Urinary transforming growth factor-β1 in children with obstructive uropathy

Aim: Obstructive uropathies (OU) in childhood constitute one of the major causes of chronic renal insufficiency. Transforming growth factor‐β1 (TGF‐β1) is considered to be the major fibrogenic growth factor. The aim of the present study was to investigate urinary TGF‐β1 levels in children with obstructive and non‐obstructive uropathies (NOU). Methods: This study involved 19 children with OU, 11 children with non‐obstructive hydronephrosis and 21 healthy children. Urinary TGF‐β1, proteinuria, microalbuminuria and urinary α1‐microglobulin were measured, and renal function was assesed. The results were statistically analyzed. Results: Mean urinary TGF‐β1 concentrations in patients with OU were significantly higher than those with NOU (4.14 ± 0.67 creatinine vs 1.80 ± 0.24 pg/mmol creatinine, P < 0.05) and healthy controls (1.66 ± 0.28 pg/mmol creatinine, P < 0.05). Positive correlations of urinary TGF‐β1 concentrations with proteinuria (r = 0.87, P < 0.0001) and urinary α1‐microglobulin (r = 0.82, P = 0.0002) were found in patients with OU. Conclusion: Children with OU have higher urinary TGF‐β1 than children with NOU. Urinary TGF‐β1 may be a useful non‐invasive tool for the differential diagnosis between OU and NOU in children. A positive correlation of TGF‐β1 with markers of renal tissue damage in patients with OU was found.     

Transforming growth factor-β1 in nephrotic syndrome treated with cyclosporine and ACE inhibitors

The aim of the study was to assess urinary transforming growth factor (TGF)-1 in children with steroid-dependent nephrotic syndrome (SDNS) treated with cyclosporine A (CyA) and ACE inhibitors (ACEI). The study involved 24 children (14 boys and 10 girls) with SDNS and signs of focal segmental glomerulosclerosis. The children were treated with prednisone, CyA, and ACEI. All children were examined four times: A during relapse of proteinuria, before treatment with CyA and ACEI, and B after 3 months, C 6 months, and D 12 months of treatment. The control group consisted of 20 healthy children of the same age. The urinary TGF-1 level was determined by ELISA (R and D Quantikine). The serum CyA level was measured by monoclonal antibody fluorescence polarization immunoassay. Prior to CyA treatment, the urinary TGF-1 level was the highest (135.61±38.31 pg/mg creatinine). During CyA treatment, TGF-1 was reduced to 117.96±81.57 after 3 months, to 80.26±49.52 after 6 months, and to 44.00±31.83 pg/mg creatinine after 12 months, but it was still higher than in the control group. At 3 months there was a positive linear correlation between urinary TGF-1 and proteinuria ( r =0.654, P <0.01). These results indicate that the urinary TGF-1 level increases in proportion to proteinuria during relapse of NS. Treatment with CyA and ACEI also influences urinary TGF-1, which is still higher after 12 months of treatment than in healthy children.     

Platelet parameters in children with upper urinary tract infection: is there a specific response?

Although complete blood count is routinely ordered in most upper urinary tract infections (UTI), and information regarding the patient's platelet indices is made available without added cost, the relationship between platelet count and mean platelet volume (MPV) and specific platelet responses to different infectious agents has not been extensively characterized in UTI. The objectives of this study were to examine platelet counts and platelet indices in children with culture-proven upper UTI to determine if there are organism-specific platelet responses. A retrospective analysis of data from all pediatric urine samples processed at Fatih University Medical School microbiology laboratory was undertaken for a period of two years (January 1, 2005, to December 31, 2006). Of the 200 patients with positive urine cultures, 146 (73%) were infected with gram-negative bacteria and 54 (27%) grew gram-positive bacteria. The platelet count during the episode of upper UTI and the incidence of thrombocytosis was significantly higher with the gram-positive infections than with the gram-negative infections or controls (p < 0.05). A statistically significant higher MPV was detected in the subjects with upper UTI (p < 0.05). Also, our data showed a statistically significant increase in MPV with gram-positive infections compared with the other groups (p < 0.05). In conclusion, based on the importance of the hemostatic component in the pathophysiology of infections, our findings of platelet count and MPV and predictivity of the type of the organism would suggest the usefulness of the routine measurements in children with upper UTI.     

