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Background Detectable levels of cardiac troponins are common in individuals with chronic kidney disease (CKD), even in the absence of symptomatic cardiovascular disease. Abnormal cardiac troponin values are associated with coronary artery disease and left ventricular hypertrophy (LVH) and predict poor clinical outcomes. Elevated levels of fibroblast growth factor 23 (FGF-23) contribute to LVH in CKD. We investigated the association of FGF-23 and hs-cTnI (high-sensitivity cardiac troponin I) and hs-cTnT (high-sensitivity cardiac troponin T) levels in CKD and examined the role of LVH in this association. Study Design Cross-sectional observational study. Setting & Participants 153 stable outpatients with non–dialysis-dependent CKD. Predictor The primary predictor was FGF-23 level. Outcomes hs-cTnI, hs-cTnT. Measurements FGF-23, hs-cTnI, hs-cTnT; left ventricular mass index (LVMI) assessed by echocardiography; coronary artery calcification (CAC) measured by computed tomography. LVMI and CAC were evaluated as potential mediators of the effect of FGF-23 on hs-cTnI/T. Results Mean age was 64 ± 12 (SD) years, mean estimated glomerular filtration rate was 34 ± 11 mL/min/1.73 m 2 , median FGF-23 level was 120 (25th-75th percentile, 79-223) reference unit (RU)/mL, median hs-cTnI level was 6.5 (25th-75th percentile, 3.5-14.5) pg/mL, and median hs-cTnT level was 16.8 (25th-75th percentile, 11.1-33.9) pg/mL. cTnI and cTnT concentrations were higher than the 99th percentile of a healthy population in 42% and 61% of patients, respectively. In unadjusted and multivariable-adjusted analyses, hs-cTnI/T levels were associated significantly with FGF-23 levels. Adjusting for LVMI, but not CAC, weakened the association of FGF-23 and hs-cTnI/T levels. Limitations Vitamin D levels were not measured. The prevalence of coronary artery disease may have been underestimated because it was ascertained by self-report. Conclusions Minimally elevated cTnI and cTnT levels, detectable by high-sensitivity assays, are associated with elevated FGF-23 levels in stable outpatients with CKD. FGF-23–associated LVH may contribute to detectable hs-cTnI/T levels observed in non–dialysis-dependent patients with CKD.

BACKGROUND Quantification and comparison of high-sensitivity (hs) cardiac troponin I (cTnI) and cTnT concentrations in chronic kidney disease (CKD) have not been reported. We examined the associations between hs cTnI and cTnT, cardiovascular disease, and renal function in outpatients with stable CKD. METHODS Outpatients (n = 148; 16.9% with prior myocardial infarction or coronary revascularization) with an estimated glomerular filtration rate (eGFR) of RESULTS The median (interquartile range) concentrations of cTnI and cTnT were 6.3 (3.4–14.4) ng/L and 17.0 (11.2–31.4) ng/L, respectively; 38% and 68% of patients had a cTnI and cTnT above the 99th percentile, respectively. The median CAC score was 80.8 (0.7–308.6), LV mass index was 85 (73–99) g/m2, and LVEF was 58% (57%–61%). The prevalences of prior coronary disease events, CAC score, and LV mass index were higher with increasing concentrations from both hs cardiac troponin assays (P < 0.05 for all). After adjustment for demographics and risk factors, neither cardiac troponin assay was associated with CAC, but both remained associated with LV mass index as well as eGFR and UACR. CONCLUSIONS Increased hs cTnI and cTnT concentrations are common in outpatients with stable CKD and are influenced by both underlying cardiac and renal disease.

The relationship between acute renal failure (ARF) and long-term renal function remains unknown. We therefore undertook a study of patients at the Baltimore VA Hospital to examine the effects of a bout of acute renal injury on long-term renal function. We retrospectively reviewed the relationship between serum creatinine and time of observation for 6058 individuals who had values greater than 1.4 mg/dL in any two consecutive years. Individuals were stratified according to total years of observation with a minimum of two years. Severity of acute renal injury was divided into mild, moderate, and severe with elevations in baseline creatinine of50%, 50-300% and300% respectively. Sporadic elevations in creatinine were evident in 8-15% of the population. There were a total of 1328 episodes of acute renal failure in 916 patients that were suitable for analysis. Mild ARF on a substrate of normal or mildly abnormal renal function resolved without long-term sequelae. Moderate and severe ARF occurred more frequently on a background of reduced renal function but baseline function was retained in at least 60% of patients. We conclude that ARF is more frequent in patients with chronic kidney disease but it is not invariably associated with an accelerated course to end-stage renal disease or death. Overall, the majority of ARF events resolved without adverse long-term effects suggesting appropriate management in the majority of instances.

