Carbonic-adsorbent AST-120 reduces overload of indoxyl sulfate and the plasma level of TGF-β1 in patients with chronic renal failure

Background We previously reported a significant increase in plasma TGF-β1 in patients with chronic renal failure (CRF). Progression of CRF may be caused by persistent renal production of TGF-β1. In CRF rat models, an oral carbonic absorbent (AST-120) reduces the expression of the TGF-β1 gene in the kidney, and delays the progression of CRF, in part by alleviating the overload of indoxyl sulfate. The aim of this study was to evaluate the effect of AST-120 on plasma levels of indoxyl sulfate and TGF-β1 in CRF patients. Methods Ten CRF patients (aged 59.3 ± 9.5 years, 5 men, serum creatinine 4.37 ± 1.72 mg/dl) were enrolled in this study. All patients maintained a regular dietary therapy and the same medication throughout the study. AST-120 was added at a dose of 6 g/day. Parameters including the slope of the reciprocal of the serum creatinine – time plot, plasma indoxyl sulfate level, and plasma and urinary levels of TGF-β1 were compared before and after the treatment with AST-120. The mean observation periods before and after the treatment were 9.7 ± 2.8 and 6.5 ± 2.9 months, respectively. Results Administration of AST-120 significantly reduced the plasma levels of indoxyl sulfate (1.42 ± 1.50 vs. 1.26 ± 1.40 mg/dl, P < 0.05) and TGF-β1 (17.9 ± 7.2 vs. 10.6 ± 4.7 ng/ml, P < 0.05) and improved the slope of the reciprocal of serum creatinine (−0.061 ± 0.041 vs. −0.032 ± 0.055 dl/mg/year, P < 0.05). Conclusions These results support the notion that indoxyl sulfate and TGF-β1 may be involved in the progression of CRF, and that the oral adsorbent AST-120 may suppress the progression, at least in part, by reducing overproduction of TGF-β1.     

Expression of fibrosis-associated molecules in IgA nephropathy treated with cyclosporine

Cyclosporine (CsA) treatment in immunoglobulin A nephropathy (IgAN) is controversial and has not been widely studied. The aim of this study was to investigate the effects of CsA on renal histology and the expression of interstitial fibrosis-associated molecules in childhood IgAN. The subjects were 18 children (age 4.2–13.9 years; male:female 13:5) who had been treated with CsA for 8 or 12 months and who had renal biopsies before and after treatment. Renal biopsies were assessed by routine histology and immunohistochemistry against osteopontin (OPN), transforming growth factor-β (TGF-β), CD68, and CD34. The degree of proteinuria and mesangial IgA deposits decreased or disappeared after treatment in all cases, and the percentage of patients with diffuse mesangial proliferation decreased from 44.4 to 22.2%. However, interstitial fibrosis developed or was aggravated in nine patients (50%) after treatment and was associated with an increased degree of interstitial inflammation in five patients. Tubular OPN expression (45.3 ± 23.4 vs. 37.6 ± 19.3%) and the degree of CD68-positive macrophage infiltration (136.1 ± 88.2 vs. 132 ± 86.0/mm²) were not increased after CsA treatment, but TGF-β expression was significantly increased (6.4 ± 4.2 vs. 13.3 ± 9.9%; p = 0.025). Microvascular density was increased and peritubular capillaries were of small caliber in inflamed areas. We conclude that increased levels of TGF-β and the development of interstitial fibrosis limit the long-term use of CsA in IgAN patients. Osteopontin and macrophages may be indirectly involved in renal fibrosis by prolonging interstitial inflammation rather than by directly increasing TGF-β expression.     