Variation in urine composition in the human urinary tract: evidence of urothelial function in situ?

PURPOSE: An increased awareness of the concept that the urothelium has a significant transport function led us to question whether urine composition changes as it passes along the human lower urinary tract. MATERIALS AND METHODS: Urine samples from the bladder and renal pelvis were collected from 30 adults who underwent percutaneous nephrolithotomy (27) or ureteral stent insertion before lithotripsy (3). Urine was obtained from the 2 renal pelves (operative and contralateral sides) in 6 patients (24%). Urine pH was measured using an ultra-thin glass pH electrode. Urinary osmolality, Na and K were measured by micro-osmometry and flame photometry, respectively. Comparison of data sets was achieved using conventional nonparametric statistical methods. RESULTS: Median bladder urine pH in 30 patients, osmolality in 16, Na in 16 and K in 15 were significantly higher than in the renal pelvis at 6.76 (IQR 6.23 to 6.99), 469 mOsm. kg.1 (IQR 349 to 553), 132 (IQR 100 to 154) and 45 mM. (IQR 30 to 64) versus 6.08 (IQR 5.84 to 6.89), 308 mOsm. kg.1 (IQR 248 to 465), 90 (IQR 69 to 115) and 17 mM. (IQR 10 to 47), respectively (p < or = 0.05). There was no significant difference in these parameters in the urine of the paired renal pelves. CONCLUSIONS: Bladder urine pH, osmolality, Na and K significantly differ from values in the renal pelvis in moderately hydrated humans. Our data show that urine composition is modified in the lower urinary tract, supporting the concept of a dynamic urothelium. We propose that urothelial-urinary interactions and urinalysis need reappraisal, particularly in investigations of urinary stone formation and sensory bladder function.     

Association of transforming growth factor-β1 polymorphisms with the risk of chronic kidney diseases

Abstract

The association of transforming growth factor-β1 (TGF-β1) polymorphisms with the risk of chronic kidney diseases (CKD) remains elusive. We aimed to perform a meta-analysis to evaluate the relationship between TGF-β1 polymorphisms and the susceptibility to CKD. Association studies were searched according to a defined criteria using electronic databases. The strength of association between TGF-β1 polymorphisms and CKD risk was evaluated by odds ratio (OR) with the corresponding 95% confidence interval (CI). Nine case–control studies were identified. T allele at the +869 T/C polymorphism was associated with a lower risk of CKD in Asians (p?=?0.003). TT genotype at the +869 T/C polymorphism was associated with a lower risk of CKD in overall populations and Asians (p?=?0.007 and <10^?4, respectively). CC genotype at the +869 T/C polymorphism was associated with the risk of CKD in Asians (p?=?0.002). T allele at the ?509 T/C polymorphism was associated with the risk of CKD in overall populations and Asians (p?=?0.044 and 0.050, respectively). TT genotype at the ?509 T/C polymorphism was associated with CKD risk in overall populations, Caucasians and Asians (p?<?10^?4, <10^?4, and <10^?4, respectively). No evidence of significant publication bias was noted. In conclusion, T allele at the +869 T/C polymorphism may be a protective factor against CKD risk in Asians. TT genotype at the +869 T/C polymorphism may be an indicator of lower risk of CKD in overall populations and Asians. CC genotype at the +869 T/C polymorphism may predict the susceptibility to CKD in Asians. T allele at the ?509 T/C polymorphism may be an indicator of CKD risk in overall populations and Asians. TT genotype at the ?509 T/C polymorphism was a risk factor for CKD onset in overall populations, Caucasians and Asians.     

What is the effect of circumcision on risk of urinary tract infection in boys with posterior urethral valves?

Purpose

Boys with posterior urethral valves (PUV) have increased risks of urinary tract infection (UTI) voiding dysfunction and ongoing renal damage. Circumcision has been shown epidemiologically to reduce UTIs, but no trial has yet confirmed this in PUV. Circumcision is not routinely performed in boys with PUV in our unit, but one quarter of our patients are circumcised for religious reasons. It may be hypothesized that circumcision reduces the risk of subsequent urinary tract infection in boys with PUV. This study aims to test this hypothesis by comparing the risk of UTI, and subsequent renal outcome, in PUV in uncircumcised boys with those who were circumcised.

