Critical timing of ACEi initiation prevents compensatory glomerular hypertrophy in the remaining single kidney.

Increasing evidence suggests that single in kidney states (e.g., kidney transplantation and living donation) progressive glomerulosclerosis limits kidney lifespan. Modeling shows that post-nephrectomy compensatory glomerular volume (GV) increase drives podocyte depletion and hypertrophic stress resulting in proteinuria and glomerulosclerosis, implying that GV increase could serve as a therapeutic target to prevent progression. In this report we examine how Angiotensin Converting Enzyme inhibition (ACEi), started before uninephrectomy can reduce compensatory GV increase in wild-type Fischer344 rats. An unbiased computer-assisted method was used for morphometric analysis. Urine Insulin-like growth factor-1 (IGF-1), the major diver of body and kidney growth, was used as a readout. In long-term (40-week) studies of uni-nephrectomized versus sham-nephrectomized rats a 2.2-fold increase in GV was associated with reduced podocyte density, increased proteinuria and glomerulosclerosis. Compensatory GV increase was largely prevented by ACEi started a week before but not after uni-nephrectomy with no measurable impact on long-term eGFR. Similarly, in short-term (14-day) studies, ACEi started a week before uni-nephrectomy reduced both GV increase and urine IGF-1 excretion. Thus, timing of ACEi in relation to uni-nephrectomy had significant impact on post-nephrectomy "compensatory" glomerular growth and outcomes that could potentially be used to improve kidney transplantation and live kidney donation outcomes.

Detection and in vitro metabolism of the confiscated peptides BPC 157 and MGF R23H.

A new peptide, body protecting compound (BPC), BPC 157, and a variant of mechano-growth factor (MGF), MGF R23H, were identified in confiscated vials. BPC 157 has the amino acid sequence, GEPPPGKPADDAGLV, and is currently under investigation for the promotion of healing and recovery in a variety of tissues. In vitro metabolism experiments in plasma demonstrate that MGF R23H has good stability and should be detectable in urine, while BPC 157 forms a stable metabolite that should be detectable in urine. A weak cation exchange solid phase extraction method was validated for detection of BPC 157 in urine. The method has a limit of detection of 0.1 ng/mL, precision of less than 20%, and good linearity, r2 0.998. BPC 157 was stable in urine for at least 4 days. The specificity of the method is improved by measurement of a potential BPC metabolite along with the parent peptide. Copyright © 2016 John Wiley & Sons, Ltd.

Serum and urine insulin-like growth factor-1 as biochemical growth maturity indicators.

INTRODUCTION: Biochemical markers are agents directly involved in bone growth and remodeling and can be quantitatively evaluated from various biologic fluids. The aim of this study was to assess the changes in the levels of insulin-like growth factor-1 (IGF-1) in serum and urine as a growth maturity indicator and to compare them with the cervical vertebral maturation radiographic stages. METHODS: The study was conducted with 72 female subjects aged 8 to 20 years. Cervical vertebral maturation stages, and serum and urine IGF-1 levels were recorded for all subjects, and the subjects were equally divided into the 6 cervical vertebral maturation groups. Median values of IGF-1 for each stage of cervical vertebral maturation were calculated and statistically compared with those of the other stages. RESULTS: The levels of serum and urine IGF-1 at stage 4 of cervical vertebral maturation were significantly higher than those from the other stages (P <0.01). Stage 4 corresponded to a mean age of 13.67 years. A significant correlation was observed between serum and urine IGF-1 (P <0.001). CONCLUSIONS: Urine IGF-1 follows the growth curve similar to serum IGF-1. Thus, urine IGF-1 may be regarded as a promising noninvasive tool for growth assessment. Further research is necessary to validate these results in a different population and with a larger sample.

Decreased 6-Sulfatoxymelatonin Excretion in Male GH-Deficient Children and Adolescents.