Urinary levels of TGF β-1 and of cytokines in patients with prenatally detected nephrouropathies

This study aimed to identify noninvasive biomarkers of clinically significant nephrouropathies in patients with antenatal renal and/or urinary tract alterations. Spot-urine levels of interleukin-6 (IL-6), transforming growth factor-β1 (TGF-β1) and tumor necrosis factor-α (TNF-α) were measured in 100 patients with antenatal detected nephrouropathies. Patients were divided in idiopathic hydronephrosis (n = 47), urinary tract malformations (n = 35), and dysplastic kidneys (n = 18). Urinary concentrations of TGF-β1, IL-6, and TNF-α were compared between groups according to clinical and image findings. Receiver-operating characteristic (ROC) curves were analyzed for the overall diagnostic accuracy of TGF-β1, IL-6, and TNF-α levels in discriminating infants with nephrouropathies. No significant differences in urinary TGF- β1, IL-6, and TNF-α levels were found in the comparison between the groups. TGF-β1 levels tended to be higher in patients with renal hypodysplasia compared to idiopathic hydronephrosis (p = 0.07). Twenty-nine patients had reduced DMSA uptake. In these cases, absolute urinary concentration of TGF-β1 and levels standardized for creatinine were significantly higher than in patients with normal DMSA uptake, while IL6 and TNF-α did not differ between groups. Urinary cytokine measurements were not useful as a screening test for clinically significant nephrouropathies. Conversely, increased concentrations of TGF-β1 pointed out to renal damage as indicated by reduced DMSA uptake.     

Neutrophil gelatinase-associated lipocalin as a biomarker of disease activity in pediatric lupus nephritis

We hypothesized that neutrophil gelatinase-associated lipocalin (NGAL) is an early predictive biomarker of disease activity in lupus nephritis. NGAL in serial plasma (PNGAL) and urine (UNGAL) samples was measured by enzyme-linked immunosorbent assay (ELISA) in 85 participants with pediatric systemic lupus erythematosus (pSLE), healthy children (n = 50), and children with juvenile idiopathic arthritis (JIA) (n = 30). Disease activity was measured by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Plasma and urinary NGAL were significantly increased in subjects with pSLE compared with those with JIA or with healthy controls (all p < 0.03), and unrelated to subjects’ age, weight, or height. Plasma and urinary NGAL were stable in pSLE subjects with unchanged disease activity. The pSLE subjects with worsening global or renal disease activity had a mean ± standard error (SE) increase of UNGAL (in ng/ml) of 11.5 ± 6.4 or 36.6 ± 12.1 (p < 0.01), corresponding to a 156% or 380% increase, respectively. PNGAL increased with worsening disease but to a much lesser degree than UNGAL [global disease activity (mean ± SE): 7.3 ± 6.2 or 21%; renal disease activity: 20.2 ± 6.0 or 51%; both p = not significant]. In conclusion, NGAL in urine but not in plasma represents a novel biomarker for renal disease activity in pSLE.     

Assesment of Serum Neutrophil Gelatinase-associated Lipocalin Versus Protienuria in Chronic Kidney Diseases

Serum neutrophil gelatinase-associated lipocalin (NGAL) represents a novel biomarker for early identification of various renal injuries. The purpose of the study was to evaluate the level of NGAL in some renal diseases with proteinuria. The study included 120 patients with proteinuria and 60 healthy subjects of matched age and sex. The following tests were performed: fasting blood glucose, serum creatinine, creatinine clearance, serum cholesterol, urinary albumin/creatinine ratio, 24-h urinary albumin, and NGAL. Abdominal sonography, fundus examination, and renal biopsy were done. The NGAL of a total of 120 patients (54 males and 66 females, mean age 44.6 ± 16.3 years) with renal diseases demonstrated a highly significant difference, being higher in patients with various renal diseases than in healthy controls (P < 0.001). This study confirms the existence of a raised circulating level of NGAL in CKD patients with proteinuria, which may be used as a sensitive marker for patients with proteinuria.     

