METHODS

A high proportion of tuberculosis (TB) patients experience delayed wound healing after surgery, and the specific reasons for this phenomenon are not yet clear.

OBJECTIVE

To analyse cytokine and growth factors at surgical sites to determine their contribution to delayed wound healing in patients with cervical lymph node TB (CLNT).

METHODS

We measured levels of interleukin (IL) 1β, IL-6, IL-10, IL-22, epidermal growth factor, fibroblast growth factor-2, interferon-gamma, tumour necrosis factor alpha and vascular endothelial growth factor A (VEGF-A) in cervical wound drainage fluid from 36 CLNT patients who underwent lymphadenectomy and in 24 thyroidectomy patients.

RESULTS

Wound drainage fluid from CLNT patients showed higher IL-6 (P = 0.007) and VEGF-A (P < 0.001) levels than control thyroidectomy patients (P < 0.05). CLNT patients with a delayed healing time (>5 days; n = 18) showed higher IL-6 (P = 0.002) and VEGF-A (P < 0.001) levels in wound drainage fluid than CLNT patients with normal healing times (5 days; n = 18). IL-6 (OR 11.280, 95%CI 1.413-90.028; P = 0.022) and VEGF-A (OR 13.510, 95%CI 2.168-84.182; P = 0.005) can independently and significantly predict wound healing time in CLNT patients.

CONCLUSIONS

These findings demonstrate that high IL-6 and VEGF-A levels in the post-operative wound fluid of CLNT patients correlate with delayed wound healing.

We describe a presentation of Autosomal Dominant Hypophosphatemic Rickets (ADHR) in a <em>22</em>-year-old female with normal pubertal <em>growth</em> and development and a negative family history in first-degree relatives. The patient presented with a 2-year history of upper and lower extremity proximal muscle pain and weakness and bilateral femoral neck and pubic bone insufficiency fractures. She had a normal serum calcium but a low phosphate as well as 25-hydroxyvitamin D (25(OH)D) levels leading initially to a diagnosis of osteomalacia. Urine phosphate reabsorption was low confirming a phosphate wasting disorder. She had an elevated <em>Fibroblast</em> <em>Growth</em> <em>Factor</em> 23 (FGF23) level. After Tumor-Induced Osteomalacia was ruled out by extensive imaging, she was sent for genetic testing for hereditary rickets which showed a previously reported missense variant in <i>FGF23</i>. Subsequently, she found out that her father's maternal aunt and grandfather had 'bone disorder' and were wheelchair-bound in adulthood. After replenishment of vitamin D, treatment with calcitriol and phosphate leads to complete resolution of patient's symptoms and laboratory abnormalities.

Tenascin is an extracellular matrix glycoprotein which plays a role in cell attachment, proliferation and migration. To elucidate the function of tenascin in the proliferation of endometrial carcinoma, we studied tenascin expression in the endometrial carcinoma of 36 cases. In <em>22</em> of the carcinomas, tenascin expression was intense in the entire extracellular space, especially at the front of muscle invasion. Furthermore, in cases with metastases, deep invasion into muscles and vascular invasion, the rate of tenascin expression was significantly high. Immunoelectron microscopy revealed the tenascin reaction product in the stroma around <em>fibroblasts</em> located some distance from the basal lamina of cancer cells. On the other hand, tenascin expression was found in a high proportion of cases showing weak or no expression of estrogen receptor, and intense expression of transforming <em>growth</em> <em>factor</em>-beta 1. These results suggest that tenascin not only promotes cell proliferation and invasion but also inhibits further proliferation of carcinoma.

<em>Fibroblast</em> <em>growth</em> <em>factor</em> receptor 3 (FGFR3) protein is aberrantly expressed in approximately 15% of cases of plasma cell myeloma as a result of t(4;14)(p16.3;q32), and FGFR3 expression in myeloma is associated with an adverse prognosis. Novel, recently developed therapeutic agents that target the FGFR3 pathway are currently in clinical trials for myeloma. Although extensively studied in plasma cell neoplasms, there is little information in the literature regarding FGFR3 expression in malignant lymphomas, and it is unclear whether lymphoma patients may also benefit from anti-FGFR3 therapy. We, therefore, examined the expression of FGFR3 by immunohistochemistry in 70 cases of malignant lymphoma using tissue microarrays and whole paraffin sections. Of <em>22</em> cases of plasma cell myeloma analyzed as controls, FGFR3 positivity was seen in 3/5 t(4;14) positive cases (60%) and in none of 17 t(4;14) negative cases (P=0.006). Weak-to-moderate cytoplasmic FGFR3 staining was seen in 6/43 (12%) B-cell non-Hodgkin lymphomas and 2/17 (12%) T-cell lymphomas, with staining observed at low incidence in many histologic types. Cytoplasmic staining was also seen infrequently in classic Hodgkin lymphoma (1/10, 10%). Membrane staining was not observed in any case of malignant lymphoma. These results indicate that aberrant FGFR3 staining may be seen infrequently in many forms of malignant lymphoma, although dysregulation of this pathway does not seem to contribute to lymphomagenesis in the majority of cases. Moreover, these findings suggest at least most patients with malignant lymphomas are unlikely to benefit from novel agents targeting the FGFR3 pathway.

Bile acid transporters maintain bile acid homeostasis. Little is known about the functions of some transporters in cholestasis or their regulatory mechanism. We investigated the hepatic expression of solute carrier organic anion transporter family member 3A1 (SLCO3A1, also called OATP3A1) and assessed its functions during development of cholestasis.

