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.

Background: Umbilical cord (UC) connective tissues contain plastic-adherent, colony forming unit-fibroblasts (CFU-Fs) amenable to culture expansion for potential therapeutic use. Recently, UC-derived allograft products have been made available to practitioners in orthopaedics and other specialties, by companies purporting "stem cell"-based healing. However, such marketing claims conflict with existing regulations for these human tissues, generating questions over the cellular and protein composition of current commercially available UC allograft products.

Purpose: To evaluate commercial UC allograft products for viable cells, CFU-Fs, and protein makeup.

Study design: Descriptive laboratory study.

Methods: Five commercial UC allograft products claiming to contain viable, undescribed "stem cells," 2 obtained from UC blood (UCB) and 3 from UC tissue (UCT), were analyzed. Image-based methods were used to measure cell concentration and viability, a traditional CFU-F assay was used to evaluate in vitro behavior indicative of a connective tissue progenitor cell phenotype often referred to as mesenchymal stem/stromal cells, and quantitative immunoassay arrays were used to measure a combination of cytokines and growth factors. Bone marrow concentrate (BMC) and plasma derived from the blood and bone marrow of middle-aged individuals served as comparative controls for cell culture and protein analyses, respectively.

Results: Viable cells were identified within all 5 UC allograft products, with those derived from UCB having greater percentages of living cells (40%-59%) than those from UCT (1%-22%). Compared with autologous BMC (>95% viability and >300 million living cells), no CFU-Fs were observed within any UC allograft product (<15 million living cells). Moreover, a substantial number of proteins, particularly those within UCB allograft products, were undetectable or present at lower concentrations compared with blood and bone marrow plasma controls. Interestingly, several important growth factors and cytokines, including basic fibroblast growth factor, hepatocyte growth factor, interleukin-1 receptor antagonist, and osteoprotegerin, were most prevalent in 1 or more UCT allograft products as compared with blood and bone marrow plasma.

Conclusion: CFU-Fs, often referred to as stem cells, were not found within any of the commercial UC allograft products analyzed, and clinicians should remain wary of marketing claims stating otherwise.

Clinical relevance: Any therapeutic benefit of current UC allograft products in orthopaedic medicine is more likely to be attributed to their protein composition (UCT > UCB) or inclusion of cells without colony forming potential (UCB > UCT).

Keywords: bone marrow concentrate (BMC); colony forming unit–fibroblast (CFU-F); growth factors; platelet-rich plasma (PRP); stem cells; umbilical cord allograft product.

Background: Phosphorus is essential for bone mineralization in broilers, however, the underlying mechanisms remain unclear. We aimed to investigate whether bone phosphorus retention and bone development might be regulated by related hormones and local bone-derived regulators in broilers.

<strong class="sub-title"> Methods: </strong> Broilers were fed diets containing different levels of non-phytate phosphorus (NPP) 0.15%, 0.25%, 0.35%, 0.45% and 0.55% or 0.15%, 0.<em>22</em>%, 0.29%, 0.36% and 0.43% from 1 to 21 or <em>22</em> to 42 days of age. Serum and tibia samples were collected for determinations of bone phosphorus retention and bone development parameters, related hormones and local bone-derived regulators of broiler chickens on d 14, 28 and 42, respectively.