Urinary ?2-microglobulin in very preterm neonates with chorioamnionitis

It is important to identify premature infants with prenatal inflammation as it contributes to short- and long-term complications. Our object was to study how prenatal inflammation affects the urinary β₂-microglobulin (β₂-MG) level. Preterm neonates were divided based on the presence of chorioamnionitis (CAM) into the CAM (n = 100) and non-CAM groups (n = 117). These were further subdivided into five groups each: 30 preterm neonates of 23–26; 42 neonates of 27–28; 54 neonates of 29–30; 51 neonates of 31–32; and 40 neonates of 33–34 weeks’ gestation. The urinary β₂-MG level within 48 h of birth was significantly higher in the CAM group than in the non-CAM group among the neonates of 23–26 weeks’ gestation (18.3 ± 6.9 vs 10.0 ± 5.6 × 10⁴ μg/gCr, p = 0.0018) and the neonates of 27–28 weeks’ gestation (16.2 ± 10.8 vs 8.8 ± 3.3 × 10⁴ μg/gCr, p = 0.0101). However, there was no difference in urinary β₂-MG level between the CAM and the non-CAM group among the neonates ≥ 29 weeks ’gestation. Moreover, the elevated urinary β₂-MG level in the neonates ≤ 28 weeks ’ gestation with CAM had disappeared by 1 week after birth. The reasons for the increase in urinary β₂-MG level within 48 h of birth in very preterm neonates (≤ 28 weeks’ gestation) with CAM are believed to be not only prematurity, but also prenatal inflammation. It is suggested that the urinary β₂-MG level during the early postnatal period can identify prenatal inflammation.     

Increased Expression and Activity of Hepatic Lipase in the Liver of Morbidly Obese Adult Patients in Relation to Lipid Content

Background  The types and sources of lipid deposition in the liver of most patients with morbid obesity, as well as the effects of bariatric surgery, are discussed. Methods  In 26 patients with morbid obesity who underwent bariatric surgery, we analyzed different kinds of lipids and hepatic lipase (HL) from both plasma and liver biopsies performed 12–18 months after surgery. Results  The HL activity and HL-mRNA in morbidly obese (MO) livers were high (258 ± 17 mU/g, and 4.5-fold, respectively); after surgery, the activity decreased (137 ± 15 mU/g, p < 0.001) but not the levels of HL-mRNA (4.3-fold). Plasma HL activity was also high (4.31 ± 0.94 mU/mL plasma), and it decreased during weight loss (2.01 ± 0.29 mU/mL, p < 0.01); moreover, it correlated (r = 0.3694, p < 0.05) with decreased liver HL activity. Adrenocorticotropic hormone in MO was higher (27 ± 3 pg/mL) than after surgery (13 ± 1 pg/mL, p < 0.001). All hepatic and plasma lipids were significantly increased in MO patients, but, after bariatric surgery, most of those parameters recovered or normalized. Liver HL activity correlated with total and esterified cholesterol (r = 0.4399, p < 0.001 and r = 0.4395, p < 0.01, respectively). Conclusion  High HL in MO patients could allow for liver intake of cholesterol that could be re-exported to steroidogenic organs to synthesize steroidal hormones. A decrease of plasma HL during weight loss could be a good index for improvement of liver disease.     

Association of macroalbuminuria with oxidized LDL and TGF-β in type 2 diabetic patients: a case–control study