We measured levels of OATP3A1 protein and messenger RNA and localized the protein in liver tissues from <em>22</em> patients with cholestasis and 21 patients without cholestasis, using real-time quantitative polymerase chain reaction, immunoblot, and immunofluorescence analyses. We performed experiments with Slco3a1-knockout and C57BL/6J (control) mice. Mice and Sprague-Dawley rats underwent bile duct ligation (BDL) or a sham operation. Some mice were placed on a 1% cholic acid (CA) diet to induce cholestasis or on a control diet. Serum and liver tissues were collected and analyzed; hepatic levels of bile acids and 7-α-C4 were measured using liquid chromatography/mass spectrometry. Human primary hepatocytes and hepatoma (PLC/PRF/5) cell lines were used to study mechanisms that regulate OATP3A1 expression and transport.

Hepatic levels of OATP3A1 messenger RNA and protein were significantly increased in liver tissues from patients with cholestasis and from rodents with BDL or 1% CA diet-induced cholestasis. Levels of fibroblast growth factor 19 (FGF19, FGF15 in rodents) were also increased in liver tissues from patients and rodents with cholestasis. FGF19 signaling activated the Sp1 transcription factor and nuclear factor κB to increase expression of OATP3A1 in hepatocytes; we found binding sites for these factors in the SLCO3A1 promoter. Slco3a1-knockout mice had shorter survival times and increased hepatic levels of bile acid, and they developed more liver injury after the 1% CA diet or BDL than control mice. In hepatoma cell lines, we found OATP3A1 to take prostaglandin E2 and thyroxine into cells and efflux bile acids.

We found levels of OATP3A1 to be increased in cholestatic liver tissues from patients and rodents compared with healthy liver tissues. We show that OATP3A1 functions as a bile acid efflux transporter that is up-regulated as an adaptive response to cholestasis.

Background: We sought to identify biomarkers that indicate low turnover on bone histomorphometry in chronic kidney disease (CKD) patients, and subsequently determined whether this panel identified differential risk for fractures in community-dwelling older adults.

Methods: Among CKD patients who underwent iliac crest bone biopsies and histomorphometry, we evaluated candidate biomarkers to differentiate low turnover from other bone disease. We applied this biomarker panel to 641 participants in the Health Aging and Body Composition Study (Health ABC) study with estimated glomerular filtration rate (eGFR) less than 60 mL/min/1.73 m2 who were followed for fracture. Cox proportional hazards models evaluated the association of bone mineral density (BMD) with fracture risk and determined whether biomarker-defined low bone turnover modified fracture risk at any level of BMD.

<strong class="sub-title">Results:</strong> In 39 CKD patients age 64 ± 13 years, 85% female, with mean eGFR 37 ± 14 mL/min/1.73 m2 who underwent bone biopsy, lower <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF)-23, higher ɑ-Klotho, and lower parathyroid hormone (PTH) indicated low bone turnover in accordance with bone histomorphometry parameters (individual area under the curve = 0.62, 0.73, and 0.55 respectively; sensitivity = <em>22</em>%, specificity = 100%). In Health ABC, 641 participants with CKD were age 75 ± 3 years , 49% female, with mean eGFR 48 ± 10 mL/min/1.73 m2. For every SD lower hip BMD at baseline, there was an 8-fold higher fracture risk in individuals with biomarker-defined low turnover (hazard ratio 8.10 [95% CI, 3.40-19.30]) vs a 2-fold higher risk in the remaining individuals (hazard ratio 2.28 [95% CI, 1.69-3.08]) (Pinteraction = .082).

Conclusions: In CKD patients who underwent bone biopsy, lower FGF-23, higher ɑ-Klotho, and lower PTH together had high specificity for identifying low bone turnover. When applied to older individuals with CKD, BMD was more strongly associated with fracture risk in those with biomarker-defined low turnover.

Keywords: bone mineral density; bone turnover; chronic kidney disease; fibroblast growth factor (FGF)-23; fracture; parathyroid hormone; α-Klotho.

