BACKGROUND

Living kidney donors (LKDs) experience reduction in kidney function, however serum phosphate (sPi) levels are lower compared to patients with chronic kidney disease matched for reduced kidney function. Mineral metabolism adaptations that occur in LKDs have not been adequately investigated. To evaluate the effect of nephrectomy on markers of mineral metabolism in LKDs compared to healthy volunteers (HV) over 12 months.

METHODS

Mineral parameters were evaluated in twenty-one adult LKDs and twenty HVs. Parameters included sPi, intact parathyroid hormone, fibroblast growth factor-23 (FGF23), soluble Klotho (sKl) and urinary phosphate, measured prior to donation (T0), 1 month (T1), 6 months (T6) and 12 months (T12) post-kidney donation. Statistical analyses were conducted on normalized variables and changes were assessed using 2-way analysis of variance.

RESULTS

Mean ages of LKDs and HVs were 54.1 ± 14.7 and 52.6 ± 8.0 years, respectively. There were no baseline clinical or biochemical differences between LKDs and HVs. In LKDs at T1, serum creatinine increased (from 75 ± 12 to 114 ± 22 μmol/L), FGF23 increased (52 ± 15 to 70 ± 19 pg/mL) and sKl decreased (564 [469-662] to 424 [375-523] pg/mL), all P less than 0.001. Changes were sustained at T12. After donation, LKDs consistently demonstrated lower sPi compared with T0, with the maximal sPi change at T6 (-0.19 mmol/L difference, P < 0.001). Other markers of mineral metabolism were unchanged in LKDs. There were no mineral differences in HVs over 12 months.

CONCLUSIONS

Prospective evaluation of mineral metabolism parameters in LKDs provides valuable insight into compensatory mechanisms after reduction in kidney function. Further reduction of sPi at T6 despite early alterations in FGF23 and sKl suggest adaptation of mineral metabolism continues long-term in LKDs.

The aim of this study is to analyze the concentrations of cytokines in tear of hospitalized COVID-<em>19</em> patients compared to healthy controls. Tear samples were obtained from 41 healthy controls and 62 COVID-<em>19</em> patients. Twenty-seven cytokines were assessed: interleukin (IL)-1b, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL9, IL-10, IL-12, IL-13, IL-15, IL-17, eotaxin, <em>fibroblast</em> <em>growth</em> <em>factor</em> basic, granulocyte colony-stimulating <em>factor</em> (G-CSF), granulocyte-monocyte colony-stimulating <em>factor</em> (GM-CSF), interferon (IFN)-γ, interferon gamma-induced protein, monocyte chemo-attractant protein-1, macrophage inflammatory protein (MIP)-1a, MIP-1b, platelet-derived <em>growth</em> <em>factor</em> (PDGF), regulated on activation normal T cell expressed and secreted, tumor necrosis <em>factor</em>-α and vascular endothelial <em>growth</em> <em>factor</em> (VEGF). In tear samples of COVID-<em>19</em> patients, an increase in IL-9, IL-15, G-CSF, GM-CSF, IFN-γ, PDGF and VEGF was observed, along with a decrease in eotaxin compared to the control group (p < 0.05). A poor correlation between IL-6 levels in tear and blood was found. IL-1RA and GM-CSF were significantly lower in severe patients and those who needed treatment targeting the immune system (p < 0.05). Tear cytokine levels corroborate the inflammatory nature of SARS-CoV-2.

Keywords: COVID; Cytokine; Inflammation; Tear.

Background: Chronic kidney disease (CKD) is associated with cardiovascular disease (CKD), mineral and bone disorder (CKD-MBD) and high mortality. Bone-related factors such as osteopontin (OPN), osteocalcin (OC), osteoprotegerin (OPG) and fibroblast growth factor 23 (FGF23) were linked to cardiovascular complications of CKD and are expected to have predictive value in CKD patients.

Purpose: The aim of this study was to assess the relationship of OPN, OC, OPG and FGF23 to clinical characteristics and to evaluate their ability to predict mortality in patients with different CKD stages.

Methods: The following study groups were enrolled: subjects with end-stage renal disease (38 ESRD), CKD stages 3 and 4 (19 CKD3-4) and non-CKD controls (19), respectively. Blood was withdrawn once to perform the measurements and cardiac computed tomography was used to evaluate coronary calcium score (CS). Patients were followed for 5 years for the ascertainment of their all-cause mortality.

Results: Serum OPN, OC and OPG concentrations increased significantly along with the progression of renal disease. We found a significant positive correlation among these proteins. Additionally, OPN and OPG were significantly and positively correlated to CS. Serum OPG revealed the strongest correlation to the calcium turnover markers of GFR decline and was significantly associated with an increased risk of death in subjects with CKD3-4 or ESRD (HR 5.8, CI 95%).

Conclusion: Single measurement of osteoprotegerin is associated with 5-year all-cause mortality in patients with CKD3-4 or ESRD. We suggest assessing its concentration, preferably in combination with calcium score, to stratify mortality risks in CKD patients.