Results: Tibia ash phosphorus, total phosphorus accumulation in tibia ash (TP_TA), bone mineral concentration (BMC), bone mineral density (BMD), bone breaking strength (BBS), and ash on d 14, 28 or 42, serum 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) on d 28 and 42, mRNA expressions of tibia fibroblast growth factor 23 (FGF23) and dentin matrix protein 1 (DMP1) on d 14 and 28 increased linearly or quadratically (P < 0.05), while serum parathyroid hormone (PTH) on d 28, tibia alkaline phosphatase (ALP) on d 14, 28 and 42, bone gal protein (BGP) on d 14, and mRNA expression of tibia phosphate-regulating gene with homologies to endopeptidases on the X chromosome (PHEX) on d 14 and 28 decreased linearly or quadratically (P < 0.04) as dietary NPP level increased. TP_TA, BMC, BMD, and ash on d 28 and 42, BBS on d 28, and ash phosphorus on d 42 were positively correlated (r = 0.389 to 0.486, P < 0.03) with serum 1,25(OH)₂D₃. All of the above parameters were positively correlated (r = 0.380 to 0.689, P < 0.05) with tibia DMP1 mRNA expression on d 14, 28 and 42, but negatively correlated (r = - 0.609 to - 0.538, P < 0.02) with serum PTH on d 28, tibia ALP on d 14, 28 and 42, and BGP on d 14. TP_TA, BMC and ash on d 14 and BMD on d 28 were negatively correlated (r = - 0.397 to - 0.362, P < 0.03) with tibia PHEX mRNA expression, and BMD on d 28 was positively correlated (r = 0.384, P = 0.04) with tibia FGF23 mRNA expression.

Conclusions: These results suggested that bone phosphorus retention and bone development parameters had moderate to strong correlations with serum PTH and 1,25(OH)₂D₃ and tibia DMP1, PHEX, FGF23, ALP and BGP in broilers during the whole growth period, and thus they might be partly regulated by these related hormones and local bone-derived regulators.

Keywords: Bone; Broiler chicken; Hormone; Local bone-derived regulator; Phosphorus.

<em>Fibroblast</em> <em>growth</em> <em>factors</em> (FGFs) are a large family of secretory molecules that act through tyrosine kinase receptors known as FGF receptors. They play crucial roles in a wide variety of cellular functions, including cell proliferation, survival, metabolism, morphogenesis, and differentiation, as well as in tissue repair and regeneration. The signaling pathways regulated by FGFs include RAS/mitogen-activated protein kinase (MAPK), phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)-protein kinase B (AKT), phospholipase C gamma (PLCγ), and signal transducer and activator of transcription (STAT). To date, <em>22</em> FGFs have been discovered, involved in different functions in the body. Several FGFs directly or indirectly interfere with repair during tissue regeneration, in addition to their critical functions in the maintenance of pluripotency and dedifferentiation of stem cells. In this review, we summarize the roles of FGFs in diverse cellular processes and shed light on the importance of FGF signaling in mechanisms of tissue repair and regeneration.

Keywords: FGF; FGFR; cell differentiation; cell proliferation; tissue regeneration; tissue repair.

Cellular signaling proteins maintain the basic activities of cell and communication, between the cells for normal <em>growth</em> and development and pathological situation as well. <em>Fibroblast</em> <em>growth</em> <em>factor</em> (FGF) and <em>fibroblast</em> <em>growth</em> <em>factor</em> receptors (FGFRs) have a comparatively huge part to play in the cellular communication processes. Human FGF has <em>22</em> members, 18 ligands, and 4 tyrosine kinase receptors for binding and is expressed in a wide range of cells. Any alteration in these <em>factors</em> would disrupt their normal function, leading to various abnormalities. The aim of this systematic analysis, is to understand the FGFs, the physiological and pathological role of FGF in oral diseases, and to predict the use of FGF in the predilection toward odontogenic cyst and tumors. This review helps confer the role of FGF in various physiological and pathological aspects in systemic diseases and analyzes its role in diagnosis and prognosis of odontogenic cysts and tumors.

Keywords: Fibroblast growth factor; mutations; odontogenic tumors and cyst; receptor signaling.