<AbstractText>The aims of this study were to measure changes in <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF-23), neutrophil (elastase, lactoferrin)/platelet activation marker (mean platelet volume-to-platelet count ratio [MPR]), and angiogenin according to the stage of chronic kidney disease (CKD), and to evaluate the association of FGF-23, elastase, lactoferrin, MPR, and angiogenin with arterial stiffness using brachial-ankle pulse wave velocity (ba-PWV) in CKD patients.</AbstractText><AbstractText>According to the estimated glomerular filtration rate (eGFR) calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation, the patients were allocated to five groups: (1) normal controls (eGFR ≥90 mL/min/1.73 m2 without pathologic, urine [proteinuria], blood [electrolyte], and imaging abnormalities; n = <em>22</em>); (2) CKD stage 2 (eGFR 60-89 mL/min/1.73 m2; n = 17); (3) CKD stage 3 (eGFR 30-59 mL/min/1.73 m2; n = <em>22</em>); (4) CKD stage 4 (eGFR 15-30 mL/min/1.73 m2; n = 17); and (5) CKD stage 5-hemodialysis (HD) (n = 30). All the patients were free of clinically apparent cardiovascular disease. Serum FGF-23, elastase, lactoferrin, and angiogenin concentrations and the MPR were measured to study the association of the above parameters with the clinical (age, sex, presence of diabetes mellitus, and blood pressure), biochemical (calcium, phosphorus, uric acid, intact parathyroid hormone [PTH], low-density lipoprotein cholesterol, and high-sensitivity C-reactive protein), and ba-PWV values of the CKD patients.</AbstractText><AbstractText>(1) The mean ba-PWV values were 1,497.2 ± 206.4 cm/s in the controls, 1,649.0 ± 247.9 cm/s in the CKD stage 2 group (p < 0.05 vs. controls), 1,655.8 ± 260.3 cm/s in the CKD stage 3 group (p < 0.05 vs. controls), 1,823.0 ± 402.4 cm/s in the CKD stage 4 group (p < 0.05 vs. controls and CKD stages 2 and 3), and 1,905.2 ± 374.1 cm/s in the CKD stage 5-HD group (p < 0.05 vs. controls and CKD stage 2). (2) The mean log10(FGF-23) concentration values were 0.77 ± 0.27, 0.97 ± 0.48, 1.10 ± 0.35 (p < 0.05 vs. controls and CKD stage 2), 1.35 ± 0.48 (p < 0.05 vs. controls and CKD stages 2 and 3), and 2.12 ± 0.82 (p < 0.05 vs. controls and CKD stages 2-4); the mean angiogenin levels were 230.6 ± 70.5 pg/mL, 283.0 ± 53.5 pg/mL (p < 0.05 vs. controls), 347.3 ± 76.9 pg/mL (p < 0.05 vs. controls and CKD stage 2), 445.9 ± 90.6 pg/mL (p < 0.05 vs. controls and CKD stages 2 and 3), and 370.9 ± 142.4 pg/mL (p < 0.05 vs. controls and CKD stages 2 and 3). (3) In the stage 3-4 CKD/HD patients, the mean elastase-to-neutrophil and lactoferrin-to-neutrophil ratios were significantly lower than in the controls and the stage 2 CKD patients. (4) Our multivariate linear regression analyses showed that age, pulse pressure, mean arterial pressure, PTH, and FGF-23 were independently associated with ba-PWV values.</AbstractText><AbstractText>Circulating FGF-23 and angiogenin concentrations gradually increased as CKD advanced, whereas neutrophil activation markers were significantly lower in the stage 3-4 CKD/HD patients than in the controls and stage 2 CKD patients. FGF-23 was weakly associated with ba-PWV values in patients with CKD/HD and no previous cardiovascular disease.</AbstractText>

Background: Dietary supplemental nicotinamide is used to treat hyperphosphatemia in humans. However, the mechanisms of its impact on body phosphorus homeostasis remain unclear.

Objective: This study was to determine effects and molecular mechanisms of 3 dietary nicotinamide concentrations on body phosphorus homeostasis in laying hens.

Methods: Hy-Line Brown layers (total = 21; 40 wk old; body weight: 1,876 ± 24 g) were individually housed (n = 7) and fed a corn-soybean meal-based diet supplemented with nicotinamide at 20 (N20), 140 (N140), and 1000 (N1000) mg/kg for 21 d. Serum phosphorus and fibroblast growth factor 23 (FGF23) concentrations, phosphorus and calcium excretion, and mRNA and/or protein of type II sodium-phosphate co-transporters (NPt2a, NPt2ab) and FGF23 and FGF23 receptors were measured in the intestines, calvaria, kidney, and liver.

Results: Hens in the N1000 group had a 16% lower serum phosphorus concentration and 22% greater phosphorus excretion than those in the N20 or N140 group (P ≤ 0.05). Compared with hens in the N20 group, hens in the N140 and N1000 groups, which did not differ, had 15-21% lower serum FGF23 concentrations, 19-22% greater calcium excretion, 43-56% lower ileum NPT2b protein production, and 1.5- to 1.6-fold greater kidney NPT2a protein production, respectively (all differences at P ≤ 0.05).

Conclusions: Supplementing high concentrations of nicotinamide in diets for laying hens led to accelerated phosphorus and calcium excretions and decreased serum phosphorus and FGF23 concentrations, which were associated with downregulated intestinal NPt2b protein production. Our findings exclude kidney NPt2a protein production as a primary mechanism for the nicotinamide-induced body phosphorus loss.

Keywords: fibroblast growth factor 23; laying hen; nicotinamide; phosphorus excretion; type II sodium-phosphate co-transporter.

OBJECTIVE

In an attempt to improve in vitro embryo production, we investigated the effect of fibroblast growth factor 10 (FGF10) during in vitro maturation on the developmental capacity of bovine oocytes.

METHODS

Cumulus-oocyte complexes (COCs) were aspirated from follicles of 3-8 mm diameter. After selection, the COCs were matured in medium with or without 0.5 ng/mL of FGF10. The effect of FGF10 during in vitro maturation (IVM) on nuclear maturation kinetics and expansion of the cumulus cells was investigated. Oocyte competence was assessed by the production and development speed of embryos and the relative expression of genes associated with embryo quality.

RESULTS

FGF10 delayed the resumption of meiosis from 8 h onwards, but did not affect the percentage of oocytes reaching metaphase II, nor did it increase cumulus expansion at 22 h of maturation. We found no difference between treatments regarding embryo production, developmental speed, and gene expression.

CONCLUSIONS

In conclusion, the presence of FGF10 during IVM had no effect on embryo production, developmental speed, and gene expression.