The pathogenesis of diabetic nephropathy, mainly characterized by macroalbuminuria, is still poorly understood, but it is reported that transforming growth factor-β (TGF-β) plays a key role. In vitro evidence suggests that administration of oxidized LDL (ox-LDL) can lead to upregulation of TGF-β by human glomerular mesangial cells. This study aimed to evaluate the association between macroalbuminuria, ox-LDL, and TGF-β in diabetic patients. A total of 77 type 2 diabetic patients with macroalbuminuria (albumin excretion rate: AER ≥ 300 mg/24 h) and 66 patients with normoalbuminuria (AER ≤ 30 mg/24 h) were recruited. Fasting blood samples were obtained and serum levels of ox-LDL and TGF-β were determined. Ox-LDL and TGF-β were significantly higher in patients with macroalbuminuria than in those with normoalbuminuria (98.93 ± 3.99 vs. 72.45 ± 2.48 U/l; P < 0.001 and 6.46 ± 0.74 vs. 2.49 ± 0.39 ng/ml; P < 0.001, respectively). In patients with macroalbuminuria, there was a significant correlation between Ox-LDL and TGF-β (r = 0.376; P < 0.01). AER was significantly correlated to ox-LDL (r = 0.302; P < 0.05) and TGF-β (r = 0.306; P < 0.05) in macroalbuminuric patients. This association remained significant after adjustment for potential confounders. Adjustment for TGF-β (ox-LDL), attenuated the association of ox-LDL (TGF-β) with AER. In conclusion, this study demonstrated the association of TGF-β and ox-LDL with albuminuria in macroalbuminuric type 2 diabetic patients, and suggested that this relationship is highly mediated through the correlation between TGF-β and ox-LDL.     

Urinary interleukin-6 is useful in distinguishing between upper and lower urinary tract infections

This study was designed to determine whether the measurement of interleukin (IL)-6 in urine is useful for distinguishing between acute pyelonephritis and lower urinary tract infection. This observational study was carried out at León Hospital (Spain) on 35 patients (ten boys) aged between 0 and 14 years with urinary tract infection. Urinary levels of IL-6 were determined with enzyme-linked immunosorbent assay (ELISA) at diagnosis and after recovery. Renal dimercaptosuccinate acid (DMSA) scan was performed on all patients to discard or confirm acute pyelonephritis. The mean urinary concentration [x ± standard deviation (SD)] of IL-6 at diagnosis was 20.3 ± 23.3 and 5.3 ± 9.7 pg/ml in patients with acute pyelonephritis and lower urinary infection, respectively [95% confidence interval (CI): 2.6–27.4; p < 0.01]. Specificity for a value of IL-6 >15 pg/ml, was 94.1% (95% CI: 91.1–97.1). Positive predictive value for IL-6 >15 pg/ml was 87.5% (95% CI: 81.1–93.8). IL-6 was undetectable in the urine of both groups of patients at the time of recovery. Urinary levels of IL-6 are useful in differentiating between upper and lower urinary tract infection in children. In this clinical setting, a value >15 pg/ml is a strong indicator of acute pyelonephritis.     

Urinary prostacyclin metabolites in patients with IgA nephropathy

Background. Urinary 6-keto-prostaglandin F_1α (6kPGF) is an index of renal prostacyclin (PGI₂) synthesis, while urinary 2,3-dinor-6-keto-prostaglandin F_1α (23d6kPGF) reflects extrarenal PGI₂ synthesis. We therefore examined the relationship between the urinary excretion of PGI₂ metabolites and the deterioration of renal function and histopathological findings. Methods. We measured these PGI₂ metabolites in 24-h collected urine from 32 patients with IgA nephropathy (IgAN) and 24 normal controls. Results. The urinary 6kPGF excretion in all patients (790 ± 395 pg/mg creatinine [Cr]) did not differ significantly from that in the normal controls (896 ± 351 pg/mgCr), but it was decreased in the patients with histopathological deterioration (661 ± 281 pg/mgCr). There were no such findings for 23d6kPGF. In the IgAN patients, the urinary 6kPGF excretion demonstrated a positive correlation to the 1/creatinine slope, which indicated deterioration of renal function (r = 0.43; P < 0.03). Conclusions. Renal PGI₂ production is reduced in IgAN patients with histopathological deterioration, and 24-h urinary 6kPGF excretion may be an effective marker to evaluate the renal injury. Received: June 22, 1998 / Accepted: December 19, 1999     

NGAL is an early predictive biomarker of contrast-induced nephropathy in children

