Background: The blood pressure-lowering effects of the Dietary Approaches to Stop Hypertension (DASH) dietary pattern and reduced sodium intake are well established. The effects on other biomarkers related to vascular health are of interest and might assist in explaining the effects of the DASH diet and sodium reduction.

Objectives: We hypothesized that a low-sodium DASH diet improves (lowers) biomarkers of inflammation [C-reactive protein (CRP) and soluble urokinase plasminogen activator receptor (suPAR)] and mineral metabolism [phosphorus and fibroblast growth factor-23 (FGF23)].

Methods: We conducted a secondary analysis of the DASH-Sodium trial using frozen serum samples. This controlled feeding study randomly assigned 412 adults (≥22 y) with elevated blood pressure (120-159/80-95 mmHg) to consume either a DASH diet or control diet. Within each arm, participants received 3 sodium levels [low (1150 mg), intermediate (2300 mg), high (3450 mg)] in random sequence, each for 30 d. To maximize contrast, samples collected at the end of the low-sodium DASH (n = 198) and high-sodium control (n = 194) diets were compared. Between-diet differences in serum CRP, suPAR, phosphorus, and FGF23 concentrations were assessed using linear regression adjusted for age, sex, race, income, education, smoking status, and BMI.

Results: CRP concentrations did not differ between groups (P = 0.83), but suPAR was higher after the low-sodium DASH diet than the high-sodium control [geometric mean 2470 pg/mL (95% CI: 2380, 2560 pg/mL), compared with 2290 pg/mL (95% CI: 2210, 2380 pg/mL); P = 0.006]. Phosphorus was higher after the low-sodium DASH diet [geometric mean 3.50 mg/dL (95% CI: 3.43, 3.57 mg/dL)] compared with the high-sodium control diet [geometric mean 3.39 mg/dL (95% CI: 3.33, 3.46 mg/dL); P = 0.04]. FGF23 was also higher after the low-sodium DASH diet [geometric mean 35.3 pg/mL (95% CI: 33.3, 37.3 pg/mL) compared with 28.2 pg/mL (95% CI: 26.6, 29.8 pg/mL); P < 0.001].

Conclusions: Contrary to our hypothesis, biomarkers of inflammation and mineral metabolism were increased or unchanged by a low-sodium DASH diet compared with a high-sodium control diet in adults with elevated blood pressure.

Keywords: C-reactive protein; DASH; cardiovascular; fibroblast growth factor-23; inflammation; phosphorus; soluble urokinase plasminogen activator receptor.

Scope: Despite its beneficial properties, higher adiponectin concentrations are paradoxically associated with mortality in advanced age. Several mechanisms are being discussed. However, little is known about postprandial regulation of adiponectin in older adults. We assessed age-specific differences of the adiponectin response to different test meals considering potential determinants.

<strong class="sub-title"> Methods and results: </strong> Older (n = 20) and younger (n = <em>22</em>) women were randomized to a dextrose (DEX) or high-fat (HF) dietary challenge. Postprandial adiponectin and <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) concentrations were measured before and 60, 120, 240 min after ingestion. We assessed postprandial changes and group differences using linear mixed models controlled for possible determinants. In younger women, postprandial adiponectin remained stable after both test meals. In contrast, adiponectin increased following DEX and decreased after HF in older women, irrespective of control variables. Postprandial adiponectin was positively associated with malondialdehyde and inversely associated with interleukin-6 following DEX and also negatively associated with metabolic parameters after both test meals. In older women, elevated postprandial FGF21 concentrations were associated with a higher adiponectin response (β = 30.7, 95%CI 10.6; 50.8, p = 0.007).

Conclusions: Adiponectin response is associated with type of dietary challenge, age and FGF21 response. Age-group differences are partly attributable to metabolic parameters and oxidative stress. This article is protected by copyright. All rights reserved.

Keywords: FGF21; adiponectin; inflammation; oxidative stress; postprandial.

Apert syndrome (AS) is caused by the heterozygous presence of 1 of 2 specific missense mutations of the fibroblast growth factor receptor 2 (FGFR2) gene. The 2 adjacent substitutions, designated p.Ser252Trp (S252W) and p.Pro253Arg (P253R), account for more than 98% of cases. Previous research has identified elevated hearing difficulties and incidence of cleft palate in this population. However, the influence of FGFR2 genotype on the speech, language, and communicative participation of children with AS has yet to be examined.

Methods: A retrospective case note analysis was completed for all patients with a genetically-confirmed Apert mutation who attended the Oxford Craniofacial Unit over a 43-year period (1978-2020). Medical records were analyzed for speech, language, hearing, and communication data in detail. The therapy outcome measures, based on the World Health Organization International Classification of Functioning, Disability, and Health was used to classify patient's communicative participation.

Results: The authors identified 55 AS patients with genetically-confirmed mutation of the FGFR2 gene. One patient with a S252F mutation was excluded. There were 31 patients with the S252W mutation (male = 14; female = 17), age range of last hearing assessment (1-18 years), 64% (18/28) of patients had a cleft palate (including bifid uvula), 15 patients had conductive hearing loss, 1 patient had mixed hearing loss, 18 had otitis media with effusion (4 of whom had a cleft palate); 88% (21/24) of patients had receptive language difficulties, 88% (22/25) of patients had expressive language difficulties, 96% (27/28) of patients had a speech sound disorder. There were 23 patients with the P253R mutation (male = 13; female = 10); age range of last hearing assessment (1-13 years), 35% (8/23) patients had a cleft palate (including bifid uvula), 14 patients had a conductive hearing loss, 17 had otitis media with effusion (2 of whom had a cleft palate). Results indicated that 85% (17/20) of patients had receptive language difficulties, 80% (16/20) had expressive language difficulties, 100% (21/21) had a speech sound disorder. The S252W mutation was significantly-associated with the presence of cleft palate (including bifid uvula) (P = 0.05).Data about the cumulative impact of all of these factors for communicative participation using the therapy outcome measures were available for 47 patients: (30 S252W; 17 P253R). Patients with a S252W mutation had significantly more severe difficulties with communicative participation when compared to individuals with a P253R mutation (P = 0.0005) Cochran-Armitage trend test.