Lung adenocarcinoma is one of the most common cancers worldwide. However, the molecular mechanisms of lung adenocarcinoma development are still unclear. This study aimed to investigate the expression profiles of anti-lung cancer target genes in different cancer stages and to explore their functions in tumor development. Lung adenocarcinoma transcriptome and clinical data were downloaded from Genomic Data Commons Data Portal, and the anti-lung cancer target genes were retrieved from the Thomson Reuters Integrity database. The results showed that 16 anti-lung target genes were deregulated in all stages. Among these target genes, <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> showed the most important role in transcription regulatory networks. Further analysis revealed that APC, BRIP1, and PTTG1 may regulate <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> and subsequently influence MAPK signaling pathway, Rap1 signaling pathways, and other tumorigenic processes in all stages. Moreover, high <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> expression leads to poor overall survival (hazard ratio = 1.55, P = .019). These findings provide valuable information for the pathological research and treatment of lung adenocarcinoma. Future studies are needed to verify these results.

FR901464 (FR) was first described as an anticancer drug and later identified as a modulator of splicing <em>factor</em> 3B subunit 1 (SF3B1). Although the effectiveness of splicing modulators has been investigated in colorectal cancer (CRC) cells, their usefulness in animal experiments has not been confirmed. The association of <i>SF3B1</i> with CRC progression and the influence of FR on transcriptional activity in CRC has not been fully elucidated. FR showed strong cytotoxicity against CRC cell lines, <i>SF3B1</i>-mutated cancer cell lines, and human <em>fibroblasts</em> with IC₅₀ values less than 1 ng/ml. FR-resistant clones derived from HCT116, DLD1, Lovo, and CT26 cells showed IC₅₀ values greater than 100 ng/ml. <i>SF3B1</i> sequencing demonstrated low frequencies of <i>SF3B1</i> mutations in CRC and mutations in codon 1074 of exon <em>22</em> in all FR-resistant clones. Unlike hematological malignancies, <i>SF3B1</i> expression was not associated with CRC progression. Although FR showed significant <em>growth</em> inhibition in a xenograft model of RKO cells, severe toxicity was also induced. These data indicated CRC might be a suitable target of FR unless toxicity occurs. Microarray analysis and real-time quantitative PCR demonstrated downregulation of genes associated with Fanconi anemia (<i>BRCA1 and BRCA2</i>) and 28 driver oncogenes. These data suggested combination treatment of FR with other anticancer drugs whose sensitivity is associated with genes affected by FR treatment. Combination treatment with PARP1 inhibitor olaparib, whose sensitivity was enhanced by <i>BRCA</i> 1/2 deficiency, showed synergistic effects in CRC cells. Our data indicates the potential of FR in combination therapy rather than monotherapy for CRC treatment.

A systematic review and meta-analysis was conducted of studies that address the association of bile acid (BA) with obesity and of studies on the effects of treatment in patients with obesity on BA metabolism, assessed from systemic BA, <em>fibroblast</em> <em>growth</em> <em>factor</em> 19 (FGF19), 7α-hydroxy-4-cholesten-3-one (C4) level, and faecal BA. We searched PubMed, Embase, and the Cochrane Library from inception to 1 August 2019 using the keywords obesity, obese, body mass index, and overweight with bile acid, FGF19, FXR, and TGR5. Two reviewers independently searched, selected, and assessed the quality of studies. Data were analysed using either fixed or random effect models with inverse variance weighting. Of 3771 articles, 33 papers were relevant for the association of BA with obesity of which <em>22</em> were included in the meta-analysis, and 50 papers were relevant for the effect of obesity interventions on BA of which 20 were included in the meta-analysis. Circulating fasting total BA was not associated with obesity. FGF19 was inversely and faecal BA excretion was positively associated with obesity. Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy (SG) modulated BA metabolism, ie, increased BA and FGF19. Our results indicate that BA metabolism is altered in obesity. Certain bariatric surgeries including RYGB and SG modulate BA, whether these underlie the beneficial effect of the treatment should be investigated.

Objective: The aim of this cross-sectional study was to estimate the prevalence of XLH in Paraná, a state in southern Brazil, and report the clinical features and complications of the disease.

<strong class="sub-title"> Methods: </strong> We invited all endocrinologists (n = 205), nephrologists (n = <em>22</em>1), orthopedic surgeons (n = 1020), and pediatricians (n = 1000) in Paraná to fill out an electronic survey with information on patients with X-linked hypophosphatemia (XLH), and searched the records of the state's health department for all calcitriol prescriptions in 2018.

<strong class="sub-title"> Results: </strong> In all, 244 (10%) specialists responded to the email, of whom 18 (7.4%) reported to be taking care of patients with XLH and answered the online survey. A total of 57 patients with XLH were identified (prevalence 5 per million inhabitants). The median age at diagnosis was <em>22</em> years, and 42.2% were children and adolescents. Fifteen patients had genetic testing showing a <i>PHEX</i> mutation. Overall, 91.2% had bone deformities, 30.8% had a history of fragility fractures, and <em>22</em>.4% had renal complications.

Conclusion: This study demonstrated a prevalence of XLH of 5 cases per million inhabitants in the state of Paraná, a rate lower than the one reported in other countries. Manifestations of renal calcification and bone fragility were frequent among the patients. This is the first epidemiological study evaluating the prevalence and clinical presentation of XLH in Latin America.

