Prospective Study on Several Urinary Biomarkers as Indicators of Renal Damage in Children with CAKUT.

PURPOSE: The aim of the study was to investigate urinary levels of monocyte chemotactic protein-1 (MCP-1), epidermal growth factor (EGF), ß-2-microglobulin (ß2M), and FAS-ligand (FAS-L) in children with congenital anomalies of kidney and urinary tract (CAKUT) disease at risk of developing glomerular hyperfiltration syndrome. For this reason, we selected patients with multicystic kidney, renal agenesia and renal hypodysplasia, or underwent single nephrectomy. MATERIALS AND METHODS: This prospective, multicentric study was conducted in collaboration between the Pediatric Surgery Unit in Foggia and the Pediatric Nephrology Unit in Bari, Italy. We enrolled 80 children with CAKUT (40 hypodysplasia, 22 agenetic; 10 multicystic; 8 nephrectomy) who underwent extensive urological and nephrological workup. Exclusion criteria were recent urinary tract infections or pyelonephritis, age > 14 years, presence of systemic disease, or hypertension. A single urine sample was collected in a noninvasive way and processed for measuring by enzyme-linked immunosorbent assay urine levels of MCP-1, EGF, ß2M, and FAS-L. As control, urine samples were taken from 30 healthy children.Furthermore, we evaluated the urinary ratios uEGF/uMCP-1 (indicator of regenerative vs inflammatory response) and uEGF/uß2M (indicator of regenerative response vs. tubular damage). RESULTS: These results suggest that urinary levels of MCP-1 are overexpressed in CAKUT patients. Furthermore, our findings clearly demonstrated that both uEGF/uMCP-1 and uEGF/uß2M ratios were significantly downregulated in all patient groups when compared with the control group. CONCLUSION: These findings further support that CAKUT patients may, eventually, experience progressive renal damage and poor regenerative response. The increased urinary levels of MCP-1 in all groups of CAKUT patients suggested that the main factor responsible for the above effects is chronic renal inflammation mediated by local monocytes.

Time- and dose-dependent effects of chronic wound fluid on human adult dermal fibroblasts.

BACKGROUND: Wound healing is a biologic process that is altered in patients affected by chronic venous ulcers. The wound microenvironment is reflected in the chronic wound fluid (CWF), an exudate containing serum components and tissue-derived proteins. OBJECTIVES: We investigated the effects of increasing doses of CWF collected from patients suffering from chronic venous ulcers on human adult dermal fibroblasts cultured in vitro and the relationship among CWF effects and treatment length. METHODS: Fibroblasts were treated with 60, 240, and 720 microg/mL CWF for 3 and 7 days. We evaluated cell proliferation and viability by MTT and Trypan blue assay, cell morphology by light microscopy, F-actin microfilaments organization by tetramethylrhodamine B isothiocyanate-conjugated phalloidin, alpha-smooth muscle actin expression by immunofluorescence, and senescence-associated beta-galactosidase activity. RESULTS: CWF induced an increase in cell proliferation in the first 3 days of treatment. In contrast, at 7 days, a strong decrease in cell viability was observed. These changes were related to a cytoskeletal F-actin reorganization and not to fibroblast-myofibroblast differentiation nor to changes in cellular senescence. CONCLUSIONS: This study shows a dose-dependent and biphasic effect of CWF on dermal fibroblasts, suggesting that a continuous exposure to chronic wounds microenvironment may induce late cellular dysfunctions possibly involved in the delayed wound healing.

Urinary epidermal growth factor, monocyte chemotactic protein-1, and ß2-microglobulin in children with ureteropelvic junction obstruction.

BACKGROUND/PURPOSE: We demonstrated down-regulation of epidermal growth factor (EGF) and up-regulation of monocyte chemotactic protein-1 (MCP-1) in the renal parenchyma in children who underwent pyeloplasty for ureteropelvic junction obstruction (UPJO). These findings were paralleled by urinary levels of EGF and MCP-1 before and after surgery. The aim of this study is to evaluate the urinary excretion of these cytokines and ß2-microglobulin (ß2M) in children with urine flow impairment at the ureteropelvic junction or who underwent pyeloplasty. METHODS: Seventy-six patients with UPJO and 30 normal children (CTRL) were enrolled in the study. The UPJO patients were divided into obstructive (12), functional (36), and operated (28). Epidermal growth factor, MCP-1, and ß2M urinary levels were measured by enzyme-linked immunosorbent assay and normalized to urine creatinine. RESULTS: Urinary ß2M and MCP-1 increased significantly in the UPJO groups compared with the CTRL and significantly improved in the operated group. The obstructive group displayed reduced EGF excretion compared with the CTRL group. The urinary (u)EGF/uMCP-1, and uEGF/uß2M ratios significantly decreased in both untreated groups. In the operated group, these ratios improved significantly. CONCLUSIONS: The present study substantiates the role of urinary EGF, MCP-1, and ß2M as markers of tubulointerstitial damage in human obstructive nephropathy. Furthermore, it suggests that surgical intervention is effective in the management of children with UPJO.

Transcription factor decoy against promoter C of estrogen receptor alpha gene induces a functional ER alpha protein in breast cancer cells.

This study addresses the hypothesis that transfection of oligonucleotide mimicking a negative regulatory sequence of promoter C of estrogen receptor alpha (ER alpha) gene is sufficient for its re-expression in ER-negative human cancer cell lines. Even if the negative transcription regulator subtracted by the transcription factor decoy is not yet been identified, we demonstrated that after this decoy treatment, the cells produced a functional ER alpha protein able to respond to 17-beta-estradiol and to transactivate a transfected estrogen response element (ERE)-regulated reporter gene. The effects of reactivated ER alpha protein and its estrogen dependence on endogenous target gene expression level, such as ER beta, have been also assessed. The proliferation of the cells transfected with low levels of decoy was significantly increased by estrogen and not by tamoxifen, suggesting that the levels of reactivated ER alpha in these decoy conditions confers a certain hormone sensitivity. On the contrary, high-level expression of ER alpha obtained at high doses of transfected decoy molecule produced a progressive decrease of cell proliferation. Since ER alpha is important in the transcription of different genes and its loss is involved in several pathological processes including neoplastic and chronic diseases, our findings may be of relevance for a possible new therapeutical approach of such diseases.