Methods

A retrospective cross-sectional case note review of boys with PUV was performed, and the following were documented: age at presentation, method of diagnosis, method of treatment, initial renal status, and timing of treatment; use and timing of urinary tract diversion; timing of circumcision; and UTIs—date, organism, and treatment.

Results

Seventy-eight patients were identified, mean age 6.7 years (range, 1-18). These boys experienced 78 UTIs in the uncircumcised state. Subsequently, 27 were circumcised, experiencing 8 UTIs. Eighteen boys were diverted. The incidence of UTI was reduced from 0.50 ± 0.14 (mean ± SEM) UTIs annually uncircumcised to 0.09 ± 0.02 (mean ± SEM) circumcised (P < .01, Student's t test).

Conclusion

In PUV, circumcision reduces the incidence of UTI by 83%, every circumcision prevents 1 UTI on average. Early circumcision in all PUV is beneficial, but a larger randomised control trial should be considered to confirm this.     

Managing children under 36 months of age with febrile urinary tract infection: a new approach

Background  

Recent guidelines on urinary tract infection (UTI) agree on reducing the number of invasive procedures. None of these has been validated by a long-term study. We describe our 11-years experience in the application of a diagnostic protocol that uses a reduced number of invasive procedures.     

Transforming growth factor-β in renal disease with glycogen storage disease I

We report a 14-year-old patient with Japanese glycogen storage disease I (GSD-I) who was found to have proteinuria. Renal biopsy revealed massive tubular atrophy and interstitial fibrosis with mononuclear cell infiltration, but the glomeruli were almost normal. The epithelial cells of tubules contained periodic acid-Schiff-positive glycogen deposits digested by diastase. In an immunohistological study, transforming growth factor (TGF)- expression was increased in tubular epithelial cells compared with a normal control kidney specimen. These data suggest that increased TGF- expression is involved in the pathophysiology of renal interstitial fibrosis in a patient with GSD-I.     

Plasma transforming growth factor-β1 levels in patients with erectile dysfunction

Aim: To evaluate the plasma TGF-β1 level in erectile dysfunction (ED) patients of various causes. Methods: Sixty-two patients with ED and 26 potent men were subjected to the study. Based on multidisciplinary work-ups, including medical history, physical examinations, blood tests with lipid profile and hormones, penile duplex Doppler ultrasonogram and neurophysiological tests, causes for ED were classified as psychogenic (n=15), neurogenic (n=16) and vasculogenic (n=31). The plasma TGF-β1 level was measured by the ELISA method. Results: The plasma TGF-β1 level was significantly increased in the ED group (6.7 ± 4.9 ng/mL), compared to the control (4.0±2.1 ng/mL) (P <0.01). In the ED groups, there was a significant increase in the vasculogenic group (9.0 ± 5.5 ng/mL), compared to the psychogenic (3.8 ± 1.8 ng/mL) and neurogenic groups (4.8 ± 3.2 ng/mL) (P<0.01). Of the vascular risk factors, both the smoking (7.5 ± 4.7 ng/mL) and dyslipidemia groups (7.4 ± 4.4 ng/mL) showed significantly increased     

Change of autophagy in the transforming growth factor-β1-induced activation of renal fibroblasts in vitro