Conclusions: Speech, language, communicative participation, and hearing difficulties are pervasive in patients with AS. The severity and functional impact of these difficulties are magnified in patients with the S252W mutation. Results reinforce the importance of considering patients with AS according to genotype.

The histopathology slide seminar "Classic Examples in Toxicologic Pathology XXVII" was held on February 21 and <em>22</em>, 2020, at the Department of Pathology at the University of Veterinary Medicine in Hannover, Germany, with joint organization by the European Society of Toxicologic Pathology. The goal of this annual seminar is to present and discuss classical and actual cases of toxicologic pathology. This article summarizes the presentations given during the seminar, including images of representative lesions. Ten actual and classical cases of toxicologic pathology, mostly induced by a test article, were presented. These included small intestine pathology and transcriptomics induced by a γ-secretase modulator, liver findings in nonhuman primates induced by gene therapy, drug-induced neutropenia in dogs, device-induced <em>growth</em> plate lesions, polycystic lesions in CAR/PXR double knockout mice, inner ear lesions in transgenic mice, findings in Beagle dogs induced by an inhibitor of the myeloid leukemia cell differentiation protein MCL1, findings induced by a monovalent <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 antagonist, kidney lesions induced by a mammalian target of rapamycin inhibitor in combination therapy, and findings in mutation-specific drugs.

Keywords: CAR/PXR double knockout; DOTA; EGFR; FGF23; FGFR1; HER2; KRAS; Notch; classic examples; everolimus; growth plate; mTOR; mechanical growth modulation; mineralization; nitinol; otoconial dysgenesis; polycystic kidney disease; small intestine; vitamin D; γ secretase modulator.

Background We report a Phase 1 study of LY3076<em>22</em>6, an antibody-drug conjugate composed of human IgG1 monoclonal antibody against the human FGFR3 attached with a cleavable linker to the maytansine derivative DM4 in patients with advanced or metastatic cancer. Methods This study was comprised of two parts: (A) dose escalation in patients with advanced or metastatic cancer and (B) dose expansion in patients with urothelial carcinoma with locally determined FGFR3 alterations. The dose range of LY3076<em>22</em>6 tested was 0.2-5.0 mg/kg as an intravenous infusion on Day 1 of each 21-day cycle. The primary objective was to determine a recommended phase 2 dose (RP2D). Results Twenty-five patients were enrolled (Part A: <em>22</em>, Part B: 3) and received ≥ 1 dose of LY3076<em>22</em>6. No dose-limiting toxicities were reported. LY3076<em>22</em>6 was generally well tolerated; most of the toxicities were Grade 1 or 2. Two patients experienced treatment-related Grade 3 toxicity (embolism and decreased platelet count). Four patients experienced serious adverse events (not treatment-related), all in Part A. Dose-proportional exposure was observed, with an estimated half-life of 2-7 days. No responses were seen with LY3076<em>22</em>6 treatment. Stable disease persisting for > 6 months was observed in 1 patient receiving 3.2 mg/kg of LY3076<em>22</em>6. Conclusion The study demonstrates acceptable safety and tolerability of LY3076<em>22</em>6 up to the 5.0 mg/kg dose. Recruitment was stopped due to pipeline prioritization. Dose escalation of LY3076<em>22</em>6 beyond 5.0 mg/kg in patients with advanced tumors may be possible. The trial was registered on August 19, 2015 under identifier <a href="http://clinicaltrials.gov/show/NCT02529553" title="See in ClinicalTrials.gov">NCT02529553</a> with ClinicalTrials.gov.

<strong class="sub-title"> Keywords: </strong> Advanced cancer; Antibody–drug conjugate; Fibroblast growth factor receptor 3; LY3076<em>22</em>6; Metastatic cancer.

OBJECTIVE

The aim of this study was to understand whether or not the protective effect of green tea after fasting-induced damage in the jejunal mucosa of rat is dependent on cell proliferation and the stimulation of specific growth factors.

METHODS

Sixty adult male Wistar rats were used in this study. The animals were divided randomly into 5 groups, with 12 in each group (G1-5). The animals in G1 (control group) were fed a rat chow diet and water ad libitum. The animals in G2 (fasting group) were fasted for 3 days. The animals in the G3, G4, and G5 groups were fasted for 3 days as G2, but were given water (G3), green tea (G4), or a vitamin E (G5) solution, respectively, for another 7 days. The animals were euthanized, and the jejunum was removed and processed for histological and immunohistochemical analysis.

RESULTS

Compared to the G3 group, the jejunal mucosa of G4 rats showed a 70.6% higher level (p < 0.001) of expression of proliferating cell nuclear antigen and 98% higher level (p = 0.0001) of the expression of transforming growth factor-β1 (TGF-β1), whereas the level of fibroblast growth factor-1 (FGF-1) and insulin-like growth factor-1 (IGF-1) expression was 22 and 11% lower, respectively, in G4 animals as compared to G3 rats. These differences in the expression of FGF-1 and IGF-1 in G4 animals were not statistically significant.