Keywords: PHEX mutation; Rickets; X-linked hypophosphatemia; fibroblast growth factor 23, human; hypophosphatemia.

♦

BACKGROUND

Fibroblast growth factor 23 (FGF23) is a phosphate regulating protein. Several studies demonstrated that elevated FGF23 is independently associated with mortality for early-stage chronic kidney disease and incident hemodialysis (HD) patients. However, little is known about the significance of elevated FGF23 in peritoneal dialysis (PD) patients. Here, we analyzed the association of FGF23 with cardiovascular (CV) events, all-cause mortality, residual renal function (RRF), and CV parameters in PD patients. ♦

METHODS

The present study is a single-center, retrospective study. Patients who started PD at Seoul National University Hospital between January 2005 and July 2011 and whose baseline serum samples were available were enrolled. C-terminal FGF23 was measured. Subjects were divided into 2 groups; lower 2 tertiles (FGF23 ≤ 119.0 RU/mL) and top tertile (FGF23>> 119.0 RU/mL). The primary outcome was time to fatal or non-fatal CV events. In the subgroup analysis, the associations of FGF23 with aortic stiffness or with vascular calcification were analyzed. ♦

RESULTS

A total of 205 incident PD patients were analyzed. Mean duration of follow-up was 41.6 ± 20.0 months. The baseline median FGF23 level was 78.6 RU/mL (inter-quartile range [IQR], 34.1 - 155.0). At baseline, subjects in the higher FGF23 group were younger, and had a lower RRF, lower prevalence of diabetes mellitus (DM), and cerebrovascular disease. During follow-up, 22 of the 205 patients (10.7%) reached primary outcome. After adjustment for age, DM, pre-existing coronary artery disease, cerebrovascular disease, congestive heart failure, and left ventricular mass index, the higher FGF23 group exhibited significantly higher risk of primary outcome, compared with the lower group (hazard ratio [HR], 2.54; 95% confidence interval [CI], 1.05 - 6.12; p = 0.045). There were no significant differences in all-cause mortality and development of anuria between the 2 FGF23 groups. In the subgroup analysis, FGF23 groups were not associated with pulse wave velocity and abdominal aortic calcification score. ♦

CONCLUSIONS

Elevated FGF23 is associated with higher risk of adverse CV outcome for incident PD patients.

OBJECTIVE

Clefts of lip and palate belong to the most common birth defects worldwide. Growth factors and genes play an important role in tissue growth, differentiation and induction and upregulation of growth factors, apoptosis and matrix metalloproteinases might be involved in pathogenesis of facial clefts. The aim of this study was investigation of palate tissue in children with unilateral cleft lip palate for detection of local tissue growth factors, barx1 and apoptosis.

METHODS

We investigated soft and hard palate tissue from 36 children with complete unilateral cleft lip and palate from cleft area.14 children were in age before and primary dentition, but 22 children were in mixed dentition period. We examined the localization of barx1, FGFR1, NGFR, TGFbeta, BMP2/4, MMP2, PGP 9,5 by immunohistochemistry. TUNEL method was performed for detection of apoptotic cells.

RESULTS

Abundance of FGFR1 positive cells was seen almost in all cases. FGFR richly stained cells of soft and hard palate tissue. Abundance of NGFR positive cells was detected in basal epithelium, hair follicles, nerve fibers in wall of blood vessels and subepithelium, and was more often seen in children before mixed dentition. TGFbeta has showed intensive expression in epithelium, cartilage and bone in both dentition ages. Chondrocytes, fibroblasts and macrophages expressed MMP2 predominant before mixed dentition. Regional expression of barx1 was observed in epithelium before the mixed dentition, while during mixed dentition gene appeared in hyaline cartilage. TUNEL discovered apoptosis in both dentition ages.

CONCLUSIONS

FGFR1 and TGFbeta are main tissue stimulating growth factors in both dentition ages. Expression of barx1 appears in cleft lip palate affected structures mainly in mixed dentition ages. NGFR and neuropeptides-containing structures are mainly characteristic in cleft tissue before mixed dentition. Distribution of genes, GF and apoptosis seem to correlate rather with dentition age than to type of CLP.

<AbstractText>Herpes simplex virus-1 (HSV-1) infections of the central nervous system (CNS) can result in HSV-1 encephalitis (HSE) which is characterized by severe brain damage and long-term disabilities. Different cell types including neurons and astrocytes become infected in the course of an HSE which leads to an activation of glial cells. Activated glial cells change their neurotrophic <em>factor</em> profile and modulate inflammation and repair. The superfamily of <em>fibroblast</em> <em>growth</em> <em>factors</em> (FGFs) is one of the largest family of neurotrophic <em>factors</em> comprising <em>22</em> ligands. FGFs induce pro-survival signaling in neurons and an anti-inflammatory answer in glial cells thereby providing a coordinated tissue response which favors repair over inflammation. Here, we hypothesize that FGF expression is altered in HSV-1-infected CNS cells.</AbstractText><AbstractText>We employed primary murine cortical cultures comprising a mixed cell population of astrocytes, neurons, microglia, and oligodendrocytes. Astrocyte reactivity was morphometrically monitored by an automated image analysis algorithm as well as by analyses of A1/A2 marker expression. Altered FGF expression was detected by quantitative real-time PCR and its paracrine FGF activity. In addition, HSV-1 mutants were employed to characterize viral <em>factors</em> important for FGF responses of infected host cells.</AbstractText><AbstractText>Astrocytes in HSV-1-infected cortical cultures were transiently activated and became hypertrophic and expressed both A1- and A2-markers. Consistently, a number of FGFs were transiently upregulated inducing paracrine neurotrophic signaling in neighboring cells. Most prominently, FGF-4, FGF-8, FGF-9, and FGF-15 became upregulated in a switch-on like mechanism. This effect was specific for CNS cells and for a fully functional HSV-1. Moreover, the viral protein ICP0 critically mediated the FGF switch-on mechanism.</AbstractText><AbstractText>HSV-1 uses the viral protein ICP0 for the induction of FGF-expression in CNS cells. Thus, we propose that HSV-1 triggers FGF activity in the CNS for a modulation of tissue response upon infection.</AbstractText>