AIM: To evaluate melatonin secretion in a group of untreated and treated male growth hormone (GH)-deficient children and adolescents. METHODS: We studied 44 male subjects: 8 untreated GH-deficient patients (GHDnt), 16 treated GH-deficient patients (GHDt) and 20 healthy children and adolescents as control group (CG). We measured urinary 6-sulfatoxymelatonin (6-SM) in total (24-hour samples), nocturnal (18.00-8.00 h) and diurnal samples (8.00-18.00 h). Levels of 6-SM were expressed as micrograms excreted per time interval and x0394; values (difference between nighttime and daytime values). RESULTS: Significant differences were observed among the 3 groups of pediatric subjects studied for total 6-SM (p < 0.0001), nocturnal 6-SM (p < 0.0001) and x0394; values (p < 0.0001). Subsequent analysis showed significantly higher levels for total 6-SM, nocturnal 6-SM and nighttime-daytime x0394; in the CG versus the GHDnt (p < 0.01) and in the CG versus the GHDt group (p < 0.01). No significant correlations were found between 6-SM excretion and insulin-like growth factor-1 values in the children and adolescents studied. CONCLUSIONS: GH-deficient patients showed lower levels of 6-SM. Our findings provide a different insight to a further understanding of some chronobiological disorders involved in GH deficiency in children.

Insulin-like growth factors and kidney disease.

Insulin-like growth factors (IGF-1 and IGF-2) are necessary for normal growth and development. They are related structurally to proinsulin and promote cell proliferation, differentiation, and survival, as well as insulin-like metabolic effects, in most cell types and tissues. In particular, IGFs are important for normal pre- and postnatal kidney development. IGF-1 mediates many growth hormone actions, and both growth hormone excess and deficiency are associated with perturbed kidney function. IGFs affect renal hemodynamics both directly and indirectly by interacting with the renin-angiotensin system. In addition to the IGF ligands, the IGF system includes receptors for IGF-1, IGF-2/mannose-6-phosphate, and insulin, and a family of 6 high-affinity IGF-binding proteins that modulate IGF action. Disordered regulation of the IGF system has been implicated in a number of kidney diseases. IGF activity is enhanced in early diabetic nephropathy and polycystic kidneys, whereas IGF resistance is found in chronic kidney failure. IGFs have a potential role in enhancing stem cell repair of kidney injury. Most IGF actions are mediated by the tyrosine kinase IGF-1 receptor, and inhibitors recently have been developed. Further studies are needed to determine the optimal role of IGF-based therapies in kidney disease.

Current trends in mass spectrometry of peptides and proteins: Application to veterinary and sports-doping control.

Detection of misuse of peptides and proteins as growth promoters is a major issue for sport and food regulatory agencies. The limitations of current analytical detection strategies for this class of compounds, in combination with their efficacy in growth-promoting effects, make peptide and protein drugs highly susceptible to abuse by either athletes or farmers who seek for products to illicitly enhance muscle growth. Mass spectrometry (MS) for qualitative analysis of peptides and proteins is well-established, particularly due to tremendous efforts in the proteomics community. Similarly, due to advancements in targeted proteomic strategies and the rapid growth of protein-based biopharmaceuticals, MS for quantitative analysis of peptides and proteins is becoming more widely accepted. These continuous advances in MS instrumentation and MS-based methodologies offer enormous opportunities for detection and confirmation of peptides and proteins. Therefore, MS seems to be the method of choice to improve the qualitative and quantitative analysis of peptide and proteins with growth-promoting properties. This review aims to address the opportunities of MS for peptide and protein analysis in veterinary control and sports-doping control with a particular focus on detection of illicit growth promotion. An overview of potential peptide and protein targets, including their amino acid sequence characteristics and current MS-based detection strategies is, therefore, provided. Furthermore, improvements of current and new detection strategies with state-of-the-art MS instrumentation are discussed for qualitative and quantitative approaches.

Mass spectrometric characterization of a biotechnologically produced full-length mechano growth factor (MGF) relevant for doping controls.