Although it is anticipated that most patients with renal insufficiency will progress towards end-stage renal disease (ESRD) there have been few population-based studies to validate this assumption. We examined serial creatinines from 3,874 anonymous patients at an urban VA medical center who had a baseline creatinine of 1.4 mg/dl or greater to estimate the frequency of deterioration in renal function (DRF). DRF was defined as the first Cr (1stCr) value being lower than the last Cr (LCr) for each patient. The median follow-up was 48.3 ± 0.5 months with 18 ± 0.5 creatinine values per patient. The median 1stCr was 1.6 ± 0.1 mg/dl with 32.2% of the patients having a 1stCr greater than or equal to 1.7 mg/dl. In the study group, 1,723 (44.4%) had DRF including 1,089 (41.4%) of those patients with a 1stCr of 1.4–1.7 mg/dl. However, 45 (36.6%) of those with a 1stCr value 3.0–5.0 mg/dl did not have DRF, the percent with stable creatinine in this group did not vary with length of follow-up. Over the study period, 299 (7.7%) of all the patients had a creatinine rise to 7.0 mg/dl, with 104 (4%) of those with a 1stCr of 1.4–1.7 mg/dl reaching this endpoint. Conclusion: A majority, but not all, patients with renal insufficiency lose renal function over time and those with even mild hypercreatinemia are at risk for deterioration in renal function. Hypercreatinemia, however, does not accurately discriminate between those renal insufficiency patients who are stable versus those at high risk for ESRD.

Aim: To examine the response of the insulinlike growth factor (IGF) system in the kidney during a state of extreme growth. Methods: We studied the mRNA expression of IGF-I, IGF-I receptor, and IGF-binding proteins (BP) using sensitive RNase protection assays following subcutaneous implantation of growth hormone pituitary cells (GH3) in rats. Results: Within 5 weeks, the serum GH levels increased from 18.0 ± (SE) 5.0 ng/ml in control animals to 389.8 ± 30.3 ng/ml in GH3 rats (n = 5, p < 0.001). The circulating IGF-I levels were also elevated. The kidney weights increased from 0.74 ± 0.01 g in controls to 1.06 ± 0.03 g in GH3 animals (n = 5, p < 0.001). Similar changes were observed at week 10. The renal IGF-I mRNA averaged 1.0 ± (SD) 0.33 relative densitometry units in controls (n = 4) and increased to 2.11 ± 0.13 relative densitometry units in GH3 rats (n = 5, p < 0.001). On the other hand, mRNA for the type I IGF receptor decreased in hypersomatotropic rats. Messenger RNAs for IGFBP-1 and IGFBP-4, which have been localized to renal tubules, both decreased significantly following growth induction, while IGFBP-3, the mRNA of which has an interstitial localization, was increased at week 10. Conclusion: These data suggest that there is a dynamic relationship between tubular and interstitial compartments with regard to the IGF system in the kidney which may be important in the regulation of the cell mass.

Purpose: The etiology of interstitial cystitis is unknown. We previously identified an interstitial cystitis urine factor, antiproliferative factor, that inhibits proliferation of bladder epithelial cells in vitro and complex changes in epithelial growth factor levels, including profound decreases in heparin-binding epidermal growth factor-like growth factor (HB-EGF). Bladder and renal pelvic catheterization of patients with interstitial cystitis indicated that the antiproliferative factor is made and/or activated in the distal ureter or bladder. Therefore, we determined whether bladder epithelial cells from interstitial cystitis cases produced the antiproliferative factor and whether purified antiproliferative factor could alter production of growth factors known to be abnormal in interstitial cystitis.Materials and Methods: Antiproliferative factor activity was determined by 3H-thymidine incorporation into primary bladder epithelial cells. The antiproliferative factor was purified by size fractionation ...

We studied gene expression for transforming growth factor (TGF)-alpha, epidermal growth factor (EGF), heparin binding (HB) EGF, and the EGF receptor following acute renal failure induced by mercuric chloride administration to gain insight into potential mechanisms of renal repair. Twenty four hours after HgCl2, 2 mg/kg, creatinine increased from 0.3+/-0.01 mg/dl in controls to 2.2+/-0.26 mg/dl in injured rats (n = 5, p0.01). Similar changes were observed after 3 days. Messenger RNA expression for EGF was decreased at 24 hours in HgCl2 treated rats and remained depressed for at least 3 days. On the other hand steady state mRNA for TGF-alpha increased nearly 2 fold at day 3 in HgCl2 treated rats 4 mg/kg. Heparin binding EGF was increased early, by day one in injured kidneys and gene expression for the EGF receptor was increased as well. Immunohistochemistry documented an increase in expression of TGF-alpha in injured kidneys at distal nephron sites. These studies suggest that TGF-alpha along with HB EGF may be important ligands for the EGF receptor during repair from renal injury.

Objectives. A low molecular weight urine factor that inhibits the proliferation of normal bladder epithelial cells in vitro was previously shown to be present significantly more often in the urine of patients with interstitial cystitis (IC) than in the urine of asymptomatic age-, race-, and sex-matched control subjects. We sought to determine the specificity of this finding for IC by determining whether the urine of patients with other urogenital inflammatory disorders also contains a factor that inhibits bladder epithelial cell proliferation. Methods. Urine was collected from women with IC, acute bacterial cystitis, or vulvovaginitis, as well as from asymptomatic control women. The proliferation of primary normal adult bladder epithelial cells was determined by measuring 3 H-thymidine incorporation in vitro. Results. Osmolality- and pH-corrected urine specimens from 50 (86%) of 58 women with IC significantly inhibited human bladder epithelial cell proliferation compared with 3 (8%) of 36 asymptomatic control women, 7 (12%) of 58 women with bacterial cystitis, and 0 (0%) of 12 women with vulvovaginitis (P , 0.001 for the comparison of mean percent change in 3 H-thymidine incorporation with IC urine versus urine from each of the control groups). Optimal sensitivity and specificity values of 91.4% and 90.6%, respectively, were achievable at a cutoff of 25% inhibition of 3 H-thymidine incorporation, using all three control groups. Conclusions. The measurement of urine antiproliferative activity may be a useful noninvasive means for diagnosing IC in women. UROLOGY 52: 974‐978, 1998. © 1998, Elsevier Science Inc. All rights reserved.