CONCLUSIONS

In this study, green tea repaired the fasting-induced damage in the jejunal mucosa of rats, mainly by inducing a significant expression of TGF-β1 in the jejunal mucosa.

The aim of the study was to evaluate the role of morphogenetic proteins--<em>fibroblast</em> <em>growth</em> <em>factor</em>-23 (FGF-23) and extracellular form (alpha) of Klotho protein, present in the sera of patients with chronic renal disease (CRD) as markers of cardiovascular risk. The study included 130 patients (64 men and 66 women) with stage I-VD CRD. The patients'age was 20-65 (mean 41 ± 6.7) years. 30 of them had chronic glomerulonephritis, 23 chronic tubulointerstitial nephritis, 28 hypertensive nephrosclerosis, <em>22</em> polycystic kidneys, 27 type 2 diabetes mellitus. Inclusion and exclusion criteria were standardfor clinical studies of CRD patients. Control group contained 30 healthy volunteers matched for age and sex. The patients were uniformly distributed by stages of CRD in terms of age and sex (18-20 per group). All of them were followed up during 1 year Standard clinical and laboratory examination was supplemented by the measurement of the parathyroid hormone (PTH), Ca and P levels. Serum FGF-23 and Klotho levels were determined by ELISA before and 1 year after onset of the study. Blood pressure including brachial (peripheral) and aortic (central) one as swell as the pulse wave velocity was measured by a Sphygocorr apparatus (Australia). Other studies included ECG, EchoCG, and X-ray of abdominal aorta in the lateral projection using the Kaupilla method. Comparison of FGF-23 and Klotho levels in patients at different stages of CRD revealed their decrease with decreasing glomerular filtration rate that started before (at IIIA stage) a rise in the serum P and PTG levels (IV-V stage). Negative relationship was documented between the Klotho level and the degree of cardiac calcinosis estimated from a semi-quantitative scale (r = 0.64; p < 0.01). Serum FGF-23 significantly correlated wiht myocardial remodeling (r = 0.612; p < 0.01). Multiple regression analysis showed that patients with elevated FGF-23 and P levels, high central systolic pressure and pulse wave velocity had greater left ventricular myocardium mass. ROC-analysis demonstrated that FGF-23 level over 412 pg/ml is indicative of left ventricular hypertrophy with sensitivity 80% and specificity 76%. Patients undergoing hemodialysis who died within 1 year after the onset of the treatment had a higher FGF-23 level than survivals on hemodialysis. The risk of death during the first year on hemodialysis correlated with the FGF-23 level (r = 0.564, p < 0.01). ROC-analysis showed that Klotho levels below 387 pg/ml suggested an increased risk of myocardiym calcification with sensitivity 80% and specificity 75%. It is concluded that morphogenetic proteins, <em>fibroblast</em> <em>growth</em> <em>factor</em> and Klotho, not only play an important role in mineral metabolism in patients with CRD but also produce pleiotropic effects on the development of cardiovascular complications (via involvement in cardiac and vascular calcification and remodeling). It provides a basis for the use of these proteins as early markers of cardiovascular risk in CRD patients.

A series of novel 3-(thiophen-2-ylthio)pyridine derivatives as insulin-like <em>growth</em> <em>factor</em> 1 receptor (IGF-1R) inhibitors was designed and synthesized. IGF-1R kinase inhibitory activities and cytotoxicities against HepG2 and WSU-DLCL2 cell lines were tested. For all of these compounds, potent cancer cell proliferation inhibitory activities were observed, but not through the inhibition of IGR-1R. Selected compounds were further screened against various kinases. Typical compound <em>22</em> (50% inhibitory concentration [IC₅₀ ] values, HepG2: 2.98 ± 1.11 μM and WSU-DLCL2: 4.34 ± 0.84 μM) exhibited good inhibitory activities against <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor-2 (FGFR2), FGFR3, epidermal <em>growth</em> <em>factor</em> receptor, Janus kinase, and RON (receptor originated from Nantes), with IC₅₀ values ranging from 2.14 to 12.20 μM. Additionally, the cell-cycle analysis showed that compound <em>22</em> could arrest HepG2 cells in the G1/G0 phase. Taken together, all the experiments confirmed that the compounds in this series were multitarget anticancer agents worth further optimizing.

<AbstractText>This study assessed the impact of iron administration on serum <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF23) levels.</AbstractText><AbstractText>Of 123 hemodialysis (HD) patients treated with erythropoiesis-stimulating agents, <em>22</em> received once-weekly intravenous iron and 17 received daily oral iron with iron-containing phosphate binders. Intact FGF23 and biomarkers of iron metabolism were measured from blood samples drawn before each HD session, at baseline and on days 3, 5, 7, and 14.</AbstractText><AbstractText>Phosphate levels did not differ among the 3 groups during the 14-day period. Ferritin levels were significantly increased in both iron treatment groups compared with the non-iron treatment group, but changes in transferrin saturation levels were similar in the intravenous iron and non-iron groups. However, intact FGF23 levels were continuously higher in the intravenous iron group than those in the other groups.</AbstractText><AbstractText>Intravenous iron administration may influence intact FGF23 levels in HD patients independently of phosphate and iron metabolism.</AbstractText>