OBJECTIVE: Since Goldspink and colleagues identified the expression of the mRNA of an insulin-like growth factor 1 (IGF-1) isoform in response to mechanical stress in 1996, substantial research into the so-called mechano growth factor and its modus operandi followed until today. Promising preclinical results were obtained by using the synthetic, 24-amino acid residues spanning peptide translated from the exons 4-6 of IGF-1Ec (which was later referred to as the mechano growth factor (MGF) peptide), particularly with regard to increased muscle myoblast proliferation. Consequently, the MGF peptide represented a promising drug candidate for the treatment of neuromuscular disorders; however, its misuse potential in sport was also identified shortly thereafter, and the substance (or class of substances) has been considered prohibited according to the regulations of the World Anti-Doping Agency (WADA) since 2005. While various MGF peptide versions have been known to sports drug testing authorities, the occurrence of a 'full-length MGF' as offered via illicit channels to athletes or athletes' managers was reported in 2014, arguably being undetectable in doping controls. METHODS: An aliquot of the product was obtained and the content characterized by state-of-the-art analytical approaches including gel electrophoretic and mass spectrometric (top-down and bottom-up) sequencing approaches. Upon full characterization, its implementation into modified routine doping controls using ultrafiltration, immunoaffinity-based isolation, and nanoliquid chromatography-high resolution/high accuracy mass spectrometry was established. RESULTS: A protein with a monoisotopic molecular mass of 12264.9 Da and a sequence closely related to IGF-1Ec (lacking the signal- and propeptide moiety) was identified. The C-terminus was found to be modified by the elimination of the terminal lysine and a R109H substitution. With the knowledge of the compound's composition, existing doping control assays targeting peptide hormones such as IGF-1 and related substances were assessed as to their capability to detect the full-length MGF. The analyte was detectable at concentrations of 0.25 ng/mL using adapted routine test methods employing immunoaffinity purification followed by nanoscale liquid chromatography-high resolution/high accuracy (tandem) mass spectrometry. CONCLUSIONS: A potentially performance enhancing 'full-length' MGF derivative was identified and successfully implemented into sports drug testing protocols. Future tests are indicated probing for optimized/dedicated detection methods and assessment of efficacy and elimination kinetics of the substance.

Determination of IGF-1 and IGF-2, their degradation products and synthetic analogues in urine by LC-MS/MS.

Peptide analysis in doping controls by means of nano-UPLC coupled high resolution/high mass accuracy mass spectrometry provides the state-of-the-art technique in modern sports drug testing. The present study describes a recent application of this technique for the qualitative determination of different urinary insulin-like growth factor (IGF) related peptides. After simultaneous isolation by solid phase extraction and magnetic particle-based immunoaffinity purification, target analytes (IGF-1, IGF-2, Des1-3-IGF-1, R(3)-IGF-1 and longR(3)-IGF-1) were separated by nano-liquid chromatography prior to mass spectrometric detection. Endogenously produced IGF-1 and IGF-2, as well as the degradation product Des1-3-IGF-1, were frequently detected in urine samples from healthy volunteers in a concentration range of 20-400 pg mL(-1). The impact of IGF binding proteins (IGFBPs), being also present in urine, was potentially estimated by an additional ultrafiltration step in the sample preparation procedure. The synthetic analogue longR(3)-IGF-1, which is assumed to be subject to misuse by cheating athletes, was also analysed and detected in fortified urine samples. Besides the intact molecule, an N-terminally truncated degradation product Des1-10-longR(3)-IGF-1 was identified as the more stable target for doping controls using urine samples. The method was validated for qualitative purposes considering the parameters specificity, limit of detection (20-50 pg mL(-1)), recovery (10-35%), precision (<20%), linearity, robustness and stability.

Effect of physical fitness and endurance exercise on indirect biomarkers of growth hormone and insulin misuse: Immunoassay-based measurement in urine samples.

Indirect biomarkers of recombinant human growth hormone (rhGH), insulin-like growth factor-I (IGF-I), insulin-like growth factor-II (IGF-II), insulin-like growth factor binding proteins (IGFBP-2 and IGFBP-3) and insulin (C-peptide) were measured together with urinary parameters of renal damage (beta(2)-microglobulin and proteinuria) by immunoassays, in house validated for the purpose, in 61 subjects (36 elite athletes, 18 recreational athletes and 7 sedentary individuals) with different levels of physical fitness and endurance exercise. Validation parameters were good for the evaluated assays, excluding a high inter-assay imprecision and inaccuracy of 24 and 26% obtained for GH assay. The range of concentrations found in urine samples under investigation was generally covered by the calibration curves of the studied immunoassays. However, for the samples below or above the calibration curve, opportune dilution or concentration were performed. Particularly, C-peptide samples had to be diluted 1:5 and beta(2)-microglobulin ones assayed using a triple sample volume, to fall within the calibration range. Urinary C-peptide was the only biomarker statistically higher in samples of elite athletes when compared to recreational athletes and sedentary individuals. Among elite athletes, tae-kwon-do athletes showed the highest IGF-II basal values while weightlifting athletes showed the lower IGF-I and IGFBP-3 basal values. The trend observed in weightlifters' basal samples was confirmed in their training samples: IGF-I, IGF-II, IGFBP-3 and beta(2)-microglobulin were lower in with respect to those from synchronised swimming. Over the training season, within athlete variability was observed for IGFBP-3 for weightlifting athletes. In the studied subjects, no direct associations were found between biomarkers of GH or insulin misuse and urinary parameters of renal damage, eventually due to high-workload endurance training. The variations observed in different biomarkers should be taken in consideration in the hypothesis of setting reference concentration ranges for doping detection.