We studied the renal expression of the insulin-like growth factor (IGF) system to gain a better perspective of its potential role in the hyperplastic adaptation of the distal nephron to potassium deficiency. Rats were pair fed 1% or 0.002% potassium diets for periods up to 10 days. IGF-I mRNA was diminished in potassium-deficient rats within 4 days, whereas mRNA for IGF binding protein-1 (IGFBP-1), a collecting duct-associated protein, was increased by day 7. At day 10 mRNA for IGFBP-1 in potassium-deficient animals averaged 2.07 +/- 0.53 (mean +/- SD, relative densitometry units) compared with 0.89 +/- 0.26 in control rats (n = 4, P = 0.002). Conversely, IGFBP-3, a binding protein whose mRNA has been localized to the interstitial compartment, averaged 2.40 +/- 0.02 in potassium-deficient rats and 4.77 +/- 0.05 in controls (n = 4, P < 0.03) at day 10 of treatment. Immunohistochemistry performed using a specific IGFBP-1 antibody revealed hyperplasia of distal nephron segments along with an increase in IGFBP-1 in potassium-depleted rats. These data suggest that IGFBP-1 may play an important role in the control of cellular adaptations in the hypokalemic rat kidney either directly by influencing cell migration or indirectly by localizing IGF-I to the distal nephron.

To characterize the expression of transforming growth factor-alpha (TGF-alpha) in various histologic types of renal cell carcinomas.Immunohistochemistry of renal cell carcinoma and adjacent normal tissue was performed on formalin-fixed tissue using a specific monoclonal antibody to TGF-alpha.Clear and distinct staining was present in normal distal convoluted tubules and collecting ducts. The growth factor was not observed in the glomerulus or the proximal tubule. In tumors composed of clear cells, staining was evident only in endothelial cells but not in the tumor cells themselves. In granular cell type tumors, the tumor cells as well as endothelial cells stained for TGF-alpha. When mixed cell type tumors were studied, a heterogenous pattern of growth factor expression was found. Endothelial cells and granular cells but not clear cells demonstrated positive staining.These studies suggest that TGF-alpha is likely to play a major role in neovascularization of clear cell carcinomas and that the growth factor may be more important in supporting proliferation of granular cell type tumors.

The cellular effects of insulin-like growth factor I (IGF-I) are modified by a family of binding proteins (IGFBPs) that act as reservoirs in serum for the growth factor and are produced locally by tissues, including the kidney. Because regulation of these proteins may influence renal repair, either directly or by their interactions with IGF-I, we studied gene expression during the recovery from renal failure induced by folic acid and during the compensatory increase in renal function following uninephrectomy (UNX). Expression of IGF-I, the IGF-I receptor (IGF-IR), and all six IGFBPs was detected using an ribonuclease protection assay. IGFBP-5 was the most abundant binding protein mRNA present in kidney, whereas IGFBP-2 and -6 were the least abundant. During regeneration following folic acid-induced acute renal failure, IGF-I, IGFBP-3, and IGFBP-5 mRNAs declined in abundance approximately two- to threefold. On the other hand, IGF-IR, IGFBP-1, and IGFBP-2 were increased (approximately 2-, 6-, and 6-fold, respectively) in the first 24 h. IGFBP-1 mRNA remained elevated for at least 3 days. Despite the known increase in cellular RNA content following UNX, little difference in specific expression of mRNAs was observed. Because IGFBP-1 has been shown to stimulate cell migration and has previously been localized to the distal nephron, the site of greatest injury in the folic acid model, these data are compatible with the notion that this protein may function either directly to affect cellular repair or act as a reservoir for IGF-I under conditions of cellular damage.

Insulin-like growth factor (IGF)-binding proteins and the IGF-I receptor in rat kidney were studied to gain perspective on their potential roles in the control of renal cell mass. Thirty days after nephrectomy (UNx), membranous whole-kidney binding of IGF-I averaged 9.5 +/- 1.0 pmol/kidney, whereas binding to sham-operated controls (SNx) averaged 6.3 +/- 0.8 pmol/kidney (N = 6; P0.01). Scatchard analysis of IGF-I binding per milligram of protein to glomerular membranes or proximal tubule basolateral membranes (BLM) at Day 30 did not reveal significance differences in Bmax or KD between SNx and UNx; however, protein per glomerulus increased from 361 +/- 33 ng/glomerulus in SNx animals to 826 +/- 123 ng/glomerulus in UNx rats (N = 6; P0.002). Histomorphometrics documented an increase in proximal tubule circumference per cell and an increase in the basolateral/basement membrane ratio. IGF-I affinity labeling studies demonstrated three proteins in both glomerular membranes and proximal tubule BLM; molecular weight approximately 140,000 d, the alpha subunit of the IGF-I receptor, a protein200,000 d, and a protein approximately 31,000 d that was immunostained with IGF-binding protein-5 antibodies. Differences in expression between SNx and UNx were not observed at Day 30 in either glomeruli or BLM. These studies suggest that cytosolic hypertrophy of proximal nephron structures is accompanied by membrane hypertrophy with a fixed density of IGF-I receptor and IGF-binding protein-5.