A <em>growing</em> number of public health bodies, regulators and governments around the world consider electronic vapor products a lower risk alternative to conventional cigarettes. Of critical importance are rapid new approach methodologies to enable the screening of next generation products (NGPs) also known as next generation tobacco and nicotine products. In this study, the activity of conventional cigarette (3R4F) smoke and a range of NGP aerosols (heated tobacco product, hybrid product and electronic vapor product) captured in phosphate buffered saline, were screened by exposing a panel of human cell-based model systems using Biologically Multiplexed Activity Profiling (BioMAP® Diversity PLUS® Panel, Eurofins Discovery). Following exposure, the biological activity for a wide range of biomarkers in the BioMAP panel were compared to determine the presence of toxicity signatures that are associated with specific clinical findings. NGP aerosols were found to be weakly active in the BioMAP Diversity PLUS Panel (≤3/148 biomarkers) whereas significant activity was observed for 3R4F (<em>22</em>/148 biomarkers). Toxicity associated biomarker signatures for 3R4F included immunosuppression, skin irritation and thrombosis, with no toxicity signatures seen for the NGPs. BioMAP profiling could effectively be used to differentiate between complex mixtures of cigarette smoke or NGP aerosol extracts in a panel of human primary cell-based assays. Clinical validation of these results will be critical for confirming the utility of BioMAP for screening NGPs for potential adverse human effects.

Keywords: ACM, aerosol collected mass; AhR, Aryl hydrocarbon receptor; Alternative methods; COPD, Chronic obstructive pulmonary disease; EGFR, epidermal growth factor receptor; ELISA, enzyme-linked immunosorbent assay; EVP, Electronic vapor product; HDFn, Human neonatal dermal fibroblasts; HTP, Heated Tobacco Product; HUVEC, Human umbilical vein endothelial cells; HYB, Hybrid product containing e-liquid drawn through a tobacco plug; IL, interleukin; ISO, International Organization for Standardization; In vitro assays; MOA, Mechanism of action; M−CSF, Macrophage colony-stimulating factor; NGP, Next generation product; NRC, National Research Council; NRF2, Nuclear factor erythroid 2-related factor 2; Next generation products; PBMC, Peripheral blood mononuclear cells; PBS, Phosphate buffered saline; Panel; Phenotypic screening; SRB, Sulforhodamine B; TCR, T cell receptor; TF, Tissue factor; TLR, toll-like receptor; TNFα, tumor necrosis factor alpha; TPM, Total particulate matter; Toxicity signature; bPBS, Bubbled phosphate buffered saline; mTOR, mechanistic target of rapamycin.

Objective: This retrospective radiographic controlled study investigates the dental phenotype in patients with Crouzon syndrome to determine if differences are observed as suggested by the FGFR2^C342Y/+ Crouzon mouse models, and whether these models could be of interest to study the role of this mutation in tooth development.

<strong class="sub-title"> Design: </strong> We assessed dental phenotype using dedicated linear measurements in <em>22</em> children with Crouzon syndrome and compared tooth morphology in both primary and permanent dentitions to an age-matched control group. Descriptive statistics were performed with "Sex" and "Age" as covariates for the permanent tooth models and "Sex" only for the primary tooth models, to take into account potential confounding <em>factors</em>.

Results: We showed that permanent but not primary tooth dimensions were globally reduced in Crouzon syndrome, without microdontia. In permanent dentition, crown height, mesiodistal and faciolingual cervical diameters were reduced by 6.3%, 5.7% and 5.5% respectively (p < 0.05).

Conclusion: Our results underline the implication of Fibroblast Growth Factor Receptor 2 (FGFR2) in dental development of humans and contribute to support FGFR2^C342Y/+ Crouzon mouse models as partial replicas of this condition, including in the oral region.

Keywords: Craniofacial dysostosis; Crouzon syndrome; Dental morphology; Morphometrics; Receptor Fibroblast Growth Factor, type 2; Tooth phenotype.

Background and aims: Protein profiling in patients with inflammatory bowel diseases (IBD) for diagnostic and therapeutic purposes is underexplored in IBD. This study analysed the association between phenotype, genotype and the plasma proteome in IBD.