IGF-1, IGFBP-3 and ALS in adult patients with chronic kidney disease.

BACKGROUND: Insulin-like growth factor I (IGF-1) is for the most part bound in a ternary complex with IGF-binding protein-3 (IGFBP-3) and acid-labile subunit (ALS). This ternary complex is a storage form of IGF-1 in blood and passes not through the renal glomerulus. Little information is available in regard to the components of the ternary complex in adult renal disease. OBJECTIVE: To investigate levels of serum IGF-1, IGFBP-3 and ALS in relation to renal function and extent of proteinuria. DESIGN AND PATIENTS: We measured IGF-1, IGFBP-3 and ALS concentrations in 137 patients who were investigated due to proteinuria and/or haematuria and/or renal impairment. The patients received renal biopsies and the histological diagnosis was documented. Urinary albumin excretion and relevant clinical parameter were evaluated. RESULTS: IGF-1 showed a highly positive correlation to IGFBP-3 and ALS, and the latter to IGFBP-3. IGF-1, IGFBP-3 and ALS decreased with increasing age. IGF-1 and IGFBP-3 showed no significant change depending on the creatinine clearance. However, ALS decreased with decreasing renal function. In patients with heavy proteinuria ALS levels, but not IGF-1 and IGFBP-3 levels, decreased significantly. Patients with chronic ischaemic renal damage and diabetic glomerulopathy showed higher IGF-1 and IGFBP-3 levels compared to patients with thin glomerular basement membrane disease despite their older age. CONCLUSIONS: IGF-1 and IGFBP-3 levels seem to be independent of renal function and severity of proteinuria. However, ALS levels are altered in renal failure and nephrotic syndrome, which may be due to increased renal loss or diminished hepatic production or both.

[Puberty, pregnancy, gender and hormonal status--putative risk factors for diabetic nephropathy]. / Pubertatea, sarcina, sexul si statusul hormonal--posibili factori de risc pentru nefropatia diabetica.

Contemplation of non-genetic risk factors that are influencing the onset and development of diabetic nephropathy (diabetic kidney disease--DKD) is very important. This article is integrative, assessing the existent data about several possible risk factors for DKD. Because the age of onset and postpubertal duration of diabetes seems to be strongly correlated with DKD, it is feasible for puberty to be another independent risk factor. Data analysis regarding puberty and possible explanatory mechanisms to link it with DKD, as the connection with DKD of other situations, with special hormonal status (like pregnancy), is also part of this article. Summing up the data about hormonal status, we can conclude that ANF levels are a risk factor for diabetic nephropathy because they are implicated in diminution of urinary Na elimination and hypertension and subsequent urinary albumin excretion (UAE) in case of inadequate glycaemic control. The evidences regarding GH are indicating that it is a risk factor for DKD and that he is probably implicated in glomerular hypertrophy onset at puberty. The urinary elimination levels of GH are very strong correlated with UAE being putative early marker for DKD. Also the GH deficiency seems to be a protective mechanism for DKD apparition. GH is strongly correlated with IGF-1 that has very high urinary levels in microalbuminuric patients. These levels are very well related to UAE, kidney volume--important markers for glomerular hypertrophy. The evidences accumulated until now regarding the role of masculine gender, testosterone and estrogens in DKD are inarticulate.

Relationship between low levels of anabolic hormones and 6-year mortality in older men: the aging in the Chianti Area (InCHIANTI) study.