Keywords: calcium score; chronic kidney disease; osteocalcin; osteopontin; osteoprotegerin.

Wound healing is a complex biological process that requires coordinated cell proliferation, migration, and extracellular matrix production/remodeling, all of which are inhibited/delayed in chronic wounds. In this study, a formulation was developed that marries a fibrin-based, provisional-like matrix with collagen mimetic peptide (CMP)/PDGF gene-modified collagens, leading to the formation of robust gels that supported temporally controlled PDGF expression and facile application within the wound bed. Analysis employing in vitro co-gel scaffolds confirmed sustained and temporally controlled gene release based on matrix metalloproteinase (MMP) activity, with ~30% higher PDGF expression in MMP producing <em>fibroblasts</em> as-compared with non-MMP-expressing cells. The integration of fibrin with the gene-modified collagens resulted in co-gels that strongly supported both <em>fibroblast</em> cell recruitment/invasion as well as multiple aspects of the longer-term healing process. The excisional wound healing studies in mice established faster wound closure using CMP-modified PDGF polyplex-loaded co-gels, which exhibited up to 24% more wound closure (achieved with ~2 orders of magnitude lower <em>growth</em> <em>factor</em> dosing) after 9 days as compared to PDGF-loaded co-gels, and <em>19</em>% more wound closure after 9 days as compared to CMP-free polyplex loaded co-gels. Moreover, minimal scar formation as well as improved collagen production, myo<em>fibroblast</em> activity, and collagen orientation was observed following CMP-modified PDGF polyplex-loaded co-gel application on wounds. Taken together, the combined properties of the co-gels, including their stability and capacity to control both cell recruitment and cell phenotype within the murine wound bed, strongly supports the potential of the co-gel scaffolds for improved treatment of chronic non-healing wounds.

Keywords: Collagen mimetic peptide; MMP; PDGF; gene delivery; temporal control; wound healing.

<em>Fibroblast</em> <em>growth</em> <em>factors</em> (FGF) <em>19</em>, 21, and 23 have been reported as functional <em>factors</em> in human metabolic diseases and malignancies. We performed a prospective survey to compare circulating FGF levels in urothelial carcinoma (UC) patients and normal controls. Between 2016 and 2017, 39 patients with UC of the urinary bladder or upper urinary tract who received surgical intervention were included. All the serum samples were obtained before surgeries. The control group included 28 healthy volunteers. Analysis of the circulating FGF<em>19</em>, 21, and 23 levels among all 67 subjects, as well as a subgroup analysis of the 39 UC patients were performed. The median levels of serum FGF<em>19</em>, 21, and 23 in the UC patients were 84.2, 505.3, and 117.6 pg/mL, respectively, which were statistically different from levels found in the healthy controls (P = 0.015, <0.001 and < 0.001, respectively). In the subgroup analysis, the FGF<em>19</em> and FGF21 levels were significantly higher in end-stage renal disease UC patients, while FGF21 was also higher in the UC patients with cardiovascular diseases and history of recurrent UC. In the receiver operating characteristic (ROC) curve analysis, FGF<em>19</em>, 21, and 23 were all significant predictors of UC [area under the curve (AUC)] 0.674, P = 0.015; AUC 0.918, P < 0.001; AUC 0.897, P < 0.001, respectively). In UC patients, serum FGF<em>19</em> level was significantly lower, while FGF21 and 23 were significantly higher, than respective levels in healthy controls. All three markers may serve as good predictors of UC occurrence, and FGF21 level was associated with disease recurrence. © 2018 BioFactors, 2018.

MicroRNAs have emerged as critical mediators of cerebral ischaemia/reperfusion injury. Recent studies have demonstrated that microRNA-302b-3p (miR-302b-3p) plays an important role in regulating apoptosis and oxidative stress in various cells. However, whether miR-302b-3p is involved in regulating cerebral ischaemia/reperfusion injury-induced neuronal apoptosis and oxidative stress remains unknown. In the present study, we explored the potential function and molecular mechanism of miR-302b-3p in oxygen-glucose deprivation/re-oxygenation (OGD/R)-induced neuronal injury, using an in vitro model of cerebral ischaemia/reperfusion injury. We found that miR-302b-3p expression was up-regulated by OGD/R treatment in neurons. The inhibition of miR-302b-3p improved cell viability, and reduced apoptosis and the production of reactive oxygen species, showing a protective effect against OGD/R-induced injury. Interestingly, miR-302b-3p was shown to target and modulate murine fibroblast growth factor 15 (FGF15). Moreover, our results showed that miR-302b-3p down-regulation contributed to the promotion of nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE)-mediated antioxidant signaling associated with the inactivation of glycogen synthase kinase-3β. However, the knockdown of FGF15 significantly reversed the miR-302b-3p inhibition-mediated protective effect in OGD/R-treated neurons. Overall, these results demonstrated that miR-302b-3p inhibition confers a neuroprotective effect in OGD/R-treated neurons by up-regulating Nrf2/ARE antioxidant signaling via targeting FGF15, providing a novel target for neuroprotection in cerebral ischaemia/reperfusion injury.