We hypothesized that neutrophil gelatinase-associated lipocalin (NGAL) is an early predictive biomarker of contrast-induced nephropathy (CIN). We prospectively enrolled 91 children (age 0–18 years) with congenital heart disease undergoing elective cardiac catheterization and angiography with contrast administration (CC; Ioversol). Serial urine and plasma samples were analyzed in a double-blind fashion by NGAL enzyme-linked immunosorbent assay (ELISA). CIN, defined as a 50% increase in serum creatinine from baseline, was found in 11 subjects (12%), but detection using increase in serum creatinine was only possible 6–24 h after CC. In contrast, significant elevation of NGAL concentrations in urine (135 ± 32 vs. 11.6 ± 2 ng/ml without CIN, p < 0.001) and plasma (151 ± 34 vs. 36 ± 4 without CIN, p < 0.001) were noted within 2 h after CC in those subjects. Using a cutoff value of 100 ng/ml, sensitivity, specificity, and area under the receiver-operating characteristic (ROC) curve for prediction of CIN were excellent for the 2-h urine NGAL (73%, 100%, and 0.92, respectively) and 2-h plasma NGAL (73%, 100%, and 0.91, respectively). By multivariate analysis, the 2-h NGAL concentrations in the urine (R ² = 0.52, p < 0.0001) and plasma (R ² = 0.72, p < 0.0001) were found to be powerful independent predictors of CIN. Patient demographics and contrast volume were not predictive of CIN.     

Effect of Roux-en Y Gastric Bypass on Bone Metabolism in Patients with Morbid Obesity: Mansoura Experiences

Background  Roux-en-Y gastric bypass (RYGBP) has been found to be the most efficient way to lose weight and maintain the weight loss in morbid obesity. However, with the formation of a new stomach and the modification of intestinal anatomy, there are significant changes on bone metabolism. The objectives of this study were to evaluate effects of weight loss on bone metabolism after Roux-en Y gastric bypass in patients with morbid obesity. Methods  Our study included 70 patients with morbid obesity; RYGB was done for all patients. Daily postoperative oral supplementation with 1,000 mg of calcium and 800 IU of vitamin D was done for each patient. Body weight (BW), body mass index (BMI), total body fat, total lean tissue mass, bone mineral content (BMC), bone mineral density (BMD), total bone area (TBA; using dual energy X-ray absorptiometry), serum calcium, parathyroid hormone (PTH), 25-OH vitamin D, 24-h urinary calcium, and bone-specific alkaline phosphatase (BSAP) were assessed preoperatively and 1 year after surgery. Results  In our study, females comprised 70% of cases. The mean age was 35 ± 8.8 years. One year after RYGB, BW decreased significantly from 132.8 ± 26.5 to 90.3 ± 17.3 kg (p = 0.001). BMI decreased significantly from 48 ± 7.3 to 32.6 ± 4.1 kg/m² (p = 0.001). BMC decreased significantly from 2,968.6 ± 71.4 to 2,700.8 ± 45.4 g (p = 0.001). BMD decreased significantly from 1.026 ± 0.03 to 1.22 ± 0.015 g/cm² (p = 0.001). TBA decreased significantly from 2,356.2 ± 35.4 to 2,216.3 ± 43.5 cm² (p = 0.001). Serum calcium, 24-h urinary calcium, and BSAP were not significantly decreased while 25-OH vitamin D and PTH were not significantly increased after surgery. Conclusions  From this study, it is shown that RYGBP operation gives very good results as regards reduction of body weight in morbidly obese patients. Postoperative supplementation with calcium and vitamin D partially corrects osteoporosis. Thus, these patients need periodic follow-up for BMD, PTH, calcium, serum vitamin D, and markers of bone resorption and formation specially postmenopausal female.     