BACKGROUND: Aging in men is characterized by a progressive decline in levels of anabolic hormones, such as testosterone, insulinlike growth factor 1 (IGF-1), and dehydroepiandrosterone sulfate (DHEA-S). We hypothesized that in older men a parallel age-associated decline in bioavailable testosterone, IGF-1, and DHEA-S secretion is associated with higher mortality independent of potential confounders. METHODS: Testosterone, IGF-1, DHEA-S, and demographic features were evaluated in a representative sample of 410 men 65 years and older enrolled in the Aging in the Chianti Area (InCHIANTI) study. A total of 126 men died during the 6-year follow-up. Thresholds for lowest-quartile definitions were 70 ng/dL (to convert to nanomoles per liter, multiply by 0.0347) for bioavailable testosterone, 63.9 ng/mL (to convert to nanomoles per liter, multiply by 0.131) for total IGF-1, and 50 microg/dL (to convert to micromoles per liter, multiply by 0.027) for DHEA-S. Men were divided into 4 groups: no hormone in the lowest quartile (reference) and 1, 2, and 3 hormones in the lowest quartiles. Kaplan-Meier survival and Cox proportional hazards models adjusted for confounders were used in the analysis. RESULTS: Compared with men with levels of all 3 hormones above the lowest quartiles, having 1, 2, and 3 dysregulated hormones was associated with hazard ratios for mortality of 1.47 (95% confidence interval [CI], 0.88-2.44), 1.85 (95% CI, 1.04-3.30), and 2.29 (95% CI, 1.12-4.68), respectively (test for trend, P <.001). In the fully adjusted analysis, only men with 3 anabolic hormone deficiencies had a significant increase in mortality (hazard ratio, 2.44; 95% CI, 1.09-5.46 (test for trend, P <.001). CONCLUSIONS: Age-associated decline in anabolic hormone levels is a strong independent predictor of mortality in older men. Having multiple hormonal deficiencies rather than a deficiency in a single anabolic hormone is a robust biomarker of health status in older persons.

Urinary epidermal and insulin-like growth factor excretion in autistic children.

Growth factors have been implicated in the pathogenesis of autism. We have investigated daily urinary excretion of insulin-like growth factor-1 (IGF-1), epidermal growth factor, and insulin-like growth factor binding protein-3 in autistic children (n=34, age 2-5 years) and age-matched control children (n=29). The mean urinary IGF-1 level was lower in the autism group than the control group (p=0.03). Height was normal. These findings suggest altered IGF-1 metabolism in young autistic children. The cause-effect relationship should be examined by longitudinal studies and insulin-like growth factor provocation tests.

Hyperphosphatemia is prevalent among children with nephrotic syndrome and normal renal function.

The aim of the study was to analyze systematically our observation that children with severe nephrotic syndrome (NS) have hyperphosphatemia despite normal kidney function. Forty-seven children with NS and normal glomerular filtration rate (GFR) were studied [26 with steroid-sensitive nephrotic syndrome (SSNS) and 21 with persistent NS]. The plasma phosphate level was expressed as the number of standard deviations (SDs) from the mean levels in age- and gender-matched controls. In SSNS plasma phosphate concentration was elevated (+3.7+/-2.0 SDs) during relapse and normalized (-0.7+/-1.7 SDs) in remission. In persistent NS the phosphate level was +4.0+/-2.1 SDs. Patients with marked hyperphosphatemia (>4 SDs) were younger (p<0.001), had lower plasma albumin (p<0.001), and had higher urinary protein levels (p<0.05). Hyperphosphatemia did not correlate with GFR, plasma calcium, or urinary sodium levels. Children with persistent NS had decreased serum 25(OH)D(3) and insulin-like growth factor 1 (IGF-1) concentrations. Hyperphosphatemia is prevalent among children with persistent nephrotic syndrome and normal renal function, correlates with its severity, and may result from increased urinary IGF-1 wasting.

Urinary insulin-like growth factor-I measurement in an actual sport competition, an additional approach in laboratory antidoping tests.