OBJECTIVE

To determine the association of fibroblast growth factor 23 (FGF23) with left ventricular hypertrophy (LVH) through the assessment of left ventricular (LV) mass and left ventricular mass index (LVMI) in patients on hemodialysis, this study was done.

METHODS

All patients on hemodialysis who are older than 18 years and in whom hemodialysis vintage was at least 6 months were enrolled. All patients were on hemodialysis thrice a week for 4 h using low-flux dialysis filters, polysulfone membranes, reverse osmosis purified water, and bicarbonate-base hemodialysis solution. The exclusion criteria were any respiratory illness or pulmonary infection, cigarette smoking, and the presence of pericarditis or pericardial effusion. Additionally, patients with a known coronary artery disease, any form of cardiac arrhythmias, any cardiomyopathy or severe valvular heart disease diagnosed by echocardiography, acute congestive heart failure (CHF), and acute myocardial infarction were not included. Echocardiography was conducted by an experienced operator for all the enrolled patients using the ACUSON SC2000™ ultrasound system transducer (Siemens), with a frequency bandwidth of: 1.5-3.5 MHz. Patients were considered to have LVH if the LVMI was greater than 134 g/m2 for men and greater than 110 g/m2 for women.

RESULTS

A total of 61 patients (19 female and 42 male) were enrolled to the study. Mean (± SD) age of the patients was 59.6 ± 13.1 years. The median duration of hemodialysis was 23 (range: 6-120) months. The median predialysis level of FGF23 was 1,977 pg/mL (range: 155-8,870). LVH was seen in 73.8% of the patients (n = 45) and of them 66.7% were male. There was a statistically significant direct correlation between FGF23 and left ventricle diameter in end systole (LVDs) (r = 0.29, P = 0.027). However, the association of FGF23 with LV mass, LVMI, and left ventricular ejection fraction (LVEF) was not significant.

CONCLUSIONS

This study does not show the correlation between FGF23 and LV mass in stable hemodialysis patients.

OBJECTIVE

Serum levels of 32 kDa-phosphaturic hormone fibroblast growth factor 23 (FGF23) rise early in renal failure in order to keep phosphatemia within the normal range; however, this compensatory mechanism itself contributes to chronic kidney disease-mineral bone disorder. High FGF23 is also associated to left ventricular hypertrophy, vascular calcifications and thus increased cardiovascular risk. The aim of this pilot pre-post study was to evaluate the effects of a single hemodiafiltration session with acetate-free biofiltration (AFB) on FGF23 serum levels.

METHODS

Nine hemodialysis patients were enrolled; sessions were performed using the Integra® monitor (Hospal, Bologna, Italy) and a polyacrylonitrile membrane. Peripheral venous blood samples were taken before (pre-HD), at mid- and after treatment (post-HD); dialysate samples were collected by the Quantiscan™ monitoring system. FGF23 was measured by a human FGF-23 ELISA kit. Mid- and post-HD values were corrected for hemoconcentration.

RESULTS

Pre-HD FGF23 levels positively correlated with dialysis vintage (r = 0.7192; p = 0.0443). They were significantly reduced by the hemodialysis session (from 2.38 ± 1.80 to 1.15 ± 1.21 ng/ml, p = 0.0171) with a reduction ratio of 52.55 ± 28.76%. FGF23 was detected in the dialysate samples.

CONCLUSIONS

FGF23 underwent a significant reduction during AFB. Such removal was greater than that induced by conventional hemodialysis as reported in the literature (19%-decrease using modified cellulosic membranes). This difference may be attributed to the ability of AFB hemodiafiltration to efficiently remove middle molecules by convection. Whether a better clearance of FGF23 during hemodialysis may result in improved cardiovascular outcomes in the long term needs to be confirmed by randomized controlled trials.

BACKGROUND

A fascinating aspect of bile acid homeostasis is the coordination between bile acid uptake in intestine and hepatic bile acid synthesis. In response to bile acid uptake in enterocytes, farnesoid X receptor is activated and induces transcription of <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF)15 in mice, or FGF<em>19</em> in humans. FGF15/<em>19</em> is secreted into the enterohepatic circulation, and through activation of hepatic receptors, leads to repression of Cyp7a1, a rate-limiting enzyme for bile acid synthesis. Using a genetic approach, we identified a novel protein, Diet1, as a control point for FGF15/<em>19</em> production.

CONCLUSIONS

Mice with a Diet1-null mutation have reduced FGF15 secretion, causing impaired feedback repression of hepatic bile acid synthesis, and increased fecal bile acid excretion. As a result, Diet1-deficient mice constitutively convert cholesterol to bile acids and are resistant to diet-induced hypercholesterolemia and atherosclerosis. Diet1 affects FGF15/<em>19</em> production at the posttranscriptional level, and the proteins appear to have overlapping subcellular localization in enterocytes. Diet1 appears to be a control point for the production of FGF15/<em>19</em> in enterocytes, and thus a regulator of bile acid and lipid homeostasis. Studies to evaluate the role of common and rare DIET1 genetic variants in human health and disease are warranted.