<AbstractText>Vascular calcification has played a vital role in increasing the prevalence of cardiovascular disease (CVD) and mortality in maintenance haemodialysis (MHD) patients. This study is aimed at exploring the prognostic value of abdominal aortic calcification (AAC) estimated by plain lateral abdominal radiography in MHD patients.</AbstractText><AbstractText>Lateral abdominal radiography was used to determine the abdominal aortic calcification score (AACS). The serum level of <em>fibroblast</em> <em>growth</em> <em>factor</em>-23 was tested by enzyme-linked immunosorbent assay. Patients were divided into two groups: no or minor calcification group (AACS < 5) and moderate to severe calcification group (AACS ≥ 5). All patients were followed up to death or the end of the study (30 November 2016).</AbstractText><AbstractText>A total of 114 patients were enrolled in this study, including 64 males (56.1%), and the mean age was 57.42 ± 13.48 years. Seventy-six patients (66.7%) exhibited AAC. Independent predictors for moderate to severe calcification were older age (odds ratio (OR) 1.06 (1.02-1.10), P = 0.003), longer dialysis vintage (OR 1.01 (1.00-1.02), P = 0.039), presence of smoking (OR 3.01 (1.18-7.70), P = 0.021) and higher Log <em>fibroblast</em> <em>growth</em> <em>factor</em>-23 serum levels (OR 2.83 (1.01-7.94), P = 0.048). During a median follow-up of 6.0 (5.6, 6.1) years, <em>22</em> patients (19.3%) died of all-cause death, and 17 cases (14.9%) died of CVD. Kaplan-Meier survival curves showed that patients in the moderate to severe calcification group had significantly higher all-cause (28.3 vs 11.5%, P = 0.028) and CVD mortality (<em>22</em>.6 vs 8.2%, P = 0.035) than that in the no or minor calcification group. A multivariate Cox regression showed that AACS (hazard ratio 1.08 (1.01-1.15), P = 0.0<em>22</em>) was an independent predictor of CVD mortality. Compared with the no or minor calcification group, the risk of CVD mortality was increased by a <em>factor</em> of 3.14 in patients in the moderate to severe calcification group (hazard ratio 3.14 (1.04-9.44), P = 0.042).</AbstractText><AbstractText>Our data suggest that AAC is prevalent in MHD patients and could provide potential predictive information for CVD mortality. Plain lateral abdominal radiography, which is simple and cheap and involves lower radiation, might represent an appropriate screening method for evaluating vascular calcification in daily clinical practice.</AbstractText>

Our purpose was to assess the relationship between serum <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF23) and left ventricular function, carotid intima-media thickness (CIMT) and laboratory features in children on peritoneal dialysis (PD). The study population consisted of 17 patients (11 female; median age 7.83 years, range 0.66-17.75) undergoing PD for <em>22</em> months (range 2-98). Serum FGF23, serum phosphorus, calcium, intact parathyroid hormone (iPTH), 25(OH) vitamin D, 1,25(OH)2 vitamin D and Kt/V urea, left ventricular mass (LVM) and LVM index (LVMI) were assessed. Median FGF23 level was 29.92 pg/ml (<em>22</em>.7-74.76), phosphorus was 5.2 mg/dl (3.1-9.9), iPTH was 438 pg/ml (16-1446), 25(OH) vitamin D was 11 ng/ml (5-35), 1,25(OH)2 vitamin D was 11 pg/ml (2-106), Kt/V urea was 2.33 (1.01-3.84). FGF23 level was independently associated with Kt/V urea (p<0.001). We found that effective dialysis may be the leading determinant of FGF level, independent from the calcium-phosphorus-PTH axis, in pediatric PD patients.

<b>Rationale</b>: Passive heat therapy improves vascular endothelial function, likely via enhanced nitric oxide (NO) bioavailability, although the mechanistic stimuli driving these changes are unknown. <b>Objective</b>: To determine the isolated effects of circulating (serum) <em>factors</em> on endothelial cell function, particularly angiogenesis, and NO bioavailability. <b>Methods and Results</b>: Cultured human umbilical vein endothelial cells (HUVECs) were exposed to serum collected from 20 healthy young (<em>22</em> ± 1 years) adults before (0 wk), after one session of water immersion (Acute HT), and after 8 wk of either heat therapy (N = 10; 36 sessions of hot water immersion; session 1 peak rectal temperature: 39.0 ± 0.03°C) or sham (N = 10; 36 sessions of thermoneutral water immersion). Serum collected following acute heat exposure and heat therapy improved endothelial cell angiogenesis (Matrigel bioassay total tubule length per frame, 0 wk: 69.3 ± 1.9 mm vs. Acute HT: 72.8 ± 1.4 mm, p = 0.04; vs. 8 wk: 73.0 ± 1.4 mm, p = 0.03), with no effects of sham serum. Enhanced angiogenesis was NO-mediated, as addition of the NO synthase (NOS) inhibitor L-NNA to the culture media abolished differences in tubule formation across conditions (0 wk: 71.3 ± 1.8 mm, Acute HT: 71.6 ± 1.9 mm, 8 wk: 70.5 ± 1.6 mm, p = 0.69). In separate experiments, we found that abundance of endothelial NOS (eNOS) was unaffected by Acute HT serum (p = 0.71), but increased by 8 wk heat therapy serum (1.4 ± 0.1-fold from 0 wk, p < 0.01). Furthermore, increases in eNOS were related to improvements in endothelial tubule formation (r² = 0.61, p < 0.01). <b>Conclusions</b>: Passive heat therapy beneficially alters circulating <em>factors</em> that promote NO-mediated angiogenesis in endothelial cells and increase eNOS abundance. These changes may contribute to improvements in vascular function with heat therapy observed <i>in vivo</i>. <b>Abbreviations</b>: Ang-1: angiopoietin-1; ANOVA: analysis of variance; bFGF: basic <em>fibroblast</em> <em>growth</em> <em>factor</em>; CV: cardiovascular; CVD: cardiovascular diseases; eNOS: endothelial nitric oxide synthase; HSPs: heat shock proteins; HT: heat therapy; HUVECs: human umbilical endothelial cells; L-NNA: Nω-nitro-L-arginine; MnSOD: manganese superoxide dismutase; NO: nitric oxide; NOS: nitric oxide synthase; PBMCs: peripheral blood mononuclear cells; RM: repeated measures; sFlt-1: soluble VEGF receptor; SOD: superoxide dismutase; TGF-β: transforming <em>growth</em> <em>factor</em>- β; VEGF: vascular endothelial <em>growth</em> <em>factor</em>.