Determination of Clara cell protein urinary elimination as a marker of tubular dysfunction

Clara cell 16-kDa protein (CC16) is a protein expressed primarily by the bronchial cells. It is rapidly eliminated by glomerular filtration, reabsorbed almost entirely, and catabolized in proximal tubule cells. To date, normal values for urinary CC16 in healthy children have not been determined. We have studied 63 pediatric patients (mean age 8.17 ± 3.91 years) and 31 healthy children (control group; mean age 8.83 ± 3.65 years). In the control group, the CC16/creatinine ratio was 1.22 ± 1.52 μg/g. In 16 out of 31 control children, the value of the ratio was zero. Fourteen patients (22.2%) showed a high CC16/creatinine ratio; in contrast, among these same patients, the ratio N-acetyl-β-d-glucosaminidase (NAG)/creatinine was elevated in seven cases (11.1%) and the ratio β2-microglobulin/creatinine was elevated in seven cases (11.1%). The three parameters were in agreement in 51 patients (80.9%). Among the patients, the CC16/creatinine ratio was correlated with both the β2-microglobulin/creatinina ratio (r = 0.76, P < 0.001) and the NAG/creatinine ratio (r = 0.6, P < 0.001). Our findings indicate that CC16 is a good marker of proximal tubular function in childhood. The highest observed values were in children with proximal tubulopathies, in children with chronic renal failure, and in those treated with cyclosporine.     

Urinary MMP-9/NGAL complex in children with acute cystitis

The matrix metalloproteinase-9 (MMP-9) and neutrophil gelatinase associated lipocalin (NGAL) are shown to increase in an inflammatory situation. Based on our previous reports that NGAL can be detected in the urine of children with urinary tract infection (UTI), we also asked whether MMP-9/NGAL complex could be detected in the urine of children with UTI. This multicenter, prospective study was conducted between October 2009 and October 2010. Seventy-one patients with symptomatic culture proven UTI, 37 asymptomatic children with contaminated urine and 37 healthy children were recruited. Mean uMMP-9/NGAL/Cr levels were significantly higher in the UTI group than in the control group (p < 0.0001). According to ROC analysis, the optimal cut-off level was 0.08 ng/mg to predict UTI. Using a cut-off value, sensitivity and specificity were 98.6 and 97.3%, respectively. The mean levels of uMMP-9/NGAL/cr in the UTI group were also significantly higher than those in the contamination group (p < 0.0001). There was no statistically significant difference between contamination group and the control group (p = 0.21). The mean uMMP-9/NGAL/Cr in the UTI group were significantly higher before treatment than after treatment (p < 0.0001). The area under the curve was 0.997 (SE: 0.002, 95% CI: 0.993 to 1.001) for uMMP-9/NGAL/Cr. Urinary MMP-9/NGAL/Cr level was also correlated with positive urine nitrite test, positive urine leukocyte esterase reaction and renal scarring (p = 0.0001, p = 0.0001, p = 0.04, respectively) whereas was not correlated to leukocytosis and positive CRP level in serum. Urine MMP-9/NGAL/cr can be used as a diagnostic biomarker for UTI in children. Identification of NGAL-MMP-9/cr levels in the urine of suspected UTI patients may also be useful to differentiate between contamination and infection and for monitoring of treatment response in children.     

Regulation of Bone Mineral Density in Morbidly Obese Women: A Cross-sectional Study in Two Cohorts Before and After Bypass Surgery