CONCLUSIONS

Further elucidation of the Diet1-FGF15/<em>19</em> interaction will provide new insights into the intricate regulatory mechanisms underlying bile acid metabolism.

<AbstractText><em>Fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>) takes part in maintaining the balance of glycolipids and may be involved in regulating the secretory activity of islet beta cells in patients with type 2 diabetes. This study aimed to evaluate the relationship between the levels of serum FGF<em>19</em> and endogenous islet beta cell function in type 2 diabetic patients.</AbstractText><p><div><b>Methods</b></div>Samples were obtained from 271 subjects: 85 drug-naïve type 2 diabetes participants exclusively on lifestyle intervention (N-DM group), 122 type 2 diabetes subjects previously used medications (DM group) and 64 normal controls (NC group). Serum FGF<em>19</em> concentrations were measured by ELISA. The insulin sensitivity (MI), insulin secretion (AUC_ins/AUC_glu) and insulin secretion-sensitivity index-2 (ISSI-2) were also measured in the N-DM and DM.</p><p><div><b>Results</b></div>Serum FGF<em>19</em> levels decreased, in order, from the NC group [median (interquartile range), 245.03 (126.23-317.43) pg/mL] to the N-DM group [170.05 (89.01-244.70) pg/mL] and, finally, to the DM group [142.25 (55.55-187.58) pg/mL] (<i>p</i> for trend < 0.05). Among subjects in the DM group, there was a positive trend in the serum FGF<em>19</em> concentration; plasma insulin levels at 60 min, 120 min (INS60, INS120, respectively); and area under the insulin curve (AUC_ins) at two points (<i>r </i>= 0.214, <i>p</i> = 0.025; <i>r</i> = 0.189, <i>p</i> = 0.048; <i>r</i> = 0.188, <i>p</i> = 0.049). However, the differences were no longer observed among the N-DM subjects. Simultaneously, the ISSI-2 was closely related to the serum FGF<em>19</em> levels (<i>r</i> = 0.297, <i>p</i> = 0.002) among DM subjects. Furthermore, after adjusting for age, sex, duration, therapy and other clinical <em>factors</em> via multiple logistic regression analysis, ISSI-2 was a key independent <em>factor</em> in the levels of FGF<em>19</em> (<i>β </i>=<i> 0.281</i>, <i>t </i>=<i> 2.557</i>, <i>p </i>=<i> 0.013</i>).</p><AbstractText>The serum FGF<em>19</em> level has a close relation with endogenous beta cell function among DM subjects, as assessed by the ISSI-2. As ISSI-2 is higher in N-DM group, FGF<em>19</em> may be a main protector in dysfunction of beta cell.</AbstractText>

<AbstractText>Rapid tissue expansion has been attempted, aiming at shortening the period of conventional expansion. However, it has scarcely been clinically applied because of its drawbacks such as low expansion efficiency and tissue destruction. Adipose-derived stem cell transplantation is a promising therapeutic method in regenerative medicine. However, its effects on rapid expansion remain poorly understood.</AbstractText><AbstractText>Twenty-four expanders were implanted in the dorsum of 12 pigs. Rapid expansion persisted for 1 week with 20 ml of saline daily. The increased area of the expanded skin was measured. Histologic and ultrastructural analysis and cell tracking were performed. The expression of vascular endothelial <em>growth</em> <em>factor</em>, <em>fibroblast</em> <em>growth</em> <em>factor</em>-2, and epidermal <em>growth</em> <em>factor</em> was also determined.</AbstractText><AbstractText>The increased area of adipose-derived stem cell-grafted expanded skin (0.91 ± 0.06 cm) was significantly more than the non-adipose-derived stem cell-treated control (0.51 ± 0.05 cm) (p < 0.01). Enhanced tissue regeneration in the adipose-derived stem cell-grafted expanded skin was evidenced by increased skin thickness, proliferating cells, extracellular matrix, and vascularization (113 ± <em>19</em>/mm versus control 59 ± 14/mm) (all p < 0.05). Higher expression of vascular endothelial <em>growth</em> <em>factor</em> and epidermal <em>growth</em> <em>factor</em> was observed in the adipose-derived stem cell-transplanted expanded skin (p < 0.01 and p < 0.05, respectively), whereas the expression of <em>fibroblast</em> <em>growth</em> <em>factor</em>-2 was higher in the non-adipose-derived stem cell-treated control (p < 0.05). Transmission electron microscopy showed that a high density of collagen fibers could be seen in the adipose-derived stem cell-treated expanded skin. Cell tracking showed that the positively stained cells could be seen.</AbstractText><AbstractText>For rapid tissue expansion, adipose-derived stem cell transplantation may limit tissue destruction and improve the expansion efficiency by promoting tissue regeneration.</AbstractText>