Known stimulators of the calcium-sensitive phospholipid-dependent protein kinase C were investigated for their ability to regulate ornithine decarboxylase (ODC) activity in rat kidney. In the control state ODC activity averaged 84.9 +/- 13.2 pmol mg-1 min-1 in a soluble fraction (n = 4). Four hours following the intraperitoneal injection of 2.5 nmol/g body weight of phorbol 12-myristate 13-acetate (PMA) activity increased to 284.1 +/- 10.9 pmol mg-1 min-1 (n = 4; p less than 0.001). A chemically distinct stimulator of PKC, mezerein, had a similar effect on ODC in kidney and liver. Activity stimulated by PMA in kidney was dependent on the synthesis of both new mRNA and protein. Four hours following unilateral nephrectomy (UNX), ODC activity increased from 112.9 +/- 15.6 pmol mg-1 min-1 in sham-operated animals to 319.1 +/- 30.0 pmol mg-1 min-1 in animals postnephrectomy (n = 4; p less than 0.01). Activity of ODC stimulated by phorbol esters was not additive to that seen following UNX. Twelve hours following the induction of renal growth by folic acid. ODC-specific activity was at basal levels. Neither UNX nor PMA were able to stimulate ODC at this time. These data suggest that protein kinase C may be involved in the regulation of ODC in rat kidney.

The aim of this study was to explore African American patients' experiences managing their hypertension and to investigate their perspective on how technology might be used to improve hypertension self-management. We conducted and analyzed four focus groups with 32 African American participants diagnosed with hypertension to develop a culturally tailored content for a home-based telecare program aimed at improving hypertension care in African Americans. The discussion about the use of technology was well accepted, demonstrated culturally and gender specific barriers in hypertension care, and generated a comprehensive list of concepts and features to be included to a home-based computerized hypertension telemanagement system.

The association between physical activity and renal function in subjects with and without metabolic syndrome was examined.Renal function was evaluated in subjects of the Third National Health and Nutrition Examination Survey by using calculated glomerular filtration rate (GFR) and urine microalbuminuria (urine albumin-creatinine ratio). These parameters were studied as a function of physical activity by using a 1-month recall. Measures included activity variety, number of different types of exertion; activity frequency, the sum of all activity periods; and metabolic equivalent (METS), the total sum of energy expenditures. Individuals were segregated into those with metabolic syndrome and no metabolic syndrome; ages 18 to 44, 45 to 55, and older than 55 years; men and women; and 3 racial groups: Caucasians, African Americans, and Mexican Americans.Younger individuals, men, and those with higher levels of education had a lower risk for metabolic syndrome. The groups also had better renal function, measured by using GFR and urinary protein. Those without metabolic syndrome performed larger numbers of activity varieties in the unadjusted analysis (2 +/- 2 [SD]; n = 11,184) compared with those with metabolic syndrome (1 +/- 1; n = 2,569; P0.0001). Similar differences in activity frequency and METS also were observed. Conversely, GFR correlated with activity variety and METS, but negatively with activity frequency in those without metabolic syndrome after adjustment for confounders. In subjects with metabolic syndrome, GFR correlated only with activity variety. Variable observations were made among ages, sexes, and races.There is a clear association between physical activity and GFR, particularly in subjects without metabolic syndrome; however, cross-sectional analysis precludes establishing causality.

In patients with renal disease, infection remains among the most common causes of morbidity and mortality. Alterations in the function of the immune system, as well as unique exposures of this patient population, account for the increased risk. Vaccination is an invaluable tool in preventing many infectious diseases. Unfortunately, responsiveness to vaccination in patients with renal disease can be diminished. In the present review, we examine the available evidence on the use of vaccinations in adult patients at different stages of chronic kidney disease. We address efficacy, clinical outcomes, and potential costs of individual vaccinations and provide our recommendations based on the literature reviewed. We also identify areas in which additional research is needed.

Aim: We studied control of the epidermal growth factor (EGF) receptor and its ligands during kidney injury, since they may be importantly involved in repair. Methods: The folic acid model of renal injury was used in these studies. Messenger RNA (mRNA) was evaluated by solution hybridization. Immunohistochemistry of transforming growth factor alpha (TGF-α) was also performed. Results: Twenty-four hours after folic acid induced acute renal injury, creatinine increased from 0.3 ± 0.03 mg/dl in controls to 2.0 ± 0.8 mg/dl in folic acid injured kidneys (n = 4, p < 0.03). mRNA for the EGF receptor was increased nearly sevenfold by 24 h, and mRNA for the receptor was increased as early as 1 h following folic acid treatment. EGF receptor ligand caused a profound downregulation of the receptors in proximal tubule basolateral membranes, but receptors returned rapidly to the membrane surface in injured kidneys. The mRNA levels for heparin-binding EGF and TGF-α, two EGF receptor ligands, increased within 1 h of injury. TGF-α mRNA increased from 1.0 ± 0.09 (relative densitometry units) in control animals to 2.9 ± 0.13 in folic acid treated rats at 24 h (n = 4, p < 0.01), and immunohistochemical staining for TGF-α increased in injured kidneys at distal nephron sites. Conclusions: These studies indicate that upregulation of the EGF receptor is related to an increase in mRNA. The rapid return of receptors to the membrane surface following ligand stimulation may be useful in maintaining a mitogenic stimulus. Multiple EGF-like ligands may be important in activating the upregulated EGF receptor during repair from renal injury.