Objective To explore the changes of autophagy in the transforming growth factor (TGF)-β1-induced activation of renal fibroblasts in vitro. Methods (1) NRK-49F cells were cultured with 10 μg/L TGF-β1 for different times (0, 12, 24 h) in vitro. Morphological changes of the cells were observed under inverted microscope. The protein expressions of α-smooth muscle actin (α-SMA) and typeⅠcollagen (ColⅠ) in NRK-49F cells were measured by Western blotting. (2) NRK-49F cells were cultured with 0, 2, 5, 10, 15, 20 μg/L TGF-β1 for 1 hour and with 10 μg/L TGF-β1 for different times (0 min, 7 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h) in vitro. The protein expressions of microtubule-associated protein 1 light chain 3(LC3), p62, total- mammalian target of rapamycin (t-mTor), phospho-mammalian target of rapamycin (p-mTor) and Beclin 1 were detected by Western blotting. (3) NRK-49F cells were cultured with 10 μg/L TGF-β1 for different times (0, 1, 4 h) in vitro after cultured with serum-free medium for 2.5 hours. The protein expressions of LC3 and p62 in NRK-49F cells were measured by Western blotting. Results (1) The morphology of NRK-49F cells changed from stellate to spindle shape after cultured with TGF-β1. The expressions of cell activation markers α-SMA and ColⅠgradually increased as the extend of stimulation time (all P＜0.05). (2) TGF-β1 transiently increased the expressions of autophagy proteins p62 (peak value appeared after 4 h) and p-mTor (peak value appeared after 30 min), while decreased Beclin1 expression level (all P＜0.05). (3) TGF-β1 decreased the protein expression of LC3-Ⅱ in NRK-49F cells cultured with serum-free medium, whereas increased the protein expression of p62 at the same time (all P＜0.05). Conclusions The autophagy activity of renal fibroblasts is inhibited by the TGF-β1-induced cellular activation in vitro, which may contribute to the progression of renal interstitial fibrosis.     

The efficacy of ultrasound and dimercaptosuccinic acid scan in predicting vesicoureteral reflux in children below the age of 2 years with their first febrile urinary tract infection

We evaluated the efficacy of dimercaptosuccinic acid (DMSA) scan and ultrasound (US) in comparison with voiding cystourethrography (VCUG) in predicting vesicoureteral reflux (VUR) in children below the age of 2 years. Medical records and radiologic studies of children (<2 years) suffering their first febrile urinary tract infection (UTI) between January 2001 and May 2007 were retrospectively reviewed. We evaluated the sensitivity, specificity, and positive and negative predictive values of US and DMSA scans in diagnosing VUR. Among 220 children with their first febrile UTI, VUR was detected in 67 (30.4%). The detection rate of VUR by US was 41.7% and 86% in the low (I, II) and high grade (III~IV) groups, respectively. Detection rate of VUR by DMSA scan was 37.5% and 88.4% in the low and high grade groups, respectively. Combining US and DMSA scan, we found that the detection rate of high grade VUR was 95.3% and that of low grade was 62.5%. During follow up, most of the low grade VURs with normal DMSA and US scans resolved or were downgraded. Most high grade VURs could be detected by US and DMSA scan, but the prediction rate was not as high in low grade VURs. However, we can anticipate spontaneous improvement without complications in patients with either low or high grade reflux and negative findings on US and DMSA scan.     

Elevation of serum transforming growth factor-β1 Level in patients with metastatic prostate cancer

We measured serum transforming growth factor β1 (TGF-b1) levels in patients with untreated prostate cancer and compared them with other prognostic indicators, specifically serum prostate-specific antigen (PSA) and the Gleason histopathologic grading score. Prior to treatment, the Sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure TGF-b1 concentrations directly in sera from 55 patients with prostate cancer (21 localized and 34 metastatic), 13 age-matched healthy male control subjects, and 8 patients with benign prostatic hyperplasia (BPH). Serum TGF-b1 levels in patients with lymph node and/or distant metastases were significantly higher than in patients with localized disease (p = 0.0003), but did not differ significantly among localized cancers as to tumor extension. Ten of 11 prostate cancer patients whose serum TGF-b1 was higher than an arbitrary cut-off value of 60 ng/ml had lymph node or distant metastasis (specificity 95.20, although in 24 of 34 metastatic patients serum TGF-b1 levels were less (sensitivity 29.4%). The correlation between serum TGF-b1 and tumor grade as assessed by the Gleason scoring system was weak (r = 0.340). Moreover, serum TGF-b1 was not correlated with the serum PSA level. Interestingly, there was an inverse correlation between serum TGF-b1 in the prostate cancer patients and patient age at diagnosis (r = -0.406). These findings suggest that elevation of the serum TGF-b1 levels reflects certain malignant potentials associated with metastasis that are unpredictable by PSA and the Gleason scoring system.     