BACKGROUND

Hepatitis C virus (HCV) is a major cause of chronic liver disease in Egypt, leading to hepatic fibrosis, liver cirrhosis (LC), and hepatocellular carcinoma (HCC). Liver fibrosis is characterized by excessive deposition of extracellular matrix (ECM). Newly-recognized pathogenic mechanisms point to the epithelial-mesenchymal transition (EMT) of hepatocytes to matrix synthesizing (myo-) fibroblasts. Transforming growth factor-beta (TGF-β1), bone morphogenic protein (BMP)-7, and connective tissue growth factor (CTGF) are biomarkers reflecting the EMT process. YKL-40 is a glycoprotein member of ECM and plays a role in cancer cell proliferation. The purpose of this study was to determine the serum biomarkers of EMT and its impact on the fibrogenic process and tumorigenesis in HCV-genotype 4 patients.

METHODS

In this case-control study that was conducted in 2013-2014, 97 HCV-infected patients were subjected to clinical examination, laboratory investigations, and liver biopsy. According to the histopathologic examination, they were classified to F0 (14 cases), F1 (17 cases), F2 (15 cases), F3 (18 cases), F4 (22 cases), and HCC (11 cases). Fifteen age- and gender-matched subjects were included as normal controls. Serum levels of TGF-β1, BMP-7, CTGF, YKL-40 were assessed, and the TGF-β1/BMP-7 ratios were calculated. The data were analyzed by plotting the receiver operating characteristic curve (ROC), Pearson product-moment correlation coefficient, and Spearman's rank correlation coefficient (Spearman's rho).

RESULTS

Serum levels of TGF-β1, BMP-7, CTGF, and YKL-40 were significantly increased in all patient groups compared to controls (p < 0.001). LC exhibited the highest CTGF level and YKL-40 was highest in HCC. The TGF-β1/ BMP-7 ratios reflected the progression of EMT from CHC to LC, however, there was no significant difference between LC and HCC. TGF-β1/ BMP-7 ratio is considered to reflect positive correlation with CTGF in LC group (r = 0.629; p < 0.03) and YKL-40 in HCC group (r = 0.504; p < 0.04).

CONCLUSIONS

Increased TGF-β1/BMP-7 ratio and CTGF levels reflect the rate of EMT and provide information about fibrogenic activity. Also, this ratio, in association with YKL-40, can be used to predict malignant transformation in HCV-genotype 4 Egyptian patients.

Various <em>growth</em> <em>factors</em> regulate synapse development and neurogenesis, and are essential for brain function. Changes in <em>growth</em> <em>factor</em> signaling are implicated in many neuropsychiatric disorders such as depression, autism and epilepsy. We have previously identified that <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> (FGF<em>22</em>) is critical for excitatory synapse formation in several brain regions including the hippocampus. Mice with a genetic deletion of FGF<em>22</em> (FGF<em>22</em> null mice) have fewer excitatory synapses in the hippocampus. We have further found that as a behavioral consequence, FGF<em>22</em> null mice show a depression-like behavior phenotype such as increased passive stress-coping behavior and anhedonia, without any changes in motor, anxiety, or social cognitive tests, suggesting that FGF<em>22</em> is specifically important for affective behavior. Thus, addressing the precise roles of FGF<em>22</em> in the brain will help understand how synaptogenic <em>growth</em> <em>factors</em> regulate affective behavior. In the hippocampus, FGF<em>22</em> is expressed mainly by CA3 pyramidal neurons, but also by a subset of dentate granule cells. We find that in addition to synapse formation, FGF<em>22</em> also contributes to neurogenesis in the dentate gyrus: FGF<em>22</em> null mice show decreased dentate neurogenesis. To understand the cell type-specific roles of FGF<em>22</em>, we generated and analyzed CA3-specific FGF<em>22</em> knockout mice (FGF<em>22</em>-CA3KO). We show that FGF<em>22</em>-CA3KO mice have reduced excitatory synapses on CA3 pyramidal neurons, but do not show changes in dentate neurogenesis. Behaviorally, FGF<em>22</em>-CA3KO mice still show increased immobility and decreased latency to float in the forced swim test and decreased preference for sucrose in the sucrose preference test, which are suggestive of a depressive-like phenotype similar to FGF<em>22</em> null mice. These results demonstrate that: (i) CA3-derived FGF<em>22</em> serves as a target-derived excitatory synaptic organizer in CA3 in vivo; (ii) FGF<em>22</em> plays important roles in dentate neurogenesis, but CA3-derived FGF<em>22</em> is not involved in neurogenesis; and (iii) a depression-like phenotype can result from FGF<em>22</em> inactivation selectively in CA3 pyramidal neurons. Our results link the role of CA3-derived FGF<em>22</em> in synapse development, and not in neurogenesis, to affective behavior.