Interstitial cystitis (IC) is a chronic bladder disease for which the etiology is unknown. Because the bladder epithelium is often abnormal in IC, we determined whether the levels of specific urine growth factors postulated to be important for bladder epithelial proliferation are altered in IC.ELISAs were used to determine levels of epidermal growth factor (EGF), insulin-like growth factor 1 (IGF1), insulin-like growth factor binding protein 3 (IGFBP3), and heparin binding epidermal growth factor-like growth factor (HB-EGF) in urine specimens from women with IC, asymptomatic women without bladder disease, and women with bacterial cystitis.Urine HB-EGF levels were specifically and significantly decreased in IC patients as compared to asymptomatic controls or patients with bacterial cystitis, whether expressed as concentration (amount per volume of urine) or the amount relative to urine creatinine in each specimen. In contrast, urine EGF, IGF1, and IGFBP3 levels were all significantly elevated in IC patients compared to asymptomatic controls. Further, the amounts of urine EGF and IGF1 were also elevated in IC patients as compared to patients with bacterial cystitis, and urine IGFBP3 levels were significantly elevated when expressed per milligram of urine creatinine.These findings indicate that complex changes in the levels of urine epithelial cell growth factors (EGF, IGF1, and HB-EGF) and a growth factor binding protein (IGFBP3) are associated with IC. While EGF, IGF1, and IGFBP3 levels are either the same or increased in the urine of IC patients as compared to patients with bacterial cystitis or asymptomatic controls, HB-EGF levels are significantly decreased in the urine of IC patients. Understanding the reasons for these changes may lead to understanding the pathogenesis of this disorder.

Previous studies have demonstrated that the glomerulus and proximal tubule basolateral membrane possess both insulin-like growth factor (IGF) receptors and IGF-binding proteins (IGFBPs). The purpose of this study is to examine the control of these proteins in defined nephron segments during dietary protein restriction. Animals were fed isocaloric 6% (low-protein [LP]) or 40% (high-protein) protein diets for 7 to 10 days. 125 I[IGF] affinity labeling of membranes prepared from isolated glomeruli or proximal tubule basolateral membranes demonstrated two proteins on autoradiograms of 6% polyacrylamide gels with molecular weights of 140,000 d and more than 200,000 d that were blocked by 300 nmol/L unlabeled IGF-I. The lower molecular weight species has been identified previously as the alpha subunit of the IGF-I receptor and was upregulated by a 6% LP diet. The upregulation of the IGF-I receptor was evident on 12% polyacrylamide gels of both glomeruli and basolateral membranes. Another protein with a molecular weight of approximately 38,000 d also was upregulated by LP diet. The protein was evident on 125 I-IGF-I ligand blots and was immunostained with IGFBP-5 antibodies. Cytosol prepared from cortical tissue also demonstrated a 31,000-d protein that was immunostained with IGFBP-5 antibodies in animals fed a LP diet, but not in animals fed a high-protein or normal diet. RNA prepared from cortical tissue and hybridized with a IGFBP-5 cDNA probe revealed a 6.0-Kb transcript that was increase by LP diet. These data indicate that the enhanced IGF-I binding observed in proximal nephron structures following a LP diet is a result of increased expression of the IGF-I receptor as well as IGFBP-5. The increase in membrane-associated and soluble binding protein when IGF-I circulating levels are diminished suggests that the protein might function as an extravascular reservoir, limiting degradation and facilitating growth factor interaction with the receptor.

This study examines growth-regulating adaptations made by the proximal nephron in response to hypophysectomy (HYPX). Fourteen days after HYPX, circulating insulin-like growth factor I (IGF-I) levels were diminished, averaging 97 +/- 7 compared with 650 +/- 69 ng/ml in controls (n = 5, P < 0.001). Similar data were observed at day 7. Binding of 125I-IGF-I to isolated glomerular membranes and proximal tubule basolateral membranes (BLM) was increased in HYPX rats. Affinity labeling of membranes with 125I-IGF-I followed by electrophoresis on 6% polyacrylamide gels demonstrated two bands, one of approximately 140 kDa and another of > 200 kDa. The lower-molecular-mass protein, which has been identified as the alpha-subunit of the IGF-I receptor, and the higher-molecular-mass species were both upregulated by HYPX. Ligand blotting with IGF-I demonstrated a 31-kDa protein in both membranes, identified by immunostaining as IGF binding protein 5 (IGFBP-5), not IGFBP-1 or IGFBP-2. Affinity labeling documented an upregulation of this protein in both membranes after HYPX. Ligand blotting demonstrated a 31-kDa protein in HYPX cortical but not normal cortical or medullary cytosol that was immunostained with IGFBP-5 antibodies. RNA prepared from normal kidney cortical tissue demonstrated a 6.0-kb IGFBP-5 transcript, which was increased at day 14 after HYPX. Adaptations in the kidney after HYPX include an upregulation of the IGF-I receptor as well as IGFBP-5.(ABSTRACT TRUNCATED AT 250 WORDS)

