Novel Urinary Biomarkers for Diagnosis of Acute Pyelonephritis in Children.

Urinary tract infection (UTI) is common among pediatric population. Pyelonephritis, especially in young infants, is associated with a significant morbidity. Usually, clinical manifestations, laboratory findings, and imaging are used to differentiate between lower and upper UTI. Lack of specific clinical findings and commonly used nonspecific blood indices are important hamper differentiation between lower and upper UTI in early stages. Imaging techniques are neither cost benefit nor safe for detection of UTI. Recent efforts have focused on characterization of novel serum and urinary biomarkers for early detection of acute pyelonephritis in children. It seems that urinary NGAL, NAG, TNF-α and IL-8 may be used as novel markers for early diagnosis of acute pyelonephritis in children.

Symptom- and urinalysis-based approach to diagnosing urinary tract infections in children with neuropathic bladders.

BACKGROUND: Accurately diagnosing urinary tract infections (UTI) in children with neuropathic bladders can be difficult given the lack of specificity of both clinical symptoms and routine screening tests. We aimed to identify a priori unknown classes/groups of children with neuropathic bladder with respect to symptoms and UA results and examine their relationships with odds of UTI. METHODS: We used latent class analysis (LCA) to identify unobserved classes/groups of children with neuropathic bladder based on symptoms and urinalysis (UA) results, respectively. Demographic and clinical data were gathered by retrospective chart review of a cohort with neuropathic bladder. Symptoms and UA results were obtained by chart review of visits where urine culture was ordered. RESULTS: Around 193 patients were included in UA results analysis and 179 in symptom-based analysis. Two latent classes of patients were identified with respect to symptoms, labeled "pyelonephritis class" and "cystitis class," and two, with respect to UA results, were labeled "positive UA class" and "negative UA class." The pyelonephritis class had significantly higher odds of UTI compared to the asymptomatic class. While odds of UTI in cystitis class were higher than the asymptomatic class, this difference was not statistically significant. Positive UA class had significantly higher odds of UTI compared to negative UA class. CONCLUSION: Two unobserved classes/groups exist in children with neuropathic bladder with respect to symptoms, corresponding to cystitis and pyelonephritis, and two classes of UA results that correspond with either a positive or negative UA. Our results suggest a differential approach to treatments may be considered.

Association of mRNA Levels of IL6, MMP-8, GSS in Saliva and Pyelonephritis in Children.

Nowadays, saliva is a subject of growing scientific interest because of its definite advantages as diagnostic medium. The aim of our study was to investigate the diagnostic potential and reliability of messenger RNAs (mRNAs) of selected genes-interleukin-6 (IL-6), matrix metalloproteinase-8 (MMP-8) and glutathione synthetase (GSS)-as salivary markers in children with diagnosed pyelonephritis and to correlate their levels with typical urine para-clinical indicators of the disease. Analysis of the mRNA levels for IL-6, MMP-8 and GSS in 28 children hospitalized with the diagnosis of pyelonephritis was conducted applying the method of quantitative reverse transcription polymerase chain reaction (RT-qPCR). In the study group (n = 28), IL-6 mRNA levels demonstrated 64-fold increase (p < 0.001). MMP-8 and GSS mRNA levels were increased in 12 samples in patients with pyelonephritis 3.27 (p < 0.01) and 1.94 (p < 0.001) times, respectively. We found a strong and significant correlation (p < 0.001) between the investigated mRNA for IL-6 and MMP-8, IL-6 and GSS, MMP-8 and GSS. Moderate degree of correlation was established between IL-6 and the typical para-clinical indicator of leucocytes (0.43, p < 0.05) and between GSS and leucocytes (0.54, p < 0.01). Salivary IL-6, MMP-8 and GSS mRNA levels in combination with urine test analysis could be useful diagnostic tool for the very distributed disorder of pyelonephritis in childhood.

Assessment the Effect of Dexamethasone on Urinary Cytokines and Renal Scar in Children with Acute Pyelonephritis.