BACKGROUND: The insulin-like growth factor hormone (IGF-I) is an important protein hormone under investigation with physical exercise and for doping detection. Urinary IGF-I level in fact represents a relevant measurement when the postexercise proteinuria is under analysis. To verify the IGF-I level variation in the circulation and in urinary excretion in the occasion of a competition, the plasma and urine IGF-I in athletes before and after an actual competitive event were measured. METHODS: Twenty well-trained cyclists took part in a competition (102 km) and concluded the intense physical exercise in approximately 2(1/2) h. Urine and blood samples were collected from each athlete 10-20 min before and at the end of the competition. Plasma and urine total IGF-I (pIGF, uIGF), total urinary proteins (uPr), and creatinine (uCr) concentrations were measured. RESULTS: The uIGF [from 76.2+/-15.8 to 256.9+/-29.1 ng/l (p<0.001)], uPr [from 29.4+/-6.7 to 325.9+/-95.1 mg/l (p<0.005)], and uCr [from 6.3+/-1.0 to 10.0+/-0.8 mmol/l (p<0.005)] significantly increased. The pIGF was 262.6+/-14.3 and 247.3+/-11.8 microg/l before and end-exercise, respectively. A statistical correlation between uIGF and uPr was demonstrated (p<0.001). The pIGF/uIGF ratio was significantly (p<0.05) decreased comparing the end with before the competition. CONCLUSIONS: The pIGF/uIGF significantly decreased at the end, compared with before the competition, suggesting a changed uIGF excretion. This increment appeared to be increased, although not significantly, considering the ratio with uCr.

Influence of acute exercise on urinary protein, creatinine, insulin-like growth factor-I (IGF-I) and IGF binding protein-3 concentrations in children.

Insulin-like growth factor-I (IGF-I) is a polypeptide hormone and present in human urine. Insulin-like growth factor binding protein-3 (IGFBP-3) is the major form of binding protein in human circulation and functions as a carrier for IGF-I. Our goal was to determine the effects of volleyball exercise on the concentrations of urine protein, creatinine, IGF-I, and IGFBP-3 in children and to find out whether these effects differ between boys and girls. Volunteer children (13 females and 14 males), aged 10-13 years old were included in this work. Weight and height of the subjects were measured, and urine samples of their were collected before and after 2 hours of exercise. Urinary protein, creatinine, IGF-I and IGFBP-3 levels were analysed. Urinary protein, creatinine and IGF-I concentrations were increased after two hours of exercise wheres urinary IGFBP-3 concentrations did not change. In addition, no statistically significant difference in all parameters analysed was observed between boys and girls of similar age and body mass index.

Urine and serum free IGF-1 levels in patients with bladder cancer: a brief report.

Insulin-like growth factor (IGF)-1, a mitogenic and anti-apoptotic peptide, can affect the proliferation of epithelial cells, and is thought to play a role in cancer development. The free IGF-1 represents the biologically active fraction of IGF-1. We hypothesised that there is a difference in free IGF-1 levels in the urine and serum from patients with TCC and normal subjects. Urine and blood samples were collected from 30 cases of superficial TCC and an equal number control subjects without malignancy. Free IGF-1 levels were measured in duplicate by radioimmunoassay. Specimens of bladder carcinoma were staged histopathologically using the Mostoffi grading system. Statistical analyses were performed using the Mann-Whitney U-test, Pearson correlation and covariate analysis. There was no significant difference in urine and serum free IGF-1 levels between the two groups. The correlation between urine and serum free IGF-1 levels and age was not significant. There was also no significant relationship between free IGF-1 levels and histopathological grading. The results of this pilot study reveal that the free IGF-1 level does not help predict tumour marker in the patients with bladder cancer.

Effects of acute, heavy-resistance exercise on urinary peptide hormone excretion in humans.

To examine physical exercise-related changes in urinary excretion of protein/peptide hormones and to correlate modifications with the general increase in post-exercise proteinuria, urine C-peptide, insulin and insulin-like growth factor-I (IGF-I) and their plasma concentrations were measured. Plasma and urinary C-peptide, insulin and IGF-I before (Bex) and at the end (Eex) of physical exercise (a 2.5-hour competition, 102 km) were analysed in 20 young cyclists. At Eex compared with Bex, concentration of urinary C-peptide decreased slightly but significantly (21.3 +/- 2.7 vs. 13.5 +/- 1.7 nmol/l), but urinary insulin and urinary IGF-I concentrations significantly increased at Eex (92.5 +/- 4.2 vs. 131.4 +/- 15.7 pmol/l and 10.0 +/- 2.1 vs. 33.6 +/- 3.8 pmol/l, respectively). Plasma insulin and plasma C-peptide significantly decreased, whereas plasma IGF-I was unchanged. Urinary concentrations of total proteins and creatinine significantly increased. Both Eex urinary C-peptide/urinary protein and urinary C-peptide/urinary creatinine ratios were significantly reduced. The correlation between C-peptide and insulin in plasma was confirmed at Bex as well as Eex, but in urine only at Bex. An increased renal tubular reabsorption of C-peptide at the end of exercise might be suggested, but the expected values considering creatinine excretion were almost three times less. The Eex urinary insulin concentration was higher than expected, considering the circulation levels, but lower when compared with the expected concentration considering creatinine excretion. Physical exercise proteinuria, related to an increased protein filtration and a saturation of the mechanisms responsible for the reabsorption, does not appear similar for all peptide hormones.