Methods: Ninety-two (92) inflammation-related proteins were quantified in plasma of 1,028 patients with IBD (567 Crohn's disease [CD]; 461 ulcerative colitis [UC]) and 148 healthy individuals to assess protein-phenotype associations. Corresponding whole-exome sequencing and global screening array data of 919 patients with IBD were included to analyse the effect of genetics on protein levels (protein quantitative trait loci (pQTL) analysis). Intestinal mucosal RNA sequencing and fecal metagenomic data were used for complementary analyses.

<strong class="sub-title"> Results: </strong> Thirty-two (32) proteins were differentially abundant between IBD and healthy individuals, of which <em>22</em> proteins independent of active inflammation. Sixty-nine (69) proteins were associated with 15 demographic and clinical <em>factors</em>. <em>Fibroblast</em> <em>growth</em> <em>factor</em>-19 levels were decreased in CD patients with ileal disease or a history of ileocecal resection. Thirteen novel cis-pQTLs were identified and 10 replicated from previous studies. One trans-pQTL of the fucosyltransferase 2 (FUT2) gene (rs602662) and two independent cis-pQTLs of C-C motif chemokine 25 (CCL25) affected plasma CCL25 levels. Intestinal gene expression data revealed an overlapping cis-expression (e)QTL-variant (rs3745387) of the CCL25 gene. The FUT2 rs602662 trans-pQTL was associated with reduced abundances of fecal butyrate-producing bacteria.

Conclusions: This study shows that genotype and multiple disease phenotypes strongly associate with the plasma inflammatory proteome in IBD and identifies disease-associated pathways that may help to improve disease management in the future.

Keywords: Genetics; Inflammatory Bowel Disease; Proteomics.

<b>Objective:</b> The 8p11 myeloproliferative syndrome [EMS] is a rare myeloproliferative disorder which usually develops rapidly with chromosomal translocation of the <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 gene. The gene has 15 fusion partners, including the breakpoint cluster region (BCR) gene on chromosome <em>22</em>. Of all the tests available, chromosome karyotype determination is the most important for the diagnosis of EMS. Here, we describe one case of a patient characterized by marked increase of white blood cells and thrombocytopenia and diagnosed as EMS with t(8;<em>22</em>)(p11;q11) by chromosome karyotype.<b>Methods:</b> 28-year-old man was referred to our hospital. He had a onemonth history of intermittent coughing and a small amount of expectoration after catching a cold. As an outpatient, his complete blood count showed: WBC was 130.04 × 10⁹/L with 80.20% granulocytes.Hematologic investigations, bone marrow analysis and genomic DNA sequencing studies were performed.<b>Results:</b> Despite additional chromosomal abnormalities,the patient progressed rapidly with a B blast cell clone in one month. After diagnosis inthree months, the patient underwent the haplo-identical BMT of his brother, followed up for three years, and had a high rate of survival.<b>Conclusions:</b> Our report provides a definite conceptual framework for a better understanding of the characteristics of The 8p11 myeloproliferative syndrome [EMS].

Keywords: BCR/FGFR1; cell transplantation; myeloproliferative.