We investigated the heterogeneity of cells in terms of androgen responsiveness within a single tumor mass of Shionogi carcinoma SC-115 showing androgen-dependent <em>growth</em>. After cloning of the tumor by the limiting dilution method in the presence of androgen, we isolated 40 clones at random. Twenty-two clones required androgen for <em>growth</em> (androgen-dependent phenotype), 16 did not (androgen-independent phenotype), and the remaining two clones showed <em>growth</em> inhibition when androgen was added (androgen-suppressed phenotype). In addition, <em>22</em> androgen-dependent clones showed heterogeneity in <em>growth</em> <em>factor</em> sensitivity in the absence of androgen. All clones were sensitive to both acidic and basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF), 7 of <em>22</em> clones were sensitive to epidermal <em>growth</em> <em>factor</em> (EGF) and transforming <em>growth</em> <em>factor</em> (TGF)-alpha, and 2 of <em>22</em> clones were sensitive to TGF-beta. This preexisting heterogeneity may be partly responsible for the <em>growth</em> of androgen-dependent tumor under hormone-deprived circumstances. Three typical clones, SC2G, SC1G, and SC4A, were selected from androgen-dependent, -independent, and -suppressed phenotypic groups, respectively. These clones, as well as original solid tumors, were found to produce heparin-binding <em>growth</em> <em>factors</em> of heterogeneous elution positions. The molecular nature of these <em>growth</em> <em>factors</em> is not yet known. Neither anti-basic FGF antibody nor anti-EGF antibody inhibited the cell <em>growth</em> when added in cell culture, suggesting the <em>factors</em> were distinct from basic-FGF and EGF.

OBJECTIVE

Studies were conducted on oryzalin (3,5-dinitro-N,N-di(n-propyl)sulfanilamide), a widely used dinitroaniline sulfonamide herbicide, which was identified from plant extracts as an inhibitor of mitogen- and growth factor-mediated intracellular free Ca2+ ([Ca2+]i) signalling in mammalian cells.

RESULTS

Oryzalin inhibited vasopressin, bradykinin and platelet-derived growth factor [Ca2+]i signalling in Swiss 3T3 fibroblasts with IC50 values of 14, 16 and 18 microM, respectively. 45Ca2+ uptake into nonmitochondrial stores of saponin-permeabilized Swiss 3T3 cells was inhibited by oryzalin with an IC50 of 34 microM. Oryzalin inhibited colony formation of HT-29 colon carcinoma cells with an IC50 of 8 microM and inhibited the growth of a number of other cancer cell lines and primary human tumors in vitro with IC50 values in the range 3 to 22 microM. A number of oryzalin analogues were studied and an association was found between the ability to inhibit [Ca2+]i signalling and inhibition of the growth of HT-29 human colon cancer cells (P = 0.001) and of CCRF-CEM human leukemia cells (P = 0.016). Oryzalin at doses up to 600 mg/kg administered orally or subcutaneously daily to mice for 3 to 10 days beginning a day after tumor inoculation inhibited the growth of murine B16 melanoma by 63% but showed no appreciable activity when administered subcutaneously or intraperitoneally to mice beginning a number of days after tumor inoculation against a variety of human tumor xenografts. The peak plasma concentration of oryzalin following repeated subcutaneous administration of oryzalin at 600 mg/kg per day to mice was 37 microM and of its major metabolite N-depropyl oryzalin was 53 microM.

CONCLUSIONS

It is unlikely that the absence of significant antitumor activity of oryzalin is a result of the inability to achieve adequate plasma concentrations.

Objectives: To explore the expression of cytoskeletal and cell proliferation proteins in urothelial cells of patients diagnosed with various clinical subtypes of interstitial cystitis/bladder pain syndrome.

<strong class="sub-title"> Methods: </strong> Biopsy specimens from 85 interstitial cystitis/bladder pain syndrome patients were classified according to findings on cystoscopy. Cytokeratins and cell proliferation proteins detected in the specimens were evaluated with immunofluorescence staining and quantified with western blotting. A total of <em>22</em> patients diagnosed with pure stress urinary incontinence were enrolled as controls.

Results: Interstitial cystitis/bladder pain syndrome patients with Hunner's lesion and with grade 3 glomerulation hemorrhage had smaller bladder capacities than the other interstitial cystitis/bladder pain syndrome patients without Hunner's lesion. Diminished expression of CK14, CK20, cell proliferation protein tumor protein 63, sonic hedgehog, and fibroblast growth factor receptors 3 and 4, and increased expression of CK5 and BCL2-associated X protein were observed in biopsy specimens from patients with Hunner's lesion compared with those from patients without Hunner's lesion and controls. In the patients with grade 3 glomerulation hemorrhage, lower expression levels of urothelial CK20, tumor protein 63 and fibroblast growth factor receptor 4, and lower expression of CK5 and BCL2-associated X protein were detected compared with other types of NHIC.

Conclusion: A diminished expression of proliferation proteins tumor protein 63 and the mature urothelium marker CK20, and increased expression of the immature marker CK5 in specimens from both Hunner's lesion and grade 3 glomerulation hemorrhage patients can be observed. The urothelium of patients with interstitial cystitis/bladder pain syndrome might be in a state of persistent or chronic injury that could relate to the limited expression of cell proliferation proteins.

Keywords: cytoskeleton; proliferation; transcription factor; urothelium.

Background/purpose: We examined whether engineered overexpression of fibroblast growth factor-2 (Fgf2) in donor mesenchymal stem cells (MSCs) could enhance spina bifida coverage induced by transamniotic stem cell therapy (TRASCET).

Methods: Pregnant Sprague-Dawley dams (n = 24) exposed to retinoic acid for induction of fetal spina bifida were divided in three groups. An untreated group had no further manipulations. Two groups received volume-matched intra-amniotic injections into all fetuses (n = 157) of either amniotic fluid-derived MSCs (afMSC; n = 85) or afMSCs transduced with an Fgf2 transgene (Fgf2-afMSC; n = 72) on gestational day 17 (term=21-22 days). Defect coverage was categorized at term by histology and pan-cytokeratin immunohistochemistry. Statistical coverage comparisons were by logistic regression.