The measurement of insulin-like growth factor-I (IGF-I) concentration in random urine samples.

The aim of the present study was to assess a suitable expression of the urinary concentration of a protein/ peptide hormone such as insulin-like growth factor-I (IGF-I), measured in the urine of healthy individuals when the specimen collection is executed randomly. One hundred and twenty male subjects were divided by age into four groups, namely healthy sedentary young (SYA) and older (SOA) adults, older (OC) and young (YC) children. In a single urine specimen, randomly collected during the morning from each individual, total urinary IGF-I was measured by immunoradiometric method, and urinary creatinine (uCr) and total proteins (utPr) were measured by capillary electrophoresis and spectrophotometric methods, respectively. The urinary IGF-I concentrations were not significantly different in all groups investigated and they were (mean +/- SD): 82.7 +/- 82.8 ng/l, 103.5 +/- 83.3 ng/l, 80.4 +/- 64.4 ng/l in OC, SYA and SOA, respectively; only in the YC group there was a tendency to higher values (125.2 +/- 93.2 ng/l) compared with the other groups. utPr ranged from 26 to 40 mg/l and did not demonstrate significant differences between groups. The urinary IGF-I correlated with uCr and utPr, and statistical significance was observed in all measurements. The measurement of urinary IGF-I in random urine and its ratio to utPr is an innovative, useful way of investigation of urinary protein/peptide hormones.

Urinary insulin-like growth factor I in athletes, before and after physical exercise, and in sedentary subjects.

BACKGROUND: Insulin-like growth factor I (IGF-I), like growth hormone (GH), is excreted in urine in a smaller fraction than the concentration found in blood. Exercising subjects undergo post-exercise proteinuria. The present work aims to propose a method for urinary IGF-I analysis (uIGF-I) by defining urinary concentration in sedentary individuals and athletes before and after strenuous exercise. METHODS: Urine samples were collected from 30 sedentary healthy male individuals during the morning and from 30 well-trained cyclists, before and after a competition of about 3 h (150 km). uIGF-I was measured in undiluted acidified urine by radioimmunoassay (RIA) method using a purified polyclonal rabbit antibody, human 125I-IGF-I and a second anti-rabbit antiserum. The acidification of the urine samples and the excess of IGF-II addition in the incubation medium of the assay were used to dissociate the binding and to block the interference from IGF binding proteins (IGFBPs). Urinary growth hormone (uGH), total protein (utPr) and creatinine (ucr) concentrations were also measured by immunoradiometric assay (IRMA), colorimetric and capillary electrophoresis methods, respectively. RESULTS: The analysis range was 0-2500 ng/l (0-327 pmol/l), the intra- and inter-assay coefficients of variations (CVs) ranged from 2.3% to 7.8%, respectively. The detection limit was 0.6 pg/tube. The uIGF-I/creatinine (cr) ratio in healthy subjects was 70 +/- 8 pg/mg cr. The uIGF-I/creatinine ratio (pg/mg cr) was different (p<0.001) in athletes before vs. after competition 93 +/- 27 vs. 136 +/- 13. Athletes' [uIGF-I/total proteins] ratio (ng/mg tPr) before and post-exercise was 2.3 +/- 0.5 and 2.5 +/- 0.3, respectively. CONCLUSIONS: uIGF-I assay appears to be an effective way of monitoring IGF-I excretion. In the cyclists, in the pre-exercise state, uIGF-I was comparable with that measured in sedentary healthy individuals. In the cyclists, after strenuous exercise, the increased uIGF-I/cr and uGH/cr ratios suggested a relation with the post-exercise proteinuria. In conclusion, proteinuria physiologically obtained, such as post-exercise proteinuria, might be a new approach in IGF-I system investigation.