INTRODUCTION: One of the most serious complications of acute febrilepyelonephritis in children is the development of renal scar. Thisstudy aimed to investigate the effect of dexamethasone on urinarycytokine levels and renal scar in children with acute pyelonephritis. METHODS: In a double-blind randomized clinical trial, 60 childrenaged 3 months to 12 years with acute febrile pyelonephritis enrolled.The experimental group was treated with a combination of antibioticand dexamethasone, and the control group underwent treatmentwith antibiotic and placebo. The urinary levels of interleukin -6(UIL-6) and -8 (UIL-8) were measured before treatment as baselineand were repeated four days later. RESULTS: 52 cases (23 patients with mean age of 34.19 ± 30.82 monthsin the dexamethasone group, and 29 patients with mean age of50.55 ± 44.41 months in the control group) completed the study. Inthe control group, the UIL-6 and UIL-8 level became significantlylower after four days treatment (P < .05). In the dexamethasonegroup, there was a statistically significant difference between bothUIL-6 and UIL-8 levels before and after treatment (P < .05). Inpatients who had scar on DMSA scan, the mean UIL-8 and UIL-6levels were significantly high before and after treatment. CONCLUSION: Results of this study showed that dexamethasone plusantibiotic have no clear superiority to antibiotic therapy alone indecreasing inflammatory cytokines and scar formation. We foundout that patients with scar had sustained high levels of biomarkersbefore and after treatment.

Host and Bacterial Markers that Differ in Children with Cystitis and Pyelonephritis.

OBJECTIVE: To determine whether treatment for urinary tract infections in children could be individualized using biomarkers for acute pyelonephritis. STUDY DESIGN: We enrolled 61 children with febrile urinary tract infections, collected blood and urine samples, and performed a renal scan within 2 weeks of diagnosis to identify those with pyelonephritis. Renal scans were interpreted centrally by 2 experts. We measured inflammatory proteins in blood and urine using LUMINEX or an enzyme-linked immunosorbent assay. We evaluated serum RNA expression using RNA sequencing in a subset of children. Finally, for children with Escherichia coli isolated from urine cultures, we performed a polymerase chain reaction for 4 previously identified virulence genes. RESULTS: Urinary markers that best differentiated pyelonephritis from cystitis included chemokine (C-X-C motif) ligand (CXCL)1, CXCL9, CXCL12, C-C motif chemokine ligand 2, INF γ, and IL-15. Serum procalcitonin was the best serum marker for pyelonephritis. Genes in the interferon-γ pathway were upregulated in serum of children with pyelonephritis. The presence of E coli virulence genes did not correlate with pyelonephritis. CONCLUSIONS: Immune response to pyelonephritis and cystitis differs quantitatively and qualitatively; this may be useful in differentiating these 2 conditions.

Urinary neutrophil gelatinase-associated lipocalin, kidney injury molecule-1, uromodulin, and cystatin C concentrations in an experimental rat model of ascending acute kidney injury induced by pyelonephritis.

Recent evidence suggests that neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), cystatin C (CysC), uromodulin (UMOD), and some interleukins (IL-6 and IL-18) can be considered as diagnostic markers of acute kidney injury (AKI). The aim of this study was to verify the applicability of four urinary (u) markers, namely uNGAL, uKIM-1, uCysC, and uUMOD, for the diagnosis of ascending AKI induced by bacterial pyelonephritis. The study included 30 female rats that were divided into three groups (n = 10 each) and were inoculated transurethrally with various doses of Escherichia coli to induce isolated pyelonephritis (group 1, 105 CFU/ml), pyelonephritis-induced AKI (group 2, 107 CFU/ml), or AKI and urosepsis (group 3, 109 CFU/ml). The inoculate contained a highly virulent E. coli strain isolated from a patient with pyelonephritis. Urine samples were obtained prior to the inoculation and 7, 14, and 21 days thereafter. The concentrations of all assessed proteins were determined in the urine samples by ELISA. All the study groups showed elevated concentrations of uNGAL and uCysC at all study time points. The concentrations of uKIM-1 in group 1 were the same as that at the baseline, whereas it was elevated in groups 2 and 3 at all study time points. The concentrations of uUMOD in groups 1 and 2 tended to decrease with the time from inoculation, whereas it rapidly increased in group 3 at 21 days postinfection. uKIM-1 seems to be the only marker of ascending AKI associated with urinary tract infection. Elevated concentrations of uNGAL, uCysC, and uUMOD were found in both AKI and isolated pyelonephritis. Thus, it can be concluded that none of these markers can be used as a single diagnostic marker of ascending AKI, as it may produce false-negative results, leading to incorrect diagnosis, lack of adequate treatment, and increased mortality risk.

[Features of the proteome of the urine in chronic pyelonephritis.]