The importance of receptor expression and protein kinase C to epidermal growth factor (EGF)-induced cell proliferation was studied in LLC-PK1 kidney cells. These cells have both high- and low-affinity binding sites for EGF. Neither transforming growth factor-beta nor tumor necrosis factor altered EGF receptor expression. On the other hand, retinoic acid induced a concentration-dependent increase in EGF binding that was maximal at 1 mumol/L. One micromolar of retinoic acid increased EGF binding from 0.38 +/- 0.01 fmol/10(6) cells in controls to 1.10 +/- 0.03 fmol/10(6) in treated cells at 18 hours (n = 8, P < 0.001). The increase in binding was the result of an increase in the Bmax of the high-affinity receptor. The upregulation of the EGF receptor induced by retinoic acid was associated with enhanced EGF-induced growth promotion. A 45-minute incubation of cells with phorbol 12-myristate 13-acetate caused a concentration-dependent decrease in EGF binding that was prevented by a 40-hour, 2 mumol/L pre-exposure to phorbol 12-myristate 13-acetate; 10(-8) mol/L EGF also caused a downregulation of the EGF receptor that was not prevented by phorbol 12-myristate 13-acetate or retinoic acid. Downregulation of protein kinase C did not interfere in the capacity of EGF to induce growth in these cells. These studies demonstrate that EGF receptor upregulation plays an important role in the control of EGF-induced cell growth. Protein kinase C regulates EGF binding in these cells; however, it is not necessary for EGF-induced growth promotion or receptor downregulation.

1. We examined the effects of dietary protein manipulations in partially nephrectomized (one and one-third nephrectomy) and normal rats to gain perspective on the relative importance of circulating versus intrarenal (collecting tubule) insulin-like growth factor-I in the control of proximal nephron receptor density. In addition, we studied the factors that influence liver insulin-like growth factor-I secretion in partially nephrectomized rats. 2. Dietary protein restriction (6% versus 40%) lowered circulating levels of insulin and insulin-like growth factor-I in both normal and partially nephrectomized rats up to 3 weeks after institution of the diets; however, growth hormone levels were little changed. Reduced renal mass stimulated intrarenal production of insulin-like growth factor I regardless of the diet. 3. Scatchard analysis revealed that the density of insulin-like growth factor-I receptors on glomerular and proximal tubule basolateral membranes increased when circulating levels of insulin-like growth factor-I were diminished, despite raised levels of intrarenal insulin-like growth factor-I, in partially nephrectomized rats. 4. Circulating insulin-like growth factor-I, rather than the tissue level, plays the dominant role in the control of proximal nephron receptor density under physiological conditions. Insulin, but not growth hormone, may play a role in liver insulin-like growth factor-I secretion in partially nephrectomized rats during dietary manipulations.

Purpose To determine association between physical activity and renal parameters including Glomerular Filtration rate, (GFR), Serum Creatinine, Blood Urea Nitrogen (BUN) and Proteinuria (Urine Albumin to Creatinine Ratio) in subjects with and without metabolic syndrome. Methods Subjects were classified as having metabolic syndrome if they had 3 of the following 5 components: high blood pressure, high serum triglycerides, high waist circumference, high plasma glucose, and low HDL-cholesterol. Physical activity was defined as the number of physical activity sessions in the prior 30 days as reported by the subjects on the questionnaire. Renal parameters were measured in laboratory analysis, and GFR was calculated from serum creatinine using the MDRD formula. Linear regression for the cross-sectional association of physical activity with the individual renal parameters was adjusted for age, race, sex, education, hypertension, diabetes, household size, family size, hypertension, diabetes and poverty income ratio. Results Analysis was performed on 15,705 subjects, of whom 53% were female, 42% non-Hispanic White, 27% African American, and 27% Mexican American. Metabolic syndrome was present in 20%. In unadjusted regression in subjects with vs. without metabolic syndrome, physical activity had mixed association with GFR (β = 0.02; p p p = 0.0001 vs. β = 0.0007; p p = 0.21 vs. β = 0.0003; p = 0.002) and a significant positive association with BUN (β = 0.011; p = 0.04 vs. β = 0.005; p = 0.001). In adjusted regression in subjects with vs. without metabolic syndrome, physical activity had a significant positive association with GFR (β = 0.05; p p p = 0.02 vs. β = 0.0001; p = 0.11); no association with proteinuria (β = 0.0002; p = 0.62 vs. β = 0.0001; p = 0.07) and no association with BUN (β = 0.004; p = 0.43 vs. β = 0.002; p = 0.09). Conclusion Higher levels of physical activity were associated with better GFR regardless of metabolic syndrome.