The use of procedures adapted from a routinely successful method of culturing bovine bone has led to the first system for the study of dentinogenesis in vitro. Two types of cells have been grown from pulp obtained from the <em>growing</em> root tips of impacted third molars extracted from 14- to <em>19</em>-years olds: (1) epithelial-like cells that are probably derived from fragments of the epithelial root sheath and (2) odontoblast-like cells. The cultured epithelial-like cells grow out in distinctive rounded plaques while the odontoblast-like cells are tethered to and/or grow on top of the epithelial-like cells. The odontoblast-like cells produce mineralized matrix by 10 days when cultured on a defined mineralization formula containing conditioned medium obtained from fetal bovine bone cell cultures. <em>Growth</em> <em>factors</em> in this conditioned medium are important to cell proliferation and <em>growth</em> and to the synthesis of mineralized matrix. Sequential enzyme digestion in dispase and dispase/collagenase in serum-free Dulbecco's Modified Eagle's Medium is essential to obtaining adequate cell yields from the apical 3-5 mm of the developing root. Reduction of the number of <em>fibroblasts</em> by treating cultures with dispase in Tyrode's solution midway through the initial <em>growth</em> period enhances the purity of these cell cultures.

<em>Growth</em> <em>factors</em> lead to the induction of tissue regeneration in bone healing when coated on biomaterials. Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) combines osteoinduction and neoangiogenesis. This study evaluated bFGF-coated hydroxylapatite implants in two experimental groups with 10 or 100 microg (n = 5 per group) compared with uncoated control implants in the rabbit patellar groove model. We observed an unexpected ineffectiveness compared to the control groups with no significant difference of bone <em>growth</em> after 35 days. However, all samples from the 100 microg experiment (control and coated implant) showed significantly stronger <em>19</em>-25 day label than both 10 microg groups (control and coated implant). Earlier bone labels are stronger in the 10 microg group with equal observation of similarity between experiment and control site and may indicate a possible inhibitory effect of the higher dosing or osteoclast induction. This result indicates a possible systemic effect of the transient <em>growth</em> <em>factor</em> coating.

Bronchiolitis obliterans-organizing pneumonia (BOOP) is an inflammatory and fibrosing disease involving the distal bronchioles, bronchiolar ducts, and alveoli. We studied 91 patients with BOOP. Univariate analysis was used to relate age, sex, smoking, morphology, and expression of immunohistochemical markers CD68, D2-40, CD31, CD34, collagen IV, collagen III, platelet-derived <em>growth</em> <em>factor</em> receptor, and vascular endothelial <em>growth</em> <em>factor</em> (VEGF) with the response to corticosteroid therapy. Seventy-two patients had idiopathic BOOP and <em>19</em> secondary BOOP. The median age of the patients was 59.54 years. Most patients were current or former smokers. All cases had a patchy lesion consisting of small buds of fibromyxoid tissue in small bronchioles, bronchiolar ducts, and alveoli. The buds contained collagen and reticulin fibers, <em>fibroblasts</em>, macrophages, mononuclear inflammatory cells, and vessels in different proportions. We found no morphologic differences between primary and secondary BOOP. Patients younger than 38 years and nonsmokers had a significant good response to corticosteroid therapy. Favorable morphologic predictors were the presence of large bronchial plugs and mild inflammatory reaction (P = .093). By immunohistochemistry, the presence of collagen IV with the absence of collagen III, CD68-positive cells and positive VEGF were associated with a good response to corticosteroid therapy. We conclude that age, smoking, localization, and extension of proliferative intrabronchiolar plugs and positive immunostains for CD68, VEGF, and collagen IV with negative collagen III were useful to predict response to corticosteroid therapy and relapse.

<AbstractText>Biomarkers in irritable bowel syndrome (IBS) may guide targeted therapy in this multi<em>factor</em>ial disease. It has been suggested that 75% accuracy and cost <$500 categorise biomarkers as cost-effective.</AbstractText><AbstractText>To identify differences in faecal bile acids, faecal fat and fasting serum C4 (7a-hydroxy-4-cholesten-3-one) and <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>19</em> (FGF<em>19</em>) among patients with IBS-D, IBS-C and healthy controls and to determine accurate, cost-effective biomarkers for clinically relevant diarrhoea and constipation.</AbstractText><AbstractText>We assessed daily stool frequency and consistency (Bristol Stool Form Scale) from validated bowel diaries, 48 hours total and individual faecal bile acids, 48 hours faecal fat and weight, fasting serum C4 and FGF<em>19</em>, and colonic transit by scintigraphy from healthy volunteers (HV) and patients with IBS-D and IBS-C (Rome III criteria). We utilised multivariate logistic regression to determine biomarkers of clinically significant diarrhoea or constipation based on stool frequency, consistency and weight.</AbstractText><AbstractText>Among the 126 HV (44M/82F, 37.5 ± 10.9 years [SD]), 64 IBS-D (5M/59F, 41.9 ± 12.2 years), and 30 IBS-C (0M/30F, 44.6 ± 10 years) patients, there were significant differences between all groups in stool weight, frequency, and consistency; in addition, there were differences in colonic transit at 48 hours, faecal fat, and total and individual faecal bile acids between IBS-D and IBS-C. Reduced total and primary faecal bile acids and increased faecal lithocholic acid were significant predictors of decreased faecal weight, frequency and consistency with AUC > 0.82 (sensitivity >76%, specificity >72%). Total and primary faecal bile acids and faecal fat were significant predictors of increased stool weight, frequency and consistency with AUC > 0.71 (sensitivity >55%, specificity >74%).The faecal parameters had a 11.5 positive likelihood ratio in predicting elevated faecal weight.</AbstractText><AbstractText>Faecal bile acids and faecal fat are cost-effective and accurate biomarkers associated with significant bowel dysfunction among IBS-D and IBS-C patients.</AbstractText>