Clinical observation and examination of 12 patients with chronic pyelonephritis (CPN) were performed. The first group (GI) included patients with exacerbation of the disease. In the comparison group (GII)- the same patients after 1.5-3 months after completion of treatment, without clinical manifestations of exacerbation of CPN. Laboratory signs of acute renal damage were not revealed in all examined patients. Additionally, urine was collected in the afternoon after Breakfast, in the form of a freely separated 2nd fraction and its sample preparation, consisting of the stages: recovery, alkylation, protein deposition and proteolysis using trypsin. The resulting polypeptide mixture was separated by liquid chromatography in three repetitions and analyzed on a system consisting of Agilent 1100 chromatograph and ltq-FT ultra hybrid mass spectrometer. A list of proteins was obtained, indicating the number of peptides by which they were identified, and the parameters of its reliability. Most of the information about the obtained proteins was obtained from UniProt databases. Identified and analyzed 10 proteins that differ significantly in occurrence in the clinical group of patients in the period of exacerbation of PN. The appearance of these proteins in urine in 1patients with exacerbation of chronic PH allows us to consider them as potential biomarkers directly associated with inflammation and damage to the epithelial lining of the renal tubules.

Validity of Neutrophil Gelatinase Associated Lipocaline as a Biomarker for Diagnosis of Children with Acute Pyelonephritis.

PURPOSE: Novel biomarkers have been investigated for various renal disorders, including urinary tract infection (UTI). The aim of this study was to assess whether urine neutrophil gelatinase associated lipocaline (NGAL), could represent a reliable biomarker for diagnosis and treatment of children with acute pyelonephritis (APN). MATERIALS AND METHODS: A total of 37 children (32 females, 5 males) with APN were included in this prospective study. Urine NGAL was measured before and 5-7 days after antibiotic treatment in the UTI group, using ELISA kit and compared with 26 (8 females, 18 males) control group children admitted for other bacterial infections. RESULTS: Mean age of the UTI group was 39 ± 28 months, compared to 43.6 ± 31.5 months for the control group with no statistically significant difference. Median urine NGAL level was significantly higher in patients with APN than the other subjects [0.48 (interquartile range (IQR): 0.15-0.72) vs. 0.065 (0.01-0.24), P = .001], and decreased significantly after antibiotic treatment (P = .002). Using a cutoff of 0.20 ng/mL, sensitivity and specificity of urine NGAL were 76% and 77% for prediction of APN, respectively. The area under the ROC curve (AUC) for urine NGAL was 0.75 (CI= 0.61-0.88), suggesting urine NGAL as a relatively good predictive biomarker of APN. CONCLUSION: Urine NGAL is a good biomarker for diagnosis and treatment monitoring of APN in children. .

Differentiation of acute pyelonephritis from other febrile states in children using urinary neutrophil gelatinase-associated lipocalin (uNGAL).

BACKGROUND: Acute pyelonephritis is a severe disease which is sometimes difficult to recognize based on clinical symptoms and routinely available diagnostic tests, especially in young children. The aim of this study was to assess the diagnostic value of urinary neutrophil gelatinase-associated lipocalin (uNGAL) as a biomarker of acute pyelonephritis. METHODS: In this case-control study we analyzed 134 children (median age 2.5 years) who were admitted to the Pediatric Clinic of University Hospital Centre Osijek, Croatia. Eighty of them had acute pyelonephritis, while 54 children had febrile state of different etiology including cystitis and they represented the control group. uNGAL, white blood cells, C-reactive protein, urinanalysis, urine culture, kidney ultrasound and a dimercaptosuccinic acid scintigraphic scan were done for each child. uNGAL was measured using chemiluminiscent microparticle immunoassay on ARHITECT i1000SR (Abbott Diagnostics, IL, USA). RESULTS: uNGAL values were significantly higher in children with acute pyelonephritis compared to the control groups (113.6 ng/mL vs. 10.2 ng/mL, p<0.001). A receiver operating characteristic curve comparison was done for tested parameters and encouraging results were obtained for uNGAL (AUC=0.952). A cut-off value of 29.4 ng/mL had 92.5% sensitivity and 90.7% specificity. We showed that uNGAL can also serve in differentiating acute pyelonephritis from cystitis (cut-off 38.5 ng/mL), and for differentiation of cystitis from febrile states with etiology other than urinary tract infection (UTI) (cut-off 20.4 ng/mL). CONCLUSIONS: uNGAL can be a useful diagnostic biomarker in acute pyelonephritis in children, but also in differentiating cystitis from febrile states other than UTI.

[Molecular mechanisms of individual platelet reactivity in hematuria secondary to lithotripsy].