Our previous observations of increased renal protein synthesis in rats subjected to the constant intravenous reinfusion of half their urine output has suggested that the circulatory retention of renotrophic factors in urine is capable of stimulating renal growth. In the present studies, using this same model of "half-urine-reinfusion," which is designed to produce a selective halving of renal excretory function, we have demonstrated significant increases in total DNA content and the incorporation of tritiated thymidine in renal DNA. In addition, a bioassay method was developed in which an assay rat, given an intravenous infusion of urine from another rat, exhibited increases in the incorporation of thymidine into renal DNA and the incorporation of radiolabelled choline into renal phospholipid. This renotrophic activity in the urine was only minimally decreased by heating to 100 degrees C for 30 min and was confined to ultrafiltration fractions retained on a membrane with a nominal 10,000-dalton solute rejection. Removal of one kidney from the rats from which the urine was obtained led to only a modest and transient reduction in the excretion of renotrophic activity, suggesting that the urinary renotrophic factors are of circulatory, not renal, origin. Isolated renal cortical fragments incubated with an ultrafiltration retentate of urine displayed a dose-dependent increase in choline incorporation into phospholipid, suggesting a direct action of the factors on kidney tissue. Finally, no evidence of stimulation of either DNA or phospholipid synthesis could be seen in hepatic tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

The relative roles of phospholipid fatty acyl chain length and phospholipid fatty acyl chain unsaturation in the determination of rat renal brush border membrane order were examined using multilamellar liposomes. Exposure of brush border membranes to sphingomyelinase resulted in a time- and concentration-dependent decrement in sphingomyelin content. Liposomes prepared from lipid extracts of these membranes were reconstituted to defined phosphatidylcholine (PC)/sphingomyelin (SPH) ratios with pure synthetic PCs of defined chain length and degrees of unsaturation. Mixed-acid PCs from bovine liver, egg, and the rat renal brush border membrane were also examined. The steady state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH) at 37 degrees C was used to reflect acyl chain packing. The steady state anisotropy of DPH in liposomes isolated from the rat renal brush border membrane averaged 0.205 +/- 0.001, n = 8. When liposomes were reconstituted to PC/SPH ratios of 1.1, 1.6, and 2.4 with saturated PCs of acyl chain length 16 to 22, differences in anisotropy between groups were not observed. However, when PCs containing unsaturated or mixed-acid fatty acyl chains were introduced, anisotropy decreased in a concentration dependent fashion. These data suggest that phospholipid fatty acyl chain unsaturation, but not acyl chain length, has a powerful influence on renal brush border membrane order and the PC/SPH ratio is an important determinant of renal membrane order by virtue of the unsaturated fatty acids normally present with these phospholipids.

The possible role of calmodulin in solute transport was examined in the kidney of the rat. Utilizing a radioimmunoassay, calmodulin was identified and quantitated in homogenates of the cortex of the kidney. The physiologic significance of these findings was examined utilizing in vivo microperfusion techniques applied to the proximal convoluted tubule of the thyroparathyroidectomized rat. The addition of dibutyryl cyclic adenosine monophosphate (cAMP) to the luminal perfusion solution resulted in a lower rate of water absorption of 1.67 +/- 0.09 nl min-1 mm-1 as compared to 2.46 +/- 0.11 in controls. The addition of either of two compounds with affinity for calmodulin, trifluoperazine (TFP) or W-13, reversed the cAMP-induced inhibition of water absorption. In the absence of cAMP, neither agent affected water absorption. Analogs of TFP and W-13 with lower binding affinities for calmodulin had no effect on water absorption and did not reverse the cAMP effect. None of the above experimental maneuvers affected the absorption of phosphate. These results demonstrate the presence of calmodulin in the kidney of the rat and suggest that calmodulin may be involved in cAMP-associated inhibition of water and electrolyte transport in the proximal tubule of the rat.

Activity of calcium-sensitive phospholipid-dependent protein kinase C following nephron loss. The localization and activity of the calcium-sensitive phospholipid-dependent protein kinase C (PKC) were examined following the loss of 50% of functioning nephron mass. Four hours following unilateral nephrectomy in rats, soluble (100,000g supernatant) proteins in the contralateral kidney were increased by 11% compared to sham operated controls; the increase was 33% 144 hours following surgery. The specific activity of PKC did not change in the cytosol at any of the time periods examined and averaged 63.9 ± 8.2 pmol · mg−1 · min−1 in unilaterally nephrectomized animals four hours following surgery. Four hours following sham surgery total soluble PKC activity averaged 1667.0 ± 278.4 pmol · kidney−1 · min−1, whereas activity averaged 3067.7 ± 415.4 pmol · kidney−1 · min−1 in animals post-nephrectomy (N = 5, P < 0.04). Similar data was seen 144 hours following surgery. To examine the PKC activity in plasma membranes of proximal tubular cells, brush border membranes were prepared from rat kidney cortex. Twenty-four hours following unilateral nephrectomy, activity averaged 193.8 ± 14.9 pmol · mg−1 · min−1, while activity in membranes isolated from sham operated animals averaged 76.6 ± 8.0 pmol mg−1 min−1 (N = 5, P < 0.001). Similar data was evident 48 hours following surgery. A small increment in activity was seen in the basolateral membrane preparation 24 hours following unilateral nephrectomy but not at 48 hours. These data indicate that cellular PKC activity increases rapidly following reductions in renal mass, and there are selective increments in the brush border membrane of the proximal tubular cell. The localization of PKC to this membrane may have important consequences for adaptations following nephron loss.