In rodents, exercise alters the plasma concentration of exerkines that regulate white adipose tissue (WAT) browning or brown adipose tissue (BAT) metabolism. This study aims to analyse the acute and chronic effect of exercise on the circulating concentrations of 16 of these exerkines in humans. Ten young sedentary adults (6 female) performed a maximum walking effort test and a resistance exercise session. The plasma concentration of 16 exerkines was assessed before, and 3, 30, 60, and 120 minutes after exercise. Those exerkines modified by exercise were additionally measured in another 28 subjects (<em>22</em> women). We also measured the plasma concentrations of the exerkines before and after a 24-week exercise program (endurance + resistance; 3-groups: control, moderate-intensity and vigorous-intensity) in 110 subjects (75 women). Endurance exercise acutely increased the plasma concentration of lactate, norepinephrine, brain-derived neurotrophic <em>factor</em>, interleukin 6, and follistatin-like protein 1 (3 minutes after exercise), and musclin and <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (30 and 60 minutes after exercise), decreasing the plasma concentration of leptin (30 minutes after exercise). Adiponectin, atrial natriuretic peptide (ANP), β-aminoisobutyric acid, meteorin-like, follistatin, pro-ANP, irisin and myostatin were not modified or not detectable. The resistance exercise session increased the plasma concentration of lactate 3 minutes after exercise. Chronic exercise did not alter the plasma concentration of these exerkines. In sedentary young adults, acute endurance exercise releases to the bloodstream exerkines that regulate BAT metabolism and WAT browning. In contrast, neither a low-volume resistance exercise session nor a 24-week training program modified plasma levels of these molecules.<b>Trial registration:</b> ClinicalTrials.gov identifier: <a href="http://clinicaltrials.gov/show/NCT02365129" title="See in ClinicalTrials.gov">NCT02365129</a>..

Keywords: beige fat; brown fat; exerkines; thermogenesis.

We present a case of tumor-induced osteomalacia (TIO) in a young woman of <em>22</em> years. The <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 transmitting tumor in her left foot remained undetected for several years. She suffered several fractures including insufficiency fractures of both femoral necks requiring bilateral proximal femoral nailing. After phosphaturia was diagnosed any known genetic etiology was excluded. Even advanced imaging modalities were unable to detect the clinically silent tumor until an ⁶⁸Ga-DOTA-TOC-PET/CT-scan revealed a mass with paraneoplastic activity in the left foot. Complete resection of the tumor proved to cure her condition after 9 years of uncertainty and suffering. Serum phosphate levels returned to normal within days. After presentation of the case report, the current literature on published cases of TIO between 1956 and 2021 is summarized to emphasize the importance of an accurate and early diagnosis. Our case report aims to illustrate that a long latency period of diagnosis may be avoided utilizing the latest imaging techniques to spare affected patients from long treatment of symptoms instead of finding the underlying cause.

Keywords: 68Ga-DOTA-TOC-PET/CT-scan; 68Ga-DOTA-TOC-PET/CT-scan, 8Ga-DOTA(0)-Phe(1)-Tyr(3)-octreotide positron emission tomography/computed tomography; FDG-PET, Fluorodeoxyglucose positron emission tomography; FGF 23; FGF, fibroblast growth factor; PTH, parathyroid hormone; Phosphaturic tumor; TIO, tumor-induced osteomalacia; TIR, tumor-induced hypophosphatemic rickets; Tumor-induced osteomalacia.

Background: Dietary protein and phosphorus (P) restriction is the mainstay for nutritional management of chronic kidney disease (CKD). However, adequate restriction levels for cats with early CKD remain unclear.

Objectives: To investigate responses in cats with early CKD to varying dietary protein, P, and calcium (Ca) : P ratio.

Animals: Nineteen research colony cats with International Renal Interest Society stages 1-2 CKD.

<strong class="sub-title"> Methods: </strong> In an opportunistic longitudinal case study, cats were fed a low protein (59 g/Mcal), low P (0.84 g/Mcal) dry diet (LP-LP; Ca : P = 1.9) for 18 months and later transitioned onto a moderate protein (76-98 g/Mcal), moderate P (1.4-1.6 g/Mcal) dry-wet diet regimen (MP-MP; Ca : P = 1.4-1.6) for <em>22</em> months. Fold-changes in serum creatinine, total Ca (tCa) and P (primary outcomes) and <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF23) were assessed by linear-mixed models.