AIM: To investigate the mechanisms of individual platelet reactivity to ADP and adrenaline associated with the variability of hematuria after lithotripsy in patients with chronic obstructive pyelonephritis (COPN). MATERIALS AND METHODS: The study included 41 COPN patients admitted to the Department of Urology for lithotripsy (LT). The contact ultrasonic LT was performed using the Karl Storz Calcuson Ultrasonic Lithotripsy System. Postoperative hematuria was assessed by microscopic red blood cell count. Platelets were separated from the citrated peripheral blood by centrifugation. Platelet aggregation was measured by Chrono-log aggregometer using agonists (ADP, adrenaline) at a concentration of EC50 and EU10. RESULTS: There were three types of platelet functional response to ADP and adrenaline after LT (increased, unchanged and decreased aggregation), but the predominant type of individual response was increased platelet aggregation. Testing 24 hours after LT revealed 7 platelet phenotypes differing in functional activity of 2-adrenoceptor agonist and purine receptors (R2Y1 and R2Y12). Normal purine receptor activity was associated with the ability of platelets to respond to adrenaline by increasing the functional activity aimed at limiting hematuria. Reduced platelet response to ADP after LT reaching the level of hyporesponsiveness may be viewed as a predictor of severe hematuria after surgery. CONCLUSION: Individual platelet reactivity, manifested by the interaction of ADP and adrenaline agonist, determines the effectiveness of the increase in pro-aggregation capacity of platelets in developing postoperative hematuria.

Predictive value of urinary and serum biomarkers in young children with febrile urinary tract infections.

BACKGROUND: Early predictive biomarkers for the diagnosis and management of febrile urinary tract infections (UTIs) can be valuable diagnostic tools in children. METHODS: The study cohort comprised 73 pediatric patients with febrile UTIs [46 with acute pyelonephritis (APN) and 27 with lower UTIs] and 56 healthy children. Urine neutrophil gelatinase-associated lipocalin (uNGAL) and kidney injury molecule-1 (uKIM-1) levels and serum cystatin C (sCysC) levels were measured. RESULTS: The uNGAL/creatinine (Cr) and uKIM-1/Cr levels were higher in the UTI group than in the controls (P < 0.05). uNGAL/Cr and sCysC levels were higher in patients with APN than in those with lower UTIs (P < 0.05). uNGAL/Cr levels in both the APN and UTI groups decreased following the administration of antibiotics compared to those before treatment (P < 0.05). The uNGAL/Cr level was correlated with serum levels of white blood cells, C-reactive protein, CysC and with uKIM-1/Cr (P < 0.05). uKIM-1/Cr was also correlated with sCysC (P < 0.05). Receiver operating curve analyses showed good diagnostic profiles of uNGAL/Cr and uKIM-1/Cr for identifying UTIs [area under the curve (AUC) 0.9 and 0.66, respectively) and of uNGAL/Cr and sCysC for predicting APN (AUC 0.78 and 0.72, respectively). CONCLUSIONS: Our results suggest that uNGAL, uKIM-1 and sCysC levels may be useful for predicting and managing febrile UTIs in children.

[Modern principles of diagnosis and prediction of course of chronic obstructive pyelonephritis in children].

An assessment of changes in the levels of cytokines, cell adhesion molecules, acute phase proteins in serum, urine and biopsy using ELISA was performed. The evaluation was performed in 167 patients with various congenital urinary tract obstructions in the perioperative period. It is shown that serum C-reactive protein and IL-6, IL-8 urine levels have high diagnostic significance for determining the severity of inflammation in the urinary tract in the development of chronic obstructive pyelonephritis at different levels of obstruction. The calculation of the proposed index of inflammation activity based on IL-8 and IL-10 urine levels in children in the dynamics allows to estimate the phase of inflammation and efficacy of the treatment.

Urinary adrenomedullin level in children with acute pyelonephritis before and after treatment.