Background  The mechanisms by which increased body weight influence bone mass density (BMD) are still unknown. The aim of our study was to analyze the relationship between anthropometric and body composition variables, insulin growth factor-I (IGF-I), adiponectin and soluble tumor necrosis factor-α receptors (sTNFR) 1 and 2 with BMD in two cohorts of morbid obese patients, before and after bypass surgery. Methods  The first cohort included 25 women aged 48 ± 7.6 years studied before bypass surgery. The second included 41 women aged 46 ± 9.2 years, 12 months after surgery. We studied anthropometric variables obtained from whole body DEXA composition analysis. Serum IGF-I, intact serum parathyroid hormone, 25-hydroxivitamin D₃, plasma adiponectin concentrations, sTNFR1, sTNFR2 concentrations were measured. Results  In the first cohort, the BMI was 44.5 ± 3.6 kg/m², parathyroid hormone, IGF-I, and adiponectin concentrations were lower, and sTNFR1 concentrations were higher than in the second cohort. In the multiple regression analysis, BMD remained significantly associated with body fat percentage (β −0.154, p = 0.01), lean mass (β 0.057, p = 0.016) and phosphate concentration (β 0.225, p = 0.05). In the second cohort, BMI was 31 ± 5.1 kg/m². In the multiple regression analysis, BMD remained significantly associated with lean mass (β 0.006, p = 0.03). Conclusion  The inverse correlation found between body fat and BMD in the first cohort indicates morbid obesity increases the risk of osteoporosis and we found a positive correlation with lean and fat mass before bariatric surgery and with lean mass after bypass surgery.     

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.     

Bone-Resorbing Cytokines from Peripheral Blood Mononuclear Cells after Hormone Replacement Therapy: A Longitudinal Study

Conflicting results have been reported in several cross-sectional studies measuring cytokine production from adherent monocytes in pre- and postmenopausal women. Furthermore, the target cells for the action of estrogen are still debated. We therefore assessed in a longitudinal manner the cytokine production from different fractions of peripheral blood mononuclear cells (PBMC) cultured for 48 h. PBMC were obtained from 30 postmenopausal women before and after 6 months of hormone replacement therapy (HRT). Women were randomly allocated to two groups: an adherent PBMC group (n= 20) and a total PBMC group (n= 9). After 6 months of treatment, urinary pyridinoline levels were markedly decreased in both groups (353 ± 24 vs 114 ± 13 μg/mmol creatinine and 325 ± 35 vs 164 ± 31 μg/mmol creatinine respectively, p<0.01). Culture supernatants were assayed for interleukin 1β (IL-1β), interleukin 6 (IL-6), soluble IL-6 receptor (IL-6rs) and tumor necrosis factor alpha (TNF-α). In the adherent PBMC group, HRT induced a nonsignificant trend toward decreased levels of IL-1β (35 ± 10 vs 13 ± 5 pg/ml), TNF-α (333 ± 58 vs 222 ± 30 pg/ml) and IL-6 (115 ± 70 vs 17 ± 10 pg/ml). In contrast, in the total PBMC group, HRT induced a consistent and dramatic decrease in levels of IL-1β (104 ± 22 vs 25 ± 8 pg/ml), IL-6 (5950 ± 1041 vs 1011 ± 361 pg/ml), IL-6rs (148 ± 33 vs 35 ± 12 pg/ml) (p<0.01) and TNF-α (1468 ± 315 vs 585 ± 207 pg/ml, p= 0.05). We then evaluated whether HRT had the same effect in vitro. Adherent or total PBMC of 8 postmenopausal women were cultured with or without 10⁻⁸M 17β-estradiol or tibolone for 48 h. Production of IL-1β, TNF-α, IL-6 and IL-6rs was not affected by the presence of 17β-estradiol or tibolone in cultures of these cell fractions. In conclusion, our data indicate that non-adherent PBMC could mediate the response to HRT. HRT may exert its action indirectly via noncirculating cells, as suggested by the absence of an in vitro effect. Received: 11 July 2000 / Accepted: 15 January 2001     