Levels of urinary transforming growth factor β-1 in children with D+ hemolytic uremic syndrome

About 25–50% of survivors of the acute phase of postdiarrheal hemolytic uremic syndrome (D+ HUS) develop chronic renal disease. Transforming growth factor β-1 (TGFβ-1) is the main fibrogenic growth factor in humans, and there is a significant correlation between its levels and the grade of interstitial fibrosis in chronic nephropathies. We hypothesized that increased urinary TGFβ-1 may be an early indicator of sequelae in D+ HUS patients who show no sign of renal damage as determined by conventional diagnostic tests. We therefore compared the levels of TGFβ-1 in urine collected from healthy controls (HC) (n = 18) with that from patients with a past history of D+ HUS (n = 39). We found that TGFβ-1 excretion was significantly higher (p < 0.001) in the patient group (median level 73 pg/mg creatinine) than in the HC (median level 28 pg/mg creatinine). TGFβ-1 excretion did not correlate with age, white blood cell count, length of oligoanuric period, maximum creatinine at the acute stage, or length of the follow-up. Since TGFβ-1 excretion may reflect ongoing renal tissue damage, our results emphasize the need for the lifelong follow-up of patients with a past history of D+ HUS, even those showing apparent recovery. Long-term monitoring of this cohort is necessary to determine the clinical utility of our findings.     

Transforming growth factor-β pathway is activated in cholecystolithiasis

Background The etiopathogenesis of cholecystolithiasis is not well defined. Primary dysmotility of the organ, due to fibrosis of the gallbladder wall or muscular dysfunction, is suggested as a crucial factor. Transforming growth factor-beta (TGF-) and connective tissue growth factor (CTGF) are involved in several fibrotic disorders and play a critical role in fibrogenesis, thereby changing the physiological function of the organs. In the present study we analyzed the role of TGF- and its downstream target CTGF in patients with cholecystolithiasis.Methods Gallbladders were obtained from 16 individuals undergoing surgery for symptomatic cholecystolithiasis. Normal human gallbladder tissue samples from five individuals without any history of gallbladder disease were obtained through an organ donor transplantation program. Northern blot analysis, in situ hybridization, and immunohistochemistry were used to analyze the expression of TGF-1 and CTGF in the gallbladder tissue samples.Results By northern blot analysis there was an enhanced TGF-1 mRNA expression (eightfold increase; P<0.04) in the cholecystolithiasis tissue samples in comparison with normal controls. There was also a concomitant increase in CTGF (41-fold increase; P<0.01). By in situ hybridization and immunohistochemistry, CTGF mRNA was localized mainly in the mucosa layer, while intensive staining of the smooth muscle cells with TGF-1 and CTGF was observed. In addition, TGF-1 immunoreactivity was also localized in the fibroblasts and inflammatory cells. TGF-1 m-RNA levels showed a significant relationship with the degree of fibrosis in the tissue samples (P<0.04, r=0.5).Conclusion Our data indicate that TGF- and CTGF are involved in ultrastructural tissue changes in patients with cholecystolithiasis. Activation of the TGF- pathway, predominantly in the remaining mucosa and submucosal layer, indicates that extracellular matrix (ECM) synthesis with subsequent gallbladder wall fibrosis is an important step in gallbladder dysfunction in this disorder.     

Transforming growth factor-β in calcium alginate beads for the treatment of articular cartilage defects in the rabbit