Bovine colostrum (BC) has been used for nutraceutical purposes for animals and humans. Bovine colostrum is a complex heterogeneous product and its antimicrobial activity, antioxidant potential, and <em>growth</em> <em>factors</em> can vary depending on age and species of the cow as well as their environment. Bovine colostrum preparation in skimmed or whey fractions can also alter properties of BC. Our goal was to compare cumulative anti-inflammatory, antioxidant, and adipogenic properties of natural (whole) versus whey BC. We compared properties of whole and whey BC in 3T3-L1 preadipocytes permanently transfected with reporters responding to changes in inflammatory (NfκbRE/green fluorescent protein), anti-inflammatory (Nrf2/YFP), and adipogenic (Fabp4/cyan fluorescent protein) status in cells. Interleukin-6 secretion in these cells was measured by ELISA. Whole and whey BC induce IL-6 secretion from 3T3-L1 <em>fibroblasts</em>; however, whey preparation stimulated less IL-6 secretion. Cumulative inflammatory nuclear <em>factor</em> (NF)κB activation in the presence of lipopolysaccharide was reduced by both whole (-27%) and whey BC (-<em>22</em>%) compared with lipopolysaccharide-treated cells (100%). Treatment with whole BC was more effective in the reduction of NFκB activation compared with whey BC and occurred in a dose-dependent manner. In consonance with decreased NFκB activation, the Nrf2 promoter activity was also reduced in response to whole (-27%) and whey (-13%) treatments compared with nontreated cells (100%). Whole and whey BC suppressed adipogenesis, measured as induction of Fabp4, by -27 and -13%, respectively, compared with nontreated 3T3-L1 <em>fibroblasts</em> (100%). Our results showed distinct differences in properties of whey and whole BC that could be used to attain reduced adipogenic or cumulative inflammatory responses.

OBJECTIVE

In order to search for substances that reduce the neurotoxicity of paclitaxel, the sensitivity of differentiated rat neuronal PC12 cells to paclitaxel was compared to that of malignant and non-malignant cells, and the extent to which four antioxidants can alleviate paclitaxel-induced neurotoxicity was investigated.

METHODS

Viability of cells was determined by the MTT method. Cytotoxicity was evaluated as the concentration that reduced cell viability by 50% (CC50). Tumor specificity of paclitaxel was determined as the ratio of CC50 against non-malignant cells to that against malignant cells.

RESULTS

Paclitaxel was three-fold more cytotoxic towards human oral squamous cell carcinoma cell lines (Ca9-<em>22</em>, HSC-2, HSC-3. HSC-4) than human normal epithelial and mesenchymal (human gingival <em>fibroblast</em>, human periodontal ligament <em>fibroblast</em>, human pulp cell) normal cells, confirming its antitumor potential. However, paclitaxel at as low a concentration as 5 ng/ml significantly reduced neurite formation in nerve <em>growth</em> <em>factor</em>-induced differentiated PC12 cells, although complete killing of cells was not achieved even at 2,000-fold higher concentration (10 μM). Paclitaxel-induced neurotoxicity was enhanced with the prolongation of incubation time and reduction of inoculation cell density. Four antioxidants, namely docosahexaenoic acid, acetyl-L-carnitine hydrochloride, N-acetyl-L-cysteine and sodium ascorbate, only partially protected PC12 cells from paclitaxel-induced toxicity.

CONCLUSIONS

The present study suggests the involvement of both oxidative and other mechanisms in paclitaxel-induced neurotoxicity.

Immune checkpoint inhibitors (ICIs) are commonly used in patients with advanced non-small cell lung cancer (NSCLC). An unmet need remains for new biomarkers associated with ICIs. In this study, consecutive patients with advanced NSCLC treated with nivolumab or pembrolizumab were included. Plasma at ICIs initiation was prospectively collected and a multiplex ELISA assay testing 48 cytokines and <em>growth</em> <em>factors</em> was performed. Exploratory endpoints were the association between plasma biomarkers with outcome and grade III-IV immune related adverse events (irAEs). Thirty-five patients were included. Patients without clinical benefit (<i>n</i> = <em>22</em>) had higher pre-ICI soluble Hepatocyte <em>Growth</em> <em>Factor</em> (sHGF) (210.9 vs. 155.8 pg/mL, <i>p</i> = 0.010), lower pre-ICI soluble <em>Fibroblast</em> <em>Growth</em> <em>Factor</em> (sFGF) (4.0 vs. 4.8 pg/mL, <i>p</i> = 0.043) and lower pre-ICI interleukine-12 (IL-12) (1.3 vs. 2.2 pg/mL, <i>p</i> = 0.043) concentrations. Patients with early progression (<i>n</i> = 23) had higher pre-ICIs sHGF (206.2 vs. 155.8 pg/mL, <i>p</i> = 0.025) concentrations. Patients with low sHGF levels at ICIs initiation had longer progression-free survival and overall survival than those with high sHGF levels: respectively 2.5 vs. 8.0 months (<i>p</i> = 0.002), and 5.5 vs. 35.0 months (<i>p</i> = 0.001). TNF-α, IL-16, IL-12p40 and MCP3 were associated with high grade irAEs. This study shows the potential association between several plasma biomarkers with outcome and grade 3-4 IrAEs in advanced NSCLC treated with ICIs.