Although normal human astrocytes rarely express <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 1 (FGFR 1) mRNA, expression of FGFR 1 mRNA in astrocytomas increases as malignancy progresses. This result suggests that FGFR 1 can be one of important <em>factors</em> for glioblastoma cell <em>growth</em>. In our study, specific antisense oligonucleotides for FGFR 1 mRNA inhibited cell <em>growth</em> of SNB <em>19</em>, human glioblastoma cell line, while sense oligonucleotides showed no cell reduction. And Southern blot analysis demonstrated decreased expression of FGFR 1 mRNA in only antisense group. Furthermore, cross-linking analysis revealed decreased number of FGFR 1 in antisense-treated SNB <em>19</em> in protein level. These results conclude that cell <em>growth</em> inhibition was caused by suppression of both transcription and translation of FGFR 1 mRNA. Interestingly, alpha-specific antisense also inhibited expression of beta-, and gamma-type of FGFR 1 mRNA which had no binding sites for the oligonucleotides. This fact indicates that antisense oligonucleotides binds to premature mRNA, which is previous form of mature mRNA before splicing, in nucleus. It is previously reported that basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF), major ligand to FGFR 1, was overly expressed in human malignant astrocytomas. Thus, increased number of FGFR 1 may contribute to the acceleration of bFGF autocrine or paracrine mechanism in glioblastoma.

OBJECTIVE

To study the effects of transforming growth factor-β1 (TGF-β1) on the gene expression of connective tissue growth factor (CTGF) in cultured lung fibroblasts of embryonic rats in vitro.

METHODS

Wistar rats of embryonic 19 days were used for primary culture of lung fibroblasts (LFs). The cells in the experimental group were treated by different concentrations (1, 5 or 10 ng/mL) and different durations (12, 24 or 48 hrs) of TGF-β1 to stimulate the LFs. The cells in the control group were cultured in serum-free medium. RT-PCR method was applied to detect CTGF mRNA expression in LFs.

RESULTS

Compared with the control group, the levels of CTGF mRNA in LFs in the experimental group increased significantly (P<0.05). CTGF mRNA expression gradually increased with increasing concentration and duration of TGF-β1 treatment (P<0.05).

CONCLUSIONS

TGF-β1 can stimulate CTGF gene expression in LFs and increase CTGF gene expression in a dose-and time-dependent manner.

A previous publication (Leith et al., <em>19</em>92) showed that administration of basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF-2, 0.25 mg kg-1, q.i.d. x 7) to mice bearing xenografted DLD-2 human colon cancers would increase treated tumour <em>growth</em> rates as compared to control neoplasms. Additionally, at the end of the 7 day treatment period, clonogenic excision assays showed that the percentage of hypoxic cells in tumours from mice receiving FGF-2 administration was significantly decreased as compared to control neoplasms (from about 42 to about <em>19</em>%). The present study was undertaken to better define the kinetics of changes in hypoxic percentages as a function of tumour volume and FGF-2 treatment. In sham-injected control tumours, the hypoxic percentage increased from about 14% at day 15 postimplantation, (i.e. when sham- or FGF-2 injections were begun) to about 42% by day 22, and to about 75% at 29 days postimplantation (respective average volumes 220, 910, and 2810 mm3). In contrast, the hypoxic percentages in mice treated with FGF-2 remained at the levels seen in control mice on day 15, not only throughout the 7 day FGF-2 treatment schedule, but for at least 1 week after the cessation of <em>growth</em> <em>factor</em> administration. The hypoxic percentage was 16% on day 29 postimplantation, even though average tumour volumes were about 4325 mm3. These data show that the effect of FGF-2 administration on tumour <em>growth</em> rate and hypoxic percentages in xenografted DLD-2 neoplasms is rapid, and continues for some period of time even after administration is ended. Studies of tumour perfusion with injected 86RbCl at equivalent tumour volumes of about 1800 mm3 indicated that the percentage of cardiac output to FGF-2 treated tumours was 33% greater than in sham-injected control neoplasms.