<strong class="sub-title"> Results: </strong> While feeding LP-LP, mean serum creatinine decreased (0.87-fold, 95% confidence interval [CI] 0.81, 0.93, P < .001) to within reference range after 6 months, while increases in total Ca (tCa; 1.16-fold, 95% CI 1.11, 1.<em>22</em>, P < .001) and FGF23 (2.72-fold, 95% CI 1.72, 4.31, P < .001), but not in P (1.03-fold, 95% CI 0.945, 1.124, P = .94), were observed after 17 months. On MP-MP, mean creatinine, tCa and P remained within reference ranges and did not significantly change (P = .11, P = .98, and P = 1, respectively), while FGF23 significantly decreased (0.58-fold, 95% CI 0.36, 0.95, P = .02) after <em>22</em> months.

Conclusions and clinical importance: Cats with early CKD developed hypercalcemia after long-term feeding of a highly P-restricted diet. Increasing dietary P and reducing Ca : P ratio maintained renal markers, while improving Ca-P balance. Cats with early CKD could benefit from moderately protein- and P-restricted diets.

Keywords: FGF23; IRIS 1-2; feline CKD; hypercalcemia; phosphate.

Commercially available cultured epithelial keratinocyte sheets (KSs) have played an essential role in wound healing over the last four decades. Despite the initial uptake by the dermal elements, the survival rate of KS on the dermis-like tissue generated by conventional artificial dermis (AD) is low, making this method unsuitable for standard treatments. Therefore, an innovative AD such as collagen/gelatin sponge (CGS) that maintains the release of human recombinant basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) may promote wound healing. In this study, we examined whether combination therapy with KSs and CGS with bFGF (bFGF-CGS) could enhance KS survival by heterologous grafting by transplantation of human-derived KSs in an athymic nude rat wound model of staged skin reconstruction. The CGSs were implanted into skin defect wounds on athymic nude rats, which were then divided into two experimental groups: the bFGF group (CGSs containing bFGF, n = 8) and control group (CGSs with saline, n = 8). Two weeks after implantation, human epithelial cell-derived KSs were grafted onto the dermis-like tissue, followed by assessment of the survival and morphology at one week later using digital imaging, histology (hematoxylin and eosin and Masson's trichrome staining), immunohistology (von Willebrand <em>factor</em>), immunohistochemistry (cytokeratin 1-5-6, Ki-67), and immunofluorescence (collagen IV, pan-cytokeratins) analyses. The bFGF group showed a significantly higher KS survival area (86 ± 58 vs. 32 ± <em>22</em> mm2; p < 0.05) and increased epidermal thickness (158 ± 66 vs. 86 ± 40 µm; p < 0.05) compared with the control group, along with higher dermis-like tissue regeneration, neovascularization, epidermal maturation, and basement membrane development. These results indicate that the survival rate of KSs in the dermis-like tissue formed by bFGF-CGS was significantly increased. Therefore, combination treatment of bFGF-CGS and KSs shows potential for full-thickness skin defect reconstruction in clinical situations.

Objective: To identify potential biomarkers to distinguish familial Mediterranean fever (FMF) from sepsis.

<strong class="sub-title"> Method: </strong> We recruited 28 patients diagnosed with typical FMF (according to the Tel Hashomer criteria), <em>22</em> patients with sepsis, and 118 age-matched controls. Serum levels of 40 cytokines were analyzed using multi-suspension cytokine array. We performed a cluster analysis of each cytokine in the FMF and sepsis groups in order to identify specific molecular networks. Multivariate classification (random forest analysis) and logistic regression analysis were used to rank the cytokines by importance and determine specific biomarkers for distinguishing FMF from sepsis.

Results: Fifteen of the 40 cytokines were found to be suitable for further analysis. Levels of serum granulocyte-macrophage colony-stimulating factor (GM-CSF), fibroblast growth factor 2, vascular endothelial growth factor, macrophage inflammatory protein-1b, and interleukin-17 were significantly elevated, whereas tumor necrosis factor-α (TNF-α) was significantly lower in patients with FMF compared with those with sepsis. Cytokine clustering patterns differed between the two groups. Multivariate classification followed by logistic regression analysis revealed that measurement of both GM-CSF and TNF-α could distinguish FMF from sepsis with high accuracy (cut-off values for GM-CSF = 8.3 pg/mL; TNF-α = 16.3 pg/mL; sensitivity, 92.9%; specificity, 94.4%; accuracy, 93.4%).