INTRODUCTION: Adrenomedullin (AM) is a 52-amino acid peptide that causes vasodilatation by increased synthesis of nitric oxide. Its production by different cells such as cardiac myocytes, smooth muscle, endothelial, and oncogenic cells is stimulated by inflammatory processes. It has been shown that in the presence of inflammation in the urinary system, concentration of AM increases. In this study, we measured urinary AM in children with acute pyelonephritis before and after treatment and compared its level with that in healthy children. MATERIALS AND METHODS: In a case-control study, 31 children with clinical and paraclinical documentation of pyelonephritis (case group) and 30 healthy children without pyelonephritis or other infections (control group) were studied. Urinary AM were measured on spot urine samples by high-performance liquid chromatography, and creatinine was measured by spectrophotometry to report the AM-creatinine ratio. RESULTS: Urinary AM-creatinine ratios were 61.3 +/- 119.4 pg/mg and 4.26 +/- 11.4 pg/mg, respectively, in the case and control groups (P = .01). After treatment of pyelonephritis in the patients of the case group, this ratio decreased to 13.1 +/- 21.9 (P = .048). The coefficient correlation between urinary AM and leukocytes count was 0.252 (P = .17). Urinary AM levels were 1896 +/- 1748 pg/dL and 391 +/- 477 pg/dL in the patients with 4+ versus negative C-reactive protein levels, respectively (P = .008). CONCLUSIONS: Urinary AM increases in the course of pyelonephritis and decreases significantly after treatment.

Urinary concentration of cytokines in children with acute pyelonephritis.

Urinary tract infection (UTI) is a common bacterial infection among infants and children. Predicting which children with upper UTI will develop long-term sequelae remains difficult. We aimed at evaluating the predictive value of urine concentrations of interleukin-6 (UIL-6) and interleukin-8 (UIL-8) in subsequent renal scarring. In the current observational prospective study, urine samples for UIL-6 and UIL-8 were obtained from two groups: 31 children with first episode of febrile UTI and 22 febrile children of other origin. UIL-6 and UIL-8 were increased in children with febrile UTI, compared to children with fever of other origin [median and range (picograms per milliliter): (1) UIL-6, 74.46 (0-168) vs. 10.51 (0-47.50), respectively, p = 0.0001; (2) UIL-8, 2,660.38 (0-13,801) vs. 0, respectively, p = 0.0001]. Renal scarring was found in 5/31 (16 %) children with acute pyelonephritis. Initial median UIL-8 values were significantly higher in children with later renal scarring than in those without renal scarring [median and range (picograms per milliliter): 6,163 (2,021-13,801) vs. 1,490.5 (0-5,737), respectively, p = 0.018]. In conclusion, UIL-8 might serve as a predictive biomarker for renal scarring after an acute episode of pyelonephritis. Since UIL-8 emerges as a renal-specific diagnostic and prognostic marker, it may be suitable as a selective screening tool for children with febrile UTI.

[Proinflammatory cytokines and C-reactive protein in urine: prospects for diagnosis in patients with chronic pyelonephritis].

The study involved 85 patients with chronic pyelonephritis in age from 26 to 62 years. All patients determined C-reactive protein (CRP), interleukin 6 and tumor necrosis factor-alpha in the blood and urine by standard techniques. With the simultaneous study of CRP in the blood and urine of patients with chronic pyelonephritis detected high levels of this biomarker in the urine, which is several times higher than its indicators levels in blood. When studying the levels of proinflammatory cytokines, which are the main inducers excretion of CRP, we found that the increase in their concentration in urine was more pronounced than in the blood, which confirms the opinion that CRP can be synthesized in inflammation focus by lymphocytes, macrophages and endothelial. Studying the levels of cytokines and C-reactive protein in the urine can be used to assess the severity of inflammatory changes in the kidney and to monitor the effectiveness of anti-inflammatory therapy.

[Administration of vitaprost plus for prevention of infectious-inflammatory complications of transurethral prostatic resection in patients with prostatic adenoma].

The study of efficacy and safety of the drug vitaprost plus (rectal suppositories) in its use for prevention of infectious-inflammatory complications after transurethral resection (TUR) of the prostate for adenoma included 55 male surgical patients with prostatic adenoma (PA). The patients were randomized into two groups matched by age, symptoms and clinical data. The control group consisted of 20 patients (group 1), the study group--of 35 patients (group 2). TUR of the prostate was made in both groups, but patients of group 2 were given additional prophylactic treatment--vitaprost plus, one suppository before going to bed 2 days before surgery and for 8 days after TUR, the course lasted for 10 days. Postoperative blood and urine samples showed inflammatory changes more often in group 1, free of infection bacterial tests were more often in group 2. Renal microcirculation in group 1 after TUR evidenced for functional depression on perfusion intensity and renal ischemia and congestion. These changes persisted on postoperative day 8, being a potential cause of renal inflammation and ascending pyelonephritis. Group 2 patients exhibited insignificant postoperative microcirculatory disorders in the kidneys and were not registered on postoperative day 8. Infectious-inflammatory complications in group 1 patients were registered much more frequently than in group 2 patients. Thus, vitaprost plus can be recommended for prophylaxis of infectious-inflammatory complications of TUR in PA patients.