Results: Among 84 survivors with isolated spina bifida, 71 had definitive histology. Defect coverage rates in both the afMSC (38.5%) and Fgf2-afMSC (73.3%) groups were statistically significantly higher than in the untreated group (10%; p<0.001 for both). There was a significantly higher coverage rate in the Fgf2-afMSC group compared with the afMSC group (p = 0.025).

Conclusions: Fgf2 overexpression in donor mesenchymal stem cells increases defect coverage rates in a rodent model of transamniotic stem cell therapy for spina bifida. Genetic engineering of donor cells is a promising strategy for the enhancement of this emerging therapy.

Keywords: Amniotic mesenchymal stem cell; Fetal gene therapy; Fgf2; Spina bifida; TRASCET; Transamniotic stem cell therapy; bFGF.

Background: To describe ocular adverse events and retinal changes during fibroblast growth factor receptor (FGFR) inhibitor (AZD4547) anticancer therapy.

Methods: This is a sub-study examining ocular adverse effects from AZD4547 therapy (single-centre, open-label, single arm phase II clinical trial). Comprehensive ocular examinations were performed 3 weekly in 24 patients. Macular optical coherence tomography (OCT) scan (30⁰ ×25⁰ ) was obtained at each visit and OCT parameters (central 1 mm retinal thickness [CRT] and total macular volume in central 6 mm) extracted. OCT scans were subdivided into outer (ELM to RPE) and inner (ELM to ILM) layers to compare outer and inner retinal changes.

Results: In 24 patients, AZD4547 was associated with eyelash elongation (n=5, 21%) and punctate corneal erosion (n=2, 8%). One patient developed clinically significant posterior capsular opacification during the study. OCT data were available in 23 patients, retinal changes ranged from an asymptomatic increased visibility of the interdigitation zone (IDZ) (n=10, 43%) to multilobular subretinal fluid pockets (n=5, 22%), which was associated with mild visual acuity loss. In a subset of patients (n=9) with pre-AZD4547 dosing OCT baseline, CRT increased by mean (SD) of 9 (4) μm in those with IDZ change only compared to 64 (38) μm in those with other retinal changes. Retinal changes tended to be bilateral, self-limiting and improved over time without medical intervention.

Conclusions: The ocular signs and symptoms did not result in dose cessation. Posteriorly, FGFR inhibition leads to outer retinal changes ranging from increased visibility of IDZ to distinct, multiple fluid pockets.

Keywords: Anticancer; FGFR inhibitor; IDZ; OCT; retinal changes.

Desmoplastic small round cell tumor (DSRCT) is characterized by the EWSR1-WT1 t(11;<em>22</em>) (p13:q12) translocation. Few additional putative drivers have been identified, and research has suffered from a lack of model systems. Next generation sequencing (NGS) data from 68 matched tumor-normal samples, whole-genome sequencing data from 10 samples, transcriptomic and affymetrix array data, and a bank of DSRCT patient-derived xenograft (PDX) are presented. EWSR1-WT1 fusions were noted to be simple, balanced events. Recurrent mutations were uncommon, but were noted in TERT (3%), ARID1A (6%), HRAS (5%), and TP53 (3%), and recurrent loss of heterozygosity (LOH) at 11p, 11q and 16q was identified in 18%, <em>22</em>%, and 34% of samples, respectively. Comparison of tumor-normal matched versus unmatched analysis suggests overcalling of somatic mutations in prior publications of DSRCT NGS data. Alterations in <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 4 (FGFR4) were identified in 5/68 (7%) of tumor samples, while differential overexpression of FGFR4 was confirmed orthogonally using 2 platforms. PDX models harbored the pathognomic EWSR1-WT1 fusion and were highly representative of corresponding tumors. Our analyses confirm DSRCT as a genomically quiet cancer defined by the balanced translocation, t(11;<em>22</em>)(p13:q12), characterized by a paucity of secondary mutations but a significant number of copy number alterations. Against this genomically quiet background, recurrent activating alterations of FGFR4 stood out, and suggest that this receptor tyrosine kinase, also noted to be highly expressed in DSRCT, should be further investigated. Future studies of DSRCT biology and pre-clinical therapeutic strategies should benefit from the PDX models characterized in this study. Implications: These data describe the general quiescence of the desmoplastic small round cell tumor (DSRCT) genome, present the first available bank of DSRCT model systems, and nominate FGFR4 as a key receptor tyrosine kinase in DSRCT, based on high expression, recurrent amplification, and recurrent activating mutations.

<strong class="sub-title"> Background: </strong> MicroRNAs (miRNAs), a class of <em>22</em> nucleotide (nt) non-coding RNAs, negatively regulate mRNA post-transcriptional modification in various biological processes. Initiation of skin hair follicles in cashmere goats is a dynamic process involving many key signalling molecules, but the associated cellular biological mechanisms induced by these key signalling molecules have not been reported.

Results: In this study, differential expression, bioinformatics, and Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on miRNA expression profiles of Inner Mongolian cashmere goats at 45, 55, and 65 days during the foetal period, and chi-miR-370-3p was identified and investigated further. Real-time fluorescence quantification (qRT-PCR), dual luciferase reporting, and western blotting results showed that transforming growth factor beta receptor 2 (TGF-βR2) and fibroblast growth factor receptor 2 (FGFR2) were the target genes of chi-miR-370-3p. Chi-miR-370-3p also regulated the expression of TGF-βR2 and FGFR2 at mRNA and protein levels in epithelial cells and dermal fibroblasts. DNA staining, Cell Counting Kit-8 (CCK8), and fluorescein-labelled Annexin V results showed that chi-miR-370-3p inhibited the proliferation of epithelial cells and fibroblasts, but had no effect on apoptosis. Cell scratch test results showed that chi-miR-370-3p promoted the migration of epithelial cells and fibroblasts.