Conclusion: Determination of GM-CSF and TNF-α levels in combination may represent a biomarker for the differential diagnosis of FMF from sepsis, based on measurement of multiple cytokines.

Keywords: Cytokine profile; Familial Mediterranean fever; GM-CSF; TNF-α; sepsis.

Purpose: Radiation-induced pulmonary fibrosis (RIPF) is a major side effect after radiotherapy for thoracic malignancies. However, rare anti-RIPF therapeutics show definitive effect for treating this disease. Ubiquitin-specific peptidase 11 (USP11) has been reported to promote transforming growth factor β (TGFβ) signaling which plays essential role underlying RIPF. Herein, we explored the role of USP11 on RIPF.

Materials and methods: In the present study, USP11-knockout (Usp11^-/- ) mice were used to explore the effects of USP11 on RIPF. The lung tissue was obtained after receiving 30Gy X-ray irradiation. The expression of USP11, TGF-β1, and a-SMA were determined by immunohistochemical and Western Blot, respectively. γ-H₂AX foci and TUNEL positive cells were detected by fluorescent technique to assess DNA damage and apoptosis. High-throughput proteomics analysis was applied to further explore the related mechanisms. The transwell co-culture method was used to investigate bystander effects in HELF cells induced by irradiated HMEC-1 cells in vitro.

Results: Here we found that radiation activated USP11 in vivo and in vitro. Our results showed that USP11 deficiency effectively decreased serum TGF-β1 level, suppressed α-SMA expression and mitigated pulmonary fibrosis. In addition, fewer γ-H₂AX foci and decreased apoptotic cells were identified after irradiation in the primary cells isolated from the lungs of Usp11^-/- mice. High-throughput proteomics analysis results showed that 22-upregulated and 158-downregulated proteins were identified in the lung tissues of Usp11^-/- mice after irradiation. Furthermore, gene set enrichment analysis (GSEA) revealed that USP11 deficiency affect tight junction signaling pathway.

Conclusions: We verified that USP11 deficiency remarkably reinforced tight junction in the endothelial cells and alleviated TGF-β1 to inhibit fibrosis of fibroblast cells. The present study preliminarily showed that USP11-knockout mitigated RIPF via reinforcement endothelial barrier function.

Keywords: Proteomics; RIPF; TGF-β1; USP11; tight junction.

Background: Cerebral palsy (CP) associates cerebral function damages with strong locomotor defects and premature sarcopenia. We previously showed that fibroblast growth factor 19 (FGF19) exerts hypertrophic effects on skeletal muscle and improves muscle mass and strength in mouse models with muscle atrophy. Facing the lack of therapeutics to treat locomotor dysfunctions in CP, we investigated whether FGF19 treatment could have beneficial effects in an experimental rat model of CP.

Methods: Cerebral palsy was induced in male Wistar rat pups by perinatal anoxia immediately after birth and by sensorimotor restriction of hind paws maintained until Day 28. Daily subcutaneous injections with recombinant human FGF19 (0.1 mg/kg bw) were performed from Days 22 to 28. Locomotor activity and muscle strength were assessed before and after FGF19 treatment. At Day 29, motor coordination on rotarod and various musculoskeletal parameters (weight of tibia bone and of soleus and extensor digitorum longus (EDL) muscles; area of skeletal muscle fibres) were evaluated. In addition, expression of specific genes linked to human CP was measured in rat skeletal muscles.