The activity of the calcium sensitive phospholipid dependent protein kinase C (PKC) was studied in cytosol and in the proximal tubular luminal membrane of rats during growth. Cytosolic activity was elevated at 14 and 21 days of age and fell to adult levels by day 60. Luminal brush border membrane activity on the other hand was low on day 14 but reached adult levels by day 21. Changes in brush border membrane PKC activity may have important consequences for the development of electrolyte transport in proximal tubular cells.

The binding of 125I-labeled epidermal growth factor (EGF) to plasma membranes prepared from rat kidney cortex was studied following unilateral nephrectomy, a model of proximal tubule cell hypertrophy, and following the administration of folic acid, a model of proximal tubule cell hyperplasia. Binding of 125I-EGF was a linear function of basolateral membrane protein content and time of incubation. Specific binding to luminal brush-border membranes was not evident in these studies. Neither insulin nor insulin-like growth factor I could displace EGF binding, indicating that binding was specific. Scatchard analysis revealed a single binding site. The KD in sham-operated animals 48 h after surgery was 11.2 +/- 1.4 nM, whereas Bmax averaged 95.2 +/- 4.1 fmol/mg protein (n = 3). Similar values were obtained in nephrectomized animals. The Bmax of folic acid-untreated animals averaged 212.5 +/- 6.9 fmol/mg 48 h after administration, whereas that of vehicle-injected controls averaged 85.4 +/- 9.2 (n = 3, P less than 0.001). Differences in binding were not related to changes in affinity, ligand degradation by the preparations, or receptor binding of endogenous EGF. These data indicate that regeneration following folic acid administration is associated with an upregulation of proximal nephron EGF receptors that may play an important role in the mitogenic response.

The substrate specificities of calcium/phospholipid-dependent kinase-C (PKC) were examined in rat kidney cortex, and localization of the protein was studied after the induction of diabetes. The cytosolic kinase was eluted from an anion exchange resin using a linear gradient of 0-0.15 M NaCl. A sharp peak of activity was demonstrated at approximately 80 mM using histone as a substrate. The kinase demonstrated a broad pH optimum of 6.5-8.0. ATP was the preferred phosphorus donor. The Ka for ATP averaged 2.6 +/- 0.1 microM (n = 4) and was not different in diabetic animals. Lysine-rich histones, but not arginine-rich or mixed histones, were the most suitable phosphorus acceptors. Phosphatidylserine stimulated kinase activity with Ka of 4.5 +/- 0.7 microM in the presence of 20 microM diolein (n = 3). Twenty micromolar diolein in the presence of 25 microM phosphatidylserine lowered the apparent Ka for calcium from 17.2 +/- 1.4 to 3.3 +/- 1.5 microM (n = 3; P less than 0.01). Similar data were evident in diabetic animals. Diabetic renal growth was induced by the injection of streptozotocin (35 mg/kg, iv). At the end of 4 weeks, blood glucose averaged 119.6 +/- 7.4 mg/dl in vehicle-injected controls and 548.7 +/- 21.6 mg/dl in diabetic animals (n = 5; P less than 0.001). Despite reduced weight gains in diabetic animals, renal protein content was increased in this group compared to the control value. Neither cytosolic nor proximal tubule basolateral membrane PKC activity changed after the induction of diabetes; however, luminal brush border PKC activity increased from 83.8 +/- 4.6 pmol/mg.min in control animals to 107.3 +/- 55 pmol/mg.min in diabetic animals (n = 5; P less than 0.02). The increased activity in the brush border membrane may have important consequences for the growth response of the kidney in diabetes.

A sensitive enzymatic assay for the measurement of intracellular choline is described. The separation of choline from choline-containing phospholipids is accomplished by a minor modification of the Folch technique. The method is based on the specific oxidation of choline by choline oxidase. Phenol and 4-aminoantipyrine in the presence of hydrogen peroxide generated by the oxidation of choline and peroxidase form a red quinone dye which can be detected spectrophotometrically. The assay was useful between 12.5 and 100 nanomoles of choline. The recovery of standard choline in liver homogenates averaged 102 ± 1.6%. Structurally similar compounds produced minimal interference.

The possible role of progressive methylation of phosphatidylethanolamine to phosphatidylcholine on sodium transport was examined in a suspension of rabbit proximal convoluted tubules. The relation between progressive methylation and the action of parathyroid hormone on sodium transport in this nephron segment was also determined. Incubation of the suspended tubules for 10 minutes at 37 degrees C with 200 microM S-adenosyl-L-[3H]-methyl methionine, a methyl donor, revealed a significant rate of de-novo phosphatidylcholine synthesis. The inactive adenosine analogue, 3-deazaadenosine (DZA), had a significant inhibitory effect on lipid methylation. Despite the inhibition of methylation by DZA, the ouabain sensitive component of oxygen consumption, an index of sodium transport, was not affected. PTH significantly inhibited ouabain sensitive oxygen consumption but had no effect on the methylation of phosphatidylethanolamine. Inhibition of methylation by DZA, did not affect the inhibitory effect of PTH on oxygen consumption. These studies demonstrate that in the proximal convoluted tubule of the rabbit the progressive methylation pathway is present and that inhibition of this pathway does not affect sodium transport. In addition, these studies suggest that the inhibitory effect of PTH on sodium transport is not mediated by the methylation pathway.