Purpose: Articular cartilage has only limited capability for intrinsic repair. The use of growth factors has been suggested to improve the repair of cartilage after injury. Reliable delivery systems for these agents are needed. In this study we tested calcium alginate for the delivery of TGF-β in the treatment of osteochondral defects in the rabbit knee. Type of Study: Randomized trial animal study and basic science study. Methods: In vitro, to establish the kinetics of TGF-β release from the alginate, ¹²⁵I- labeled TGF-β was suspended in 1.2% sodium alginate at concentrations of 1 μg/mL and 10 μg/mL. Beads were formed from 50 μL aliquots and placed into standard culture medium by immersion in calcium chloride solution and incubated at 37° C. A gamma counter was used to measure the amount of TGF-β that was released into the medium at various time points. In vivo, osteochondral defects were created in the trochlear grooves of 32 New Zealand White rabbits. Defects were treated with plain alginate or with alginate containing TGF-β at 20 ng/mL or 2,000 ng/mL. Untreated defects served as a control. Animals were killed after 6 and 12 weeks. Knee joints were evaluated grossly with a 12-point grading scale. Histologic sections of the repair tissue were stained with Safranin O and evaluated using a 24-point grading scale by 2 independent blinded observers. Mean scores and standard deviations were calculated. P values were determined using the Student t test. Results: The TGF-β was released at a surprisingly slow but steady rate. Release rates extrapolated from the gamma counter measurements were 0.25% per hour and 0.33% per hour, for the 1 μg/mL and 10 μg/mL beads, respectively. Gross analysis scores at 6 and 12 weeks resulted in higher scores for both TGF-β groups without reaching statistical significance. The lower TGF-β concentration reached the highest scores, whereas the higher concentration (2,000 ng/mL) resulted in increased osteophyte formation. Histologic analysis at 6 weeks resulted in average scores ranging from 14.5 for empty defects and 18.1 for alginate-treated defects, to 20.0 and 20.3 for the 2,000 ng/mL and 20 ng/mL TGF-β groups, respectively (P <.05). At 12 weeks, histologic scores ranged from 14.9 for empty and 14.5 for alginate to 20.1 and 20.5 for the 2,000 ng/mL and 20 ng/mL TGF-β groups, respectively (P <.05). These results indicate a significant improvement of the quality of the repair tissue at 6 and 12 weeks with TFG-β treatment, especially at the lower concentration. Conclusions: The use of alginate allows the controlled delivery of TGF-β selectively to the site of injury, potentially avoiding systemic side effects. Furthermore, treatment with TGF-β appears to improve the repair of articular cartilage defects. Longer-term studies are needed to assess whether the benefits of the TGF-β treatment can be sustained.Arthroscopy: The Journal of Arthroscopic and Related Surgery, Vol 18, No 8 (October), 2002: pp 892–900     

Polythelia: still a marker of urinary tract anomalies in children?

OBJECTIVE: Supernumerary nipples (SNN), or polythelia, are the most common form of the accessory mammary tissue malformation. The frequency of this condition ranges from 0.2% to 5.6% depending on various factors. This condition is associated with several anomalies, although this association is often controversial. The aim of this study was to evaluate the association between SNN and kidney/urinary tract (K/UT) anomalies, where anomalies is taken to mean functional disorders, malformations and diseases. MATERIAL AND METHODS: A case-control study was performed. The study evaluated 166 children (case group) referred to the Pediatric Nephrology Unit of the Department of Pediatrics of the Catholic University of Rome and 182 children (control group) admitted to the Department of Pediatrics because of pathologies not involving the urinary tract. RESULTS: There were 11 children with SNN in the case group, and only two patients in the control group (6.62% vs 1.09%, p<0.05). CONCLUSION: The results show a high incidence of K/UT anomalies in children with SNN, and therefore K/UT should be investigated in this specific population.     

What is the efficacy of circumcision in boys with complex urinary tract abnormalities?

The risk of urinary tract infection (UTI) in normal boys is 1%. This risk is significantly increased in boys with congenital abnormalities of the urinary tract, which includes such abnormalities as vesico-ureteric reflux, obstructive megaureter (VUJO) and posterior urethral valves. UTI in these boys can lead to urosepsis, a potentially life-threatening complication, and in the longer term renal scarring complicating pyelonephritis can lead to chronic renal impairment or even end-stage renal disease. Circumcision has been shown in normal boys to reduce the risk of UTI by 90%, and potentially could be a simple intervention to reduce the risk of urosepsis and renal scarring. In order to make this decision a clinician really needs to have the answers to two questions: 1) What is the risk of UTI in this particular boy? 2) What is the evidence of efficacy of circumcision in this particular condition? This article reviews what evidence exists to make a calculation of the risk/benefit ratio for circumcision in boys with abnormalities of the urinary tract.