BACKGROUND

Basic fibroblast growth factor (FGF-2), a potent angiogenic peptide, is known to be present in gonadotropes of the anterior pituitary parenchyma of rats and mice, and has been isolated from endothelial cells of many organs. Its localization within endothelial cells has not been determined, nor the mechanisms by which it might be released from endothelial cells during normal organogenesis.

METHODS

Localization of FGF within endothelial cells of the anterior pituitary was accomplished by immunocytochemistry and studied by light- and electron microscopy. Capillaries within the anterior pituitary were studied in fetal rats from day 15 to term, and in adult rats.

RESULTS

At the onset stages of vascularization (15-18 days fetal), the cytoplasm of the endothelial cells of many of the invading, immature capillaries (thick-walled with few or no fenestrations) was intensely immunopositive for FGF. Immunoprecipitate-filled blebs and slender cytoplasmic processes projected from the endothelial cells into the presumptive pericapillary space and toward the parenchymal cells. As gestation progressed (19-20 day fetal), and an increasing number of capillaries acquired the features characteristic of capillaries in the anterior pituitary of adult animals, i.e., thin-walled and fenestrated, there were fewer capillaries demonstrating immunopositivity for FGF. Foci of released FGF, i.e., extracellular, were occasionally evident within the presumptive pericapillary spaces throughout gestation. By comparison, capillaries of the anterior pituitary of adult rats did not contain immunostainable FGF in their cytoplasm, nor were any blebs and/or processes filled with immunoprecipitate evident. However capillaries did reveal an immunopositive enhancement of their lumenal and ablumenal surfaces.

CONCLUSIONS

During vascularization of the anterior pituitary, FGF within the cytoplasm of endothelial cells is released from blebs and/or processes of endothelial cells, and after the capillary bed is stabilized postnatally, these characteristics of vascularization are absent.

OBJECTIVE

To explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (HFESCs) and the feasibility of the cultured cells as the target cells for gene transfection.

METHODS

The HFESCs were isolated by means of type IV collagen rapid adhering method. The culture medium for HFESCs was prepared according to that for human fetal <em>fibroblasts</em>. The cultured cells were identified by immunohistochemistry staining of keratin-<em>19</em> and integrin-beta1, cell cycle analysis and clone forming rate determination. Then the cultured cells were gene transfected in vitro by liposome mediating method in which eukaryon expression vector pcDNA3.1/VEGF165 containing vascular endothelial <em>growth</em> <em>factor</em> 165 (VEGF165) were transfected into cultured cells, or by virus vector mediating method in which recombinant adenovirus accompanied vector (raav) containing green fluorescent protein (GFP) (raav/GFP) were transfected into the cultured cells, respectively. The results of in vitro gene transfection of HFESCs were observed by immunohistochemisty staining and fluorescence microscope.

RESULTS

HFESCs grew well and formed large clones with higher cloning efficiency and higher ratio of G1 cells than keratinocytes. The cultured cells were strongly positive with immunohistochemistry staining of keratin-<em>19</em> and integrin-beta1. After being gene-transfected by pcDNA3.1/VEGF165, the VEGF165 of HFESCs showed positive immunohistochemistry staining property, while the HFESCs transfected by raav/GFP exhibited strong fluorescence.

CONCLUSIONS

HFESCs could be isolated and cultured in vitro by means of rapid adherence to type IV collagen. It seemed feasible that HFESCs were gene transfected with liposome or adeno-associated virus as the vector.

Tubular damage and loss associated with interstitial inflammation and fibrosis may be the most important determinants in chronic renal allograft rejection. To elucidate potential pathophysiologic mechanisms associated with tubulointerstitial lesions, we examined the expression of a fibrogenic cytokine, acidic <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF-1) and its high-affinity receptors, in both relevant renal transplant controls (n=5) and tissue from patients (n=<em>19</em>) who underwent nephrectomy after graft loss, secondary to chronic rejection. In situ hybridization and immunohistochemical analyses demonstrated minimal expression of FGF-1 mRNA and protein in the tubulointerstitial compartment of the normal human kidney. In contrast, tubulointerstitial lesions in kidney allografts experiencing chronic rejection demonstrated the exaggerated appearance of both FGF-1 protein and mRNA in resident inflammatory and tubular epithelial cells. Patterns of staining were consistent throughout tubular compartments and did not appear to be localized to any particular region. The tubulointerstitium in kidneys with findings of chronic rejection also exhibited increased immunodetection of proliferating cell nuclear antigen in the tubular epithelium, inflammatory cell infiltrate, and neovascular structures. The enhanced appearance of FGF-1 and readily detectable <em>fibroblast</em> <em>growth</em> <em>factor</em> receptors suggests that this polypeptide mitogen may serve as an important mediator of <em>growth</em> and repair responses, associated with development of angiogenesis and tubulointerstitial lesions during chronic rejection of human renal allografts.