Results: Compared to controls, CP rats had reduced locomotion activity (-37.8% of distance travelled, P < 0.05), motor coordination (-88.9% latency of falls on rotarod, P < 0.05) and muscle strength (-25.1%, P < 0.05). These defects were associated with reduction in soleus (-51.5%, P < 0.05) and EDL (-42.5%, P < 0.05) weight, smaller area of muscle fibres, and with lower tibia weight (-38%, P < 0.05). In muscles from rats submitted to CP, changes in the expression levels of several genes related to muscle development and neuromuscular junctions were similar to those found in wrist muscle of children with CP (increased mRNA levels of Igfbp5, Kcnn3, Gdf8, and MyH4 and decreased expression of Myog, Ucp2 and Lpl). Compared with vehicle-treated CP rats, FGF19 administration improved locomotor activity (+53.2%, P < 0.05) and muscle strength (+25.7%, P < 0.05), and increased tibia weight (+13.8%, P < 0.05) and soleus and EDL muscle weight (+28.6% and +27.3%, respectively, P < 0.05). In addition, it reduced a number of very small fibres in both muscles (P < 0.05). Finally, gene expression analyses revealed that FGF19 might counteract the immature state of skeletal muscles induced by CP.

Conclusions: These results demonstrate that pharmacological intervention with recombinant FGF19 could restore musculoskeletal and locomotor dysfunction in an experimental CP model, suggesting that FGF19 may represent a potential therapeutic strategy to combat the locomotor disorders associated with CP.

Keywords: Cerebral palsy; Fibroblast growth factor 19; Sarcopenia; Skeletal muscle.

Endurance exercise induces an increase in the expression of exercise-induced peptides that participate in the repair and regeneration of skeletal muscles. The present study aimed to evaluate the time course and role of exercise-induced cytokines in muscle damage and repair after a marathon race. Fifty-seven Brazilian male amateur marathon finishers, aged 30-55 years, participated in this study. The blood samples were collected 24 h before, immediately after, and 24 and 72 h after the São Paulo International Marathon. The leukogram and muscle damage markers were analyzed using routine automated methodology in the clinical laboratory. The plasma levels of the exercise-induced cytokines were determined using the Human Magnetic Bead Panel or enzyme-linked immunosorbent assays [decorin and <em>growth</em> differentiation <em>factor</em> 15 (GDF-15)]. A muscle damage was characterized by an increase in plasma myocellular proteins and immune changes (leukocytosis and neutrophilia). Running the marathon increased interleukin (IL)-6 (4-fold), IL-8 (1.5-fold), monocyte chemoattractant protein-1 (2.4-fold), tumor necrosis <em>factor</em> alpha (TNF-α) (1.5-fold), IL-10 (11-fold), decorin (1.9-fold), GDF-15 (1.8-fold), brain-derived neurotrophic <em>factor</em> (BDNF) (2.7-fold), follistatin (2-fold), and <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF-21) (3.4-fold) plasma levels. We also observed a reduction in musclin, myostatin, IL-15, and apelin levels immediately after the race (by <em>22</em>-36%), 24 h (by 26-52%), and 72 h after the race (by 25-53%). The changes in BDNF levels were negatively correlated with the variations in troponin levels (<i>r</i> = -0.36). The variations in IL-6 concentrations were correlated with the changes in follistatin (r = 0.33) and FGF-21 (<i>r</i> = 0.31) levels after the race and with myostatin and irisin levels 72 h after the race. The changes in IL-8 and IL-10 levels had positive correlation with variation in musclin (<i>p</i> < 0.05). Regeneration of exercise-induced muscle damage involves the participation of classical inflammatory mediators, as well as GDF-15, BDNF, follistatin, decorin, and FGF-21, whose functions include myogenesis, mytophagia, satellite cell activation, and downregulation of protein degradation. The skeletal muscle damage markers were not associated to myokines response. However, BDNF had a negative correlation with a myocardial damage marker. The classical anti-inflammatory mediators (IL-10, IL-8, and IL-6) induced by exercise are associated to myokines response immediately after the race and in the recovery period and may affect the dynamics of muscle tissue repair.

Keywords: endurance exercise; inflammation; muscle damage; muscle repair; myocardial damage; myokines.