Keywords: Fibroblast growth factor; biomarker; hepatocyte growth factor; immune checkpoint inhibitor; interleukine-12; non-small cell lung cancer; plasma; resistance; toxicity.

Three experiments have been conducted to investigate the effects of graded dietary levels of vitamin D3 ( VD3: ) on <em>growth</em> performance, metabolic regulation of calcium (CA), phosphorus (P), and bone development of Chinese yellow-feathered broilers during 3 <em>growth</em> phases: 1 to 21 d, <em>22</em> to 42 d, and 43 to 63 d. Dietary Ca and P in the corn-soybean-based diet were adequate. A total of 2,000 1-day-old, 1,600 <em>22</em>-day-old, and 1,600 43-day- old Lingnan yellow male broilers were randomly assigned to 1 of 8 dietary treatments with 5 replicates per treatment (50 birds per replicate for 1 to 21 d, 40 birds for both <em>22</em> to 42 d and 43 to 63 d). Dietary levels of VD3 were 100, 200, 300, 400, 500, 600, and 700 IU/kg for treatments 2 to 8 through the addition of VD3 to the basal mash diet which otherwise lacked detectable VD3. Graded doses of VD3 from 0 to 700 IU/kg in the diet produced linear (P<0.01) positive responses in ADG, ADFI, tibial weight, and breaking strength, and quadratic (P<0.01) responses in tibial length, bone density, ash, the levels of Ca and P in the ash , and the ratio of Ca to P. Serum concentrations of Ca, P, 25-hydroxycholecalciferol, osteocalcin, and calcitonin increased, and concentrations of <em>fibroblast</em> <em>growth</em> <em>factor</em> 23, Klotho protein, and parathyroid hormone all decreased with the increasing level of dietary VD3 (P<0.05). Adding VD3 improved meat color a* value and decreased shear force and drip loss of birds at 63 d (P<0.05). Considering bone characteristics and composition under the conditions of this study, it was concluded that the VD3 requirements of Chinese yellow-feathered broilers from 1 to 21 d for optimal tibial ash content were estimated from regression analysis to be 464 IU/kg from 1 to 21 d, 539 IU/kg from <em>22</em> to 42 d, and 500 IU/kg from 43 to 63 d.

OBJECTIVE

To compare cytokine and growth factor concentrations in canine autologous conditioned serum (ACS) to canine plasma.

METHODS

Experimental in vivo study.

METHODS

Client-owned, adult dogs (n=22).

METHODS

Blood collected from 16 medium to large breed dogs was used to produce ACS (Orthokine(®) vet irap 10 syringes) and citrated plasma (control). Canine-specific ELISA assays were run per manufacturers' instructions for interleukin (IL)-10, IL-4, tumor necrosis factor (TNF)-α, insulin-like growth factor (IGF)-1, fibroblast growth factor (FGF)-2, transforming growth factor (TGF)-β1, IL-1β, and interleukin-1 receptor antagonist (IL-1ra). Serum, in addition to plasma and ACS, was collected from an additional 6 dogs for TNF-α, IL-1β, and IL-1ra analysis (total of 22 dogs). Data were analyzed for differences in each cytokine concentration using pairwise comparisons between ACS, plasma, and serum using Wilcoxon signed-rank tests. Significance was set at P<.05.

RESULTS

There was a large variability in growth factor and cytokine concentrations in ACS and plasma for individual dogs. There were no significant differences in IL-10, TNF-α, IGF-1, FGF-2, and TGF-β1 concentrations between ACS, plasma, and serum. The IL-1β concentrations in ACS (median, range 46.3 pg/mL, 0-828.8) and IL-4 (0.0 pg/mL, 0-244.1) were significantly higher than plasma (36.6 pg/mL, 0-657.1 and 0.0 pg/mL, 0-0, respectively). The IL-1ra concentration in ACS (median, range 3,458.9 pg/mL, 1,243.1-12,089.0) was significantly higher than plasma (692.3 pg/mL, 422.5-1,475.6). The IL-1ra:IL-1β ratio in ACS was significantly higher than plasma (39.9 vs. 7.2).

CONCLUSIONS

IL-1ra concentrations in canine ACS were comparable to those published for people and horses and pro-inflammatory cytokines remained low in canine ACS.