OBJECTIVE

Fibroblast growth factor-2 (FGF-2) is a multifunctional protein which plays a role in smooth muscle cell growth, wound healing, tissue repair and angiogenesis. FGF-2 is also released by mechanically wounded cells. Herein, the importance of FGF-2 release from periannular tissue in the mechanism of pannus formation in obstructed mechanical prostheses was investigated.

METHODS

Between January 1993 and December 2002, 35 patients with an obstructed bileaflet prosthetic mitral valve were classified according to the nature of obstruction as either thrombus or pannus. Data were related to patient age and gender, prosthesis model and size, intraoperative and pathology findings, and interval between implant and thrombosis. FGF-2 release was monitored immunohistochemically in all cases.

RESULTS

Thrombus formation was found in 19 patients, and pannus formation in 16. Patients were reoperated on after 3.10 +/- 0.7 years in the thrombus group, and after 6.3 +/- 0.46 years in the pannus group (p = 0.04). A foreign body reaction was found 78.9% of thrombus patients and 81.2% of pannus patients (p = 0.602), chronic inflammation in 31.5% and 50%, respectively (p = 0.317), and FGF-2 release in 78.9% and 87.5%, respectively (p = 0.582).

CONCLUSIONS

As FGF-2 release was similar in both patient groups, the duration of FGF-2 release from injured periannnular tissue was considered to form part of the chronic healing process, and was not attributed to mitral valve obstruction by pannus formation.

<AbstractText>Sensory neuropathies (SNs) are often classified as idiopathic even if immunological mechanisms can be suspected. Antibodies against the intracellular domain of the <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 3 (FGFR3) possibly identify a subgroup of SN affecting mostly the dorsal root ganglion (DRG). The aim of this study was to identify the frequency of anti-FGFR3 antibodies and the associated clinical pattern in a large cohort of patients with SN.</AbstractText><AbstractText>A prospective, multicentric, European and Brazilian study included adults with pure SN. Serum anti-FGRF3 antibodies were analysed by ELISA. Detailed clinical and paraclinical data were collected for each anti-FGFR3-positive patient and as control for anti-FGFR3-negative patients from the same centres ('center-matched').</AbstractText><AbstractText>Sixty-five patients out of 426 (15%) had anti-FGFR3 antibodies, which were the only identified autoimmune markers in 43 patients (66%). The neuropathy was non-length dependent in 89% and classified as sensory neuronopathy in 64%, non-length-dependent small fibre neuropathy in 17% and other neuropathy in <em>19</em>%. Specific clinical features occurred after 5-6 years of evolution including frequent paresthesia, predominant clinical and electrophysiological involvement of the lower limbs, and a less frequent mixed large and small fibre involvement. Brazilians had a higher frequency of anti-FGFR3 antibodies than Europeans (36% vs 13%, p<0.001), and a more frequent asymmetrical distribution of symptoms (OR 169, 95% CI 3.4 to 8424).</AbstractText><AbstractText>Anti-FGFR3 antibodies occur in a subgroup of SN probably predominantly affecting the DRG. Differences between Europeans and Brazilians could suggest involvement of genetic or environmental <em>factors</em>.</AbstractText>

Abnormal fatty acid (FA) metabolism contributes to diabetes and cardiovascular disease. The FA receptor CD36 has been linked to risk of metabolic syndrome. In rodents CD36 regulates various aspects of fat metabolism, but whether it has similar actions in humans is unknown. We examined the impact of a coding single-nucleotide polymorphism in CD36 on postprandial hormone and bile acid (BA) responses.

To examine whether the minor allele (G) of coding CD36 variant rs321<em>19</em>38 (G/T), which reduces CD36 level by ∼50%, influences hormonal responses to a high-fat meal (HFM).

Obese African American (AA) women carriers of the G allele of rs321<em>19</em>38 (G/T) and weight-matched noncarriers (T/T) were studied before and after a HFM.

Two-center study.

Obese AA women.

HFM.

Early preabsorptive responses (10 minutes) and extended excursions in plasma hormones [C-peptide, insulin, incretins, ghrelin <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF)<em>19</em>, FGF21], BAs, and serum lipoproteins (chylomicrons, very-low-density lipoprotein) were determined.

At fasting, G-allele carriers had significantly reduced cholesterol and glycodeoxycholic acid and consistent but nonsignificant reductions of serum lipoproteins. Levels of GLP-1 and pancreatic polypeptide (PP) were reduced 60% to 70% and those of total BAs were 1.8-fold higher. After the meal, G-allele carriers displayed attenuated early (-10 to 10 minute) responses in insulin, C-peptide, GLP-1, gastric inhibitory peptide, and PP. BAs exhibited divergent trends in G allele carriers vs noncarriers concomitant with differential FGF<em>19</em> responses.

CD36 plays an important role in the preabsorptive hormone and BA responses that coordinate brain and gut regulation of energy metabolism.