Higher urine heat shock protein 70/creatinine ratio in type 1 diabetes mellitus

Insidious progressive renal damage caused by type 1 diabetes mellitus (T1DM) begins in childhood before it becomes manifest in adult ages. Heat shock proteins (HSPs) regulate the cell response to any hazardous factors to prevent cell structure. The aim of the study is to determine whether urine levels of HSPs increase in diabetic children with time and indicate a progressive renal injury in T1DM. Thirty-three patients with T1DM and 24 healthy children were enrolled in the study. Renal function was normal in all patients. Urine levels of HSP27, HSP40, HSP60, HSP70, and HSP90 were measured by enzyme-linked immunosorbent assay at two consecutive years (2012 and 2013). The results were evaluated as urine HSP/creatinine ratios (uHSP/Cr). Mean urine HSP27/Cr, HSP40/Cr, HSP60/Cr, HSP70/Cr, HSP90/Cr in patient group were significantly higher than in controls in 2012 (uHSP27/Cr 460.12?±?217.64 versus 270.02?±?136.83?pg/mgCr; uHSP40/Cr 180.89?±?118.59 versus 99.44?±?62.49?pg/mgCr; uHSP60/Cr 114.40?±?64.91 versus 70.50?±?43.70?pg/mgCr; uHSP70/Cr 41.17?±?28.42 versus 16.47?±?7.32?pg/mgCr; uHSP90/Cr 175.64?±?102.22 versus 107.61?±?75.85?pg/mgCr) (p?p?=?0.001), whereas uHSP60/Cr level decreased and uHSP27/Cr, uHSP40/Cr, uHSP90/Cr levels remained stable (p?>?0.05). Area under the curve (AUC) for uHSP70/Cr (0.957) was significantly higher than the others. Using a cutoff 22.59?pg/mgCr for uHSP70/Cr to predict of diabetic damage, sensitivity and specificity were 85% and 96%, respectively. Our results suggest that uHSP70/Cr increases over time and may indicate early phases of progressive kidney damage in diabetic children.     

Regression of target organ damage in children and adolescents with primary hypertension

We assessed the effects of 12 months of non-pharmacological and pharmacological therapy on 24-h ambulatory blood pressure, regression of target organ damage (TOD) and metabolic abnormalities in 86 children (14.1 ± 2.4 years) with primary hypertension. Twenty-four hour systolic and diastolic blood pressure (BP) decreased (130 ± 8 vs 126 ± 8, 73 ± 7 vs 70 ± 7, p = 0.0001 and 0.004 respectively). Body mass index (BMI) did not change, but waist-to-hip (0.85 ± 0.07 vs 0.83 ± 0.05, p = 0.01) and waist-to-height ratio (WHtR; 0.49 ± 0.07 vs 0.48 ± 0.05, p = 0.008) decreased. Left ventricular mass index (LVMi; 38.5 ± 10.7 vs 35.2 ± 7.5 g/m^2.7, p = 0.0001), prevalence of left ventricular hypertrophy (46.5% vs 31.4%; p = 0.0001), carotid intima-media thickness (cIMT; 0.44 ± 0.05 vs 0.42 ± 0.04 mm, p = 0.0001), wall cross sectional area (WCSA; 7.5 ± 1.3 vs 6.9 ± 1.2 mm², p = 0.002), hsCRP (1.1 ± 1.0 vs 0.7 ± 0.7 mg/l, p = 0.002), and LDL-cholesterol (115 ± 33 vs 107 ± 26 mg/dl, p = 0.001) decreased. Patients who had lowered BP had a lower cIMT at the second examination (0.41 ± 0.04 vs 0.43 ± 0.04 mm, p = 0.04) and lower initial hsCRP values (0.9 ± 0.7 vs 1.5 ± 1.3 mg/l, p = 0.04) in comparison to non-responders. Regression analysis revealed that the main predictor of LVMi decrease was a decrease in abdominal fat expressed as a decrease in waist circumference (WC) (R ² = 0.280, β = 0.558, p = 0.005), for WCSA-SDS a decrease in WC (R ² = 0.332, β = 0.611, p = 0.009) and for a cIMT-SDS decrease the main predictor was a decrease in hsCRP concentrations (R ² = 0.137, β = 0.412, p = 0.03). Standard antihypertensive treatment lowered BP and led to regression of TOD in hypertensive children. Lean body mass increase and decrease in abdominal obesity correlated with TOD regression.