Fibrotic remodeling is a highly conserved protective response to tissue injury and it is essential for the maintenance of structural and functional tissue integrity. Also hepatic fibrosis can be considered as a wound-healing response to liver injury, reflecting a balance between liver repair and scar formation. In contrast, pathological fibrosis corresponds to impaired wound healing. Usually, the liver regenerates after acute injury. However, if the damaging mechanisms persist, the liver reacts with progressive and uncontrolled accumulation of extracellular matrix proteins. Eventually, excessive fibrosis can lead to liver cirrhosis and hepatic failure. Furthermore, cirrhosis is the major risk <em>factor</em> for the development of hepatocellular cancer (HCC). Therefore, hepatic fibrosis is the most critical pathological <em>factor</em> that determines the morbidity and mortality of patients with chronic liver disease. Still, no effective anti-fibrogenic therapies exist, despite the very high medical need. The regulation of <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF) signaling is a prerequisite for adequate wound healing, repair and homeostasis in various tissues and organs. The FGF family comprises <em>22</em> proteins that can be classified into paracrine, intracrine and endocrine <em>factors</em>. Most FGFs signal through transmembrane tyrosine kinase FGF receptors (FGFRs). Although FGFRs are promising targets for the treatment of HCC, the expression and function of FGFR-ligands in hepatic fibrosis is still poorly understood. This review summarizes the latest advances in our understanding of FGF signaling in hepatic fibrosis. Furthermore, the potential of FGFs as targets for the treatment of hepatic fibrosis and remaining challenges for the field are discussed.

Keywords: FGF; Fibroblast growth factors; fibrosis; hepatic stellate cells; wound healing.

<AbstractText>The development of a clinically useful <em>fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) hormone has been impeded by its inherent instability and weak FGF receptor (FGFR) binding affinity. There is an urgent need for innovative approaches to overcome these limitations.</AbstractText><p><div><b>METHODS</b></div>We devised a structure-based chimerisation strategy in which we substituted the thermally labile and low receptor affinity core of FGF21 with an HS binding deficient endocrinised core derived from a stable and high receptor affinity paracrine FGF1 (FGF1^ΔHBS). The thermal stability, receptor binding ability, heparan sulfate and βKlotho coreceptor dependency of the chimera were measured using a thermal shift assay, SPR, SEC-MALS and cell-based studies. The half-life, tissue distribution, glucose lowering activity and adipose tissue remodeling were analyzed in normal and diabetic mice and monkeys.</p><p><div><b>FINDINGS</b></div>The melting temperature of the engineered chimera (FGF1^ΔHBS-FGF21^C-tail) increased by ∼<em>22</em> °C relative to wild-type FGF21 (FGF21^WT), and resulted in a ∼5-fold increase in half-life in vivo. The chimera also acquired an ability to bind the FGFR1c isoform - the principal receptor that mediates the metabolic actions of FGF21 - and consequently was dramatically more effective than FGF21^WT in correcting hyperglycemia and in ameliorating insulin resistance in db/db mice. Our chimeric FGF21 also exerted a significant beneficial effect on glycemic control in spontaneous diabetic cynomolgus monkeys.</p><AbstractText>Our study describes a structure-based chimerisation approach that effectively mitigates both the intrinsically weak receptor binding affinities and short half-lives of endocrine FGFs, and advance the development of the FGF21 hormone into a potentially useful drug for Type 2 diabetes.</AbstractText>

Background: Dietary polyphenols including anthocyanins target multiple organs.

Objective: We aimed to assess the involvement of glucagon-like peptide 1 (GLP-1), leptin, insulin and fibroblast growth factor 21 (FGF21) in mediating metabolic beneficial effects of purified anthocyanin cyanidin-3-glucoside (Cy3G).

Methods: Intestinal proglucagon gene (Gcg; encoding GLP-1) and liver Fgf21 expression were assessed in 6-wk-old male C57BL-6J mice fed a low-fat-diet (LFD; 10% of energy from fat), alone or with 1.6 mg Cy3G/L in drinking water for 3 wk [experiment (Exp.) 1; n = 5/group]. Similar mice were fed the LFD or a high-fat diet (HFD; 60% energy from fat) with or without Cy3G for 20 wk. Half of the mice administered Cy3G also received 4 broad-spectrum antibiotics (ABs) in drinking water between weeks 11 and 14, for a total of 6 groups (n = 8/group). Metabolic tolerance tests were conducted between weeks 2 and 16. Relevant hormone gene expression and plasma hormone concentrations were assessed mainly at the end of 20 wk (Exp. 2).

Results: In Exp. 1, Cy3G administration increased ileal but not colonic Gcg level by 2-fold (P < 0.05). In Exp. 2, Cy3G attenuated HFD-induced body-weight gain (20.3% at week 16), and improved glucose tolerance (26.5% at week 15) but not insulin tolerance. Although Cy3G had no effect on glucose tolerance in LFD mice, LFD/Cy3G/AB mice showed better glucose tolerance than LFD/Cy3G mice (23%). In contrast, HFD/Cy3G/AB mice showed worse glucose tolerance compared with HFD/Cy3G mice (15%). Beneficial effects of Cy3G in HFD mice were not associated with changes in plasma leptin, insulin or GLP-1 concentrations. However, Cy3G increased hepatic Fgf21 expression in mice in Exp. 1 by 4-fold and attenuated Fgf21 overexpression in HFD mice (Exp. 2, 22%), associated with increased expression of genes that encode FGFR1 and β-klotho (>3-fold, P < 0.05).

Conclusions: Dietary Cy3G may reduce body weight and exert metabolic homeostatic effects in mice via changes in hepatic FGF21.

Keywords: Cy3G; FGF21; GLP-1; OGTT; anthocyanin; dietary intervention.