sub-title"> Keywords: </strong> <em>Fibroblast</em> <em>growth</em> <em>factor</em> receptor; <em>Fibroblast</em> <em>growth</em> <em>factor</em>19; Skeletal development; Skeletal diseases.