Conclusion: Chi-miR-370-3p inhibits the proliferation of epithelial cells and fibroblasts by targeting TGF-βR2 and FGFR2, thereby improving cell migration ability, and ultimately regulating the fate of epithelial cells and dermal fibroblasts to develop the placode (PC) and dermal condensate (DC), inducing hair follicle development.

Keywords: cashmere goat; cell migration; cell proliferation; chi-miR-370-3p; hair follicle.

Regenerative endodontic therapy intends to induce the host's natural wound-healing process, which can restore the vitality, immunity, and sensitivity of the inflammatory or necrotic pulp tissue destroyed by infection or trauma. Myriads of <em>growth</em> <em>factors</em> are critical in the processes of pulp repair and regeneration. Among the key regulatory <em>factors</em> are the <em>fibroblast</em> <em>growth</em> <em>factors</em>, which have turned out to be the master regulators of both organogenesis and tissue homeostasis. <em>Fibroblast</em> <em>growth</em> <em>factors</em>, a family composed of <em>22</em> polypeptides, have been used in tissue repair and regeneration settings, in conditions as diverse as burns, ulcers, bone-related diseases, and spinal cord injuries. Meanwhile, in dentistry, the basic <em>fibroblast</em> <em>growth</em> <em>factor</em> is the most frequently investigated. Thereby, the aim of this review is 2-fold: 1) foremost, to explore the underlying mechanisms of the bFGF in dental pulp repair and regeneration and 2) in addition, to shed light on the potential therapeutic strategies of the bFGF in dental pulp-related clinical applications.

Keywords: basic fibroblast growth factor; fibroblast growth factors; pulp regeneration; pulp repair; regenerative endodontic therapy; regenerative endodontics.

Introduction: This study assessed the performance of a new fully automated immunoassay for fibroblast growth factor (FGF) 23 (Determinar CL FGF23 CL) among healthy individuals and those with chronic hypophosphatemia compared with the previous assay (Kainos FGF23 KI).

Materials and methods: A total of 380 serum samples from healthy participants were collected to determine the reference range of FGF23 levels with CL. A total of 200 serum samples from 22 hypophosphatemic patients were collected simultaneously to compare the difference in FGF23 levels between CL and KI. The Mann-Whitney U test and linear regression analysis were adopted to assess the differences and linearity between the two assays.

Results: The median FGF23 levels among healthy individuals was 31.7 (interquartile: 26.4-37.5) pg/mL. When the reference range was calculated as the mean ± 2 standard deviation (2SD), it was 16.1-49.3 pg/mL. A total of 363 individuals (96%) among normal cases fell in this range. Among 200 samples from patients with chronic hypophosphatemic disorder, the median FGF23 levels analyzed by CL and KI were 123.0 (90.2-237.7) and 172.5 (115.8-290.7) pg/mL. KI yielded significantly higher FGF23 values than CL (p < 0.001). A linear regression model revealed the correlation between KI (x) and CL (y), which had a slope of 0.76 with a y-intercept of -0.32 and high linearity (R² = 0.99).

Conclusion: The new measurement kit yielded lower FGF23 values when compared with the previous assay. Clinicians should consider this discrepancy when they assay intact FGF23 values with CL.

Keywords: Fanconi syndrome; Fibroblast growth factor 23; Hypophosphatemia; Tumor-induced osteomalacia; X-linked hypophosphatemic rickets.

Objective: Dravet syndrome (DS) is a severe and intractable form of epilepsy with prolonged seizures which may evolve to other seizure types and associated with mild to severe intellectual disabilities. Fibroblast Growth Factor 21 (FGF-21) is a stress hormone mediating metabolic and oxidative stress and circulating level of FGF-21 had been shown to increase in some patients with impairment of oxidative phosphorylation in muscles. In DS, FGF-21 is of interest for further study as mitochondrial oxidative stress was identified previously in patients.

Methods: Plasma FGF-21 levels were compared between 22 DS patients and 22 normal controls and their clinical characteristics of DS patients at the time of plasma sampling were studied retrospectively. Besides, the relationships of FGF-21 level with intellectual development, seizure frequency, valproate treatment and types of SCN1A mutations were analyzed. Logarithmic transformation of FGF-21 levels was performed before comparison and statistical analysis.

Results: Mean of log₁₀ FGF-21 level was significantly higher in DS patients when comparing with normal controls (p = 0.0042). Mean of log₁₀ FGF-21 level was significantly higher in DS patients with normal to mild ID versus mild to severe ID (p = 0.0193) and with valproate treatment versus without valproate treatment (p = 0.015). No significant difference was shown in FGF-21 level in DS patients with missense versus truncating SCN1A variants and no correlation could be demonstrated between seizure frequency and FGF-21 level.

Significance: Significantly higher level of plasma FGF-21 was identified in DS patients. The high FGF-21 levels were shown to be associated with developmental outcome and valproate treatment. These results support further investigation on the relationship of FGF-21 with the clinical outcomes of DS and other related mechanism which is important for possible therapeutic development for this epileptic encephalopathy.

Keywords: Dravet Syndrome; FGF-21; epileptic encephalopathy; fibroblast growth factor 21; mitochondrial oxidative phosphorylation; valproate.